ANTIGEN PRESENTING SYSTEM AND METHODS FOR ACTIVATION OF T-CELLS

Abstract: The present invention relates to synthetic antigen-presenting matrices, their methods of making and their methods of use. One such matrix is cells that have been transfected to produce MHC antigen-presenting molecules and assisting molecules such as co-stimulatory molecules. The matrices can be used to activate CD8<+> T-cells to produce cytokines and become cytotoxic.

Description: ANTIGEN PRESENTING SYSTEM AND METHODS FOR ACTIVATION OF T-CELLS This invention was made with the support of the Government of the United States of America, and the Government of the United States of America has certain rights in the invention. CROSS-REFERENCE TO RELATED APPLICATION This application is a continuation-in-part of U.S. patent application Serial No. 08/400,338 filed March 8, 1995 and having the same title. TECHNICAL FIELD The present invention relates to materials and methods of activating T-cells with specificity for particular antigenic peptides, the use of activated T-cells in vivo for the treatment of a variety of disease conditions, and compositions appropriate for these uses. BACKGROUND The efficiency with which the immune system cures or protects individuals from infectious disease has always been intriguing to scientists, as it has been believed that it might be possible to activate the immune system to combat other types of diseases. Such diseases include cancer, AIDS, hepatitis and infectious disease in immunosuppressed patients. While various procedures involving the use of antibodies have been applied in those types of diseases, few if any successful attempts using cytotoxic T-cells have been recorded. Theoretically, cytotoxic T-cells would be the preferable means of treating the types of disease noted above. " However, no procedures have been available to specifically activate cytotoxic T-cells. Cytotoxic T-cells, or CD8' cells (i.e., cells expressing the molecule CD8) as they are presently, , known, represent the main line of defense against viral}. infections. CD8" lymphocytes specifically recognize and kill cells which are infected by a virus. Thus, the cost of eliminating a viral infection is the accompanying loss of the infected cells. The T-cell receptors on the surface of CD8 * cells cannot recognize foreign antigens directly. In contrast to antibodies, antigen must first be presented to the receptors. The presentation of antigen to CD8 * T-cells is accomplished by major histocompatibility complex (MHC) molecules of the Class I type. The major histocompatibility complex (MHC) refers to a large genetic locus encoding an extensive family of glycoproteins which play an important role in the immune response. The MHC genes, which are also referred to as the HLA (human leucocyte antigen) complex, are located on chromosome 6 in humans. The molecules encoded by MHC genes are present on cell surfaces and are largely responsible for recognition of tissue transplants as "non-self". Thus, membrane-bound MHC molecules are intimately involved in recognition of antigens by T-cells. MHC products are grouped into three major classes, referred to as I, II, and III. T-cells that serve mainly as helper cells express CD4 and primarily interact with Class II molecules, whereas CD8-expressing cells, which mostly represent cytotoxic effector cells, interact with Class I molecules. Class I molecules are membrane glycoproteins with the ability to bind peptides derived primarily from intracellular degradation of endogenous proteins. Complexes of MHC molecules with peptides derived from viral, bacterial and other foreign proteins comprise the ligand that triggers the antigen responsiveness of T-cells. In contrast, complexes of MHC molecules with peptides derived from normal cellular products play a role in "teaching" the T-cells to tolerate self-peptides, in the thymus. Class I molecules do not present entire, intact antigens; rather, they present peptide fragments thereof, "loaded" onto their "peptide binding groove". For many years, immunologists have hoped to raise specific cytotoxic cells targeting viruses, retroviruses and cancer cells. While targeting against viral diseases in general may be accomplished in vi vo by vaccination with live or attenuated vaccines, no similar success haε been achieved with retroviruses or with cancer cells. Moreover, the vaccine approach has not had the desired efficacy in immunosuppressed patients. At least one researcher has taken the rather non-specific approach of "boosting" existing CD8' cells by incubating them in vi tro with IL-2, a growth factor for T-cells. However, this protocol (known as LAK cell therapy) will only allow the expansion of those CD8 * cells which are already activated. As the immune system is always active for one reason or another, most of the IL-2 stimulated cells will be irrelevant for the purpose of combatting the disease. In fact, it has not been documented that this type of therapy activates anv cells with the desired specificity. Thus, the benefits of LAK cell therapy are controversial at best, and the side effects are typically so severe that many studies have been discontinued. Several novel molecules which appear to be involved in the peptide loading process have recently been identified. It has also been noted that Class I molecules without bound peptide (i.e., "empty" molecules) can be produced under certain restrictive circumstances. These "empty" molecules are often unable to reach the cell surface, however, as Class I molecules without bound peptide are very thermolabile . Thus, the "empty" Class I molecules disassemble during their transport from the interior of the cell to the cell surface. The presentation of Class I MHC molecules bound to peptide alone has generally ineffective in activating CD8 * cells. In nature, the CD8 * cells are activated by antigen-presenting cells which present not only a peptide-bound Class I MHC molecule, but also a costimulatory molecule. Such costimulatory molecules include B7 which is now recognized to be two subgroups designated as B7.1 and B7.2. It has also been found that cell adhesion molecules such as integrins assist in this process. When the CD8 * T-cell interacts with an antigen-presenting cell having the peptide bound by a Class I MHC and costimulatory molecule, the CD8 * T-cell is activated to proliferate and becomes an armed effector T-cell. See, generally, Janeway and Travers, Immunobiology, published by Current Biology Limited, London (1994) , incorporated by reference. Accordingly, what is needed is a means to activate T-cells so that they proliferate and become cytotoxic. It would be useful if the activation could be done in vi tro and the activated cytotoxic T-cells reintroduced into the patient. It would also be desirable if the activation could be done by a synthetic antigen-presenting matrix comprised of a material such as cells which not only presents the selected peptide, but also presents other costimulatory factors which increase the effectiveness of the activation. It would also be advantageous if it was possible to select the peptide so that substantially only those CD8' cells cytotoxic to cells presenting that peptide would be activated. BRIEF SUMMARY OF THE INVENTION The present invention relates to a synthetic antigen-presenting system for presenting an MHC molecule complexed to a peptide and an assisting molecule to a T-cell to activate the T-cell. In one embodiment, the system relates to a synthetic antigen-presenting matrix having a support and at least the extracellular portion of a Class I MHC molecule capable of binding to a selected peptide operably linked to the support. The matrix also includes an assisting molecule operably linked to the support. The assisting molecule acts on a receptor on the CD8 * T- cell. The MHC and assisting molecules are present in sufficient numbers to activate a population of T-cell lymphocytes against the peptide when the peptide is bound to the extracellular portion of the MHC molecule. It has been found that an antigen-presenting matrix having both an MHC molecule or a portion of a MHC molecule together with an assisting molecule, provides a synergistic reaction in activating T-cell lymphocytes against the peptide. Examples of assisting molecules are costimulatory molecules such as B7.1 and B7.2 or adhesion molecules such as ICAM-1 and LFA-3. The extracellular portion of such costimulatory molecules can also be used. Another type of costimulatory molecule is one that reacts with the CD28 molecule such as anti-CD28 antibodies or functional portions thereof, e.g. Fab portions. It has been found that a specifically effective synergistic reaction results from an antigen-presenting matrix having MHC molecules bound with a peptide, a costimulatory molecule, and an adhesion molecule. In particular, a highly effective synergistic generation of cytotoxic T-cell activity results from the combination of B7.1 and ICAM-1. The support used for the matrix can take several different forms. Examples for the support include solid support such as metals or plastics, porous materials such as resin or modified cellulose columns, rnicrobeads, microtiter plates, red blood cells and liposomes . Another type of support is a cell fragment, such as a cell membrane fragment or an entire cell. In this embodiment, the matrix is actually cells which have been transfected to present MHC molecules and assisting molecules on the cell surface to create an antigen- presenting cell (APC) . This is done by producing a cell line containing at least one expressible Class I MHC nucleotide sequence for the MHC heavy chain, preferably a cDNA sequence, operably linked to a first promoter and an expressible β- 2 microglobulin nucleotide sequence operably linked to a second promoter. The MHC heavy chain and the β- 2 microglobulin associate together form the MHC molecule which binds to the peptide. The MHC protein binds with the antigenic peptide and presents it on the surface of the cell. The cell also includes a gene for a nucleotide sequence of an assisting molecule operably linked to a third promoter. The assisting molecule is also presented on the surface of the cell. These molecules are presented on the surface of the cell in sufficient numbers to activate a population of T-cell lymphocytes against the peptide when the peptide is bound to the complexes. Other molecules on the surface of a cell or cell fragment such as carbohydrate moieties may also provide some costi ulation to the T-cells. The cell line is synthetic in that at least one of the genes described above is not naturally present in the cells from which the cell line is derived. It is preferable to use a poikilotherm cell line because MHC molecules are thermolabile . A range of species are useful for this purpose. See, for example, U.S. Patent No . 5,314,813 to Petersen et al . which discusses numerous species for this use and is incorporated by reference. It is preferred to use eukaryotic cells and insect cells in particular. In one embodiment, it is particularly preferred to have at least two assisting molecules, one being a costimulatory molecule and the other being an adhesion molecule. It has been found that this combination has a synergistic effect, giving even greater T-cell activation than either of the individual molecules combined. It has also been found to be advantageous and preferable to have at least one of the transfected genes under control of an inducible promoter. Using the present invention, it is possible to introduce the peptide to the cell while it is producing MHC molecules and allow the peptide to bind the MHC molecules while they are still within the cell. Alternatively, the MHC molecules can be expressed as empty molecules on the cell surface and the peptide introduced to the cells after the molecules are expressed on the cell surface. In this latter procedure, the use of poikilotherm cells is particularly advantageous because empty MHC molecules, those not yet complexed or bound with peptides, are ther olabile . Class I MHC molecules have been expressed in insect cells such as Drosophila melanogaster (fruit fly) cells. Since Drosophila does not have all the components of a mammalian immune system, the various proteins involved in the peptide loading machinery should be absent from such cells. The lack of peptide loading machinery allows the Class I molecules to be expressed aε empty molecules at the cell surface. Another advantage of using insect cells such as the Drosophila system is that Drosophi la cells prefer a temperature of 28 °C rather than 37 °C. This fact is very important, because empty Class I molecules are thermolabile and tend to disintegrate at 37 °C. By incubating the Class I-expressing Drosophila cells with peptides that can bind to the Class I molecule, it is possible to get virtually every Class I molecule to contain one and the same peptide. The cells are accordingly very different from mammalian cells, where the Class I molecules contain many different types of peptides, most of which are derived from our own, innocuous cellular proteins. The present invention also relates to methods for producing activated CD8 * cells in vi tro . One method comprises contacting, in vi tro, CD8' cells with one of the antigen-presenting matrices described above for a time period sufficient to activate, in an antigen-specific manner, the CD8' cells. The method may further comprise (1) separating the activated CD8' cells from the antigen-presenting matrix; (2) suspending the activated CD8 * cells in an acceptable carrier or excipient; and (3) administering the suspension to an individual in need of treatment. The antigens may comprise native or undegraded proteins or polypeptides , or they may comprise antigenic polypeptides which have been cleaved into peptide fragments comprising at least 8 ammo acid residues prior to incubation with the human Class I MHC molecules. In another variation, the invention relates to methods treating conditions in patients and specifically killing target cells in a human patient. The method comprises (1) obtaining a fluid sample containing resting or naive CD8' cells from the patient; (2) contacting, in vitro, the CD8' cells with an antigen-presenting matrix for a time period sufficient to activate, in an an t igen-specific manner, the CD8' cells; and (3) administering the activated CD8 cells to the patient. For example, the use of tumor specific peptides allows for the treatment of tumor related diseases by producing cytotoxic activated CD8 * T-cells. The invention also relates to the method of treating a medical condition by administration of an antigen-presenting matrix in a suitable suspension. In various embodiments the condition may comprise cancer, tumors, neoplasia, viral or retroviral infection, autoimmune or autoimmune-type conditions. In one embodiment, the method of administering the matrix comprises intravenous injection. BRIEF DESCRIPTION OF THE DRAWINGS Figures 1-3 diagram the construction of expression plasmids pRmHa-2 and pRmHa-3. In Figure 1, pRmHa-2 construction is shown; in Figure 2, pRmHa-3 construction is shown; and in Figure 3, the pRmHa-3 vector is illustrated, showing the restriction, polylinker, promoter, and polyadenylation sites; as well as a site at which a nucleotide sequence may be inserted for expression; Figures 4 and 5 show peptide-induced thermostabilization of HLA B27 and HLA A2.1 expressed on the surface of Drosophila cells by HIV peptides. The mean fluorescence of each cell population is shown plotted against the incubation conditions; Figure 6 illustrates data from an experiment designed to determine whether insect cells can process antigen and load it onto the Class I molecules, and whether the latter can present either endogenously or exogenously derived antigen to T-cells. Schneider 2 (SC2) or 3T3 cells transfected with A / β2 were incubated with ovalbumin protein (OvPro) or ovalbumin peptide, OVA 24 (OvPep) in isotonic (Iso) or hypertonic (Hyp) media. fMurine cell line BALB/3T3 is available from the ATCC under accession number CCL 163.) After treatment, cellε were cocultured with the T-cell hybridoma B3/CD8. B3/CD8 is a T-cell hybridoma between B3 (Carbone, et al . , J_ ^ E XP . Med. 169: 603-12 (1989)) , cytotoxic T-cell specific for ovalbumin peptide 253-276 presented by H-2K b Class I molecules, and CD8- bearing IL-2-secreting cell line. Upon antigenic εtimulation, B3/CD8 produceε IL-2, meaεured by 3 H.-thymidine incorporation in IL-2-dependent cell line CTLL (Gilliε, et al . , J. Immunol. 120: 2027 91978) ) . Thus, by measuring the amount of IL-2 produced, one can assay for T-cell recognition. The supernatant from the cocultures were analyzed for IL-2 by 3 H thymidine incorporation by the IL-2-dependent cell line CTLL (ATCC No. TIB 214) . The amount of 5 H thymidine incorporated is plotted against the initial cell treatments; Figure 7 illustrates the expression of B7.1, ICAM-1 and MHC on the surface of transfected Drosophila (fly) cells according to the present invention; Figure 8 is a graph showing resultε of a fluoreεcence-activated cell sorter experiment using recombinant L d mouse MHC linked to red blood cells,* Figure 9 is a graph showing results of a fluorescence-activated cell sorter experiment using recombinant K h mouse MHC linked to red blood cells; Figure 10 is a graph demonstrates binding of recombinant K 1, to microtiter plates by use of labeled antibodies; Figure 11 is a series of graphs showing the results from fluorescence-activated cell sorter experiments demonstrating the expression of CD69 and CD25 on CD8' 2C cells stimulated with transfected Drosophila cells ; Figure 12 is a pair of bar graphs showing IL-2-dependent proliferative responses of CD8' 2C cells elicited by peptides presented by Drosophila cells transfected with L ri only; Figure 13 is a graph showing the influence of peptide concentration on day 3 proliferative responses of CD8' 2C cells elicited by peptides presented by transfected Drosophila cells; Figure 14 iε a pair of graphs showing the influence of antigen-presenting cells dose on day 3 proliferative response and IL-2 production of CD8 * 2C cells elicited by peptides presented by transfected Drosophila cells; Figure 15 is a series of graphs showing the influence of peptide concentration on the proliferative response of CD8 * 2C cells elicited by Drosophila cells transfected with L'\B7, L''.ICAM and L .B7.ICAM; Figure 16 is a series of graphs εhowing the kineticε of the proliferative response of CD8 * and CD8 ~ 2C cells elicited by Drosophila cells tranεfected with L ,J , L d .B7, L d .ICAM and L d .B7.ICAM pluε QL9 peptide; Figure 17 iε a series of graphs showing CTL activity of CD8 * 2C cellε stimulated by Drosophila cells transfected with L''.B7, L <1 .B7.ICAM or L''.ICAM antigen- presenting cells plus QL9 peptide (10 μM) in the absence of exogenous cytokines; Figure 18 is a pair of graphs showing CTL activity of CD8' 2C cells stimulated by Drosophi la cells transfected with L''.ICAM antigen-presenting cells plus QL9 peptide (10 μM) in the absence (left) or presence (right) of exogenous IL-2 (20u/ml) ; Figure 19 is a pair of graphs εhowing the proliferative response of normal (non-transgenic) CD8' T-cells to peptides presented by transfected Drosophi la cells (left panel) and the response elicited by graded doses of N B6 and 2C B6 CDS' cells cultured with 5 x I0 r ' P.10.D2 (L' 1 ) spleen cells (2000 cGy) in the absence of peptides for 3 days without the addition of exogenous cytokines (right panel) ; Figure 20 is a graph showing stimulated mitogenesis of purified 2C+ T-cells cultured (50,000 cells per well) in plateε coated with immobilized molecules with peptide QL9 (solid line = L and antι-CD28 antibody, broken line = L d only) ,* Figure 21 is a bar graph showing stimulated mitogenesis of purified 2C+ T-cells at day 5 in culture, with various indicated peptides, cultured in 96-well plates coated with L d and anti-CD28 antibody (hatched bars = no IL-2, black bars = IL-2 added) ; Figure 22 is a graphical representation of the results of cytofluorometric analysis of cells recovered after 12 days of culture in plates coated with L' 1 and anti-CD28 antibody and exposed to peptide QL9, stained using the 2C T-cell receptor specific antibody 1B2 (M2 = positive staining cells, Ml = negative staining cells) ; Figure 23 is a graphical representation of the results of cytofluorometric analysis of cells recovered after 12 days of culture in plates coated with L d and anti-CD28 antibody and exposed to peptide p2Ca, stained using the 2C T-cell receptor specific antibody 1B2 (M2 = positive staining cells, Ml = negative staining cells) ; Figure 24 is a graphical representation of the results of cytofluorometric analysis of cells recovered after 12 days of culture in plates coated with L' 1 and antι-CD28 antibody and exposed to peptide SL9, stained using the 2C T-cell receptor specific antibody 1B2 (M2 = positive staining cells, Ml = negative staining cells) , Figure 25 is a graph showing cytolysis of target cells by activated T-cells (solid line = peptide QL9, broken line = control peptide LCMV) ; Figure 26 is a graphical representation of the cytotoxic lysis resulting from activation of human CDS T-cells with antigen-presenting cells loaded with influenza matrix peptide; Figure 27 is a graphical representation of the cytotoxic lysis resulting from activation of human CD8' T-cellε with antigen-presenting cells loaded with HIV-RT peptide; and Figure 28 is a graphical representation of the cytotoxic lysiε resulting from activation of human CD8 * T-cells with antigen-presenting cellε loaded with tyrosinase peptide. DETAILED DESCRIPTION OF THE INVENTION The present invention relateε to a synthetic antigen-presenting system which can be used to activate T-cell lymphocytes. The activated CD8 * T-cells proliferate, produce cytokines, become cytotoxic or some combination of these reεultε. In one preferred embodiment, the εyεtem activateε cytotoxic CD8' cellε which then proliferate and then are activated to εeek out and deεtroy target cells. The present invention can be used to activate T-cells in vi tro and the activated T-cells are then returned to the patient from which they were originally derived or may be used in vivo activation of T-cells. The synthetic antigen-presenting system of the present invention has two major components. The first component is at leaεt the extracellular portion of the Class I MHC molecule which is capable of binding to a selected peptide. The second major component is an assisting molecule which assists in the activation of T-cellε. In each case, an extracellular portion of a larger molecule can uεed, but in certain e bodimentε, the entire molecules are used. For ease of description, MHC molecules will be discussed generally, with the understanding that an extracellular portion of the MHC molecule may be used. The portion of the MHC molecule necesεary for the present invention iε the part which binds to the εelected peptide and preεentε the peptide to the T-cell. The peptide is selected to activate the appropriate T-cell, depending on the treatment to be conducted. For example, in the treatment of particular cancers, certain antigenic peptides are presented on the surface of the cancer cells which will react with activated T-cells. Thus, it is appropriate to use a peptide selected to activate the appropriate T-cells that will then bind with and destroy the cancer cells. The present invention allows the MHC molecules to be produced by cells with the peptide already complexed with the MHC molecule or to produce empty MHC molecules which do not yet have a peptide complexed with them. Thiε latter embodiment iε particularly useful εince it allowε the peptide to be chosen after the MHC molecules are prepared. A Class I MHC molecule includes a heavy chain, sometimes referred to as an alpha chain, and a β- 2 microglobulin. As discuεεed herein, the extracellular portion of the Claεε I MHC molecule is made up of an extracellular portion of an MHC heavy chain together with the β- 2 microglobulin. In preparing the extracellular portions of MHC to be linked to a support, soluble molecules are prepared as discussed below. These molecules generally lack the transmembrane and cytoplas ic domain m the MHC molecule. The asεisting molecule helps facilitate the activation of the T-cell when it is presented with a peptide/MHC molecule complex. The present invention includes two major categories of assiεting moleculeε. The firεt category is composed of costimulatory molecules such aε B7.1 (previously known as B7 and also known as CD80) and B7.2 (also known as CD86) which binds to CD28 on T-cells. Other costimulatory molecules are anti-CD28 antibodies or the functional portions of such antibodies, e.g. Fab portions that bind to CD28. The other major category of aεsisting molecules of the present invention are adhesion moleculeε. These include the various ICAM molecules, which include ICAM-1, ICAM-2, ICAM-3 and LFA-3. It has been found that the combination of a peptide bound to an MHC molecule used in conjunction with one of these assisting molecules activates the T-cells to an extent previously not seen. An even greater εynergistic reaction has been achieved by using a peptide-bound MHC molecule in conjunction with both a costimulatory molecule and an adhesion molecule. This has been found to be particularly effective in producing cytotoxic CD8 * cells. In accordance with the present invention, the MHC molecule and the assisting molecule are operably linked to a support such that the MHC and asεiεting moleculeε are preεent in εufficient numberε to activate a population of T-cellε lymphocyteε againεt the peptide when the peptide iε bound to the extracellular portion of the MHC molecule. The peptide can be bound to the MHC molecule before or after the MHC molecule is linked to the εupport . The support can take on many different forms. It can be a solid εupport εuch aε a plaεtic or metal material, it can be a porous material εuch as commonly uεed in εeparation columnε, it can be a lipoεome or red blood cell, or it can even be a cell or cell fragment. As discussed in more detail below, in the case where a cell serveε aε a support, the MHC and assiεting molecules can be produced by the cell. The MHC molecule iε then linked to the cell by at least the transmembrane domain if not alεo the cytoplasmic domain which would not be present in a soluble form of MHC. The extracellular portions of MHC molecule and asεiεting molecule can be linked to a εupport by providing an epitope which reactε to an antibody immobilized on the support. In addition, the MHC or assisting molecules can be produced with or linked to (His),, which would react with nickel in forming part of the support. Other means to immobilize or link MHC molecules to a support are well known in the art. As diεcussed above, the εupport can be a cell membrane or an entire cell. In such a case, an eukaryotic cell line is modified to become a synthetic antigen-presenting cell line for use with T-cell lymphocyteε . For ease of deεcription, antigen-presenting cells (APC) will also be called stimulator cells. Because empty MHC molecules are thermolabile, it is preferred that the cell culture be poikilotherm and variouε cell lineε are diεcuεεed in detail below. A culture of cellε iε firεt eεtablished. The culture is then transfected with an expresεible Class I MHC heavy chain gene which is operably linked to a promoter. The gene is chosen εo that it is capable of expressing the Class I MHC heavy chain. The cell line is also transfected with an expresεible β - 2 microglobulin gene which is operably linked to a second promoter. The gene iε chosen such it is capable of expreεsing β- 2 microglobulin that forms MHC molecules with the MHC heavy chain. In the case of soluble extracellular portions of MHC molecules to be used with solid supports and the like, a truncated MHC heavy chain gene is used as discuεεed in more detail below. The culture iε also transfected with an expressible assisting molecule gene operably linked to a third promoter. The assisting molecule gene is capable of being expresεed aε an assisting molecule which interacts with the molecule on the T-cell lymphocytes. As discussed below, each of these genes can be transfected uεing variouε methodε, but the preferred method iε to uεe more than one plasmid. The cell line tranεfected iε choεen becauεe it lackε at leaεt one of the genes being introduced. It has been found that insect cellε are advantageous not only because they are poikilother ic, but because they lack theεe geneε and the echaniεmε which would otherwise produce MHC molecules bound to peptides. This allows for greater control over the production of peptide-bound MHC molecules, and the production of empty MHC molecules. The MHC heavy chain is preferably from a different specieε, more preferably, a homeotherm εuch aε mammals and, optimally, humans. The preferred cell line is a stable poikilotherm cell line that has the firεt promoter being mducible to control the expreεεion of the MHC heavy chain. It is preferred that the cell assembleε empty MHC moleculeε and preεents them on the cell surface so that the peptideε can be chosen as desired. The resulting MHC molecules bind to the peptide and are present in sufficient numbers with the assisting molecules on the surface of the cell to activate a population of T-cell lymphocytes against the peptide when the peptide is bound to the MHC cellε. In a further embodiment, a εecond assisting molecule gene is also transfected into the cell culture In this case, the first assisting molecule gene can be for a costimulatory molecule and the second assisting molecule gene can be for an adhesion molecule It is preferred that at least one of the genes and, in particular, the MHC heavy chain gene be linked to an mducible promoter. This allows control over the production of MHC molecules εo that they are only produced at a time when the peptide of interest is available and presented in the culture to react with the produced MHC molecules. This minimizes undesirable MHC molecule/peptide complexes. Where the cell line already produceε one or more of the deεired moleculeε, it iε only neceεsary to transfect the culture with an expreεεible gene for the gene which is lacking in the cells. For example, if the cells already present the MHC moleculeε on their surface, it is only necessary to transfect the culture with an expresεible gene for the aεsisting molecule. The peptide can be introduced into the cell culture at the time the cells are producing MHC molecules. Through methodε εuch aε oεmotic shock, the peptides can be introduced in the cell and bind to the produced MHC molecules. Alternatively, particularly in the case poikilotherm cell lines, the MHC molecules will be presented empty on the cell surface. The peptide can then be added to the culture and bound to the MHC molecules as desired. After the cells are produced having MHC and asεisting molecules on their surfaces, they can be lyophilized and the fragments of the cellε used to activate the population of T-cell lymphocytes. Transfected cultures of cells can be used to produced extracellular portions of MHC molecules and asεisting moleculeε. The uεe of extracellular portions in conjunction with εupports such as solid supportε has certain advantageε of production. Where living cellε are uεed to provide a εynthetic antigen-presenting cell, at least three genes, two to produce the MHC molecule and one for the asεiεting molecule must be introduced to the cell. Often, additional genes such as for antibiotic resistance are also transfected. Where a solid εupport syεtem is being used, one cell line can produce the extracellular portions of MHC molecules while another cell line produces the extracellular portion of the asεisting molecule. The MHC molecule portionε and the aεsisting molecule portions can then be harveεted from their reεpective cultures. The moleculeε are then linked to an appropriate support in sufficient numbers to activate a population of T-cell lymphocytes against a peptide when the peptide is bound to the extracellular portion of the MHC molecule. From a production standpoint, two different cultures can be used, but it is also poεsible to use the same culture, however, requiring that the culture be transfected with the additional gene for the extracellular portion of the assisting molecule. A further modification of this embodiment is to provide a third culture of cells which is transfected with an expressible second asεiεting molecule gene. In thiε example, the εecond culture of cellε produceε extracellular portions of the costimulatory molecule while the third culture of cells produce an extracellular portion of an adhesion molecule. The adhesion molecule portions are harvested and linked to the εupport. The present invention also relates to a method for activating CD8" T-cells against a selected peptide. The method relateε to providing a cell line preεenting MHC moleculeε binding a peptide and aεεisting molecules on their surfaces. Naive CD8' T-cells can be obtained by removal from a patient to be treated. The cultured cells are then contacted with the CD8 * T-cells for a εufficient period of time to activate the CD8' T-cell lymphocyteε resulting in proliferation and transforming the T-cells into armed effector cells. The activated CD8' T-cells can then be separated from the cell line and put into a suspension in an acceptable carrier and administered to the patient. Λn alternative method involves the uεe of the synthetic antigen-presenting matrix to activate the CD8' cellε. It iε preferred that human geneε are uεed and, therefore, human molecule analogε are produced. Aε εhown in prior U.S. Patent No. 5,314,813, murine systems provide particularly useful models for testing the operation of T-cell activation and demonεtrate the applicability .of the procesε for human εyεtemε . See alεo Sykulev et al . , Immunity 1 : 15-22 (1994) . 1. Human Class I MHC Molecules Clasε I MHC molecules compriεe a heavy chain and a β-microglobulin protein. A human Clasε I MHC heavy chain of the present invention is selected from the group comprising HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G, and more preferably, from the group comprising HLA-A, HLA-B, and HLA-C. The heavy chains are uεeful in either εoluble or insoluble form. In the εoluble ("sol") form, a stop codon is engineered into the nucleotide sequence encoding the HLA molecule of choice preceding the transmembrane domain. While it iε possible to isolate nucleotide sequences encoding human Clasε I MHC heavy chainε from known, eεtabliεhed cell lineε carrying the appropriate variantε -- e.g., tranεformed cell lineε JY, BM92, WIN, MOC, and MG -- it is more practical to syntheεize the nucleotide sequence from a portion of the gene via polymeraεe chain reaction (PCR) , uεing the appropriate primer . Thiε method haε been εuccessfully used to clone full-length HLA cDNA; for example, the sequenceε for HLA-A25, HLA-A2, HLA-B7, HLA-B57, HLA-B51, and HLA-B37 are depoεited in the GenBank databaεe under acceεεion nos. M32321, M32322, M32317, M32318, M32319 and M32320, respectively. Known, partial and putative HLA amino acid and nucleotide sequences, including the consensuε sequence, are publiεhed (see, e.g., Zemmour and Parham, Immunocrenetics 33: 310-320 (1991)) , and cell lines expressing HLA variants are known and generally available as well, many from the American Type Culture Collection ("ATCC") . Therefore, using PCR, it iε poεsible to syntheεize human Class I MHC-encoding nucleotide sequences which may then be operatively linked to a vector and used to transform an appropriate cell and expressed therein. Particularly preferred methods for producing the Clasε I MHC heavy chain, β- 2 microglobulin proteinε and asεiεting molecules of the present invention rely on the use of preselected oligonucleotides as primers in a polymerase chain reaction (PCR) to form PCR reaction products as described herein. Gene preparation is typically accomplished by primer extenεion, preferably by primer extension in a polymerase chain reaction (PCR) format . If the genes are to be produced by (PCR) amplification, two primers, i.e., a PCR primer pair, muεt be uεed for each coding strand of nucleic acid to be amplified. (For the sake of εimplicity, εyntheεis of an exemplary HLA heavy chain variant sequence will be discussed, but it is expresεly to be understood that the PCR amplification method described is equally applicable to the synthesis of β- 2 microglobulin, costimulatory molecules, adhesion molecules, and all HLA variants, including those whoεe complete sequences are presently unknown. ) The first primer becomes part of the antisense (minus or complementary) εtrand and hybridizeε to a nucleotide εequence conserved among HLA (plus or coding) strands. To produce coding DNA homologs, first primers are therefore chosen to hybridize to (i.e. be complementary to) conserved regionε within the MHC geneε, preferably, the conεenεuε εequence or similar, conserved regions within each HLA group -- i.e., consensuε sequences within HLA-A, HLA-B, HLA-C, and the less-polymorphic groups, HLA-E, -F, and -G. Second primerε become part of the coding (pluε) εtrand and hybridize to a nucleotide εequence conεerved among minus strands. To produce the HLA-coding DNA homologs, second primers are therefore chosen to hybridize with a conserved nucleotide sequence at the 5' end of the HLA-coding gene such as in that area coding for the leader or first framework region. In the amplification of the coding DNA homologs the conserved 5 ' nucleotide sequence of the second primer can be complementary to a sequence exogenously added using terminal deoxynucleotidyl transferase as described by Loh et al., Science 243: 217-220 (1989) . One or both of the first and second primers can contain a nucleotide sequence defining an endonuclease recognition site. The site can be heterologous to the immunoglobulin gene being amplified and typically appearε at or near the 5' end of the primer. The high turn over rate of the RNA poly eraεe amplifieε the εtarting polynucleotide as has been described by Chamberlin et al . , The Enzv es. ed. P. Boyer, PP. 87-108, Academic Press, New York (1982) . Another advantage of T7 RNA polymerase is that mutations can be introduced into the polynucleotide synthesis by replacing a portion of cDNA with one or more mutagenic oligodeoxynucleotides (polynucleotideε) and tranεcribing the partially-miεmatched template directly aε haε been previouεly described by Joyce et al . , Nuc . Acid Reε . 17 : 711-722 (1989) . Amplification εyεtemε based on transcription have been deεcribed by Gingeras et al . , in PCR Protocols. A Guide to Methods and Applications, pp 245-252, Academic Press, Inc., San Diego, CA (1990) . PCR amplification methodε are deεcribed in detail in U.S. Patent Noε. 4,683,192, 4,683,202, 4,800,159, and 4,965,188, and at least in several texts including "PCR Technology: Principles and Applications for DNA Amplification", H. Erlich, ed., Stockton Press, New York (1989) ; and "PCR Protocols: A Guide to Methods and Applications", Innis et al . , edε . , Academic Preεs, San Diego, California (1990) . Various preferred methods and primers used herein are described hereinafter and are also described in Nilsson, et al . , Cell 58: 707 (1989) , Ennis, et al . , PNAS USA 87: 2833-7 (1990) , and Zemmour, et al . , Immunogenetics 33: 310-20 (1991) , for example. In particular, it iε preferred to deεign primerε from co pariεon of 5' and 3' untranεlated regionε of HLA alleleε (e.g., -A, -B, -C, -E, -F, or -G alleles) , with εelection of conεerved sequences. Restriction siteε may also be incorporated into the 5' and 3' primers to enable the amplification products to be subcloned into sequencing or expression vectors. It may also be helpful to place a -base spacer sequence proximal to the restriction site to improve the efficiency of cutting amplification products with enzymes. The following primers are preferred for amplification of HLA-A, -B, -C, -E, -F, and -G cDNA, preferably in separate reactions. Resulting cDNAs may then be cloned and sequenced as deεcribed herein. These primers are appropriate for use in amplifying all known and presently unknown types of HLA. HLA A 5' primer: 5' CC ACC ATG GCC GTC ATG GCG CCC 3' (SEQ ID NO 1) 3' primer: 5' GG TCA CAC TTT ACA AGC TCT GAG (SEQ ID NO 2) HLA B 5' primer: 5' CC ACC ATG CTG GTC ATG GCG CCC 3 ' (SEQ ID NO 3) 3' primer: 5' GG ACT CGA TGT GAG AGA CAC ATC 3' (SEQ ID NO 4) HLA C 5' primer: 5' CC ACC ATG CGG GTC ATG GCG CCC 3' (SEQ ID NO 5) 3' primer: 5' GG TCA GGC TTT ACA AGC GAT GAG 3' (SEQ ID NO 6) HLA E 5' primer: 5' CC ACC ATG CGG GTA GAT GCC CTC C 3' (SEQ ID NO 7) 3' primer: 5' GG TTA CAA GCT GTG AGA CTC AGA (SEQ ID NO 8) HLA F 5' primer: 5' CC ACC ATG GCG CCC CGA AGC CTC (SEQ ID NO 9) 3' primer: 5' GG TCA CAC TTT ATT AGC TGT GAG A (SEQ ID NO 10) HLA G 5' primer: 5' CC ACC ATG GCG CCC CGA ACC CTC (SEQ ID NO 11) 3' primer: 5 ' GG TCA CAA TTT ACA AGC CGA GAG (SEQ ID NO 12) In preferred embodiments only one pair of first and second primers is used per amplification reaction. The amplification reaction products obtained from a plurality of different amplifications, each using a plurality of different primer pairs, are then combined. However, the present invention also relates to DNA homolog production via co-amplification (using two pairs of primerε) , and multiplex amplification (uεing up to about 8, 9 or 10 primer pairε) . In preferred embodimentε, the PCR process is used not only to produce a variety of human Class I-encoding DNA molecules, but also to induce mutations which may emulate those observed in the highly-polymorphic HLA loci, or to create diversity from a single parental clone and thereby provide a Clasε I MHC molecule-encoding DNA "library" having a greater heterogeneity. In addition to the mutation inducing variationε deεcribed in the above referenced U.S. Patent No. 4,683,195 and εuch aε diεcuεsed in U.S. Patent No. 5,314, 813. 2. DNA Expression Vectorε A vector of the present invention is a nucleic acid (preferably DNA) molecule capable of autonomous replication in a cell and to which a DNA εeg ent , e.g. , gene or polynucleotide, can be operatively linked εo aε to bring about replication of the attached εegment . One of the nucleotide εegmentε to be operatively linked to vector εequenceε encodeε at leaεt a portion of a mammalian Claεε I MHC heavy chain. Preferably, the entire peptide-coding εequence of the MHC heavy chain is inεerted into the vector and expreεεed; however, it iε alεo feaεible to conεtruct a vector which alεo includes some non-coding MHC sequenceε aε well. Preferably, non-coding εequenceε of MHC are excluded. Alternatively, a nucleotide εequence for a soluble ("sol") form of an Clasε I MHC heavy chain may be utilized; the "sol" form differs from the non-sol form in that it contains a "εtop" codon inεerted at the end of the alpha 3 domain or prior to the transmembrane domain. Another preferred vector includes a nucleotide sequence encoding at least a portion of a mammalian β- 2 microglobulin molecule operatively linked to the vector for expresεion. Still another preferred vector includeε a nucleotide εequence encoding at leaεt a portion of a mammalian assiεting molecule operably linked to the vector for expreεsion. It is also feaεible to conεtruct a vector including nucleotide sequenceε encoding a Class I MHC heavy chain and a β- 2 microglobulin and an asεisting molecule, or some combination of theεe. A preferred vector compriseε a caεεette that includes one or more translatable DNA sequences operatively linked for expreεεion via a sequence of nucleotides adapted for directional ligation. The cassette preferably includes DNA expression control sequences for expressing the polypeptide or protein that is produced when a translatable DNA sequence is directionally inserted into the cassette via the sequence of nucleotides adapted for directional ligation. The casεette also preferably includes a promoter sequence upstream from the translatable DNA sequence, and a polyadenylation sequence downstream from the mammalian MHC heavy chain sequence. The casεette may also include a selection marker, albeit it is preferred that such a marker be encoded in a nucleotide sequence operatively linked to another expresεion vector εequence. The choice of vector to which a caεεette of thiε invention iε operatively linked depends directly, as is well known in the art, on the functional properties desired , e.g., vector replication and protein expression, and the host cell to be tranεformed, these being limitations inherent in the art of constructing recombinant DNA molecules. In various embodiments, a vector is utilized for the production of polypeptides useful in the present invention, including MHC variants and antigenic peptides. Exemplary vectors include the plasmidε pUC8, pUC9, pUC18, pBR322, and pBR329 available from BioRad Laboratorieε (Richmond, CA) , pPL and pKK223 available from Pharmacia (Piscataway, NJ) , and pBS and M13mpl9 (Stratagene, La Jolla, CA) . Other exemplary vectors include pCMU (Nilsεon, et al . , Cell 58: 707 (1989)) . Other appropriate vectorε may alεo be εyntheεized, according to known methodε; for example, vectorε pCMU/K 1 ' and pCMUII uεed in various applications herein are modifications of pCMUIV (Nilsson, et al . , supra ) . In addition, there iε preferably a εequence upstream of the translatable nucleotide sequence' encoding a promoter sequence. Preferably, the promoter is conditional (e.g., inducible) . A preferred conditional promoter used herein is a metallothionein promoter or a heat shock promoter. Vectorε may be conεtructed utilizing any of the well-known vector conεtruction techniques. Thoεe techniqueε, however, are modified to the extent that the tranεlatable nucleotide εequence to be inserted into the genome of the host cell is flanked "upstream" of the sequence by an appropriate promoter and, in some variations of the present invention, the translatable nucleotide sequence is flanked "downstream" by a polyadenylation site. Thiε iε particularly preferred when the "hoεt" cell iε an inεect cell and the nucleotide sequence iε tranεmitted via tranεfection. Tranεfection may be accompliεhed via numerous methods, including the calcium phosphate method, the DEAE-dextran method, the stable tranεfer method, electroporation, or via the lipoεome mediation method. Numerouε texts are available which εet forth known tranεfection methodε and other procedureε for introducing nucleotides into cells; see, e.g., Current Protocols in Molecular Biology. John Wiley & Sons, NY (1991) . The vector itself may be of any suitable type, such as a viral vector (RNA or DNA) , naked straight-chain or circular DNA, or a vesicle or envelope containing the nucleic acid material and any polypeptides that are to be inserted into the cell. With respect to vesicles, techniques for construction of lipid vesicleε, such as liposomes, are well known. Such lipoεomeε may be targeted to particular cellε uεing other conventional techniques, such as providing an antibody or other specific binding molecule on the exterior of the liposome. See, e.g., A. Huang, et al . , J. Biol. Chem. 255: 8015-8018 (1980) . See, e.g., Kaufman, Meth. Enzvmol. 185: 487-511 (1990) . In a preferred embodiment, the vector also contains a selectable marker. After expression, the product of the translatable nucleotide sequence may then be purified using antibodies against that sequence. One example of a selectable marker is neo ycin reεiεtance. A plasmid encoding neomycin resiεtance, εuch as phεhεneo, phεneo, or pcopneo, may be included in each transfection such that a population of cells that expreεε the gene(s) of choice may be ascertained by growing the transfectants in selection medium. A preferred vector for uεe according to the preεent invention iε a plaεmid; more preferably, it is a high-copy-number plasmid. It is also desirable that the vector contain an inducible promoter sequence, aε inducible pro oterε tend to limit selection pressure against cellε into which εuch vectorε (which are often constructed to carry non-native or chimeric nucleotide sequences) have been introduced. It is also preferable that the vector of choice be best suited for expression in the chosen hoεt . If the host cell population is a Drosophila cell culture, then a compatible vector includes vectors functionally equivalent to those such as p25-lacZ (see Bello and Couble, Nature 346: 480 (1990) ) or pRmHa-1, -2 , or -3 (see Bunch, et al . , Nucl . Acids Res. 16: 1043-1061 (1988)) . In the preferred embodiment, the vector is pRmHa-3, which is shown in Figure 3. Thiε vector includes a etallothionein promoter, which is preferably upεtream of the site at which the MHC sequence is inserted, and the polyadenylation site is preferably downstream of εaid MHC εequence. Inεect cellε and, in particular, Drosophila cellε are preferred hosts according to the present invention. Drosophila cells such as Schneider 2 (S2) cells have the necesεary tranε-acting factorε required for the activation of the promoter and are thuε even more preferred. The expreεεion vector pRmHa-3 iε based on the bacterial plasmid pRmHa-1 (Figure 2) , the latter of which is based on plasmid pUC18 and is deposited with the American Type Culture Collection (ATCC, Rockville, MD) , having the accession number 37253. The pRmHa-3 vector contains the promoter, the 5' untranslated leader sequence of the metallothionein gene (sequences 1-421, SEQ ID NO 13) with the RI and Stu sites removed; see Figure 3) . It also contains the 3' portion of the Drosophila ADH gene (sequence #6435-7270, SEQ ID NO 14) including the polyadenylation site. Therefore, cloned DNA will be transcriptionally regulated by the metallothionein promoter and polyadenylated. Construction of the pRmHa-1 plasmid is described in Bunch, et al . , Nucl. Acids Res. IG: 1043-1061 (1988) . Construction of the pRmHa-3 and pRmHa-2 plasmids (the latter of which has a metallothionein promoter sequence that may be removed aε an Eco RI fragment) iε illustrated in Figures 1, 2, and 3. With regard to pRmHa-3, a preferred plasmid for use according to the present invention, Pst I, Sph I and Hind III are in the promoter fragment and therefore are not unique. Xba is in the ADH fragment (4 bases from its 3' end) and iε also not unique. The following restriction εiteε are, however, unique in pRmHa-3 : Eco RI, Sac I, Kpn I, Sma I, Bam HI, Sal I, Hinc 2, and Ace I. A cassette in a DNA expreεεion vector of this invention is the region of the vector that forms, upon insertion of a translatable DNA sequence, a sequence of nucleotides capable of expresεing, in an appropriate hoεt, a fusion protein of thiε invention. The expression-competent sequence of nucleotides is referred to as a cistron. Thus, the cassette preferably compriseε DNA expression control elementε operatively linked to one or more tranεlatable DNA εequenceε. A ciεtron iε formed when a translatable DNA sequence is directionally inserted (directionally ligated) between the control elements via the sequence of nucleotides adapted for that purpose. The resulting translatable DNA sequence, namely the mεerted εequence, lε, preferably, operatively linked in the appropriate reading frame. DNA expression control sequences comprise a set of DNA expresεion εignalε for expreεεing a εtructural gene product and include both 5' and 3' elementε, aε is well known, operatively linked to the cistron such that the cistron is able to express a εtructural gene product. The 5' control εequences define a promoter for initiating transcription and a riboεome binding εite operatively lιnκed at the 5' terminus of the upεtream tranεlatable DNA εequence . Thuε, a DNA expreεεion vector of thiε invention provides a system for cloning translatable DNA sequences in t o the cassette portion of the vector to produce a ciεtron capable of expressing a fusion protein of this invention. 3. Cell Lines A preferred cell line of the present invention is capable of continuous growth in culture and capable of expressing mammalian Class I MHC molecules and asεiεting moleculeε on the εurface of its cells. Any of a variety of transformed and non-transformed cellε or cell lineε are appropriate for thiε purpoεe, including bacterial, yeaεt, insect, and mammalian cell lines. (See, e.g., Current Protocols in Molecular Biology. John Wiley & Sons, NY (1991) , for summaries and procedures for culturing and using a variety of cell lines, e.g., E. coli and S. cerevisiae . ) Preferably, the cell line is a eukaryotic cell line. More preferably, the cell line is poikilothermic (i.e., less sensitive to temperature challenge than mammalian cell lines) . More preferably, it is an insect cell line. Various inεect cell lineε are available for use according to the present invention, including moth (ATCC CCL 80) , armyworm (ATCC CRL 1711) , mosquito larvae (ATCC lines CCL 125, CCL 126, CRL 1660, CRL 1591, CRL 6585, CRL 6586) and εilkworm (ATCC CRL 8851) . In a preferred embodiment, the cell line is a Drosophila cell line such as a Schneider cell line (see Schneider, J. Embrvol. Exp. Morph. 27: 353-365 (1972) ) ; preferably, the cell line is a Schneider 2 (S2) cell line (S2/M3) adapted for growth in M3 medium (see Lindquist, et al . , Drosophila Information Service 58: 163 (1982) ) . Schneider cells may be prepared substantially as follows. Drosophila elanogas ter (Oregon-R) eggs are collected over about a 4 hour interval and are dechlorinated in 2.5% aqueous sodium hypochlorite and surface-εterilized by immerεion in 70% ethanol for 20 minuteε, followed by an additional 20 minuteε in 0.05% HgCl-, in 70% ethanol. After being rinsed thoroughly in sterile distilled water, the eggs are tranεferred to petri dishes containing sterile Metricel black filters backed with Millipore prefilters, both previously wetted with culture medium. The eggs are placed overnight in a 22 °C incubator and removed for culturing when 20-24 hours old. The embryos are each cut into halves or thirds, then placed in 0.2% trypsin (1:250, Difco) in Rinaldini's salt solution (Rinaldini, Nature (London) r73: 1134-1135 (1954)) for 20-45 minutes at room temperature. From 100-300 embryoε are used to initiate each culture. After the addition of fetal bovine serum (FBS) , the fragments are centrifuged at 100 X g for 2-3 minutes, resuspended in 1.25ml culture medium and seeded into glass T-9 flasks. The cultures are maintained at about 22-27 °C ± 0.5 °C, with a gaseouε phaεe of ambient air. Schneider' culture medium (Schneider, J. Exp. Zool . 156 : 91-104 (1964) , * Schneider, J. Embrvol. Exp. Morph. 15: 271-279 (1966)) containing an additional 500mg bacteriological peptone per 100ml medium and supplemented with 15% inactivated FBS is preferably used. The pH (preferably 6.7-6.8) is monitored with 0.01% phenol red. The cell lines are preferably maintained by subculturing every 3-7 days. The cells readily attach to the glass but not so firmly aε to require trypsin treatment; typically, simple pipetting is adequate to flush most of the cells from the bottom of the flaεks. The morphological appearance of the cellε is described in Schneider, J. Embrvol. Exp. Morph. 27: 353-365 (1972) . They are essentially epithelial-like in appearance and range from about 5-llμm in diameter and ll-35μm in length. Small pockets containing rounded cells may be dispersed randomly throughout the other cells. Preferably, the Schneider 2 (S2) cells are maintained in Schneider's Drosophila medium plus 10% FBS including penicillin (100 unit/ml) and streptomycin (lOOmg/ml) . It is preferable to keep the cells at a density of more than 0.5 X lO ml, and to grow them at a 24-30 °C temperature range. The cells tend to double in fewer than 24 hours and grow to high cell denεity, i.e., about 2 X lOV l or greater. The cellε may alεo be frozen in 90% FBS and 10% DMSO, for later use or analysis. One may place the cells at -70 °C and then store in liquid nitrogen. A preferred cell line according to the present invention, identified as Schneider 2 (S2) cells, has been deposited pursuant to Budapest Treaty requirements with the American Type Culture Collection (ATCC) , Rockville, MD, on February 18, 1992, and was asεigned accession number CRL 10974. Cells of the present invention are transfected with cDNAs encoding (human) MHC heavy chains, β- 2 microglobulin and one or more assisting molecules, which have each been inserted into (i.e. , operatively linked to) an expresεion vector. In a more preferred embodiment, the vector comprises Drosophi la expression plasmid pRmHa-3, into which expreεsible nucleotide sequences encoding human Clasε I MHC heavy chains, human β- 2 microglobulin or human assisting molecules have been inserted using techniques discloεed herein. Preferably, the cDNAs encoding MHC heavy chains, those encoding β - 2 microglobulin and those encoding assisting molecules are operatively linked to separate expreεεion plaεmids and are cotransfected into the cultured cellε. Alternatively, the cDNAε encoding MHC heavy chainε, β- 2 microglobulin and assisting molecules may be operatively linked to the same expresεion plasmid and cotransfected via that same plasmid. In another variation, cDNAε encoding MHC heavy chains, β- 2 microglobulin, assisting moleculeε, and a cytokine such as IL-2 are operatively linked to expression plasmids and are cotransfected into a cell line of the present invention. Selection of HLA genes, construction of appropriate vectors and primer selection are described in greater detail above. Succesεfully transformed cells, i.e., cellε that contain an expressible human nucleotide sequence according to the present invention, can be identified via well-known techniques. For example, cells resulting from the introduction of a cDNA or rDNA of the present invention can be cloned to produce monoclonal colonies. Cellε from thoεe colonieε can be harveεted, lyεed, and their DNA content examined for the presence of the rDNA uεing a method such as that deεcribed by Southern, J. Mol. Biol. 98: 503 (1975) . In addition to directly assaying for the presence of rDNA, successful transformation or tranεfection may be confirmed by well-known immunological methodε when the rDNA iε capable of directing the expreεsion of a subject chimeric polypeptide. For example, cells succeεεfully transformed with an expression vector may produce proteins displaying particular antigenic properties which are easily determined using the appropriate antibodies. In addition, succeεεful tranεformation/tranεfection may be ascertained via the use of an additional vector bearing a marker sequence, such aε neomycin resistance, as described hereinabove. It iε also preferable that the culture be stabile and capable of sustained growth at reduced temperatures. For example, it is preferred that the culture be maintained at about room temperature, e.g. , about 24-27 °C. In other embodiments, the culture is maintained at higher temperatures, particularly during the process of activating CD8 * cells. It is thus preferred that a culture according to the present invention be capable of withstanding a temperature challenge of about 30 °C to about 37 °C. Addition of β- 2 microglobulin to a culture stabilizes the Claεε I MHC to at least a 30 °C challenge; addition of β- 2 microglobulin and peptideε results in greater thermostability at higher temperatureε, i.e., at 37 °C. In order to prepare the culture for expreεεion of empty -- or more preferably, peptide-loaded -- MHC moleculeε, the culture may firεt require εtimulation, e.g., via CuS0 4 induction, for a predetermined period of time. After a εuitable induction period -- e.g., about 12-48 hours, peptides may be added at a predetermined concentration (e.g., about lOOμg/ml) . Peptides may be prepared as discusεed below. After a further incubation period -- e.g., for about 12 hourε at 27 °C -- the culture iε ready for uεe in the activation of CD8' cellε. While this additional incubation period may be shortened or perhaps omitted, the culture tends to become increasingly stable to temperature challenge if it is allowed to incubate for a time prior to addition of resting or naive CD8' cellε. For example, cultureε according to the preεent invention to which peptide has been added are capable of expreεεing εignificant amounts of peptide-loaded Class I MHC molecules even when incubated for extended periods of time at 37 °C. Nutrient media useful in the culturing of tranεformed hoεt cells are well known in the art and can be obtained from numerous commercial sources. In embodiments wherein the host cell is mammalian, a "serum-free" medium is preferably used. 4. Human β- 2 microglobulin and Assisting Molecules In order to establiεh a cell line capable of producing therapeutically uεeful amounts of surface-expressed human Class I MHC molecules, it is preferable to cotransfect a cell line of the preεent invention with a vector operably linked to a nucleotide εequence encoding β- 2 microglobulin in order to effect appropriate levels of expresεion of human MHC moleculeε in the cell line. While the nucleotide εequence encoding mammalian β- 2 microglobulin such as mouse β- 2 microglobulin increases the stability of the human Class I MHC molecules expressed in the cell lines of the present invention, it is preferable to cotransfect the cell line with a vector operably linked to an expresεible nucleotide sequence encoding a human β- 2 microglobulin. As discuεεed above, a preferred vector according to the present invention includes a nucleotide εequence encoding at leaεt a portion of a mammalian β- 2 microglobulin molecule operatively linked to the vector for expresεion. The gene for the aεεisting molecules can be linked to the εame or another vector. It iε alεo feaεible to construct a vector including nucleotide sequences encoding both a Clasε I MHC heavy chain and a β- 2 microglobulin. The εequencing and primerε used for the assisting molecules are discussed in more detail below. However, the protocols are similar. A human β - 2 microglobulin cDNA sequence haε been publiεhed (εee Suggε, et al . , PNAS 78: 6613-17, 1981) and the εequence was used as a template for a polymerase chain reaction (PCR) uεing the following primerε : 5' primer: ' GCTTGGATCCAGATCTACCATGTCTCGCTCCGTGGCCTTAGCTGTGCT CGCGCTACTCTC 3 ' (SEQ ID NO 15) 3' primer 5 ' GGATCCGGATGGTTACATGTCGCGATCCCACTTAAC 3' ( SEQ ID NO 16 ) The primers are used in a standard PCR reaction (see above and references cited therein) . The reaction products are extracted with phenol, purified uεing a Geneclean kit (Bio 101, San Diego, CA) , digested with Bam HI and cloned into the Bam HI site of pBS (Stratagene, La Jolla, CA) . After verification of the sequence, thiε Bam HI fragment iε cloned into the Bam HI εite of an appropriate expreεεion vector. In the preferred embodiment, human β- 2 microglobulin cDNA iε synthesized and operably linked to expression vector pRmHa-3. 5. Peptides Virtually all cellular proteins in addition to viral antigens are capable of being used to generate relevant peptide fragmentε that εerve aε potential Class I MHC ligand. In most mammalian cells, then, any particular MHC peptide complex would represent only a small proportion of the total MHC encoded moleculeε found on the cell surface. Therefore, m order to produce surface-expressed human Clasε I MHC molecules that have an increased capacity to specifically activate CD8' cells, it is preferable to iεolate and load peptide fragmentε of appropriate εize and antigenic characteriεticε onto Claεε I moleculeε. The peptides of the present invention bind to Clasε I MHC moleculeε. The binding occurs under biological conditions which can be created in vi vo aε well aε in vi tro . The exact nature of the binding of the peptides need not be known for practice of the invention. In a preferred embodiment, the peptides to be loaded onto the Class I MHC molecules are antigenic. It is alεo preferred that the peptides be of a uniform size, preferably 8-mers or 9-mers, and moεt preferably, 8-merε. It is alεo preferable that the peptideε prepared for loading onto the MHC moleculeε be of a εingle species; i.e., that all peptides loaded onto the MHC be identical in size and sequence. In this manner, it is possible to produce monoantigenic peptide-loaded MHC moleculeε. Peptideε may be presented to the cells via various means. Preferably, peptides are presented in a manner which allows them to enter an intracellular pool of peptides. For example, peptides may be preεented via oεmotic loading. Typically, peptideε are added to the culture medium. The peptideε may be added to the culture in the form of an intact polypeptide or protein which is subsequently degraded via cellular procesεeε, e.g., via enzymatic degradation. Alternatively, the intact polypeptide or protein may be degraded via εome other meanε εuch aε chemical digeεtion (e.g. cyanogen bromide) or proteaεeε (e.g. chymotrypεin) prior to itε addition to the cell culture. In other embodiments, the peptides are presented in smaller segmentε which may or may not compriεe epitopic amino acid εequenceε. In a preferred embodiment, a εufficient amount of protein (ε) or peptide (ε) iε added to the cell culture to allow the Claεε I MHC moleculeε to bind and subεequently present a large density of the peptide -- preferably, with the same kind of peptide attached to each MHC -- on the surface of human Class I MHC-expressing cellε of the preεent invention. It iε alεo preferred to allow the human Claεs I MHC heavy chains and human β- 2 microglobulin to bind -- i.e., to form heterodimers -- prior to preεenting peptide to the MHC moleculeε intracellularly. In another embodiment of the invention, peptideε are added to tranεfected cells of the present invention in order to enhance the thermostability of the MHC moleculeε expressed by the cells. As noted above, peptides are preferably added to the culture medium. Antigenic peptides that bind to the Class I molecules serve to thermostabilize the MHC molecules and also increase the cell surface expression. Cultures with added peptides which bind to the MHC molecules are thus significantly less susceptible to temperature challenge than cultures without added peptide. In one embodiment of the present invention, antigenic peptides are presented to the transformed/transfected cell line in various forms. For example, an entire protein or other antigenic polypeptide may be degraded chemically or enzymatically, for example, and added to the cell line in this form. For example, a protein of interest is degraded with chymotrypsin and the resultant mixture of peptide "fragments" is added to a transformed or transfected cell culture; these cells are then allowed to "choose" the appropriate peptides (which are often smaller peptides, preferably 8mers or 9mers) to load onto the Clasε I MHC molecules. Alternatively, an entire protein or polypeptide sequence may be cloned into an appropriate vector and inserted into a procaryotic cell, whereby the cell generates significant amounts of the antigenic polypeptide which are then harvested, purified, and digested into peptides which are then added to the transformed/tranεfected eukaryotic cell culture. The cells again would be allowed to "choose" the peptides to load onto the expreεεed MHC. 6. Iεolation of Resting or Precursor CD8' cells Resting (or naive or precursor) CD8' cellε -- i.e. , T-cellε that have not been activated to target a εpecific antigen -- are preferably extracted from the patient prior to incubation of the CD8' cells with the transformed cultures of the present invention. It is alεo preferred that precurεor CDS " cellε be harvested from a patient prior to the initiation of other treatment or therapy which may interfere with the CD8' cellε' ability to be specifically activated. For example, if one is intending to treat an individual with a neoplasia or tumor, it iε preferable to obtain a εample of cellε and culture εame prior to the initiation of chemotherapy or radiation treatment. Methodε of extracting and culturing lymphocytes are well known. For example, U.S. Patent No. 4,690,915 to Rosenberg describes a method of obtaining large numbers of lymphocytes via lymphocytopheresiε . Appropriate culturing conditions uεed are for mammalian cells, which are typically carried out at 37 °C. Various methods are also available for separating out and/or enriching cultures of precursor CD8 * cells. Some examples of general methods for cell separation include indirect binding of cells to specifically-coated surfaces In another example, human peripheral blood lymphocytes (PBL) , which include CD8' cells, are isolated by Ficoll-Hypaque gradient centrifugation (Pharmacia, Piscataway, NJ) . PBL lymphoblastε may be uεed immediately thereafter or may be stored in liquid nitrogen after freezing in FBS containing 10% DMSO (Sigma Chemical Co., St. Louis, MO) , which conserves cell viability and lymphocyte functions Alternative methods of separating out and/or enriching cultures of precursor cells include both positive and negative selection procedureε. For positive selection, after lymphocyte-enriched PBL populations are prepared from whole blood, sub-populations of CD8 * lymphocyteε are lεolated therefrom by affmity-based separation techniques directed at the presence of the CD8 receptor antigen. These affmity-based techniques include flow microfluorimetry, including fluorescence-activated cell sorting (FACS) , cell adhesion, and like methods. (See, e.g., Scher and Mage, in Fundamental Immunology. W.E. Paul, ed. , pp. 767-780, River Press, NY (1984) .) Affinity methods may utilize anti-CD8 receptor antibodies as the source of affinity reagent. Alternatively, the natural ligand, or ligand analogs, of CD8 receptor may be used as the affinity reagent. Various anti-T-cell and anti-CD8 monoclonal antibodies for use in these methods are generally available from a variety of commercial sources, including the American Type Culture Collection (Rockville, MD) and Pharmingen (San Diego, CA) . Negative selection procedures are utilized to effect the removal of non-CD8 from the CD8' population. This technique reεults in the enrichment of CD8 * cells from the T- and B-cell population of leucophoreεed patients. Depending upon the antigen designation, different antibodieε may be appropriate. (For a diεcuεεion and review of nomenclature, antigen deεignation, and aεsigned antibodies for human leucocytes, including T-cells, see Knapp, et al . , Immunology Today 10: 253-258 (1989) and Janeway et al . , Immunobiology, supra . ) For example, monoclonal antibodies OKT4 (anti-CD-I , ATCC No. CRL 8002) OKT 5 (ATCC Nos. CRL 8013 and 8016) , OKT 8 (anti-CD8, ATCC No. CRL 8014) , and OKT 9 (ATCC No. CRL 8021) are identified in the ATCC Catalogue of Cell Lines and Hybridomas (ATCC, Rockville, MD) aε being reactive with human T lymphocyteε, human T-cell εubεetε, and activated T-cells, respectively. Various other antibodieε are available for identifying and iεolating T-cell εpecieε. In a further embodiment, CD8' cells can be isolated by combining both negative and positive selection procedures. (See, e.g. Cai and Sprent , J . Exp. Med. 179: 2005-2015 (1994) ) . Preferably, the PBLε are then purified. For example, Ficoll gradientε may be utilized for thiε purpose. The purified PBLs would then be mixed with syngeneic Drosophila cells preincubated with the appropriate antigenic peptides. 7. In Vi tro Activation of CD8' Cells In order to optimize the in vi tro conditions for the generation of specific cytotoxic T-cells, the culture of antigen-presenting cells iε maintained in an appropriate medium. In the preferred embodiment, the antigen-presenting cells are Drosophi la cells, which are preferably maintained in serum-free medium (e.g. Excell 400) . Prior to incubation of the antigen-presentmg cells with the cells to be activated, e.g., precursor CD8 * cells, an amount of antigenic peptide is added to the antigen-presenting cell culture, of sufficient quantity to become loaded onto the human Class I molecules to be expreεsed on the surface of the antigen-presenting cells. According to the present invention, a sufficient amount of peptide is an amount that will allow about 200 to about 500,000 and preferably about 200 to 1,000 or more, human Clasε I MHC molecules loaded with peptide to be expreεsed on the surface of each antigen-presentmg cell. Preferably, the antigen-presentmg cells are incubated with >20μg/ml peptide. Resting or precursor CD8' cells are then incubated in culture with the appropriate antigen-presenting cells for a time period sufficient to activate and further enrich for a population of CD8' cells. Preferably, the CD8' cells shall thus be activated in an antigen-specific manner. The ratio of resting or precursor CD8' (effector) cells to antigen-presentmg cells may vary from individual to individual and may further depend upon variables εuch as the amenability of an individual's lymphocyteε to culturing conditions and the nature and severity of the disease condition or other condition for which the within-described treatment modality is used. Preferably, however, the lymphocyte :antigen-presenting cell (e.g. Drosophila cell) ratio is preferably in the range of about 30:1 to 300:1. For example, in one embodiment, 3 X 10 7 human PBL and 1 X 10 c live Drosophila cells were admixed and maintained in 20 ml of RPMI 1640 culture medium. The effector/antigen-presenting culture may be maintained for as long a time aε is necesεary to activate and enrich for a population of a therapeutically useable or effective number of CD8' cellε. In general termε, the optimum time is between about one and five days, with a "plateau" -- i.e. a "maximum" specific CD8' activation level -- generally being observed after five days of culture. In one embodiment of the present invention, in vi tro activation of CD8' cellε iε detected within a brief period of time after tranεfection of a cell line. In one embodiment, tranεient expreεsion in a transfected cell line capable of activating CD8' cells is detectable within 48 hourε of tranεfection. This clearly indicates that either stable or transient cultures of transformed cells expresεing human Claεs I MHC molecules are effective in activating CD8 4 cells. Preferably, the enrichment and concordant activation of CD8' cellε iε optimal within one week of exposure to antigen-preεenting ceilε. Thereafter, in a preferred embodiment, the enriched and activated CD8 ' cellε are further purified by iεolation procedures including site restriction, roεetting with antibody-red blood cell preparationε, column chromatography and the like. Following the purification, the resulting CD8 * cell preparation is further expanded by maintenance in culture for a period of time to obtain a population of 10 J activated CD8 * cells. This period may vary depending on the replication time of the cells but may generally be 14 days. Activation and expansion of CD8 * cells has been described by Riddell et al., Curr. Qpin. Immunol., 5: 484-491 (1993) . 8. Separation of CD8 * Cells from Drosophila Cells Activated CD8 * cells may be effectively separated from the stimulator (e.g., Drosophila ) cells using one of a variety of known methods. For example, monoclonal antibodies specific for the stimulator cells, for the peptides loaded onto the stimulator cells, or for the CD8' cells (or a segment thereof) may be utilized to bind their appropriate complementary ligand. Antibody-tagged cells may then be extracted from the stimulator-effector cell admixture via appropriate means, e.g., via well-known immunoprecipitation or immunoassay methodε . 9. Adminiεtration of Activated CD8' Cells Effective, cytotoxic amounts of the activated CDS' cellε can vary between in vi tro and in vi vo uses, as well aε with the amount and type of cells that are the ultimate target of these killer cellε. The amount will alεo vary depending on the condition of the patient and εhould be determined via consideration of all appropriate factors by the practitioner. Preferably, however, about 1 X 10' to about 1 X 10 ,:' , more preferably about 1 X 10" to about 1 X 10 , and even more preferably, about 1 X 10 " to about 1 X 10 1 " activated CD8' cellε are utilized for adult humanε, compared to about 5 X 10" - 5 X 10 ' cells used in mice . Preferably, as discuεεed above, the activated CD6' cells are harvested from the Drosophila cell culture prior t o administration of the CD8' cellε to the individual being treated. It iε important to note, however, that unlike other preεent and propoεed treatment modalities, the present method useε a cell culture εyεtem (i.e., Drosophila cellε) that are not tumorigenic . Therefore, if* complete εeparation of Drosophila cells and activated CD8 * cells is not achieved, there is no inherent danger known to be associated with the administration of a small number of Drosophila cells, whereas administration of mammalian tumor-promoting cells may be extremely hazardous. Methods of re-introducing cellular components are known in the art and include procedures such as those exemplified in U.S. Patent No. 4,844,893 to Honsik, et al. and U.S. Patent No. 4,690,915 to Rosenberg. For example, administration of activated CD8 * cells via intravenous infusion is appropriate. 10. HLA Typing As noted previously, HLA haplotypes/allotypes vary from individual to individual and, while it is not essential to the practice of the present invention, it is often helpful to determine the individual's HLA type The HLA type may be determined via εtandard typing procedureε and the PBLε purified by Ficoll gradients The purified PBLs would then be mixed with syngeneic Drosophila cellε premcubated with the appropriate antigenic peptideε -- e g., in therapeutic applications relating to viral infections, cancers, or malignancies, peptideε derived from viral- or cancer-specifie proteins Continuing to use viral or malignant conditions aε an example, in thoεe instances in which specific peptideε of a particular viral- or cancer-εpecifie antigen have been characterized, the εyntheεized peptideε encoding theεe epitopes will preferably be used. In caseε in which the preferred antigenic peptideε have not been preciεely determined, protease digests of viral- or cancer-specifie proteins may be used As a source for such antigen, cDNA encoding viral- or cancer-specific proteins iε cloned into a bacterial expression plasmid and used to transform bacteria, e.g., via methods disclosed herein. After HLA typing, if Drosophila cells expresεing the preferred HLA are not available, cDNAε encoding the preferred HLA may be cloned via use of the polymeraεe chain reaction. The primerε diεclosed in section B.l. above (SEQ ID NO 1 through SEQ ID NO 12) may be used to amplify the appropriate HLA-A, -B, -C, -E, -F, or -G cDNAs in separate reactions which may then be cloned and sequenced as described in the methods discloεed for HLA A2.1 below. Stable cell lineε expressing the cloned HLA may then be establiεhed in the Drosophila cells. Alternatively, a population of insect cells transiently expressing a bulk population of cloned recombinant molecules from the PCR reaction may be used for in vi tro CD8' activation. EXAMPLES The following examples are intended to illustrate, but not limit, the preεent invention. Example 1 Expression of Human Claεs I MHC Molecules A. Preparation of pRmHa-3 Expresεion Vector The pRmHa-3 expreεεion vector for uεe in expreεεing MHC proteinε in Drosophila Schneider 2 (S2) cells as deεcribed in thiε invention was constructed by ligating a Sph I linearized pRmHa-1 DNA expression vector with a DNA fragment resulting from a Sph I restriction digest of a pRmHa-2 expression vector as deεcribed below. The ligating of pRmHa-1 with the pRmHa-2 fragment in this manner was performed to remove one of two Eco RI restriction endonuclease cloning εiteε preεent in pRmHa-1. Thuε, the reεultant pRmHa-3 expreεεion vector contained only one Eco RI reεtriction site in the multiple cloning εite (polylinker) into which variouε MHC-encoding DNA fragmentε were inserted aε described in the Examples. 1. Preparation of pRmHa-1 Expression Vector The pRmHa-1 expression vector, containing a metallothionein promoter, metal reεponεe consensus sequenceε (deεignated MT) and an alcohol dehydrogenase (ADH) gene containing a polyadenylation signal isolated from Drosophila melanogaster, was constructed aε deεcribed by Bunch et al . , Nucl. Acids Res. 16: 1043-61 (1988) . A schematic of the final pRmHa-1 construct is shown in Figure 2. The plasmid expresεion vector, pUC18, having the ATCC acceεsion number 37253, was used as the source vector from which subsequent vectors described herein were derived. The pUC18 plasmid contains the following restriction sites from 5' to 3' in the multiple cloning site, all of which are not illustrated in the schematic representationε of the pUC18-derived vectorε in Figure 1: Eco RI ; Sac I; Kpn I; S a I and Sma I located at the εame poεition; Bam HI; Xba I; Sal I, Ace I and Hinc II located at the same position; Pst I; Sph I and Hind III. The pUC18 vector waε firεt digeεted with Hind III to form a linearized pUC18. Blunt ends were then created by filling in the Hind III ends with DNA polymerase I large fragment aε deεcribed by Maniatis et al . , Molecular Cloning: A Laboratory Manual, edε . Cold Spring Harbor Laboratory, New York (1982) . The reεultant linearized blunt-ended pUC18 vector was ligated with a 740 base pair (bp) Hinf I fragment from the Drosophila melanogaster ADH gene containing a polyadenylation signal. The ligated ADH allele was first isolated from the plasmid pSACI , described by Goldberg et al . , PNAS USA 77: 5794-5798 (1980) , by digestion with Hinf I followed by blunt ending with Klenow resulting in the nucleotide sequence listed in SEQ ID NO 14. The pSACI vector containing the ADH allele was constructed by subcloning into pBR322 (ATCC accession number 31344) a 4.7 kilobase (kb) Eco RI fragment of Drosophila DNA selected from a bacteriophage lambda library containing random, high molecular weight (greater than 15 kb) . The 5' Hinf I restriction site occurred naturally in the ADH gene at position 1770 as described by Kreitman, Nature 304 : 412-417 (1983) . The 3' Hinf I site waε derived from the pUC18 vector into which the ADH gene had been cloned. Thiε poεition waε four bases 3' to the Xba I site at position 2500 of the ADH gene. The ADH segment extended from the 35 bp upstream of the polyadenylation/cleavage sequence in the 3' untranslated portion of the ADH mRNA to 700 bp downstream of the polyadenylation signal. The resultant pUC18-derived vector containing the ADH gene fragment was designated pHA-1 as shown in Figure 1. The 421 bp Eco Rl/Stu I MT gene fragment was obtained from a clone containing DNA of approximately 15.3 kb in a Drosophila melanogaster genomic DNA library. The library, prepared with a Mbo I partial digestion of imaginal DNA, waε cloned in the lambda derivative EMBL4. The fragment contained the MT promoter and metal response consenεuε elements of the Drosophi la MT gene (Maroni et al. , Genetics 112: 493-504 (1986) ) . Thiε region, containing the promoter and tranεcription εtart site at nucleotide 1+, corresponded to position -370 to nucleotide position +54 of the MT gene (SEQ ID NO 13) . The resultant fragment was then ligated into pHA-1 expresεion vector prepared above that waε previouεly linearized with Eco RI and Sma I. The 3' blunt end in MT created by the Stu I digeεt waε compatible with the blunt end in pHA-1 created by the Sma I digeεt. The reεultant pUC18-derived vector containing a 5' Drosophila MT gene fragment and a 3' ADH gene fragment was designated pRmHa-1. The pRmHa-1 expression vector, shown in Figure 2, contained the origin of replication (ori) and the beta-lactamase gene conferring resiεtance to ampicillin (Amp') from pUC18 as shown in Figure 1 on the pHa-1 vector. The diagram of pRmHa-1 also showε the 5' to 3' contiguouε positions of the MT gene fragment, the multiple cloning site and the ADH gene fragment. The pRmHa-1 vector was used as described in c. below in the construction of the pRmHa-3 expresεion vector. 2. Preparation of pRmHa-2 Expreεεion Vector The conεtruction of pRmHa-2 iε shown in Figure 1. For constructing the pRmHa-2 expression vector, the MT fragment prepared above was inεerted into the pUC18-derived vector pHA-1 aε deεcribed for conεtructing pRmHa-1 above with a few odificationε . An Eco RI linker waε added to the Stu I εite of the Eco Rl/Stu I-isolated MT gene fragment prepared above to form a metallothionein fragment having Eco RI restriction siteε on both endε . The reεultant fragment was then ligated into the ADH fragment-containing pUClβ expression vector that was previously linearized with Eco RI . The reεultant pUClδ-derived vector containing a 5' Drosophila MT gene fragment and a 3' ADH gene fragment having two Eco RI restriction εiteε 5' to the multiple cloning εite waε deεignated pRmHa-2. The pRmHa-2 expression vector, shown in Figure 1, contained the origin of replication (ori) and the beta-lactamase gene conferring resistance to ampicillin (Amp 1 ) from pUClθ. The diagram of pRmHa-2 also εhows the 5' to 3 ' contiguous positionε of the MT gene fragment, the multiple cloning εite and the ADH gene fragment. The pRmHa-2 vector waε uεed along with pRmHa-1 as described in c. below in the construction of the oRmHa-3 expression vector. . Preparation of pRmHa-3 Expression Vector To prepare the pRmHa-3 expresεion vector that had only one Eco RI reεtriction εite, a fragment from pRmHa-2 waε ligated into pRmHa-1. For thiε conεtruction, pRmHa-2, prepared in b. above, waε firεt digested with Sph I . The resultant Sph I fragment beginning in the middle of the MT gene and extending to the Sph I site in the multiple cloning site was first isolated from the pRmHa-2 vector and then ligated into pRmHa-1 prepared in A.l. above. The pRmHa-1 vector was previously modified to remove the Eco RI restriction site 5' to the MT gene fragment then linearized with Sph I . This procesε is schematically illustrated in Figure 2. To remove the Eco RI site in pRmHa-1, the vector was first digested with Eco RI to form a linearized vector, then blunt ended with Mung Bean nuclease and religated. The pRmHa-1 vector lacking an Eco RI site was then digested with Sph I to remove the region corresponding to the Sph I fragment insert from pRmHa-2 and form a linearized pRmHa-1 vector. The Sph I fragment from pRmHa-2 waε then ligated into the Sph I linearized pRmHa-1 to form the pRmHa-3 expression vector. A schematic of the pRmHa-3 vector is shown in Figure 3. The relative positions of the various restriction sites from the pUClδ vector from which pRmHa-3 was derived are indicated on the figure. In addition, the relative positions and lengths of the MT and ADH gene fragments separated by the multiple cloning site (polylinker) into which the MHC gene of intereεt iε cloned are indicated on the figure. The pRmHa-3 vector, being derived from pUC18, containε the pUClδ origin of replication and beta-lactamase gene conferring ampicillin resistance. Thus, MHC encoding DNA fragments as prepared in this invention and cloned into the multiple cloning site of pRmHa-3 were transcriptionally regulated by the MT promoter and polyadenylated via the ADH gene. B. cDNA Synthesis Detailed descriptions of the cDNA of Clasε I MHC molecules of various HLA groups can be found in U.S. Patent No. 5,314,813 to Peterson et al . which has been incorporated by reference. cDNAs encoding any preferred HLA may be cloned via use of the polymerase chain reaction. The primers discloεed in section B.l. above (SEQ ID NO 1 through SEQ ID NO 12) may be used to amplify the appropriate HLA-A, -B, -C, -E, -F, or -G cDNAs in separate reactions which may then be cloned and sequenced as deεcribed in the methodε disclosed for HLA A2.1 above. Preparation of cDNA from human cells is carried out as described in Ennis, et al . , PNAS USA 87: 2833-2837 (1990) . Briefly, a blood sample is obtained from the individual and cells are collected after centrifugation and used to prepare total RNA. First strand cDNA is εynthesized by using oligo(dT) and avian myeloblastosiε virus reverse transcriptaεe. The resulting cDNA is used in a PCR amplification reaction utilizing the appropriate primer (s) as noted in section B.l. above, and a GeneAmp kit and thermal cycler (Perkin-El er/Cetus) . Reaction conditions are preferably as follows. lOOng cDNA template and 50 picomoles of each oligonucleotide primer are used. Thirty cycles are run as followε: (a) one minute at 94 °C; (b) one minute at 60 °C; and (c) one minute, 30 εecondε at 72 °C. The PCR reaction iε then heated to 100 °C for 10 minuteε to kill the Taq polymeraεe and the endε of the DNA made blunt by T4 polymerase (Stratagene, San Diego, CA) . To syntheεize HLA A2.2 , cDNA encoding a complete A2.2 (εee Holmeε, et al . , J. Immunol. 139: 936-41 (1987) , for the publiεhed sequence) is cloned into an M13mpl9 plasmid, a commercially available bacteriophage vector (Stratagene, La Jolla, CA) . cDNA is synthesized by PCR using primers derived from the published sequence of A2. The cDNA is released from an M13mpl9 clone as a Not I (overhang filled with Klenow) /Eco RI fragment. (Klenow fragments are part of the E. coli DNA polymerase I molecule, produced by the treatment of E. coli DNA pol I with subtilisin. They are used to "fill out" 5' or 3' overhangs at the ends of DNA molecules produced by restriction nucleases . ) The Not I/Eco RI fragment iε inεerted into pSP64T digeεted with Bg III (endε filled with Klenow) and Eco RI . pSP64T iε an SP6 cloning vector designed to provide 5' and 3' flanking regions from an mRNA which is efficiently translated (β-globin) to any cDNA which contains itε own initiation codon. Thiε translation SP6 vector was constructed by digesting pSP64-X3m with Bal I and Bst EII, filling in the εtaggered ends with T4 DNA polymerase and adding a Bgl II linker by ligation. Bal I cutε the S-globin cDNA two baεeε upstream of the ATG (start codon) and Bst EII cutε eight baεeε upεtream of the TAA (εtop codon) . There iε only one Bgl II site in pSP64T so that restriction enzymes cutting in the polylinker fragment, from Pst I to Eco RI can εtill be uεed to linearize the plaεmid for tranεcription. (See Kreig and Melton, Nucleic Acid Res. 12 : 7057-7070, (1984) , which also describeε the conεtruction of the plaεmid pSP64-X3m.) The reεulting plaεmid iε cleaved with Eco RI (end filled with Klenow) and Hind III which iε cloned into the pCMUII polylinker between Hind III (5') and Stu I (3') . (See Paabo, et al . , EMBO J. 5 : 1921-1927 (1986) .) The entire cDNA iε removed aε a Hind III (end filled with Klenow) Bam HI fragment which iε cloned into pRmHa-3 cleaved with Sma I and Bam HI . HLA A2.2 soluble form was prepared by engineering a stop codon into the above-described A2.2 cDNA immediately preceding the transmembrane domain. The modification is achieved by cleaving the A2.2 cDNA cloned in the eukaryotic expression vector pCMUII between Hind III 5' and Stu I 3' (see above) with Mbo II and Bam HI inserting the following oligonucleotides : 5' primer: 5' GGAGCCGTGACTGACTGAG 3' (SEQ ID NO 17) 3' primer: 5' CCCTCGGCACTGACTGACTCCTAG 3' (SEQ ID NO 18) The resulting recombinant plasmid is cleaved with Hind III, the overhanging end filled with Klenow, then cut with Bam HI releasing a restriction fragment which is cloned into pRmHa-3 in the same way as A2.2 full length. 1. Construction of Murine ICAM-1 Expression Vector Spleen cells were isolated from Balb/c mice. The spleen cells were stimulated with conA; RNA was isolated using the FastTrack kit (Invitrogen, San Diego, CA) according to the manufacturerε' instructions. cDNA was syntheεized from the mRNA uεing AMV reverεe transcriptase kit (Promega, Madison, WI) according to the manufacturers' instructions. Baεed on the publiεhed cDNA nucleotide εequence (Siu, G. et al . , J. Immunol. 143, 3813-3620 (1989) the following oligonucleotides were syntheεized aε PCR primers: 5' : TTTAGAATTCAC CATGGCTTCA ACCCGTGCCA AG 3 ' : TTTAGTCGACTC AGGGAGGTGG GGCTTGTCC The cDNA syntheεized waε subjected to PCR using theεe primerε. The product waε cleaved with the reεtriction enzymeε Eco RI and Sal I and ligated into pRmHa-3, which had been digeεted with the reεtriction enzymeε Eco RI and Sal I . 2. Conεtruction of Murine B7.1 Expreεεion Vector Spleen calls were isolated from Balb/c mice and stimulated with conA. Messenger RNA was isolated using the FastTrack kit (Invitrogen, San Diego, CA) according to the manufacturer's instructions. cDNA was syntheεized from the mRNA using AMV reverse transcriptase kit (Promega, Madison, WI) according to the manufacturer' ε inεtructions. Based on the published cDNA nucleotide sequence (Freeman, et al . , J. Exp. Med. 174: 625-631 (1991) ) the following oligonucleotides were synthesized aε PCR primerε : 5' : TTTAGAATTCAC CATGGCTTGC AATTGTCAGT TG 3 ' : TTTAGTCGACCT AAAGGAAGAC GGTCTGTTC The cDNA synthesized was subjected to PCR using these primers. The product was cleaved with the restriction enzymes Eco RI and Sal I and ligated into pRmHa-3, which had been digested with the restriction enzymes Eco RI and Sal I. 3. Construction of Murine B7.2 Expresεion Vector IC-21 cellε (obtained from ATCC) were propagated in RPMI 1640 medium containing 10% Fetal Calf Serum. mRNA waε isolated from these cells using the FaεtTrack kit (Invitrogen, San Diego, CA) according to the manufacturer's instructions. cDNA was synthesized from the mRNA using AMV reverse transcriptaεe kit (Promega, Madiεon., WI) according to the manufacturer's instructions. Based on the published cDNA nucleotide sequence (Freeman, et al . , J. Exp. Med. 178: 2185-2192 (1993)) the following oligonucleotides were syntheεized aε PCR primers: 5 ' : TTTAGAATTCAC CATGGACCCC AGATGCACCA TGGG 3 ' : TTTAGTCGACTC ACTCTGCATT TGGTTTTGCT GA The cDNA syntheεized was subjected to PCR uεing theεe primerε. The product was cleaved with the reεtriction enzymes Eco RI and Sal I and ligated into pRmHa-3, which had been digested with the restriction enzymes Eco RI and Sal I. The above expression constructε were tranεfected into Drosophila S2 cellε uεing the calcium phosphate method as listed in Table 1. Stable cell lines were selected by including 500 μg/ml Geneticin in the cell culture medium. TABLE 1 1 A 12 12 2 B 8 8 3 C 8 8 4 c 8 8 8 5 D 6 6 6 6 E 6 6 6 6 7 F 6 6 6 6 8 G 4.8 4.8 4.8 4.8 4.8 Human accessory and costimulatory moleculeε were cloned from human cell lines demonstrated to express theεe proteinε by FACS analyεiε with monoclonal antibodieε εpecific for the particular proteinε. Adheεion moleculeε belonging to the integrin family, ICAM-I (CD54) and LFA-3 (CD58) , were cloned from human cell lineε K562 and HL60, respectively. The K562 cells, originated from human chronic myelogenouε leukemia, were obtained from ATCC (CCL-243) and cultured under conditionε recommended (i.e., RPMI with 10% fetal calf serum at 37 degrees C with 5% C0 . HL60 cells, originated from a human promyelocytic leukemia, and were obtained from ATCC (CCL-240) and were cultured according to ATCC's recommendationε . Costimulatory molecules B7.1 and B7.2 were also cloned from K562 and HL60 cells respectively. 4. cDNA Messenger RNA samples were prepared from each cell line from RNA isolated by the modified guanidiniu thiocyanate method (Chromczynski, et al . Anal. Biochem. 162 : 156-159, 1987) followed by poly A+ RNA selection on oligo (dt) -cellulose columns (Sambrook, J., et al, Molecular Cloning: A Laboratory Manual, Second Edition, 6.22-6.34, Cold Spring Harbor laboratory, CSH, NY) , Induction of HL60 cells with vitamin D3 (usually required to express some cell surface molecules) waε not required to obtain the B7.2 and LFA-3 moleculeε, the proteinε were expressed in the absence of induction. cDNA was prepared using AMV reverse transcriptase kit according to the manufacturers' instructionε (Promega, Madison WI) . 5. PCR Primers PCR primers were deεigned and εyntheεized after obtaining copieε of the known εequenceε from the GENEWORKS databaεe (Intelligeneticε) and conεidering the endε needed to clone into the appropriate vectorε . They are aε follows with the top sequence of each protein the 5 'primer and the bottom one the 3 'primer: B7.1 5' -ACCCTTGAAT CCATGGGCCA CACACGGAGG CAG-3' 5 ' -ATTACCGGAT CCTTATACAG GGCGTACACT TTCCCTTCT-3' B7.2 5 ' -ACCCTTGAGC TCATGGATCC CCAGTGCACT ATG-3' 5' -ATTACCCCCG GGTTAAAAAC ATGTATCACT TTTGTCGCAT GA-3' LFA-3 5' -ACCCTTGAGC TCATGGTTGC TGGGAGCGAC GCGGGG-3' 3 ' -ATTACCGGAT CCTTAAAGAA CATTCATATA CAGCACAATA CA-3' ICAM-1 5' -ACCCTTGAAT TCATGGCTCC CAGCAGCCCC CGGCCC-3' 3' -ATTACCGGAT CCTCAGGGAG GCGTGGCTTG TGTGTTCGG-3' 6. Expression of DNA Fragment The cDNA preparations from each of the cell lines was used to clone the desired proteins. The polymerase chain reaction was used to generate cDNA fragments utilizing the appropriate PCR primer (see above) . The appropriate DNA fragmentε were cloned into the Droεophila fly vector pRMHA-3. Plasmid preparations have been prepared from all of the preparations and are now ready for transfection into the fly cells. Human β- 2 microglobulin cDNA is prepared using a published partial cDNA sequence (see Suggs, et al . , PNAS 78 : 6613-17, 1981) is used as a template for a polymerase chain reaction (PCR) with the following primers : 5' primer 5' GCTTGGATCCAGATCTACCATGTCTCGCTCCGTGGCCTTAGCTGTGCTCGC GCTACTCTC 3' (SEQ ID NO 15) 3' primer 5' GGATCCGGATGGTTACATGTCGCGATCCCACTTAAC 3' (SEQ ID NO 16) The primerε are used in a standard PCR reaction (see Nilεεon, et al . , Cell 58: 707 (1989) ) . The reaction productε are extracted with phenol, purified uεing a Geneclean kit (Bio 101, San Diego, CA) , digeεted with Bam HI and cloned into the Bam HI εite of pBS (Stratagene, La Jolla, CA) . After verification of the εequence, thiε Bam HI fragment iε cloned into the Bam HI εite of pRmHa-3. Aε noted in the Exampleε, murine Claεε I cDNA waε utilized in variouε inεtanceε. Murine Claεε I cDNA waε prepared aε followε. H-2K 1 ': cDNA encoding a complete K molecule is obtained from an expression plasmid pCMU/K h conεtructed aε followε. A partial H-2K 1 ' cDNA missing the leader εequence and most of the alpha I domain iε prepared according to the method of Reyes, et al . , PNAS 79 : 3270-74 (1982) , producing pH202. This cDNA is used to generate a full-length molecule. The missing sequence is provided using a genomic clone encoding H-2K b (Caligan, et al., Nature 291: 35-39, 1981) as a template in a PCR reaction, using a 5' primer flanked by a Not I site, followed by 21 nucleotides encoding the last seven amino acids of the leader sequence and 18 nucleotides complementary to the beginning of the alpha I domain and a 3' primer complementary to the region encompassing the Sty I site. The resulting fragment is ligated with pH202 at the Sty I site. The 5' sequence encoding the remainder of the signal sequence is obtained form the D cDNA (see below) as a Bam Hi/Not I fragment. The entire coding sequence is cleaved from the expresεion plaεmid aε a Bam HI fragment and cloned into pRmHa-3 cleaved with Bam HI . H-2L d : cDNA encoding a complete L' 1 molecule iε obtained from an expreεεion plaεmid pCMUIV/L (see Joly and Oldεtone, Gene 97: 213, 1991) . The complete cDNA iε cleaved from a eukaryotic expreεεion vector pCMU IV/L as a Bam HI fragment and cloned into pRmHa-3 aε K b . Aε noted previouεly, the pCMU vector (pCMUIV) iε derived from eukaryotic expression vector pC81G as described in Nilsson, et al . , supra . Vector pC81G, in turn, iε derived from pA81G (Paabo, et al . , Cell 33 : 445-453 (1983) ) according to the method diεclosed in Paabo, et al., EMBO J. 5: 1921-7 (1986) . H-2D 1 ': cDNA encoding a complete D 1, molecule is obtained from expresεion plaεmid pCMUIV/D 1, (see Joly and Oldstone, Science 253 : 1283-85, 1991) . The complete cDNA is cleaved from a eukaryotic expression vector pCMUIV/D ! as a Bam HI fragment and cloned into pRmHa-3 as K* . Murine β-2 microglobulin: full-length murine β- 2 microglobulin cDNA is obtained as a Hind III (5') (filled with Klenow) /Bgl II (3') fragment from pSV2neo (ATCC No. 37*149) mouse β- 2 microglobulin cDNA and cloned into pRmHa-3 cleaved with Sma I and Bam HI. Vector phshsneo confers neomycin (G418) resistance and is a derivative of phsneo (pUChsneo) with an additional heat-shock promoter (hs) sequence, which may be synthesized from commercially-available pUC8 as described in Steller, et al . , EMBO J. 4 : 167 (1985) . The heat shock promoter contained in these vectorε iε the hsp70 promoter. Other uεeful vectorε conferring neomycin resistance (G418 resistance) include cosmid vector smart2 (ATCC 37588) , which is expressed under the control of Drosophila hsp70 promoter, and plasmid vector pcopneo (ATCC 37409) . C. Insertion of Genes into Expression Vectorε The reεtriction productε are εubjected to electrophoreεiε on a 1% agaroεe gel (Maniatiε, et al . , Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory (1982)) . The reεtriction fragmentε encoding the cDNAε are excised from the gel and purified away from the agarose uεing "Geneclean", according to manufacturerε' directionε (Bio 101, San Diego, CA) . The expreεεion plaεmid pRmHa-3 (Figure 3) iε cleaved with the appropriate reεtriction enzymeε in One Phor All buffer according to the manufacturer's directions (Pharmacia, Piscataway, NJ) and treated with alkaline phosphatase aε described in the manufacturer's literature (Boehringer Mannheim, Indianapolis, IN) . One hundred ng of cleaved and phosphatased pRmHa-3 vector is mixed with 300ng of agaroεe gel purified Claεε I MHC heavy chain cDNA or β- 2 microglobulin cDNA and ligated uεing T4 DNA ligaεe and One Phor all buffer aε described in the manufacturers' literature. After incubation at 16 °C for five hours, the ligation mixture is used to transform competent E. coli JM83 (Maniatis, et al., supra (1982)) . Methods disclosed in Maniatis, et al . , supra are used to prepare the cDNA needed. The presence of the MHC heavy chain cDNA and its orientation in the vector is determined by restriction mapping. Bacteria containing the vector with the cDNA in the correct orientation relative to the metallothionein promoter are used for large scale preparation of DNA using the alkaline lysiε method and cesium chloride gradient purification. The amount of DNA obtained is determined spectrophotometrically. D. Tranεfection and Labeling of S2 Cells S2 cells are grown in Schneider medium (Gibco/BRL, Grand Island, NY) supplemented with 10% fetal calf serum (heat treated for one hour at 55 °C) , 100 units/ml penicillin, lOOmg/ml streptomycin, and ImM glutamine. (For convenience, this supplemented medium is hereinafter referred to as Schneider medium.) Cellε are grown at 27 °C and typically passaged every seven dayε by diluting 1:17 in freεh medium. Cellε are converted to growth in εerum free media (Excell 400 or 401 εupplemented with 100 unitε/ml penicillin, 100 mg/ml εtreptomycin, ImM glutamine, and 500μg/ml G418 (JRH Bioεcienceε, Lenexa, KS) by initial dilution at 50% Schneider/50% Excell 401. One week later, cellε may be paεεaged into 10% Schneider medium/90% Excell 401 and one week later into 100% Excell 401. Cellε are maintained in thiε medium and paεsaged every seven days by diluting 2:17 in fresh medium. 15 X 10' S2 cells at a concentration of 10' cellε per ml are plated out in 85mm petri diεheε. Twelve hourε later, calcium phoεphate/DNA precipitates, prepared as described below (1 ml) are added dropwise to the cells. After 48 hours, the supernatant is carefully removed and the cellε transferred to a 175cm 2 flask in a total volume of 50 ml in Schneider medium containing 500μg/ml Geneticin (G418) (Gibco/BRL, Grand Island, NY) . After 21 days, 20 ml of the culture is removed to a fresh flask containing 30 ml of Schneider medium containing 500μg/ml G418. Ten days later, a stable population of cells that weakly adhered to the flask and grew with a doubling time of approximately 24 hours is obtained and these cells are subsequently cultured and paεεaged in the εelection media aε described above. Frozen aliquots of these cells are prepared by collecting 5-20 X I0 β cells by centrifugation and reεuεpending them in 1 ml of cell freezing media (93% fetal calf εerum/7% dimethylsulfoxide) . Aliquots are then placed at -70 °C for one week and subsequently transferred to liquid nitrogen storage. Calcium phosphate precipitates are prepared as described by Paabo, et al. (EMBO J. 5: 1921-27 (1986)) , except that 25μg of DNA is used per transfection. The following combinations of DNA are used to prepare the indicated tranεfectant : (a) MHC Claεε I heavy chain alone: 23μg heavy chain expreεεion vector DNA + 2μg of phshsneo DNA. (b) MHC Clasε I heavy chain + β - 2 microglobulin: 11.5μg heavy chain expression vector DNA + 11.5μg of β - 2 microglobulin (human or mouεe) expreεεion vector DNA + 2μg of phshsneo DNA. Other combinations of mouse genes are presented in Table 1. Twenty-four hours prior to metabolic labeling, cellε are plated out at a cell density of 3-5 X 10" cells/ml (10 ml/85mm petri diεh) in Schneider medium containing ImM CuSO, . Thirty minutes prior to labelling the medium iε aεpirated from the dishes and the cells are washed with 2 X 10 ml of PBS and then incubated in Graces insect medium minus methionine and cysteine (special order from Gibco/BRL, Grand Island, NY) for 20 minuteε, and then in 1 ml of this medium containing 0. lmCi 35 S Trans label (New England Nuclear; duPont, Boston, MA) . After the labelling period, the labelling solution is aspirated and the cells are either lysed immediately on ice, with ice cold PBS/1% Triton X100 (1 ml) or after a chase period in the presence of methionine containing Schneider or Excell 400 medium (5 ml) (JRH Biosciences) . The chase medium is collected if εoluble Claεε I MHC moleculeε are being analyzed. The following operationε are all carried out with the lysates kept cold (leεε than 8 °C) . The lysates were collected into Eppendorf tubeε, centrifuged in a microfuge tube for 15 minuteε at 13,000 X g, transferred to a fresh tube containing lOOμl of a 10% slurry of protein A sepharose and placed on an end-over-end rotator for two hourε. Following a further centrifugation in the microfuge for 15 minuteε, the cell lysates are ready for analysiε. In experimentε utilizing murine MHC, S2 cellε were tranεfected with the murine MHC recombinantε deεcribed above uεing the CaPO,, precipitation method; each heavy chain iε tranεfected either alone or as a 50:50 mix with the vector encoding β- 2 microglobulin. A plasmid encoding neomycin resiεtance, phshsneo DNA, iε included in each transfection εuch that a population of cellε that εtably expreεεed MHC Claεε I could be obtained by growing the tranεfectantε in selection medium (Geneticin G418-sulphate, Gibco/BRL, Grand Island, NY) . E. Peptide Generation Antigenic peptideε according to the preεent invention may be obtained from naturally-occurring εourceε or may be synthesized using known methods. In various examples discloεed herein, peptideε are synthesized on an Applied Biosystems synthesizer, ABI 431A (Foster City, CA) and subsequently purified by HPLC. Isolation or synthesis of "random" peptides may also be appropriate, particularly when one is attempting to ascertain a particular epitope in order to load an empty MHC molecule with a peptide most likely to stimulate precursor CD8 * cells. One may produce a mixture of "random" peptides via use of proteasomeε (see, e.g., Example 2.B.6) or by subjecting a protein or polypeptide to a degradative process -- e.g. , digestion with chymotrypεin -- or peptideε may be εyntheεized. While we ' have observed that the cell lines of the preεent invention are able to degrade proteinε and polypeptideε into smaller peptides capable of being loaded onto human Class I MHC molecules, it is preferable to introduce εmaller peptideε -- e.g., 8-merε and 9-merε -- directly into the cell culture to facilitate a more rapid loading and expreεsion proceεε . If one iε synthesizing peptides, e.g., random 8-, 9- and 18-amino acid peptides, all varieties of amino acidε are preferably incorporated during each cycle of the synthesis. It should be noted, however, that various parameters --e.g., εolvent incompatibility of certain amino acidε -- may reεult in a mixture which containε peptideε lacking certain amino acidε. The proceεε should thus be adjusted as needed -- i.e., by altering solvents and reaction conditions -- to produce the greatest variety of peptides. As noted hereinabove, murine heavy chains complexed with human β- 2 microglobulin were stable at temperatures approximately 6-8 degrees higher than if complexed with murine β2 . It was also observed that the stabilitieε imparted by peptide and xenogeneic β - 2 microglobulin are additive. A large increaεe in the thermostability of the Class I moleculeε occurs if 8-9 mers are used, as compared to 12-25 ers; indeed, the difference between the εtabilization imparted by the 8-9 merε compared with the larger peptideε might be even greater than what waε obεerved previouεly, for even though the peptides have been purified by HPLC, it is likely that there is some contamination of the larger peptides by 8-9 mers. The thermostability of a Class I molecule is apparently dependent on: (1) the origin of β- 2 microglobulin; (2) the presence of peptide,- and (3) the length and sequence of this peptide. Previous work (U.S. Patent No. 5,314,813 to Peterson et al . ; Jackson et al . , PNAS USA 89 : 12117-12121 (1992)) has shown that Class I MHC heavy chains can bind peptide either alone or when they are asεociated with β- 2 microglobulin. Surface expreεεion of peptide-loaded human Class I MHC, however, appears to be beεt facilitated by loading the moleculeε with peptide after the heavy chainε have complexed with β- 2 microglobulin. 1. Expreεεion of Human MHC Once we determined that the thermoεtability of a Claεε I moleculeε iε dependent on the origin of β - 2 microglobulin, the presence of peptide, and the length and sequence of thiε peptide, we utilized thiε information in the creation of cell lineε capable of εpecifically activating CD8 * cellε via the expreεεion of peptide-loaded human Claεε I MHC molecules. Thermolability appears to be an inherent property of Clasε I moleculeε; it haε preεumably evolved to enεure that Claεs I molecules which contain either no peptide or a peptide of poor binding properties (that confers little thermostability) self-deεtruct . In this way, the cell minimizeε the number of empty Class I molecules on its εurface, for εuch a εituation would preεumably be dangerouε in that exogenouεly derived peptides could be bound and presented. Human Clasε I molecules expressed in inεect cellε with human β2 are not stable to extended incubation at 37 °C; neither are human Clasε I molecules expressed in the mutant cell line T2 which has been shown to be deficient in peptide loading onto the Class I molecules (Hosken and Bevan, Science 248: 367-70 (1990) ; Cerundolo, et al . , Nature 345: 449-452 (1990)) . Thus, it seems that the affinity between the heavy chain and ,5-2 microglobulin has been carefully conserved through co-evolution of the molecules such that empty Class I molecules, or those carrying poorly-binding peptides, self-destruct at the body temperature of the "host" organism. Human Class I MHC molecules were expressed in S2 cells. Cell lines co-expressing human β- 2 ' microglobulin and HLA A2.2Y, HLA A2.1, HLA B7, or HLA B27 were established using previously-described methods. Briefly, cDNAs encoding the above proteinε were cloned into the Drosophila expreεεion vector pRmHa-3 and cotranεfected with a human β- 2 microglobulin-containing plaεmid and phεhεneo plaεmid into S2 cellε via methodε diεcloεed herein. Three to four weeks later, the population of G418-resiεtanT-cellε waε diluted 1:5 with freεh εelection media. Once a healthy growing population of cellε waε obtained, CuSO. waε added to an aliquot of cellε and 24 hourε later, cellε were analyzed via flow cytometry uεing a monoclonal antibody W6/32 (ATCC HB95, Betheεda, MD) which recognizeε a onomorphic determinant of human Claεε I heavy chainε when they are in asεociation with β- 2 microglobulin. (See Barnεtable, et al. , Cell 14: 9 (1976) .) High levels of surface expression of each of the human Clasε I molecules were induced by the addition of CuSO,, (data not shown) . These stable populations were sorted for high expresεing cells using cytofluorimetry as described below. It is theεe sorted populations of cells which were used for all subsequent experiments . Twenty-four hours prior to FACS analysiε, CuS0 4 iε added to the εtably transfected S2 cells (3-4 X 10 6 cells/ml) to a final concentration of ImM, thereby "switching on" expression from the transfected genes. Cells are plated out in 24-well cluster dishes (2 ml per well) . Eight hours prior to FACS analysiε, the CuSO., medium is replaced with fresh medium (1 ml) with or without peptide at a concentration of 50μg/ml. 37 °C temperature challenges are carried out by transferring the dishes onto a flat surface in a 37 °C room at various time intervals prior to harvesting the cells for analysis . To analyze surface expresεion of Class I MHC on the S2 cells, aliquots of cells (5 X 10 ) are tranεferred into tubes on ice, collected by centrifugation (1,000 X g for 4 minutes) , resuεpended in 3 ml of PBS/1% BSA, 0.02% εodium azide, collected by centrifugation and reεuεpended in PBS/BSA (0.5 ml) containing the appropriate primary antibody (ascites fluids Y3 , 28:14:8S, 30.5.7, W6/32, diluted 1:200) . Rabbit antisera are diluted 1:500 and B22.293 hybridoma εupernatant iε uεed directly. After a one hour incubation on ice, cellε are waεhed twice in 3 ml of PBS/BSA and resuspended in 0.5 ml of PBS/BSA containing FITC labelled εecondary antibody (Cappell, Durham, NC) and 1 ng/ml propidiu iodide. After a 30 minute incubation on ice, cellε are waεhed once with PBS/BSA and reεuεpended in thiε buffer at a concentration of 1 X lO'/ml. Sampleε are then analyzed by FACS 440 (Becton Dickinεon) . Dead cellε εtained with propidium iodide, are excluded by including a live gate in the analysis . For cell sorting, the same procedure outlined above is used, except that all staining operationε are carried out in a sterile hood. Solutions, including antibodies, are filter-sterilized, and Schneider media or Excell 400 is used in place of PBS/BSA. Cells that specifically bound the primary antibody are sorted uεing a Becton Dickinεon cell sorter. Sorted cells (2-6 X 10 5 ) are washed once in medium before plating out at a concentration of 2 X 10 5 cells/ml. F. Loading of Membrane-Bound Empty MHC Molecules by in vi tro Incubation with Peptides In order to demonstrate that the human Class I molecules expressed on the surface of the Drosophila cells were empty, the cells were incubated at 37 °C for two hours and the cell surface expresεion waε analyzed by cytofluorimetry. The surface expresεion of both HLA B27 and A2.1 iε greatly reduced if cellε are incubated at 37 °C for 2 hourε; however, preincubating the cellε in HIV peptideε known to bind to the Claεε I moleculeε affordε εignificant thermal εtability to the Class I, while peptideε that do not bind have little effect (εee Figure 4) . (A 9-amino acid peptide ILKEPVHGV (SEQ ID NO 42) from the POL protein of HIV bindε and stabilizes HLA A2.1. A nine-amino-acid peptide from the Vpr protein of HIV bindε and εtabilizeε B27 (FRIGCRHSR; SEQ ID NO 41) . Theεe data εhow that the human Claεε I molecules expreεsed on the surface of Drosophi la cells are empty and can be εtabilized by binding εpecific HIV peptideε. Figureε 4 and 5 εhow peptide-induced thermoε t abilization of HLA B27 and HLA A2.1 expressed on the surface of Drosophila cells by HIV peptideε. Drosophila cells expresεing either HLA B27 or A2.1 were incubated with peptideε where indicated and then either maintained at 28 °C or incubated at 37 °C for two hourε prior to analyεis of the εurface expreεεion of the Claεε I moleculeε by uεe of the antibody W6/32 ( from ATCC HB95) and cytofluorimetry. The mean fluoreεcence of each cell population is shown plotted against the incubation conditions The HIV POL peptide (ILKEPVHGV, SEQ ID NO 42) stabilizes A2.1 but not B27 (Figure 4) , while the HIV Vpr peptide (FRIGCRHSR, SEQ ID NO 41) stabilizes B27, but not A2.1 (Figure 5) . Example 2 Preparation of Synthetic Antigen-Presenting Cells A. Osmotic Loading Osmotic loading of SC2 and 3T3 cells with ovalbumin protein was carried out aε deεcribed by Moore, et al., Cell 54: 777-785 (1988) . The aεεay procedure lε aε followε. In a 96-well diεh, 1 X 10 5 Drosophila cellε (with or without peptide/prote loaded) or 3T3 cellε were cocultured with 1 X 10 5 B3/CD8 T-cell hybridoma cells in 200μl of RPMI media supplemented with 10% fetal bovine serum. After 24 hours of incubation, lOOμl of the supernatant from these cultures was added to lOOμl of RPMI containing 5,000 CTLL cellε The cells were cocultured for 24 hourε at 37 °C when lμCi of J H thymidine (Amerεham) waε added. After a further incubation of 15 hourε at 37 °C, the incorporation of radiolabel into the CTLL cells was determined by scintillation counting. Asεayε conducted with murine MHC also verified that the insect cells are capable of loading peptide onto the Clasε I moleculeε. Cellε expressing as few as 200-500 MHC molecules containing a particular antigen can be detected by a T-cell. As the Drosophi l a cells do not accumulate chromium, an antigen presentation aεεay baεed on B3/CD8, a T-cell hybridoma, waε uεed. B3/CD8 lε a hybridoma between B3 , cytotoxic T-cell εpecific for ovalbumin peptide 253-276 preεented by H-2K' Claεε I moleculeε, and CD8- bearing IL-2-εecretmg cell line (see Carbone, et al . , supra, 1989) . Upon antigenic stimulation, B3/CD8 produces IL-2, measured by H thymidine incorporation in IL-2-dependent T-cell line CTLL (Gillis, et al., J. Immunol. 120: 2027 91978)) . Thuε, by measuring the amount of IL-2 produced, one can assay for T-cell recognition. In order to provide an intracellular pool of ovalbumin protein from which OVA peptides can be derived, ovalbumin (Sigma Chem. Co., MO) was osmotically loaded into the cells as described by Moore, et al, supra (1988) . Immediately after loading, the cells were mixed with the T-cell hybridoma. After two days' incubation, the medium was removed and asεayed for IL-2. The amount of IL-2 was determined by the ability of the medium to support the growth of the IL-2-dependenT-cell line CTLL (Gillis, et al . , supra. 1978) , and growth was quantitated by the amount of radioactive thymidine incorporated into the cells. S2 or 3T3 cells transfected with K''/?2 were incubated with ovalbumin protein (OvPro) or ovalbumin peptide, OVA 24 (OvPep) in isotonic (Iεo) or hypertonic (Hyp) media. (Murine cell line BALB/3T3 is available from the ATCC under accession number CCL 163. ) After treatment, cells were cocultured with the T-cell hybridoma B3/CD8. B3/CD8 iε a T-cell hybridoma between B3 (Carbone, et al . , J. Exp. Med. 169: 603-12 (1989) ) , cytotoxic T-cell εpecific for ovalbumin peptide 253-276 preεented by H-2K Claεs I moleculeε, and CD8- bearing IL-2-secreting cell line. Upon antigenic stimulation, B3/CDδ produceε IL-2, meaεured by 3 H thymidine incorporation in IL-2-dependent cell line CTLL (Gillis, et al., J. Immunol. 120: 2027 91978) ) . Thuε, by meaεuring the amount of IL-2 produced, one can aεεay for T-cell recognition. The εupernatant from the cocultureε were analyzed for IL-2 by H thymidine incorporation by the IL-2-dependent cell line CTLL (ATCC No. TIB 214) . The amount of 3 H thymidine incorporated iε plotted againεt the initial cell treatments. It can be seen in Figure 6 that the T-cells responded well to the Drosophila cells if the ovalbumin peptide was added to the culture medium, but no recognition occurred if the cells were loaded with the ovalbumin protein. The MHC Class I molecules expressed on the cell surface of the insect cell are fully functional in that they can bind peptide if it is added to the culture medium and can preεent it in the correct context for it to be recognized by a T-cell. B. Optimization of In Vi tro Conditions For the optimization of in vi tro conditions for the generation of specific cytotoxic T-cellε, the culture of Drosophila cell stimulator cells is preferably maintained in serum-free medium (e.g. Excell 400) . Drosophila cell stimulator cells are preferably incubated with >20μg/ml peptide. The effector: stimulator ratio (lymphocyte : Drosophila cell ratio) is preferably in the range of about 30:1 to 300:1. The maximum εpecific CD8' iε generally obεerved after five dayε of culture. The culture of target cellε for killing aεεay is preferably maintained in a serum-free medium. Example 3 Stimulation of Proliferation and Differentiation of Armed Effector T-Cells We have found that Drosophila S2 cellε tranεfected with MHC claεε I moleculeε and εpecific aεsisting molecules are able to stimulate primary responseε from T-cellε in vitro. We preεent data below in thiε example from a mouse model syεte . In this example, constructs coding for mouse MHC class I (L ci ) , β2 microglobulin, specific asεiεting moleculeε were uεed and teεted with CD8 * cellε from lymph nodeε of T-cell receptor tranεgenic mice. The data in Figure 7 provideε evidence that the tranεfected Drosophila S2 cells express the protein products of the transfected murine genes. Flow cytometry uεing a fluorescence-activated cell sorter (FACS) and fluorescently labelled antibodies were used to demonstrate the expression of L (MHC molecule which includes heavy chain and β2 ) and the specific asεisting molecules B7.1 (CD80) and ICAM-1 (CD54) moleculeε by tranεfected Drosophila S2 cells. Transfected cells were separated with a FACS to obtain cells expressing L d molecules and were then maintained in vitro. The transfection of Drosophila S2 cellε is summarized in Table 2. The data show L d , B7.1 and ICAM-1 expression measured by flow cytometry on the cell lines after induction with CuS0 4 It is apparent that, relative to the control antibody (ctr Ab) , all of the transfectants express L d molecules on the cell surface. Likewise, cellε cotransfected with L and B7.1 (L d .B7) expresε B7.1 but not ICAM-1, whereaε cells cotransfected with L d and ICAM-1 (L .ICAM) express ICAM-1 but not B7.1 ; triple transfection with L d , B7.1 and ICAM-1 (L .B7.ICAM) led to expresεion of all three moleculeε. Uεing a εtandard tiεεue culture εystem (Cai, Z. and Sprent, J. (1994) J. Exp. Med. 179: 2005-2015) , doεeε of 5 x 10' 1 purified CD8' 2C lymph node (LN) cellε were cultured at 37 °C with doεes of 3 x 10 ε transfected fly cells ± peptides (10 μM final concentration) . Peptides were εyntheεized by R. W. Johnεon Pharmaceutical Reεearch Inεtitute (Sykulev, et al . (1994) Immunity 1: 15-22. Proliferative reεponses were meaεured by adding 'HTdR (1 μCi/well) 8 hourε prior to harvest. IL-2 production was meaεured by removing supernatants from the cultureε at 48 hourε and adding 50 μl supernatant to an IL-2 responsive indicator cell line (CTLL) , * proliferation of the indicator line was measured by addition of 3 HTdR. The data shown in Table 2 are the means of triplicate cultures. The transfected Drosophila S2 cells die rapidly at 37 °C and fail to incorporate 3 HTdR at this temperature. The data in Table 2 demonstrate that the transfectants are able to stimulate primary responses of mouse T-cells. Table 2. Capacity of transfected fly cells to stimulate primary proliferative responseε and IL-2 production by CD8 * lymph node cellε from 2C T-cell receptor transgenic mice. TABLE 2 3 HTdR incorporation (cpm x 1 0 3 ) with transfected fly cells expressing: The 2C T-cell receptor (TCR) is strongly reactive to A molecules complexed with certain peptideε, e.g. p2Ca (SEQ ID NO 46) or QL9 (SEQ ID NO 47) . These two peptides have moderate to high affinity for soluble L d molecules, 4 x 10' M τ for p2Ca, and 4 x 10" M "1 for QL9 (Sykulev. et al . ) . When complexed to soluble L' 1 molecules, the two peptides also have high binding affinity for soluble 2C TCR moleculeε. However, in both TCR binding and L" binding, the QL9 peptide clearly has a higher affinity than the p2Ca peptide. Table 2 shows that proliferative responseε and IL-2 production by the reεponder 2C cellε to the weaker peptide, p2Ca, requires that the stimulator L d -transfected cells coexpress both B7.1 and ICAM-1 ; a mixture of cells expreεεing either L d + B7.1 or L d + ICAM-1 is nonstimulatory. By contrast, with the stronger peptide, QL9, L d .fly cells expresεing either B7 or ICAM elicit clearly-significant responses, although combined expression of B7 and ICAM generates much higher responses. In contrast to these findingε on T-cell proliferation, IL-2 production in reεponse to the QL9 peptide requires joint expresεion of B7 and ICAM ; expression of these molecules on separate cellε iε ineffective. The reεults show that Drosophila cells transfected with murine class I molecules and costimulatory molecules induce murine T-cells to mount primary proliferative responεeε and lymphokine (IL-2) production in reεponεe to peptide antigens. The system iε alεo applicable to human T-cells and could be used to stimulate unprimed (or primed) T-cells specific for tumor-specific antigens in vitro,* in vivo infuεion of clonally-expanded T-cellε εpecific for tumor-εpecific antigenε might be therapeutic for patientε with cancer. Infuεion of T-cells specific for viral antigenε would be uεeful in patients with viral infectionε, e.g. HIV. Example 4 Immobilization of Biotinylated MHC Moleculeε on Avidin-Coated Red Blood Cellε NHS-LC-biotin, neutravidin and biotin-BMCC were purchaεed from Pierce (Rockford, IL) . Sheep red blood cellε were obtained from the Colorado Serum Company (Denver, CO) . Drosophila S2 cellε expreεεing L and recombinant L d were prepared aε deεcribed in Exampleε 1 and 2. Monoclonal antibodieε 30.5.7 (anti-L d ) and 1B2 (anti-clonotypic antibody to the 2C T-cell receptor) were uεed aε hybridoma cell culture supernatants . The protocol used is described by Muzykantov and Taylor (Anal. Biochem. (1994) 223, 142-148) . Briefly, SRBC were washed 4 times in phosphate buffered saline (PBS) , biotinylated using NHS-LC-biotin, washed again 4 times in PBS, incubated with neutavidin, and finally washed 4 times and stored at 4 °C in PBS containing 3% fetal calf serum and 0.02% sodium azide. Recombinant L d was biotinylated using biotin-BMCC, a maleimide-coupled biotin which reacts with thiol groups. L d displays a free thiol group, the side chain of cystein 121, which is not in the peptide binding site. Biotinylation was performed as recommended by the manufacturer. Unreacted biotin was removed using Centricon 10. Biotinylated L was immobilized by incubation, at a final concentration of 0.2 mg/ml with avidin-coated SRBC for 30 minuteε followed by waεhing in DMEM containing 10% fetal calf serum. SRBC with attached L d were used immediately. T-cells expressing the 2C TCR transgene from lymph nodeε of mice were purified by magnetic depletion. Purified T-cellε were conεiεtently 97-98% poεitive for staining in flow cytofluorometry uεing the anti-clonotypic antibody 1B2. Immobilization of biotinylated L d on avidin-coated SRBC waε done aε indicated above. Attachment waε aεεeεεed using flow cytofluorometry using anti-L antibody 30.5.7. A typical experiment is represented in Figure 8. The negative control (cellε minuε antibody) iε εhown in dotted lineε. The filled peak comprises cells labeled with fluorescent antibody. 99.78% of the cells were labeled. Fluorescence intensity was in the same range than the highest levels of intensity that we observed for L d on synthetic antigen-presenting cells. K b (MHC molecule which includes heavy chain and β2 ) was also biotinylated using the same procedure. We could immobilize biotinylated K h on avidin-coated SRBC as assesεed by flow cytofluorometry (Figure 9) . 99.88% of the cells were labeled. Rosetting experiments verified that the attached MHC moleculeε interacted functionally with T-cells. Drosophila S2 cells expressing L d , L d -coated SRBC were incubated with QL9 peptide (0.02 mM) or an irrelevant peptide (MCMV, 0.02mM) for 30 minutes on ice; 2C+ T-cellε were then added, the proportion being 10 2C+ T-cellε for 1 Drosophila S2 cell, or 10 SRBC for 1 2C+ T-cell; the mixture waε pelleted and kept on ice for at least 30 min. Cells were then carefully resuspended and rosettes were counted, a rosette being a Drosophila S2 cell bound to at least 3 2C+ T-cells, or a 2C+ T-cell bound to at least 3 SRBC. Rosetteε were obεerved in all cases . Typically, 30-40% of the lymphocyteε were included in roεetteε when QL9 peptide was added. No rosette was obεerved in the presence of the irrelevant peptide, although occasional attachment of a few single cellε was obεerved. Theεe examples describe a new method to immobilize high amounts of MHC clasε I moleculeε on various surfaceε (fly cellε, red blood cellε, latex beadε) in native conformation aε judged by monoclonal antibody binding and roεetting experimentε (T-cell receptor binding) . Thiε method can be extended to other synthetic surfaces including artificial phospholipid membranes. phoεphatidylethanola ine aε well aε avidin-coupled phoεpholipidε are particularly relevant to our εtudieε. These phospholipids are commercially available from Lipex Biomembrane Inc., Vancouver, BC, Canada . Example 5 Immobilization of Biotinylated MHC Molecules on Avidin-Coated Latex Beads Six micron diameter latex sulfate beads were purchased from Interfacial Dynamics Corporation (Portland, OR) and biotinylated according to the protocol described in Example 4. Avidin-coated latex beads were prepared using a 1% suspension of the latex beads incubated in PBS containing lmg/ml of neutravidin for one hour at room temperature. An equal volume of PBS containing 10% fetal calf serum was then added. After one hour of incubation at room temperature, the beads were washed 3 timeε and uεed for binding of recombinant biotinylated L d . Recombinant biotinylated L waε immobilized by incubation at a final concentration of 0.2 mg/ml with avidin-coated latex beadε for 30 minuteε followed by waεhing in DMEM containing 10% fetal calf εerum. SRBC with attached L d were uεed immediately. Rosetting experiments verified that the attached MHC molecules on latex beads interacted functionally with T-cells. Drosophi la S2 cells expressing recombinant L and L d -coated latex beads were incubated with QL9 peptide (0.02 mM) or an irrelevant peptide (MCMV, 0.02mM) for 30 minutes on ice, * 2C+ T-cells were then added, the proportion being 10 2C+ T-cells for 1 Drosophila S2 cell, or L d -coated latex beads for 1 2C+ T-cell ; the mixture was pelleted and kept on ice for at least 30 min. Cells were then carefully reεuεpended and rosettes were counted, a rosette being a Drosophila S2 cell bound to at least 3 2C+ T-cells, or a 2C+ T-cell bound to at least 3 latex beads. Rosetteε were obεerved in all cases. Typically, 30-40% of the lymphocyteε were included in rosettes when QL9 peptide was added. No rosette was observed in the presence of the irrelevant peptide, although occasional attachment of a few single cells was observed. Example 6 Immobilization and Detection of Recombinant Protein Bound to Various Solid Supportε Such as Plastic Microwell Plates The MHC molecules were immobilized by direct binding to microtiter plates (Corning) and detected as follows : MHC K b molecules were diluted to desired concentration in PBS, e.g. O.OOlmg/ml for 100 ng /well. lOOμl of diluted K b was added to each well on the plastic microtiter plate. The plate was incubated for 1 hour at room temperature. After incubation, the plate was washed once with PBS and 200μl 2% bovine εerum albumin (BSA) in PBS + (.05%) and Tween (PBST) waε added, and incubated for another hour at room temperature. The plate was waεhed three timeε with PBST and biotinylated anti-K 1 mAb waε added (1:2500) in 2% BSA in PBS. The plate waε incubated another hour at room temperature and waεhed three timeε with PBST. Avidin conjugated HRP waε added (1 : 2500) in 2% BSA in PBS. Following another hour of incubation at room temperature, the plate waε waεhed three timeε with PBST and H 2 0 2 or thophenyldiamine waε added. The reaction was stopped with H 2 S0, . Reaction product was detected colorimetrically at 490 nm. Figure 10 showε the reεultε of detecting the preεence of MHC K 1 moleculeε uεing three different monoclonal antibodies. Recombinant MHC K 1 ' molecules can alternatively be bound through biotin-avidin linked interactions with the substrate. In this embodiment, the microwell plates were coated with lOOμl avidin diluted in PBS to a concentration of O.OOlmg/ml. Excess avidin was removed by a PBS wash. The above procedure for presenting and detecting K b binding followed. Recombinant MHC molecules may alternatively be immobilized by a linkage based on a poly-histidine tag added to the MHC interacting with the nickel bound to the substrate . The above procedure for binding and detection is followed using nickel chelate coated microwell plates (Xenopore) and recombinant MHC molecules with a poly-histidine tag expressed using vector pRmHa/His 6 described above . Example 7 Direct Binding of Peptide to Soluble. Empty Class I MHC Molecules In Vi tro A. Procedures H-2K b : prepared as described above in Example l.B. H-2K b Sol: K b sol cDNA is a derivative of K b , encoding the extracellular portion of the Clasε I MHC molecule. K b εol cDNA may be produced by PCR according to known methodε, εuch as those described in Ennis, et al. , PNAS USA 87: 2833-7 (1990) and Zemmour, et al . , Immunogeneticε 33: 310-20 (1991) . Specifically, cDNA encoding a truncated K molecule with a εtop codon inserted at the end of the alpha 3 domain at amino acid poεition + 275 iε excised from the pCMU expression plasmid as a Bam HI fragment and cloned into pRmHa-3 as K b cDNA. The K b sol cDNA is a derivative of the complete K b cDNA (εee above) which iε uεed as a template in a PCR reaction uεing a 5' oligonucleotide that encompaεsed the Sty I εite, and the following 3' oligonucleotide: 5' ATATGGATCCTCACCATCTCAGGGTGAGGGGC 3 ' (SEQ ID NO 43) The resulting PCR fragment is blunt-end cloned into the Sma I site of pBS (Stratagene, La Jolla, CA) , sequenced, and the remaining 5' sequence of K cloned into the Sty I site. A cDNA encoding the complete K b sol protein could be obtained as a Bam HI restriction fragment. H-2D b and H-2L d are prepared as discussed in Example l.B. above. The cDNAs encoding K b αlα2c.3 domains (274 residues) and murine β- 2 microglobulin (99 residueε) were respectively cloned into the unique Bam HI site of an expression vector harboring the metallothionein promoter pRMHa-3 (Bunch, et al . , Nucleic Acid Res. 16: 1043-1061 (1988) ) . Drosophila S2/M3 cells were tranεformed with these recombinant plasmids in addition to plasmid phshsneo (containing a neomycin-resistance gene) by the calcium-phosphate precipitation method described previously. The transformed cells selected against neomycin-analog antibiotics G418 were grown at 27 °C in serum-free medium and soluble heavy-chain K b and β- 2 microglobulin were co-expresεed by the addition of 0.7mM CuSO, . The εoluble, assembled heterodimer of K b was purified from the culture supernatants by affinity chromatography using anti-K b monoclonal antibody Y3 , followed by ion-exchange chromatography on a Pharmacia Mono Q FPLC column according to the inεtructionε of the manufacturer (Pharmacia, Piscataway, NJ) . SDS-polyacryla ide gel electrophoresiε (SDS-PAGE) of the K'' preparation followed by εtaining with Coomaεεie blue εhowed only one band of relative molecular maεε (Mr) at about 32,000 and one band of Mr at about 12,000 with nc detectable impuritieε. The highly-purified K 1 ' waε dialyzed againεt phoεphate-buffered saline (PBS) , filter-sterilized, and used for further study. Extinction coefficient of the soluble K b ("K b sol") protein (43.2 kDa) is 69,200 M^cm "1 at 280 nm. The purified K b sol (0.3μM) in PBS with or without 1% TX-100 were exposed to varying temperatureε (i.e., 4°, 23°, 32°, 37°, 42°, and 47°C) for one hour. The proteinε were then immunoprecipitated by incubating with the monoclonal antibody Y3 and protein A εepharoεe beadε (Pharmacia, Piscataway, NJ) at 4 °C for two hours, respectively. The samples were analyzed by 12.5% SDS-PAGE, followed by staining with Coomassie blue. The two thick bands on the gel are heavy and light chains of antibody Y3. In another procedure, K b sol (0.3μM) were incubated with 50μM of peptides in PBS at 23 °C for two hours to allow for K sol-peptide complex formation. After the addition of 1% TX-100, the sampleε were exposed to 12 °C, 37 °C, or 47 °C temperatures for one hour. The complexeε were immunoprecipitated and analyzed by SDS-PAGE aε deεcribed above. In a third procedure, K b εol (2.7μM) were incubated with 50μM of OVA-8, VSV-8 or SEV-9 peptideε, reεpectively, at 23 °C for two hourε. The εampleε were applied on a 5% polyacrylamide IEF gel. IEF waε run from pH 5-7 and the gel waε εtained with εilver. Next, VSV-8 peptide waε radioiodinated uεing the chloramine-T method (Hunter, et al . , Nature 19 : 495-6 (1962)) and free 1 "'I was removed by C._ column (OPC cartridge, Applied Biosyεtemε, Foεter City, CA) . The labelled peptide waε further purified by C 18 reverse-phase HPLC. After elution, the labelled peptide was lyophilized and resuεpended in PBS. The εpecific activity of [ 1 I] VSV-8 (about 250 Ci/mmole) waε determined εpectrophotometrically by uεing extinction coefficient of tyroεine at 274nm (1420 M-'cm 1 ) . Firεt, K''sol (0.5μM) waε mixed with [ ι:!5 I] VSV-8 (1.5nM) and unlabelled VSV-8 (50nM) at 23 °C for 16 hourε to allow for complex formation. A portion of the εample was analyzed by gel filtration (Superose 12, Pharmacia, Piscataway, NJ) in PBS. After elution, radioactivity contained in each fraction (0.05ml) was measured. Protein was monitored by absorbance at 280nm. In a second procedure, [ l 5 I]VSV-8 (0.39nM) was mixed with various concentrations of K b sol in PBS containing 1% bovine serum albumin (BSA) . After incubation at 23 °C for 2-16 hourε, K b sol-peptide complexes were separated from free peptide by small gel filtration (Bio-Gel P30, BioRad, Richmond, CA) in PBS. P30 gel filtration permitted over 95% separation of bound and free peptide within about 5 minutes. Radioactivity of bound and free peptides was measured and the data were analyzed by linear regression. At maximal levels of K b sol offered, about 65% of the total labelled peptides were bound. Thiε maximal binding capacity of labelled peptide to K b sol protein deteriorated over time, presumably due to radiation by 125 I bound to VSV-8. In a third procedure, each sample contained 0.39nM of [ 125 I]VSV-8 (about 18,000 cp ) , unlabelled peptides at the indicated concentration, and 30nM of K b sol that gives about 50% of the [ 12t, I] VSV-8 binding in the absence of unlabelled peptide at a final volume of 72μl. All components were dissolved and diluted in PBS containing 1% BSA. After incubation for 2-16 hours at 23 °C, 50μl sampleε were analyzed by P30 gel filtration aε deεcribed above. The dissociation constantε for unlabelled peptideε were determined from molar concentrations of [ 12 I]VSV-8 and unlabelled peptides giving 50% inhibition of [ 12B I] VSV-8 binding to K h sol as described. (See Muller, et al . , Meth. Enzymol . 92: 589-601 (1983) . ) K b sol (0.3μM) and [ 12 I]VSV-8 (0.39nM) were then incubated at 4° C, 23 °C, and 37 °C, and the association was determined at various times by P30 gel filtration. Murine ,3-2 microglobulin was added, when necessary, before the incubation at the indicated concentration. The murine β- 2 microglobulin was prepared by affinity chromatography using anti-0-2 microglobulin polyclonal antibody K355 from culture supernatants of the recombinant Drosophila cells. (See also Logdberg, et al., Molec. Immun. 14: 577-587 (1979) .) In another experiment, K b sol ( 0.3μM or l.βμM) and [ 125 I] VSV-8 (2.4nM) were incubated at 23 °C for two hours, and the peptide-K b sol complexes were isolated by P30 gel filtration. The samples contained very small amounts of [ 125 I]VSV-8 and K b sol complexes (at the maximum, 2.4nM) and empty K b sol at final concentration of about 50 to 300nM. To some samples, 3μM of β- 2 microglobulin, 3μM of j β -2 microglobulin plus 20μM of unlabelled VSV-8, 20μM of unlabelled VSV-8, or 1% TX-100 were added. The sampleε were incubated for variouε times at 37 °C and the degree of disεociation waε determined by paεεage over P30 columns . B . Discuεεion Class I MHC molecules present antigenic peptideε to cytotoxic T lymphocyteε. Direct binding of peptide to Claεε I moleculeε in vi tro has been hampered by either the presence of previously bound peptideε at the binding εite (Chen and Perham, Nature 337: 743-5 (1989) ) or the lack of binding εpecificity. (See, e.g., Frelinger, et al . , J. Exp. Med. 172: 827-34 (1990) ; Choppin, et al . , J. Exp. Med. 172: 889-99 (1990) ; Chen, et al., J. EXP. Med. 172: 931-6 (1990) .) In vi tro analyεiε of peptide binding to εoluble, empty Claεε I moleculeε purified from Drosophila cellε tranεformed with truncated H-2K εol and murine β- 2 microglobulin geneε is diεcloεed herein. The results demonstrate that peptide binding is very rapid and naturally processed peptides (octapeptides; εee, e.g., Van Bleek, et al . , Nature 348: 213-6 (1990) ; Falk, et al . , Nature 351: 290-6 (1991)) have the highest affinitieε to K b sol of the nanomolar range and indicate that K b sol complexed with octapeptides are εtable, whereaε thoεe complexed with slightly shorter or longer peptides are short-lived. Interactions between free heavy chain and β- 2 microglobulin is basically reversible in the absence of detergent. Peptides spontaneously bind to empty Class I molecules without dissolution of β- 2 microglobulin. However, excess β- 2 microglobulin apparently promotes the binding of peptide to empty Class I as a consequence of reassociation of free heavy chain with β-2 microglobulin under conditions where the heterodimers are unstable. Soluble H-2K b molecules (composed of the αlα2c.3 domain of heavy chain) and murine β- 2 microglobulin, were purified from the culture supernatantε of Drosophila cellε which were concomitantly tranεformed with the truncated heavy chain and β- 2 microglobulin geneε. Preliminary examinations suggested that Drosophila cells expresε Class I MHC molecules devoid of endogenous peptides on the cell surface. Some of the properties of empty Claεε I moleculeε include the obεervation that they are less stable at 37 °C and their structure is stabilized by the binding of peptide. (See, e.g., Schumacher, et al . , Cell 62: 563-7 (1990) ; Ljunggren, et al . , Nature 346: 476-80 (1990) .) To confirm that purified soluble K h are also empty, their thermal stability in detergent-free solution was examined. Surprisingly, the proteins heated for one hour at 47 °C were well recovered by immunoprecipitation using a confor ational antibody, Y3. This unexpected result led us to add detergent, 1% Triton X-100 (polyoxyethylene (9) octyl phenyl ether) , to the protein solution, since similar experiments to test the stability of Clasε I moleculeε have always been conducted in detergent lysateε (See Schumacher, et al . , cited supra ) . The results obtained in the presence of detergent show that the purified K b sol is now unεtable at 37 °C. Thiε and other lines of evidence suggest that K b sol heterodimer disassembles into the heavy chain and β- 2 microglobulin at elevated temperatures and that detergent may prevent ,3-2 microglobulin from reassociating with dissociated free heavy chain (see below) . Second, the possibility of stabilizing purified K b sol with peptides was studied. The reεults of the first-described examination demonstrated that the proteins can be stabilized only when they are mixed with octapeptide (vesicular stomatitis virus nucleocapsid protein [VSV-8] , see Table 3 below) which is shown to be a naturally processed peptide (see Van Bleek, et al . , cited supra ) . These obεervations are consiεtent with the characteriεticε of empty Class I molecules mentioned above. Independent support that the purified K b sol moleculeε are empty iε provided by iεoelectric focuεing (IEF) under native conditionε (data not εhown) . The εoluble K b purified from Drosophila cellε exhibited a much εimpler pattern than HLA-A2 moleculeε purified from human lymphoblaεtoid cell lines (see Figure 3 in Silver, et al., Nature 350: 619-22 (1991)) . The complicated pattern of HLA-A2 on IEF is presumed to be the result of the presence of heterogeneous peptides bound to the molecules. The simple band of purified K l, sol indicateε the abεence of endogenouε peptideε. In addition, the incubation of K b sol with antigenic peptides caused the distinct εhiftε of band on IEF gel, reflecting the change in iεoelectric point of K h εol due to the peptide binding. It should be noted that such band-shifting was not obεerved in HLA-A2 molecules when they were simply mixed with peptides, unlesε HLA-A2 are incubated with peptideε in "reconstituting conditions" after removal of previously bound endogenous peptides. Taken together, these observations on native IEF also indicate that soluble K b purified from Drosophila cellε are empty. The association of 125 I-labelled VSV-8 with K b sol was demonstrated by gel filtration (not εhown) . The radioactivity of high molecular weight materialε corresponds to peptide-K b sol complexes, while that of low molecular weight materials represents free peptides. Unlabelled VSV and ovalbumin (OVA) peptides could compete with the labelled VSV-8 (see below) , arguing that [ 125 I]VSV-8 is bound specifically to K b sol moleculeε. Reversed-phase HPLC revealed that K b -bound [ 125 I]VSV-8 haε the identical retention time to the input peptide. The binding to K b sol of the labelled VSV-8 was saturable, exhibiting a dissociation constant (K D ) of about 33nM (not shown) . From the x-axis of the Scatchard plot, it waε noted that about 65% of the labelled VSV-8 iε able to bind to K b . To determine affinitieε of variouε peptideε to K b , competitive radioimmunoaεεayε (RIA) using [ 12 I] VSV-8 were carried out (data not εhown) . The inhibitory peptides used for the RIA are listed in Table 3. K r for each peptide is summarized in Table 3 as well. TABLE 3 Various Antigenic Peptideε* Used in Present Studieε Code Sequence VSV-7 GYVYQGL VSV-8 RGYVYQGL VSV-9N LRGYVYQGL VSV-ION DLRGYVYQGL VSV-9C RGYVYQGLK VSV-IOC RGYVYQGLKS OVA-8 SIINFEKL OVA-9N ESIINFEKL 8.9 X 10 ' OVA-ION LESIINFEKL 2.8 X 10" 7 OVA-9C SIINFEKLT 1.1 X 10' 8 OVA-IOC SIINFEKLTE 1.4 X 10" e OVA-24 EQLESIINFEKLTEWTSSNVMEER 7.1 X 10 "5 SEV-9 FAPGNYPAL 2.7 X 10 "9 VSV-8: Vesicular stomatitis virus nucleocapsid protein 52-59 (Van Bleek, et al . , Nature 348: 213-216 (1990)) OVA-8: Ovalbumin 257-264 (Carbone, et al., J. Exp.Med. 169: 603-12 (1989) ) ; SEV-9: Sendai virus nucleoprotein 324-332 (Schumacher, et al., Nature 350: 703-706 (1991)) * All peptides were purified by C 18 reversed-phase HPLC to exclude contaminating shorter peptides with different binding properties. The 3-letter code designations and SEQ ID NO for each peptide are given below. VSV-7 GlyTyrValTyrGlnGlyLeu (SEQ ID NO 40, residue nos. 4-10) VSV-8 ArgGlyTyrValTyrGlnGlyLeu (SEQ ID NO 40, residue nos. 3-10) VSV-9N LeuArgGlyTyrValTyrGlnGlyLeu (SEQ ID NO 40, residue nos. 2-10) VSV-10N AspLeuArgGlyTyrValTyrGlnGlyLeu (SEQ ID NO 40) VSV-9C ArgGlyTyrValTyrGlnGlyLeuLys (SEQ ID NO 44, residue nos. 1-9) V SV-10C ArgGlyTyrValTyrGlnGlyLeuLysSer (SEQ ID NO 44) OVA-8 SerllelleAsnPheGluLyεLeu (SEQ ID NO 39, reεidue noε . 5-12) OVA-9N GluSerllelleAsnPheGluLysLeu (SEQ ID NO 39, residue nos. 4-12) OVA-ION LeuGluSerllelleAsnPheGluLysLeu (SEQ ID NO 39, residue nos. 3-12) OVA-9C SerllelleAsnPheGluLyεLeuThr (SEQ ID NO 39, residue nos. 5-13) OVA-IOC SerllelleAsnPheGluLysLeuThrGlu (SEQ ID NO 39, residue nos. 5-14) OVA-24 GluGlnLeuGluSerllelleAsnPheGluLysLeuThrGlu-TrpT hrSerSerAsnValMetGluGluArg (SEQ ID NO 39) SEV-9 PheAlaProGlyAsnTyrProAlaLeu (SEQ ID NO 45) The peptides of naturally procesεed εize (8mer for VSV and OVA, and 9mer for sendai virus nucleoprotein [SEV] ) had the highest and remarkably similar affinities from the range of 2.7 to 4.1 nM. this exceedingly high affinity of the natural peptides is consistent with recent observations. (See, e.g., Schumacher, et al . , Nature 350: 703-6 (1991) ; Christnick, et al . , Nature 352: 67-70 (1991) .) However, peptides that were shorter or longer by as little as one or two reεidueε lowered the affinity by a factor of from 2 to 100. Thiε reduction of the affinity is even more drastic for a much longer peptide; i.e., the affinity of 24mer peptide (OVA-24) is more than 10, 000-fold lower than that of OVA-8. These resultε help to explain why earlier reportε uεing longer peptideε claim the affinity of micromolar range. (See, e.g., Frelinger, et al . and Choppin, et al . , both cited supra.) It iε of particular intereεt that the extenεion of peptideε at the carboxyl terminus is much lesε deεtructive of the affinity than extenεion at the amino terminuε. According to the three-dimenεional εtructure of HLA-A2, the peptide-binding groove iε formed by two long a heliceε on the antiparallel β strands, and the cleft is about 25 angstromε long, which iε proposed to accommodate an extended peptide chain of about eight residues (see, e.g. , Bjorkman, et al . , Nature 329: 506-12 (1987) ) . At one end of the cleft, the αl and c.2 helices come close together tightly, while at the other end, the cleft is fairly open. It iε now εpeculated that both VSV and OVA peptide bind to the cleft in the εame orientation *and the carboxyl terminus of the peptides might interact with the relatively open end of the cleft so that the extension of peptide at the carboxyl terminus doeε not cause severe steric hindrance . Examinations were then performed to determine the rate of peptide binding to K b at 4 °C and 23 °C, respectively (not shown) . Binding was very rapid, especially at 23 °C, with a half-time of about 5 minutes even in extremely low concentrations of labelled peptides (about 0.4nM) . This contrasts with previous observations, which show a half-time of asεociation of about two hours. (See, e.g., Choppin, et al . , cited supra . ) Again, only 65% of the total labelled peptide was able to bind. The addition of excess ,3-2 microglobulin did not affect the peptide-binding kinetics at such low temperatures that K b heterodimer is stable (remained to be assembled) . This implies that exchange of β- 2 microglobulin is not a prerequisite for peptide binding; i.e. , peptides can spontaneouεly bind to empty Class I molecules without disεociation of ,3-2 microglobulin. In contraεt, excess free ,3-2 microglobulin apparently promoteε peptide binding at 37 °C (data not εhown) . Aε the concentration of added β- 2 microglobulin increased, more peptides bound to K 1, molecules. Since empty K b are unstable at 37 °C, a fraction of heterodimerε muεt be diεsociated to the heavy chain and ,3-2 microglobulin and thereby, the heterodimer must be in equilibrium with free heavy chain and free β - 2 microglobulin. Then, the addition of ,3-2 microglobulin should shift the equilibrium toward the formation of heterodimer that can bind peptides. Thiε view iε εupported by recent obεervationε that there are εubε t antial numberε of Claεε I free heavy chainε on the normal cell εurface and exogenouεly added β- 2 microglobulin facilitateε peptide binding to empty Class I molecules on cells as a consequence of the reassociation of ,3-2 microglobulin with free heavy chain. (See, e.g., Rock, et al., Cell 65: 611-620 (1991) ; Kozlowski, et al . , Nature 349: 74-77 (1991) ; Vitiello, et al., Science 250: 1423-6 (1990) .) The dissociation kinetics of peptide at 37 °C were then observed. Immediately after isolating [ 125 I] VSV-8 and K b complexes by gel filtration, the samples containing either 50 or 300 nM K b were exposed to 37 °C temperatures. Some samples were supplemented with 3μM β- 2 microglobulin and/or 20μM unlabelled VSV-8, or 1% TX-100. The diεsociation of labelled peptides from K b waε meaεured at variouε timeε (not εhown) . In the presence of a large excesε of unlabelled peptideε, the dissociation rate of peptide followed first-order kinetics with a half-time dissociation of about 36 minutes (a dissociation rate constant of 3.2 X 10 ~4 s "1 ) . This unexpected, relatively rapid disεociation of labelled peptide does not agree with some current views of stable peptide-Class I complexes. In fact, the results ascertained (not shown) demonstrated that K b and VSV-8 complexes are stable. This discrepancy must ariεe from the 10-fold lower affinity of radiolabelled VSV-8 (33 nM) compared with that of unlabelled VSV-8 (3.7 nM) . The firεt-order kineticε were alεo observed when the detergent was added instead of unlabelled peptide, indicating that the detergent makes the peptide dissociation process irreversible. In contraεt, the peptide dissociation profile did not follow the first-order kineticε in the absence of unlabelled peptide or the detergent. This suggests that the peptide association/dissociation is reversible and the binding of peptide is dependent on the concentration of heterodimer (compare the kinetics between 50 nM and 300 nM of K ) . This became more evident when excess β- 2 microglobulin was added. These results support the previous argument that interaction between the heavy chain, j β-2 microglobulin and peptide are basically reversible at 37 °C, if not entirely, in the absence of detergent. It is probable that a detergent such as TX-100 may prevent β-2 microglobulin from reassociating with free heavy chain at 37 °C. This could reasonably explain why K b once heated to elevated temperatures in the absence of detergent can be efficiently immunoprecipitated by conformational antibody (not shown) . Interestingly, the addition of ,3-2 microglobulin did not suppress the peptide dissociation in the presence of excess unlabelled peptides, indicating that labelled peptides are released from the complexes without dissociation of ,3-2 microglobulin. It should be remembered, however, that the affinity of [ 125 I]VSV-8 is about 10-fold lower than that of the natural peptides. Therefore, this is not necessarily the case for the natural peptides. The study using in vi tro peptide-binding asεay εyεtemε suggests that peptide binding to Class I molecules is a simple mass action and a ligand-receptor interaction. The approach used herein allows characterization of the peptide binding εpecificity to Class I molecules and of the interaction of peptide-Class I complexes with the T-cell receptor. Example 8 Therapeutic Applications A. Class I Molecule Bank A reservoir or "bank" of insect cell lines may be established and maintained, with each cell line expressing one of the 50 to 100 most common Class I MHC heavy chain, a ,3-microglobulin molecule, as well as at least one assisting molecule. cDNAs encoding these proteins may be cloned based on HLA variants obtained from cell lines containing same -- e.g., via the polymerase chain reaction (see Ennis, et al . , PNAS USA 87: 2833-7 (1990)) -- and inserted into the appropriate vector, such as an insect expression vector, to generate cell lines expressing each HLA variant. Testing according to the following protocol, for example, can be used to determine which peptides derived from the virus of choice bind the best to the different Class I MHC molecules. The various cultureε may appropriately be labeled or catalogued to indicate which Class I MHC molecules are best for use with particular peptides. Alternatively, transient cultureε may be established as needed. As discussed herein, after approximately 48 hours' incubation of a culture of insect cellε with a vector, that culture is apparently capable of expressing empty MHC molecules which may be loaded with the peptide (s) of choice for the purpose of activating CD8 * cells. B. Preparation of "Special" Cell Lines After HLA typing, if insect cell lines expresεing the preferred HLA are not available, cDNAε encoding the preferred HLA and assiεting moleculeε may be cloned via use of the polymerase chain reaction. The primers disclosed in section B.l. above (SEQ ID NO 1 through SEQ ID NO 12) may be used to amplify the appropriate HLA-A, -B, -C, -E, -F, or -G cDNAs in εeparate reactionε which may then be cloned and sequenced as described in the methods disclosed in Example 1, section 1 above. DNA is then purified from the PCR reaction using a Gene Clean kit (Bio 101, San Diego, CA) and ligated directly into the Sma I site of pRmHa-3. Individual clones are isolated, the sequences verified, and stable Drosophila cell lines expressing the HLA established. Alternatively, a bulk population of recombinant plasmids may be grown in large scale and DNA purified by cesium chloride gradients. The purified DNA is then used to transfect S2 cells using calcium phosphate precipitation techniques. After 24 hours, the precipitate is washed off the cells and replaced with fresh Schneider media containing ImM CuSO.. Forty-eight hours later, the bulk population of transiently transfected cells is used for in vi tro activation of CD8* after incubation with syngeneic peptides or protease digests of specific proteins. Stable cell lines expressing the cloned HLA may then be established. Alternatively, a population of insect cells transiently expressing a bulk population of cloned recombinant molecules from the PCR reaction may be used for in vi tro CD8 * activation. It is also possible to activate haplotype-specific CD8s in vi tro using insect cells expressing Class I MHC incubated with peptides where the cell line-expressed MHC is not the expreεεed element in vivo . This provides a unique opportunity to proliferate CD8 * cells which recognize a specific antigen aεεociated with a particular MHC which would not be poεεible in vi vo due to allelic restriction. For example, a peptide (NP) from the nuclear protein of Influenza virus is ordinarily restricted to the D b molecule; however, we have found that such a peptide can bind to K b (albeit more weakly than to D ) and can generate a degree of thermal stability to the K b (see Figure 3) . Furthermore, K b -expressing Drosophila cells preincubated with the NP peptide and cocultured with splenocytes from a B6 mouse results in the in vi tro activation of CD8 * which specifically recognize the K b molecule associated with the NP peptide. In addition, the reciprocal experiment using a K b -restricted peptide (OVA) derived from ovalbumin and D b -expressing Drosophila cells results in the proliferation of CD8 * which specifically recognize D containing the OVA peptide. Such CD8s are able to kill cells (EL4 OVA) transfected with cDNA encoding the ovalbumin protein, indicating that in vivo, some D b molecules are loaded with the OVA peptide . This system therefore provides a unique opportunity to proliferate CD8 * against specific antigens presented by a Class I molecule which, in vi vo, is not the restriction element for that peptide. Although enough antigen is presented in vivo by said Class I for the cell to be recognized by CD8 * and killed, it is not enough to proliferate such CD8s in vivo . By loading empty Class I molecules expressed by Drosophila cells with peptide, we are able to override the in vivo restriction by providing an excess of antigenic peptide to the Class I molecule in a non-competitive environment such that enough antigen is presented by the Class I to activate the specific CD8 * recognizing this complex. C. AIDS Treatment In vi tro activated cellε may be adminiεtered to patients for in vivo therapy. Preferably, the Class I MHC genotype (haplotype) of the individual is first determined. Conventional tissue typing is appropriate for this purpose and may be performed at the treatment center or by some appropriate commercial operation. Once the individual's HLA type(s) is (are) determined, the best combination of peptides and Class I MHC molecules suitable for the individual patient iε ascertained and prepared as noted above and the appropriate insect cell lines and peptides are provided. Resting or precursor CD8* T-cells from the blood of the patient are then stimulated with the appropriate peptide-loaded MHC produced by the insect cell culture. After activation, the CD8* cells are reintroduced into the patient's bloodstream, and the disease process in the patient continues to be monitored. Methods of removing and re-introducing cellular components are known in the art and include procedures such aε those exemplified in U.S. Patent No. 4,844,893 to Honsik, et al. and U.S. Patent No. 4,690,915 to Rosenberg. Additional treatments may be administered as necessary until the disease is sufficiently remediated. Similar treatment protocols are appropriate for use with other immunosuppressed individuals, including transplant patients, elderly patients, and the like. D. Cancer Treatment In cancer patients, a treatment procedure similar to that described above is utilized. However, in such patients, it should be anticipated that conventional therapy to reduce the tumor mass may precede the immune therapy described herein. Therefore, it is preferred that blood samples from the putative patient be obtained and stored (e.g. via freezing) prior to the commencement of conventional therapy such as radiation or chemotherapy, which tendε to deεtroy immune cellε. Since few, if any, forms of cancer arise in direct responεe to viral infection, target peptideε for immune treatment are less readily observed. However, recent studieε indicate that mutationε in the oncogeneε ras, neu , and p53 contribute to cancer in aε much aε 50% of all cancer cases. Thus, peptideε derived from these mutated regions of the molecules are prime candidates as targets for the present therapy. Pursuant to the protocols disclosed herein, the best combination of peptides and Claεε I moleculeε for the individual patient may be determined and administered. For example, many tumors express antigens that are recognized in vi tro by CD8 * cells derived from the affected individual. Such antigens which are not expressed in normal cells may thus be identified, as well as the HLA type that presents them to the CD8 * cells, for precisely targeted immunotherapy using the methods of the present invention. For example, van der Bruggen, et al. have described an antigen whose expression is directed by a specific gene and which antigen appears to be presented by HLA Al (Science 254: 1643-1647 (1991)) . As various human tumor antigens are isolated and described, they become good candidates for immunotherapeutic applications as described herein. In another, alternative therapeutic mode, it may be feasible to administer the in vi tro activated CD8 * cells of the present invention in conjunction with other im unogens. For example, the Large Multivalent Immunogen disclosed in U.S. Patent No. 5,045,320 may be administered in conjunction with activated CD8 * cells. It is also possible that cytokines such as IL-2 and IL-4, which mediate differentiation and activation of T-cells, may be administered as well, as cytokines are able to stimulate the T-cell response against tumor cells in vivo . It is believed that IL-2 plays a major role in the growth and differentiation of CD8 * precursorε and in CD8 * proliferation. The adminiεtration of IL-2 to cancer patients is frequently associated with an improved anti-tumor response which is likely related to induction of tumor-specific T-cells. However, the best therapeutic effects of IL-2 might be obtained by continuous local rather than systemic administration of IL-2, thuε minimizing the IL-2 toxicity and prolonging itε biological activity. One may achieve local delivery via transfecting tumor cells with an IL-2 gene construct. IL-2 cDNA is constructed as described by Karaεuyama and Melchers in Eur. J. Immunol. 18: 97-104 (1988) . The complete cDNA sequence of IL-2 is obtained as an Xho I fragment from the plasmid pBMGneo IL-2 (see Karasuyama and Melchers, supra) and directly ligated into the Sal I site in pRmHa-3. Recombinant pRmHa-3 plasmid with the insert in the correct orientation (determined via restriction mapping with Hind III) is purified by cesium gradients and used to cotransfect S2 cells using the calcium phosphate technique. (A mixture of plasmid DNA waε prepared for this purpose: lOμg pRmHa-3 containing IL-2 cDNA, 6μg each of pRmHa-3 plasmid containing MHC Class I heavy chain or β- 2 microglobulin and 2μg of phshsneo DNA.) Stable cell lines which are inducible via CuS0 4 to express heavy chain, β- 2 microglobulin and IL-2 were obtained by growing the transfectants in G418 medium. These stable cell lines were coated with peptide and used in the in vi tro asεay as described above. Tumor cells transfected with IL-2 are observed to enhance the CTL (CD8) activity against the parental tumor cells and bypass CD4 and T helper function in the induction of an antitumor or cytotoxic response in vivo . Therefore, increasing the potential of the Drosophila system via cotransfection with the IL-2 gene is suggested herein. Example 9 Dose Dependence of the Production of Activated T-Cellε Using the Antigen-Preεenting Svεtem Antigen-presenting cells (APC) were produced by transfecting Drosophila cells as deεcribed in Example 3 and then tested for their capacity to present antigen to T-cells from the 2C line of transgenic mice. With mouse cells aε antigen-presenting cells, this line displays strong alloreactivity for L d molecules complexed with an endogenouε 8-mer peptide, p2Ca (Leu-Ser-Pro-Phe-Pro-Phe- Asp-Leu, SEQ ID NO 46) , derived from a Krebs cycle enzyme, 2-oxoglutarate dehydrogenase (OGDH) . The p2Ca peptide is exposed naturally bound to L d on the surface of H-2 d cells such as B10.D2 cells. The p2Ca peptide has intermediate binding affinity for soluble L d molecules (4 x 10 6 M" 1 ) and high affinity for 2C TCR molecules (2 x 10 6 M" 1 to 1 x 10 7 M "1 ) . A closely-related 9-mer peptide, QL9 (Gln-Leu- Ser-Pro-Phe-Pro-Phe-Asp-Leu, SEQ ID NO 47) , has even higher affinity for these molecules (2 x 10 β M" 1 for L d and 2 x 10 7 M "1 for 2C TCR) . Except for one additional amino acid (glutamine) at the N terminus, QL9 has an identical sequence to p2Ca and, like p2Ca, forms part of the native sequence of OGDH. With p2Ca and QL9 peptides (prepared in synthetic form) , antigen-presenting cell requirements for mature unprimed 2C CD8* cells were studied in vi tro. The responder CD8 * cells were purified from pooled lymph nodes (LN) of 2C mice on a C57BL/6 (B6, H-2 b ) background by first removing CD4 * cells, class II-positive cells and B cells by mAb + complement treatment followed by positive panning. Cell suspensions were prepared from pooled cervical, axillary, inguinal and mesenteric LN of young adult mice using a tissue grinder. For cell purification, LN cells were first treated with a cocktail of mAbs (anti-CD4, anti-HSA, anti-I-A b ) pluε complement for 45 minutes at 37 °C. The surviving cells were further separated into CD8 * and CD8 " (CD4 ~ ) cells by panning at 4 °C for 60-90 minutes on petri dishes coated with anti-CD8 mAb. The attached CD8 * cells were recovered by incubation at 37 °C for 5 minutes followed by vigorous pipetting. Non-attached cells were eluted and treated with anti-CD8 mAb and complement to obtain CD8 * 1B2* 2C cells. More than 95% of the CD8 * cellε obtained were clonotype-poεitive (IB2 * ) and 98% of these cells displayed a naive (CD44 lσ ) phenotype . TCR stimulation elicited a complex pattern of intracellular events which lead to early up-regulation of CD69 and CD25 (IL-2 receptor or IL-2R) on the cell surface . These changes are apparent within a few hours of stimulation and are followed by cytokine synthesis and cell proliferation. Drosophila cells were transfected with genes for L d , L d and B7.1 (L d .B7) , L d and ICAM-1 (L d .ICAM) or L d , B7.1 and ICAM-1 (L d .37. ICAM) . Figure 11 shows CD69 and CD25 expression on purified naive CD8 * 2C cells stimulated for 12 hours with transfected Drosophila cells and p2Ca versus QL9 peptide at a concentration of 10 μM. Purified CD8 * 2C cells were incubated with various Drosophila cells plus a peptide (either p2Ca or QL9, 10 μM) in bulk (2 ml) culture for 12 hours and then stained for CD69 or CD25. Either p2Ca or QL9 presented by Drosophila cells transfected with L d .B7, L d .ICAM or L d .B7.ICAM were effective in stimulating the up- regulation of CD69 and CD25. However, non cultured 2C cells without either peptide or Drosophila cells did net show up-regulation of CD69 and CD25 (top panel) . In the presence of Drosophila cells expressing L d alone, induction of CD69 and CD25 expression on CD8' 2C cells was low, but significant, with the strong QL9 peptide but barely detectable with the weaker p2Ca peptide. With L d .B7 or : .ICAM antigen-presenting cells, both peptides elicited .arked up-regulation cf CD69 ana CD25. However, L d .B7. ICAM antigen-presenting cells induced even higher expression of these molecules . Drosophila cells transfected with L d alone failed to cause proliferation of 2C CD8 * cells to either p2Ca or QL9 peptide in the absence cf exogenous lymphokines, consistent with the minimal capacity of these cells as antigen-presenting cells to induce CD69 and CD25 expression. The results are shown in Figure 12. Responses to p2Ca (above) and QL9 (below) were meaεured by culturing 5 x 10* purified CD8 * 2C cells with 2 x 10 5 Drosophila cells in the presence or absence of the indicated concentrations of peptideε for 3 dayε. [ 3 H] TdR was added during the last 8 hours of culture; rIL-2 was added at a final concentration of 20 units/ml. The BAD ORIGINAL data are the mean of triplicate cultures. When supplemented with exogenous IL-2, however, both peptides stimulated significant proliferative responses at high doses (10 μM) . The proliferative responses elicited by QL9 were far stronger than for p2Ca (note the large difference between the scales of x-axes of the upper and the lower panel) . Dose-response relationships for proliferation on day 3 elicited by Drosophila cells transfected with L d .B7.ICAM and presenting p2Ca peptide were compared to those elicited by Drosophila cells transfected with L α .B7, L d .ICAM or L d .B7.ICAM presenting QL9 peptide. The results (means of triplicate cultures) are shown in Figure 13. Drosophila APC (2 x 10 5 cells) were cultured with 5 x 10* CD8 * 2C cells for three days in the presence or absence of peptides. [ 3 H] TdR was added during the last 8 hours of culture; no IL-2 was added to the cultures . Optimal proliferative responses elicited by L d .B7. ICAM antigen-presenting cells on day 3 required high concentrations of p2Ca peptide, e.g. 10 μM (10 ~5 M) , (Figure 13) . The results for QL9 peptide were different. The does-response curves for L°.B7 antigen-presenting cells plus QL9 and L d .I AM antigen-presenting cells plus QL9 approximated the results for L d .B7.ICAM antigen- presenting cells plus p2Ca (Figure 13) . However, with L .B7.ICAM antigen-presenting cells, proliferative responses to QL9 on day 3 were maximal with 100 nM (10 "" M) and were clearly apparent with doses as low as 10 pM (10" 11 M) (Figure 13) . At high doses, e.g. 10 μM (10 ~5 M) , QL9 inhibited the proliferative response on day 3 (Figure 13, note the log scale) . Inhibition of proliferation by L d .B7.ICAM antigen-presenting cells and QL9 peptide did not apply to IL-2 production (Figure 14, right) , and was only seen .00 with high doses of antigen-presenting cells (Figure 14, left) . L d .B7.ICAM antigen-presenting cells plus QL9 peptide was again found to be more effective in eliciting IL-2 production than L 2 .37. ICAM antigen-presenting cells plus p2Ca peptide (Figure 14, right) . The data indicates that antigen-presenting cells without peptides were ineffective in eliciting either response over the 100- fold range of antigen-presenting cells density tested (Figure 14, "-pep," open squares) . Changes in the dose-response relationships of the responses of CD8 * 2C cells on Day 3, Day 4 and Day 5 were examined using QL9 peptide with L d .B7, L d .ICAM or L = .37. ICAM antigen-presenting cells. The results are shown in Figure 15. Responses were measured with 5 x 10 4 CDS " 2C cells and 3 x 1C Ξ antigen-presenting cells at the indicated peptide concentrations. The data are the mean results of triplicate cultures. With an intermediate dose of 100 nM (10 ~7 M) QL9 peptide, proliferative responses to L = .B7.ICAM antigen-presenting cells were high en Day 3 (Figure 15, left) , reached a peak en Day 4 (Figure 15, center) and then declined to low levels on Day 5 (Figure 15, right) . With a high dose of 10 μM (10 ~5 M) QL9 peptide, however, the response was low or. Day 3 (Figure 15, left) , but then increased markedly to reach a high peak on Day 5 (Figure 15, right) . Transient inhibition of proliferation induced by QL9 on Day 3 was only seen when the avidity of T-cell/APC interaction was very high. Decreasing the avidity of T-cell/A?C interaction by using lower doseε of A?C (Figure 14, left) cr lower doεeε of QL9 peptide (Figure 15, left) augmented the Day 3 proliferative response. Reducing the avidity cf T-cell/APC interaction enhanced the early (Day 3) proliferative response but reduced the late (Day 5) response and also reduced IL-2 production (Figure 16, right) . In Figure 16 the results obtained using CD8 * 2C and CD8 " 2C cells on Days 2, 3, 4 and 5 are compared. The observations apply with the highly immunogenic L 3 .B7.ICAM antigen-presenting cells. However, with " the less immunogenic L d .B7 or L d .ICAM antigen-presenting cells, proliferative responses to QL9 peptide required high doses of peptide (Figure 13, Figure 15) and were crucially dependent upon CD8 expression by the responder cells (Figure 16) . These responses with L ≤ .B7 and L d .ICAM antigen-presenting cells reached a peak on Days 3 or 4 (rather than Day 5) and were far lower than with L d .B7.ICAM antigen-presenting cells. With low doses of QL9, e.g. 1 nM (10 ~5 M) , proliferative responses with L d .B7 and L d .ICAM antigen-presenting cells were completely undetectable (< 100 cpm) (Figure 13) . This was in striking contrast to the results seen using L d .B7.ICAM antigen-presenting cells, where 1 nM QL9 led to high responses (> 10,000 cpm) (Figure 13) . In contrast to the results with high dcseε of QL9 peptide (10 μM, Table 2) , the synergistic interaction between B7 and ICAM for proliferative responses became pronounced at low peptide doses. As can be seen, L d .B7.ICAM cells act as extremely potent antigen-presenting cells for naive 2C cells. Raising the avidity of T-cell/APC interaction to a high level inhibits the early proliferative response but potentiates the late response and IL-2 production is enhanced. For the high-affinity peptides such as QL9, the synergy between B7 and ICAM is pronounced at low doses of antigen. Example 10 Production of Cvtotoxic T-Cells Using the Antigen Presenting System Antigen-presenting cells (APC) were produced by transfecting Drosophila cells as described in Example 3 and then tested for their ability to induce CTL activity. CTL activity was tested on £1 Cr-labelled RMA.S-L 0' targets sensitized with QL9 peptide, no peptide or an irrelevant peptide (MCMV) . CD8 * 2C cells (5 x 10 5 ) were cultured with 2 x 10 6 transfected Drosophila cells in a volume of 2 ml in a 24-well culture plate. Cai, Z., and J. Sprent, (1994) . Peptides were present during the culture at a concentration of 10 mM. After 4 days, the cells were pooled and adjusted to the required number. To prepare targets, RMA-S.L d cells were labeled with Ξ1 Cr (100 mCi/1-2 x 10 6 cells) at 37 °C for 90 minutes in the presence or absence of peptides. After labeling, the cells were thoroughly washed and resuspended in medium with or without peptides. Specific 5: Cr release waε calculated according to established procedure. Id. The capacity cf L=.B7, A .37. ICAM and LMCAM antigen-presenting cells to induce CTL activity to 10 μM QL9 peptide in bulk cultures is shown in Figure 17. Strongly immunogenic L".37. ICAM antigen-presenting cells were efficient in generating QL9-specific CTL (Figure 17, center) . Significantly, L d .B7 antigen-presenting cells were also effective in generating CTL to QL9 (Figure 17, left) . The surprising finding, however, was that L = .ICAM antigen-presenting cells were totally unable to stimulate CTL generation (Figure 17, right) . This result (representative of three different experiments) was unexpected, because LMCAM antigen-presenting cellε were no less efficient than L .37 antigen-presenting cells at inducing proliferative responseε to QL9 (Figures 13 and 15) . The L d . ICAM-stimulated cultures in Figure 17 BAD ORIGINAL contained large numbers cf blast cells, and total cell yields were about 3-fold higher than the input number. The surprising finding that L d .ICAM antigen- presenting cells were totally unable to stimulate CTL generation applied to cultures not supplemented with exogenous lymphokines. However, when exogenous IL-2 was added to the cultures, L-.ICAM antigen-presenting cells induced strong CTL activity to QL9 (Figure 18, right; 20u/ml exogenous IL-2) . Example 11 Proliferation of Normal T-Cells Induced bv the Antigen-Presenting System Antigen-presenting cells (APC) were produced by transfecting Drosophila cells as described in Example 3 and then tested for their ability to induce proliferation in normal (nontransgenic) murine T-cells. The capacity cf L d .B7.ICAM Drosophila cells to induce a strong primary response cf 2C TCR transgenic CD8 * cells raised the question whether Drosophila cells could act as antigen-presenting ceils for normal (nontransgenic) CD8 " ceils. Since the 2C mice were on a B6 (H-2 b ) background, the response cf normal B6 CD8 * cells was tested. Graded doses of CD8 * cells from normal E6 mice were cultured with 10 μM QL9 peptide presented by L d .B7.ICAM Drosophila antigen-presenting cells, i.e. a situation where a diverse repertoire of T-cells was exposed to only a single alloantigen (L d + QL9) but at high concentration. As shown in Figure 19 (left) , presentation of QL9 by L i .B7.ICAM antigen-presenting cells was indeed immunogenic for normal B6 CD8 * cells and led to appreciable proliferative responses on Day 3 (80,000 cpm) with large doseε of respcnder cells (1 x 10 € ) in the absence of added cytokines; responses to an unrelated peptide, MCMV, were much lower (though significant) and no response occurred in the absence of peptide. As expected, the response of normal B6 CD8 * cells to QL9 plus L d .B7.ICAM antigen-presenting cells was substantially lower than with normal B10.D2 spleen cells as antigen-presenting cells (where the allostimulus was provided by a multiplicity of self peptides bound to L d , K and D d , but at low concentration) (Figure 19, right. Note the difference in the Y axis scales) . Example 12 Activation Of Cytotoxic T-Cells Using Immobilized Purified Recombinant MHC Class I Molecules And Assisting Molecules Except as noted, cells, materials and reagents were prepared as described in Example 4. Biotinylated anti-mouse CD28 monoclonal antibody (clone 37.51) was purchased from Pharmingen (San Diego, CA) . This antibody is also available from Caitag (South San Francisco, CA) . This antibody, which binds to CD28 co-stimulatory receptor, a ligand of 37.1 and B7.2 en the surface of T-cells, augments the proliferation of T-cells (Gross et al. J. Immunol. 149, 380-388, 1992) . IL-2 was used as a concanavalin A supernatant (10% final concentration) . Substrates were prepared as follows: Fifty microuters of PBS containing 1 microgram/ml of neutravidin were added to each well of a 96 well cell culture plate (Corning cat. #25860) . After 2 hours at room temperature, the wells were washed 3 times with PBS prior to incubation with biotinylated molecules. Avidin-coated mouse red blood cells were prepared as described for avidin-coated sheep red blood cells in Example 4. Avidin-coated latex beads were prepared as described Example 4. Biotinylation of recombinant L d was performed as described in Example 4. Biotinylated recombinant L a was immobilized on the substrate together with biotinylated anti-CD28 antibody. Avidin-coated red blood cells or latex beads were incubated with 0.2 mg/ml biotinylated L d , 0.025 mg/ml biotinylated anti-CD28, or a mixture thereof for 30 minutes at room temperature, then washed 3 times in DMEM containing 10% FCS and used immediately. Avidin-coated 96 well plates were incubated with 50 microliters per well of 2 microgram/ml biotinylated L d , 0.25 microgram/ml anti-CD28, or mixtures thereof for 30 minutes at room temperature, then washed 3 times using DMEM containing 10% FCS and used immediately. 2C+ T-cells were prepared as described in Example 4. CD8 * cells from C57BL/6 mice were prepared from the lymph ncdeε of these mice by magnetic depletion. Purified cells were consistently 90-92% positive for CD8 expression, as assessed by flow cytofluorometry. T-cell activation was performed in culture plates coated with purified L d molecules and anti-CD28 antibody. T-cells and peptide (0.C2 M final concentration) were added to 96 well plates coated with L d and/cr anti-CD28, in a final volume cf 0.2ml/well and cultured for the appropriate time at 37 C C in humid atmosphere containing 5% C0 2 . T-cell activation using red blood cells cr latex beads coated with purified L d molecules and anti- CD28 antibody was performed in uncoated culture plates. T-cells and peptide (0.02mM) were added to each well, together with 100,000 red blood cells or latex beads. Final volume was 0.2ml. Culture conditions were as described above. T-cell mitogenesis was assayed by incorporation cf a pulse of 1 μCi of tritiated thymidine per well by triplicate cell cultures. Cells were harvested 8 hours later and thymidine incorporation was determined by counting the filters in a scintillation counter. RMA.S cells (target cells) expressing L d were incubated with radiolabeled chromium for 90 minutes at 37 °C, washed 3 times and distributed in 96 well U-bottom plate (Costar cat 3799) (5000-10000 cells per well) in the presence of the appropriate peptide at 0.0ImM. Various amounts of activated T-cells (effector cells) were added to reach effector/target ratios (E/T ratios) ranging between 150 and 1. After 5 hours of incubation at 37 °C, 0.1ml of supernatant was collected from each well and counted in a gamma counter. Percent lysis was determined by a standard method (Ccligan et al . , Current Protocols in Immunology, section 3.11, Wiley, New York (1991) ) . Immobilized purified L d and anti-CD28 antibody were mitogenic for 2C+ T-cells. When QL9 peptide was used, a thymidine incorporation above 100,000 cpm per well was consistently measured by day 3-5 of culture (Figure 20) . Maximum thymidine incorporation in that same range was obtained using any cf the three methods of activation (molecules immobilized on plastic, on red blood cells or on latex beads) . When activation molecules were immobilized on plastic, L d alone immobilized on plastic induced a transient mitogenesiε (Figure 20, broken line) whereas L d plus anti-CD28 antibody induced a higher and mere sustained mitogenesis (Figure 20, solid line) . Ar.ti-CD28 antibody in addition to L was required for the induction of any mitcgenesiε in the model using red blood cells. However, anti-CD28 antibody alone did not induce any mitogenesis. QL9-L d complexes are recognized by the 2C T- cell receptor with a high affinity. Complexes cf ether peptides and L d recognized by this receptor with a lower affinity were able to activate 2C+ T-cells, • theεe included peptides p2Ca and SL9 (Figure 21) . However, IL- 2 added at day 2 of culture was required in order to observe mitogenesis using these peptides. Activation was peptide specific since LCMV peptide, a control peptide that is not recognized by the 2C T-cell receptor, did not induce activation. Mitogenesis was measurable starting with as little as 700 T-cells per well (96 plate well) , demonstrating that the method activated a very small number of T-cells in a peptide specific manner. 2C+ T-cells were mixed with total CD8 * cells from C57BL/6 mice at a 1/99 ratio and cells were cultured with immobilized L d and anti-CD28 antibody in the presence of peptide QL9. IL-2 was added at day 2 of culture, and subsequently every other day. Cell activation and proliferation was noted, as evidenced by the formation of clumps, presence cf enlarged cells that progressively spread in the well. At day 12 of culture, cells were harvested and percentage of cells expressing the 2C T-cell receptor was assessed by staining with the anticlonotypic antibody 1B2 and analysis by flow cytofluorometry: 43% of the cells cultured initially in the presence of peptide QL9 were 2C+, (Figure 22) whereas 49% of the cells expressed 2C after stimulation with peptide p2Ca (Figure 23) , and 22% with peptide SL9 (Figure 24) . This shows a considerable enrichment (43, 49 and 22 times) in specific T-cells after 12 cays of culture. Enrichment was observed even with SL9, a peptide that makes a low affinity complex with the 2C T- cell receptor. This method thus can specifically activate and enrich a small subpopulation of antigen specific T-cells out of a heterogenouε mixture of T- cellε. 2C+ T-cells, activated using L d and an i-CD28 antibody in the presence of QL9 peptide, were cultured for 5 days, then their cytotoxic capacity was assessed. The results, shown in Figure 25, demonstrated that immobilized activation molecules induced the cells to differentiate into effector cytotoxic T-cells. Lysis was specific, since it was observed in the presence of QL9 peptide but not in the presence of a control peptide (LCMV) . Resting T-cells were thus activated using immobilized molecules to differentiate into cytotoxic T-cells able to specifically kill targets. As can be seen, immobilized purified MHC Class I and assisting molecules can be used to specifically activate naive resting T-cells into cytotoxic T-cells. MHC class I and assisting molecules are sufficient for activation; no additional signal originating from antigen-presenting cells is necessary. MHC class I and assisting molecules immobilized on substrates, such as cell culture plates, provide an appropriate tool for T-cell activation. Several advantages are offered by such coated substrates. They are easy to prepare and to manipulate, they can be coated with well-controlled amounts of molecules, ensuring reproducible activation conditions. Example 13 Activation of Human Cytotoxic T-Cells A unit of human blood (450 mis) collected in heparin (10 μ/ml) was obtained through the General Clinical Reεearch Center (GCRC) at Scripps Clinic, La Jolla, CA. The blood was first processed in a Ficoll-Hypaque density gradient in a 50 cc conical centrifuge (Histopaque 1077, Sigma) according to the manufacturer's specifications. Once the buffy coat was obtained, the sample was washed in buffer 1 (D-PBS without Ca" or Mg") , then buffer 2 (RPMI with 4% fetal calf serum (FCS) ) and a final wash in buffer 3 (D-PBS + 1% human serum albumin (HSA, 25% Buminate/Baxter-Hyland) and 0.2% sodium citrate (w/v) ) . The total peripheral blood mononuclear cell (PBMCs) preparation was counted from the washed cells. This preparation was then taken through a MaxSep™ isolation procedure (Baxter) where CD8 * cells were selected by negative selection. A cocktail of mAbs to cells targeted for removal (CD19-PharMingen, CD4-Ortho-mune, CD15-PharMingen, CD56-PharMingen, CD14-PRI) was prepared at 2 μg/ml of cells. The total ?3MC cell count was diluted to 20 x 10 6 /ml in buffer 3 and the antibodies were added. The mixture (approximately 40 mis) was rotated (4 rp ) on a rotary shaker at 4 * C, so that the tube mixed the sample end over end, for a total of 30 minutes. After the sensitization phase, the cells were washed with buffer 3 and resuspended in the same buffer with magnetic Dynal beads coated with sheep-anti-mouse IgG (SAM) (Dynabeads M450 #110.02) . The stock of beads is usually 4 x 10 8 beads/ml. The final bead:target cell ratio is 10:1. To determine the final volume of beads to use, the formulas below were followed: (Total cells sensitized) x (% of population of target cells) = Total theoretical target cell number. Total theoretical target cell number x 10 = Total beads required. Total beads required/Bead concentration of stock = Volume of beads required. The volume of the final senεitized cells :beadε mixture was apprcximately 50 mis. The mixture was put into a 150 ml Fenwal Transfer Pack (#4R2001) , air added with a needle and the mixture rotating under similar conditions as above (30 minutes, 4 'C, 4 rpm, agitated end-over-end) . At the end of the incubation period, the target cells were removed with a MaxSep 1 " separation device according to the manufacture's specifications. The separated cells were transferred from the bag and counted to determine recovery. FACS analysis was performed to determine the purity of the sample. The resulting separated human CD8 * cells were stimulated with Drosophila (fly) antigen-presenting cells which were transfected and shown to express human HLA A2.1, E7.1 and/or LFA-3. The fly cells were diluted to 10 6 /rτ.l in Schneider's medium with 10% FCS serum. On the following day CuS04 was added to the cultured cells which were incubated for 24 hours at 27 "C and harvested. The harvested cells were washed and suspended in Insect X-press medium with a final peptide concentraticn of 100 μg/ml and incubated for 3 hours at 27 * C. The peptides used were KIV-RT (ILKEPVHGV) (SEQ ID NO 48) , Tyrcsiπase (YMNGTMSQV) (SEQ ID NO 49) , and Influenza Matrix (GILGFVFTL) (SEQ ID NO 50) . The control peptide was derived from the core of the hepatitis virus and had the sequence FLPSDFFPSV (SEQ ID NO 51) . The fly antigen-presenting cells were added to the CD8" cellε at a ratio cf 1:10 (APC.CD8 * T-cell) . The cellε were incubated in flat bottom wells (48 wells) for four days at 37 'C. At day 5, IL-2 (10 μ/ml) and IL-7 (10 μ/ml) (Genzy e) were added with a media change. On day 11 a CTL assay was performed. JY, an ΞBV-transformed human B cell line expressing HLA 2.1, 57 was uεed as the target cell in the chromium release asεay. Vissereu, M.J.W. et al. J. Immunol . , 154:3991-3998 (1995) . JY cells were seeded at 3 x 10 5 cells/πl in RPMI with 10% FCS, 24 hours prior to the assay. The JY cells were counted and washed once in RPMI wash solution (4% Rehautin FCS, 1% HEPES, 0.025% gentamycin in RPMI) to provide 5 x 10 6 cells per sample. The cell pellet was gently resuεpended in 100 μl of S1 C hromium stock (0.1 mCi, NEN) and placed in a 37 °C water - Ill - bath for 1 hour with agitation every fifteen minutes. Labelled JY cells were washed four times for 6 minutes each at 1400 rpm in 10 mis of RPMI wash solution. Cells were adjusted to 1 x 10 £ cells/ml in RPMI/10% Hyclone. Efficiency of target cell labelling is confirmed by standard gamma counter techniques. For peptide loading of target cells, two mis of labelled JY cells (2 x 10 £ cells) were incubated with 10 μg/ml peptide for 30 minutes at room temperature. Peptide stock solutions (lmg/ml) were stored at -70 * C. In a 96 well round bottom plate, 100 μl effector cells and 100 μl peptide loaded target cells were combined at ratios of 84:1, 17:1 and 3.4:1 (effector:target) . Controls to measure spontaneous release and maximum release of 51 Cr were included in duplicate. Samples were incubated at 37 * C for 6 hours. K562 cells at a concentration of 10 7 /ml in RPMI with 10% FCS were added at a ratio of 20:1 (unlabelled K562 :labelled JY) . This erythroleukemic cell line was used to reduce NK background cell lysis in the chromium release assay. Plates were centrifuged at 1000 rpm fcr 5 minutes and 100 μl supernatant from each εample transferred to 96 tube collection tubes. /Analysis cf cell lysiε iε determined by standard gamma counting techniqueε (Gammacell 1000, Nordion) . CTL activity produced by activating human CD8 * T-cellε with antigen-preεenting cellε loaded with influenza matrix peptide (SEQ ID NO 50) is shown in Figure 26. Prior exposure to influenza virus indicates that this CTL activity was a secondary response. Data points are the mean of values from triplicate cultures. Antigen-presenting cells expresεing A2.1, B7.1 and ICAM-1 were more effective than antigen-presenting cells expressing A2.1 and B7.1 cr antigen-presenting cells expressing A2.1, B7.1 and LFA-3. The results of activating human CD8 * T-cells with antigen-presenting cells loaded with HIV-RT peptide (SEQ ID NO 48) are presented in Figure 27. Since screening of the blood cf this patient had indicated no prior exposure to HIV, these results indicate the CTL activity was based on a primary response. In this case, only antigen-presenting cells expressing A2.1, 37.1 and ICAM-1 produced CTL activity that was significantly greater than control levels. Figure 28 shows the CTL activity of human CD8" T-cells activated by antigen-presenting cells loaded with tyrosinase peptide (SEQ ID NO 49) . Tyrosinase is a normally occurring enzyme that iε over-expressed in melanoma tumor cells. Again, the antigen-presenting cells expressing all three molecules, ICAM-1 as well as A2.1 and B7.1, were most effective, especially at the lowest e fector:target ratio tested. These results show that the antigen presenting system is capable of producing effective CTL activity in human CD8 * T-celis directed against a peptide that is derived from an endogeneus protein that s over-expresεed in tumor cells. This also demonstrates that this in vi tro stimulation of CE5 " T-cells using both 37 and ICAM will generate cytotcxic CD8 * T-cellε even against peptides that would otherwise be recognized as self. This is contrary to preεent knowledge that εuch "self" peotides could not be used to create cytotoxic T-cells. This method greatly expands the number of possible tumor specific antigens that can be used to activate CDS * T-cells against the turner. The foregoing is intended to be illustrative cf the present invention, but not limiting. Numerous variations and modifications may be effected without departing from the true spirit and scope of the invention. SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: Cai, Zeling Sprent, Jonathan Brunmar , Anders Jackson, Michael Peterson, Per A Luxembourg, Alain Leturcq, Didier Jean Moriarty, Ann M. ( ii) TITLE OF INVENTION: ANTIGEN PRESENTING SYSTEM AND METHODS FOR ACTIVATION OF T-CELLS (iii) NUMBER OF SEQUENCES: 51 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: Olson & Hierl, Ltd. (B) STREET: 20 North Wacker Drive, Suite 3600 (C) CITY: Chicago (D) STATE: Illinois (E) COUNTRY: USA (F) ZIP: 60606 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentin Release #1.0, Version #1.25 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: (B) FILING DATE: 8-MAR-1996 (C) CLASSIFICATION: (vii) PRIOR APPLICATION DATA: (A) APPLICATION NUMBER: US 08/400,338 (B) FILING DATE: B-MAR-1995 (C) CLASSIFICATION: (viii) ATTORNEY/AGENT INFORMATION: (A) NAME: Olson, Arne M. (B) REGISTRATION NUMBER: 30,203 (C) REFERENCE/DOCKET NUMBER: TSRI4711 (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: (312) 580-1180 (B) TELEFAX: (312) 580-1189 (2) INFORMATION FOR SEQ ID NO:1 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ill) HYPOTHETICAL: NO (IV) ANTI-SENSE: NO (XI) SEQUENCE DESCRIPTION: SΞQ ID NO:l: CCACCATG3C CGTCATGGCG CCC 23 (2) INFORMATION FOR SEQ ID NO:2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 23 base pairs (B) TYPE: nucleic acid ( C) STRAI EDNESS : smcle (D) TOPOLOGY: linear ( i) MOLECULE TYPE: DNA (ger.omic) (ill) HYPOTHETICAL: NO (IV) ANTI-SENSE: NO (Xl) SEQUENCE DESCRIPTION: SΞQ ID NO:2: GGTCACACTT TACAAGCTCT GAG 23 (2) INFORMATION FOR SEQ ID NO.3 : (l) SEQUENCE CHARACTERISTICS* (A) LENGTH: 23 base pairs (3) TYPE: nucleic acid (C) STRANDEDNESS : smcle (D) TOPOLOGY: linear ( i) MOLECULE TYPE: DNA (ger.omic) (ill) HYPOTHETICAL: NO (IV) ANTI-SENSE: NO (X ) SEQUENCE DESCRIPTION: SΞQ ID NO: 3 CCACCATGCT GGTCATGGCG CCC 23 (2) INFORMATION FOR SEQ ID NO: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) STRAIOEDNESS: single (D) TOPOLOGY: linear (n) MOLECULE TYPE* DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO ( i) SEQUENCE DESCRIPTION: SΞQ ID NO: GGACTCGATG TGAGAGACAC ATC 23 (2) INFORMATION FOR SEQ ID NO:5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS : single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SΞQ ID NO: 5: CCACCATGCG GGTCATGGCG CCC 23 (2) INFORMATION FOR SEQ ID NO:6 : ( ) SEQUENCE CHARACTERISTICS: (A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6 GGTCA3GCTT TACAAGCGAT GAG 23 (2) INFORMATION FOR SEQ ID NO:7 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ill) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SΞQ ID NO:7: CCACCATGCG GGTAGATGCC CTC 23 (2) INFORMATION FOR SEQ ID NO:8 : (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ill) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SΞQ ID NO: 8: GGTTACAAGC TGTGAGACTC AGA 23 (2) INFORMATION FOR SEQ ID NO: S • (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY linear (ii) MOLECULE TYPE: DNA (genomic) (111) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SΞQ ID NO: 9 : CCACCATGGC GCCCCGAAGC CTC 23 (2) INFORMATION FOR SEQ ID NO.IC: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 23 base pairs (B) TYPE* nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (11) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (XI) SEQUENCE DESCRIPTION: SEQ ID NO:10: GGTCACACTT TATTAGCTGT GAG 23 (2) INFORMATION FOR SEQ ID NO:11: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (i i) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11: CCACCATGGC GCCCCGAACC CTC 23 (2) INFORMATION FOR SEQ ID NO: 12: (1) SEQUENCE CHARACTERISTICS: (A) LENGTH: 23 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear ( i) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (XI) SEQUENCE DESCRIPTION: SEQ ID NO: 12: GGTCACAATT TACAAGCCGA GAG 23 (2) INFORMATION FOR SEQ ID NO:13: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 427 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SΞQ ID NO:13: AATTCGTTGC AGGACAGOAT GTGGTGCCCG ATGTGACTAG CTCTTTGCTG CAGGCCGTCC 60 TATCCTCTGG TTCCGATAAG AGACCCAGAA CTCCGGCCCC CCACCGCCCA CCGCCACCCC 120 CATACATATG TGGTACGCAA GTAAGAGTGC CTGCGCATGC CCCATGTGCC CCACCAAGAG 180 TTTTGCATCC CATACAAGTC CCCAAAGTGG AGAACCGAAC CAATTCTTCG CGGGCAGAAC 240 AAAAGCTTCT GCACACGTCT CCACTCGAAT TTGGAGCCGG CCGGCGTGTG CAAAAGAGGT 300 GAATCGAACG AAAGACCCGT GTGTAAAGCC GCGTTTCCAA AATGTATAAA ACCGAGAGCA 360 TCTGGCCAAT GTGCATCAGT TGTGGTCAGC AGCAAAATCA AGTGAATCAT CTCAGTGCAA 420 CTAAAGG 427 (2) INFORMATION FOR SEQ ID NO:14: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 740 base pairs (E) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANT -SENSE: NO (Xi) SEQUENCE DESCRIPTION: SΞQ ID NO: 14: ATTCGATGCA CACTCACATT CTTCTCCTA.-. TACGATAATA AAACTTTCCA TGAAAAATAT 60 GGAAAAATAT ATGAAAATTG AGAAATCCAA AAAACTGATA AACGCTCTAC TTAATTAAAA 120 TAGATAAATG GGAGCGGCTG GAATGGCGGA GCATGACCAA GTTCCTCCGC CAATCAGTCG 180 TAAAACAGAA GTCGTGGAAA GCGGATAGAA AGAATGTTCG ATTTGACGGG CAAGCATGTC 240 TGCTATGTGG CGGATTGCGG AGGAATTGCA CTGGAGACCA GCAAGGTTCT CATGACCAAG 300 AATATAGCGG TGTGAGTGAG CGGGAAGCTC GGTTTCTGTC CAGATCGAAC TCAAAACTAG 360 TCCAGCCAGT CGCTGTCGAA ACTAATTAAG TTAATGAGTT TTTCATGTTA GTTTCGCGCT 420 GAGCAACAAT TAAGTTTATG TTTCAGTTCG GCTTAGATTT CGCTGAAGGA CTTGCCACTT 480 TCAATCAATA CTTTAGAACA AAATCAAAAC TCATTCTAAT AGCTTGGTGT TCATCTTTTT 540 TTTTAATGAT AAGCATTTTG TCGTTTATAC TTTTTATATT TCGATATTAA ACCACCTATG 600 AAGTTCATTT TAATCGCCAG ATAAGCAA7A TATTGTGTAA ATATTTGTAT TCTTTATCAG 660 GAAATTCAGG GAGACGGGGA AGTTACTATC TACTAAAAGC CAAACAATTT CTTACAGTTT 720 TACTCTCTCT ACTCTAGAGT 740 (2) INFORMATION FOR SEQ ID NO:IΞ: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 60 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (ger.omic) 'iii) HYPOTHETICAL: NO ( v) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SΞQ ID NO: 15: GCTTGGATCC AGATCTACCA TGTCTCGC7C CGTGGCCTTA GCTGTGCTCG CGCTACTCTC 60 (2) INFORMATION FOR SEQ ID NO:l£: (i) SEQUENCE CHARACTERISTI S: (A) LENGTH: 36 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (ge omic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SΞQ ID NO: 16: GGATCCGGAT GGTTACATGT CGCGATCCCA CTTAAC 36 (2) INFORMATION FOR SEQ ID NO:!": (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: smcle (D) TOPOLOGY: linear ( i) MOLECULE TYPE: DNA (ger.cmic) (ill) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SΞQ ID NO: 17* GGAGCCGTGA CTGACTGAG 19 (2) INFORMATION FOR SEQ ID NO:18: (i) SEQUENCE CHARACTERIST S: (A) LENGTH: 24 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ill) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (Xl) SEQUENCE DESCRIPTION: S ZQ ID NO: IB: CCCTCGGCAC TGACTGACTC CTAG 24 (2) INFORMATION FOR SEQ ID NO:15: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 38 base pairs (3) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ill) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: GATCCTTATT AGATCTCACC ATCACCATCA CCATTGAG 36 (2) INFORMATION FOR SEQ ID NO:20: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 38 base pairs (3) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SΞQ ID NO:20: TCGACTCAAT GGTGATGGTG ATGGTGAGAT CTAATAAG 38 (2) INFORMATION FOR SEQ ID NO:21: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 3875 base pairs (B) TYPΞ: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPΞ: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (x ) SEQUENCE DESCRIPTION: SΞQ ID NO:21: TTGCAG3ACA GGATGTGGTG CCCGATGTGA CTAGCTCTTT GCTGCAGGCC GTCCTATCCT 60 CTG3TTCCGA TAAGAGACCC AGAACTCCGG CCCCCCACCG CCCACCGCCA CCCCCATACA 120 TATGTGGTAC GCAAGTAAGA GTGCCTGCGC ATGCCCCATG T3CCCCACCA AGAGcTTTGC 180 ATCCCATACA AGTCCCCAAA GTGGAGAACC GAACCAATTC TTCGCGGGCA GAACAAAAGC 240 TTCTGCACAC GTCTCCACTC GAATTTGGAG CCGGCCGGCG TGTGCAAAAG AGGTGAATCG 300 AACGAAAGAC CCGTGTGTAA AGCCGCGTTT CCAAAATGTA TAAAACCGAG AGCATCTGGC 360 CAATGTGCAT CAGTTGTGGT CAGCAGCAAA ATCAAGTGAA TCATCTCAGT GCAACTAAAG 420 GGGAATTCGA GCTCGGTACC CGGGGATCCT TATTAGATCT CACCATCACC ATCACCATTG 480 AGTCGACCTG CAGGCATGCA AGCTATTCGA TGCACACTCA CATTCTTCTC CTAATACGAT 540 AATAAAACTT TCCATGAAAA ATATGGAAAA ATATATGAAA ATTGAGAAAT CCAAAAAACT 600 GATAAACGCT CTACTTAATT AAAATAGATA AATGGGAGCG GCAGGAATGG CGGAGC TGG 660 CCAAGTTCCT CCGCCAATCA GTCGTAAAAC AGAAGTCGTG GAAAGCGGAT AGAAAGAATG "20 TTCGATTTGA CGGGCAAGCA TGTCTGCTAT 3TGGCGGATT GC33AGGAAT TGCACTG3AG 78C ACCAGCAAGG TTCTCATGAC CAAGAATAT GCGGTGAGTG AGCGGOAAGC TCGGTTTCTG 840 TCCAGATCGA ACTCAAAACT AGTCCAGCC-. GTCGCTGTCG AAACTAATTA AGTtAATGAG 900 TTTTTCATGT TAGTTTCGCG CTGAGCAACA ATTAAGTTTA TGTTTCAGTT CGGCTTAGAT 560 TTCGCTGAAG GACTTGCCAC TTTCAATCAA TACTTTAGAA CAAAATCAAA ACTCATTCTA 1020 ATAGCTTGGT GTTCATCTTT TTTTTTAATG ATAAGCATTT TGTCGTTTAT ACTTTTTATA 1080 TTTCGATATT AAACCACCTA TGAAGTctAT TTTAATCGCC AGATAAGCAA TATATTGTGT 1140 AAATATTTGT ATTCTTTATC AGGAAATTCA 3GGAGACGGG AAGTTACTAT CTACTAAAAG 1200 CCAAACAATT TCTTACAGTT TTACTCTCTC TACTCTAGAG TA3CTTGGCA CTGGCCGTCG 1260 TTTTACAACG TCGTGACTGG GAAAACCCT3 GCGTTACCCA ACTTAATCGC CTTGCAGCAC 1320 ATCCCCCTTT CGCCAGCTGG CGTAATAGCG AAGAGGCCCG CACCGATCGC CCTTCCCAAC 1380 AGTTGCGCAG CCTGAATGGC GAATGGCGCC TGATGCGGTA TTTTCTCCTT ACGCATCTGT 1440 GCGGTAT7TC ACACCGCATA TGGTGCACTC TCAGTACAAT CT3CTCTGAT GCCGCATA3T 1500 TAAGCCAGCC CCGACACCCG CCAACACCCG CTGACGCGCC CT3ACGGGCT TGTCT3CTCC 1560 CGGCATCCGC TTACAGACA*-*. GCTGTGACCC- TCTCCGGGAG CTGCATGTGT CAGAG3TTTT 162C CACCGTCATC ACCGAAACGC GCGAGACGAA AGGGCCTCGT GATACGCCTA TTTTTATAGG 1680 TTAATGTCAT GATAATAATG GTTTCTTAGA CGTCAGGTGG CACTTTTCGG GGAAATGTGC 1740 GCGGAACCCC TATTTGTTTA TTTTTCTAAA TACATTCAAA TATGTATCCG CTCATGAGAC 1800 AATAACCCTG ATAAATGCTT CAATAATA77 3AAAAAGGAA GA3TATGAGT ATTCAACATT 1860 TCCGTGTCGC CCTTATTCCC TTTTTT3C33 CATTTTGCCT TCCTGTTTTT GCTCACCCAG 1920 AAACGCTGGT GAAAGTAAAA GATGCTGAA3 ATCAGTTGGG TGCACGAGTG GGTTACATCG 1980 AACTGGATCT CAACAGCGGT AAGATCCTT3 AGAGTTTTCG CCCCGAAGAA CGTTTTCCAA 2040 TGATGAGCAC TTTTAAAGTT CTGCTATGT3 GCGCGGTATT ATCCCGTATT GACGCCGGGC 2100 AAGAGCAACT CGGTCGCCGC ATACACTATT CTCAGAATGA CTTGGTTGAG TACTCACCAG 2160 TCACAGAAAA GCATCTTACG GATGGCATGA CAGTAAGAGA ATTATGCAGT GCTGCCATAA 2220 CCATGAGTGA TAACACTGCG GCCAACTTAC TTCTGACAAC GATCGGAGGA CCGAAGGAGC 2280 TAACCGCTTT TTTGCACAAC ATGGGGGATC ATGTAACTCG CCTTGATCGT TGGGAACCGG 2340 AGCTGAATGA AGCCATACCA AACGACGAGC GTGACACCAC GATGCCTGTA GCAATGGCAA 2400 CAACGTTGCG CAAACTATTA ACTGGCGAAC TACTTACTCT AGCTTCCCGG CAACAATTAA 2460 TAGACTGGAT GGAGGCGGAT AAAGTTGCAG GACCACTTCT GCGCTCGGCC CTTCCGGCTG 2520 GCTGGTTTAT TGCTGATAAA TCTGGAGCCG GTGAGCGTGG GTCTCGCGGT ATCATTGCAG 2580 CACTGGGGCC AGATGGTAAG CCCTCCCGTA TCGTAGTTAT CTACACGACG GGGAGTCAGG 2640 CAACTATGGA TGAACGAAAT AGACAGATCG CTGAGATAGG TGCCTCACTG ATTAAGCATT 2700 GGTAACTGTC AGACCAAGTT TACTCATATA TACTTTAGAT TGATTTAAAA CTTCATTTTT 2760 AATTTAAAAG GATCTAGGTG AAGATCCTTT TTGATAATCT CATGACCAAA ATCCCTTAAC 2820 GTGAGTTTTC GTTCCACTGA GCGTCAGACC CCGTAGAAAA GATCAAAGGA TCTTCTTGAG 2880 ATCCTTTTTT TCTGCGCGTA ATCTGCTGCT TGCAAACAAA AAAACCACCG CTACCAGCGG 2940 TGGTTTGTTT GCCGGATCAA GAGCTACCAA CTCTTTTTCC GAAGGTAACT GGCTTCAGCA 3000 GAGCGCAGAT ACCAAATACT GTCCTTCTAG TGTAGCCGTA GTTAGGCCAC CACTTCAAGA 3060 ACTCTGTAGC ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG GCTGCTGCCA 3120 GTGGCGATAA GTCGTGTCTT ACCGGGTTGG ACTCAAGACG ATAGTTACCG GATAAGGCGC 3180 AGCGGTCGGG CTGAACGGGG GGTTCGTGCA CACAGCCCAG CTTGGAGCGA ACGACCTACA 3240 CCGAACTGAG ATACCTACAG CGTGAGCATT GAGAAAGCGC CACGCTTCCC GAAGGGAGAA 3300 AGGCGGACAG GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG AGGGAGCTTC 3360 CAGGGGGAAA CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC TGACTTGAGC 3420 GTCGATTTTT GTGATGCTCG TCAGGGGGGC GGAGCCTATG GAAAAACGCC AGCAACGCGG 3480 CCTTTTTACG GTCCTGGCCT TTTGCTGGCC TTTTGCTCAC ATGTCTTTCC TGCGTTATCC 3540 CCTGATTCTG TGGATAACCG TATTACCGCC TTTGAGTGAG CTGATACCGC TCGCCGCAGC 3600 CGAACcGACC GAGCGCAGCG AGTCAGTGAG CGAGGAAGCG GAAGAGCGCC CAATACGCAA 3660 ACCGCCTCTC CCCGCGCGTT GGCCGATTCA TTAATGCAGC TGGCACGACA GGTTTCCCGA 3720 CTGGAAAGCG GGCAGTGAGC GCAACGCAAT TAATGTGAGT TAGCTCACTC ATTAGGCACC 3780 CCAGGCTTTA CACTTTATGC TTCCGGCTC3 TATGTTGTGT GGAATTGTGA GCGGATAACA 3840 ATTTCACACA GGAAACAGCT ATGACATGAT TACCG 3875 (2) INFORMATION FOR SEQ ID NO:22: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 71 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22: GATCCTTATT AGATCTTACC CATACGACGT CCCAGATTAC GCTCGATCTC ACCATCACCA 60 TCACCATTGA G 71 (2) INFORMATION FOR SEQ ID NO:23: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 71 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: TCGACTCAAT GGTGATGGTG ATGGTGAGAT CGAGCGTAAT CTGGGACGTC GTATGGGTA∑. 60 GATCTAATAA G 7 1 (2) INFORMATION FOR SEQ ID NO:24: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 3908 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24: TTGCAGGACA GGATGTGGTG CCCGATGTGA CTAGCTCTTT GCTGCAGGCC GTCCTATCCT 60 CTGGTTCCGA TAAGAGACCC AGAACTCCGG CCCCCCACCG CCCACCGCCA CCCCCATACA 120 TATGTGGTAC GCAAGTAAGA GTGCCTGCGC ATGCCCCATG TGCCCCACCA AGAGcTTTGC 180 ATCCCATACA AGTCCCCAAA GTGGAGAACC GAACCAATTC TTCGCGGGCA GAACAAAAGC 240 TTCTGCACAC GTCTCCACTC GAATTTGGAG CCGGCCGGCG TGTGCAAAAG AGGTGAATCG 300 AACGAAAGAC CCGTGTGTAA AGCCGCGTTT CCAAAATGTA TAAAACCGAG AGCATCTGGC 360 CAATGTGCAT CAGTTGTGGT CAGCAGCAAA ATCAAGTGAA TCATCTCAGT GCAACTAAAG 420 GGGAATTCGA GCTCGGTACC CGGGGATCCT TATTAGATCT TACCCATACG ACGTCCCAGA 480 TTACGCTCGA TCTCACCATC ACCATCACCA TTGAGTCGAC CTGCAGGCAT GCAAGCTATT 540 CGATGCACAC TCACATTCTT CTCCTAATAC GATAATAAAA CTTTCCATGA AAAATATGGA 600 AAAATATATG AAAATTGAGA AATCCAAAAA ACTGATAAAC GCTCTACTTA ATTAAAATAG 660 ATAAATGGGA GCGGCAGGAA TGGCGGAGCA TGGCCAAGTT CCTCCGCCAA TCAGTCGTAA 720 AACAGAAGTC GTGGAAAGCG GATAGAAAGA ATGTTCGATT TGACGGGCAA GCATGTCTGC 780 TATGTGGCGG ATTGCGGAGG AATTGCACTG GAGACCAGCA AGGTTCTCAT GACCAAGAAT 840 ATAGCGGTGA GTGAGCGGGA AGCTCGGTTT CTGTCCAGAT CGAACTCAAA ACTAGTCCAG 900 CCAGTCGCTG TCGAAACTAA TTAAGTtAAT GAGTTTTTCA TGTTAGTTTC GCGCTGAGCA 960 ACAATTAAGT TTATGTTTCA GTTCGGCTTA GATTTCGCTG AAGGACTTGC CACTTTCAAT 1020 CAATACTTTA GAACAAAATC AAAACTCATT CTAATAGCTT GGTGTTCATC TTTTTTTTTA 1080 ATGATAAGCA TTTTGTCGTT TATACTTTTT ATATTTCGAT ATTAAACCAC CTATGAAGTc 1140 tATTTTAATC GCCAGATAAG CAATATATTG TGTAAATATT TGTATTCTTT ATCAGGAAAT 1200 TCAGGGAGAC GGGAAGTTAC TATCTACTA--. AAGCCAAACA ATTTCTTACA GTTTTACTCT 1260 CTCTACTCTA GAGTAGCTTG GCACTGGCCG TCGTTTTACA ACGTCGTGAC TGGGAAAACC 1320 CTGGCGTTAC CCAACTTAAT CGCCTTGCAG CACATCCCCC TTTCGCCAGC TGGCGTAATA 1380 GCGAA3AGGC CCGCACCGAT CGCCCTTCCC AACAGTTGCG CAGCCTGAAT GGCGAATGGC 1440 GCCTGATGCG GTATTTTCTC CTTACGCATC TGTGCGGTAT TTCACACCGC ATATGGTGCA 1500 CTCTCAGTAC AATCTGCTCT GATGCCGCAT AGTTAAGCCA GCCCCGACAC CCGCCAACAC 1560 CCGCTGACGC GCCCTGACGG GCTTGTCTGC TCCCGGCATC CGCTTACAGA CAAGCTGTGA 1620 CCGTCTCCGG GAGCTGCATG TGTCAGAGGT TTTCACCGTC ATCACCGAAA CGCGCGAGAC 1680 GAAAGGGCCT CGTGATACGC CTATTTTTAT AGGTTAATGT CATGATAATA ATGGTTTCTT 1740 AGACGTCAGG TGGCACTTTT CGGGGAAATG TGCGCGGAAC CCCTATTTGT TTATTTTTCT 1800 AAATACATTC AAATATGTAT CCGCTCATGA GACAATAACC CTGATAAATG CTTCAATAAT 1860 ATTGAAAAAG GAAGAGTATG AGTATTCAAC ATTTCCGTGT CGCCCTTATT CCCTTTTTTG 1920 CGGCATTTTG CCTTCCTGTT TTTGCTCACC CAGAAACGCT GGTGAAAGTA AAAGATGCTG 1980 AAGATCAGTT GGGTGCACGA GTGGGTTACA TCGAACTGGA TCTCAACAGC GGTAAGATCC 2040 TTGAGAGTTT TCGCCCCGAA GAACGTTTTC CAATGATGAG CACTTTTAAA GTTCTGCTAT 2100 GTGGCGCGGT ATTATCCCGT ATTGACGCCG GGCAAGAGCA ACTCGGTCGC CGCATACACT 2160 ATTCTCAGAA TGACTTGGTT GAGTACTCAC CAGTCACAGA AAAGCATCTT ACGGATGGCA 2220 TGACAGTAAG AGAATTATGC AGTGCTGCCA TAACCATGAG TGATAACACT GCGGCCAACT 2280 TACTTCTGAC AACGATCGGA GGACCGAAGG AGCTAACCGC TTTTTTGCAC AACATGGGGG 2340 ATCATGTAAC TCGCCTTGAT CGTTGGGAAC CGGAGCTGAA TGAAGCCATA CCAAACGACG 2400 AGCGTGACAC CACGATGCCT GTAGCAATGG CAACAACGTT GCGCAAACTA TTAACTGGCG 2460 AACTACTTAC TCTAGCTTCC CGGCAACAAT TAATAGACTG GATGGAGGCG GATAAAGTTG 2520 CAGGACCACT TCTGCGCTCG GCCCTTCCGG CTGGCTGGTT TATTGCTGAT AAATCTGGAG 2580 CCGGTGAGCG TGGGTCTCGC GGTATCATTG CAGCACTGGG GCCAGATGGT AAGCCCTCCC 2640 GTATCGTAGT TATCTACACG ACGGGGAGTC AGGCAACTAT GGATGAACGA AATAGACAGA 2700 TCGCTGAGAT AGGTGCCTCA CTGATTAAGC ATTGGTAACT GTCAGACCAA GTTTACTCAT 2760 ATATACTTTA GATTGATTTA AAACTTCATT TTTAATTTAA AAGGATCTAG GTGAAGATCC 2820 TTTTTGATAA TCTCATGACC AAAATCCCTT AACGTGAGTT TTCGTTCCAC TGAGCGTCAG 2880 ACCCCGTAGA AAAGATCAAA GGATCTTCTT GAGATCCTTT TTTTCTGCGC GTAATCTGCT 2940 GCTTGCAAAC AAAAAAACCA CCGCTACCAG CGGTGGTTTG TTTGCCGGAT CAAGAGCTAC 3000 CAACTCTTTT TCCGAAGGTA ACTGGCTTCA GCAGAGCGCA GATACCAAAT ACTGTCCTTC 3060 TAGTGTAGCC GTAGTTAGGC CACCACTTCA AGAACTCTGT AGCACCGCCT ACATACCTCG 3120 CTCTGCTAAT CCTGTTACCA GTGGCTGCTG CCAGTGGCGA TAAGTCGTGT CTTACCGGGT 3180 TGGACTCAAG ACGATAGTTA CCGGATAAGG CGCAGCGGTC GGGCTGAACG GGGGGTTCGT 3240 GCACACAGCC CAGCTTGGAG CGAACGACCT ACACCGAACT GAGATACCTA CAGCGTGAGC 3300 ATTGAGAAAG CGCCACGCTT CCCGAAGGGA GAAAGGCGGA CAG3TATCCG GTAAGCGGCA 3360 GGGTCGGAAC AGGAGAGCGC ACGAGGGAGC TTCCAGGGGG AAACGCCTGG TATCTTTATA 3420 GTCCTGTCGG GTTTCGCCAC CTCTGACTT3 AGCGTCGATT TTTGTGATGC TCGTCAGGGG 3480 GGCGGAGCCT ATGGAAAAAC GCCAGCAACG CGGCCTTTTT ACGGTCCTGG CCTTTTGCTG 3540 GCCTTTTGCT CACATGTCTT TCCTGCGTTA TCCCCTGATT CTGTGGATAA CCGTATTACC 3600 GCCTTTGAGT GAGCTGATAC CGCTCGCCGC AGCCGAACCG ACCGAGCGCA GCGAGTCAGT 3660 GAGCGAGGAA GCGGAAGAGC GCCCAATAC3 CAAACCGCCT CTCCCCGCGC GTTGGCCGAT 3720 TCATTAATGC AGCTGGCACG ACAGGTTTCC CGACTGGAAA GCGGGCAGTG AGCGCAACGC 3780 AATTAATGTG AGTTAGCTCA CTCATTAGGC ACCCCAGGCT TTACACTTTA TGCTTCCGGC 3840 TCGTATGTTG TGTGGAATTG TGAGCGGATA ACAATTTCAC ACAGGAAACA GCTATGACAT 3900 GATTACCG 3908 (2) INFORMATION FOR SEQ ID NO:25: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 41 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO ( i) SEQUENCE DESCRIPTION: SΞQ ID NO:25: GATCCTTATT AGATCTCACC ATCACCATCA CCATTGTTGA G 41 (2) INFORMATION FOR SEQ ID NO:26: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 41 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SΞQ ID NO:26: TCGACTCAAC AATGGTGATG GTGATGGTGA GATCTAATAA G 41 (2) INFORMATION FOR SEQ ID NO:27: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 3878 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27: TTGCAGGACA GGATGTGGTG CCCGATGTGA CTAGCTCTTT GCTGCAGGCC GTCCTATCCT 60 CTGGTTCCGA TAAGAGACCC AGAACTCCGG CCCCCCACCG CCCACCGCCA CCCCCATACA 120 TATGTGGTAC GCAAGTAAGA GTGCCTGCGC ATGCCCCATG TGCCCCACCA AGAGcTTTGC 180 ATCCCATACA AGTCCCCAAA GTGGAGAACC GAACCAATTC TTCGCGGGCA GAACAAAAGC 240 TTCTGCACAC GTCTCCACTC GAATTTGGAG CCGGCCGGCG TGTGCAAAAG AGGTGAATCG 300 AACGAAAGAC CCGTGTGTAA AGCCGCGTTT CCAAAATGTA TAAAACCGAG AGCATCTGGC 360 CAATGTGCAT CAGTTGTGGT CAGCAGCAAA ATCAAGTGAA TCATCTCAGT GCAACTAAAG 420 GGGAATTCGA GCTCGGTACC CGGGGATCCT TATTAGATCT CACCATCACC ATCACCATTG 480 TTGAGTCGAC CTGCAGGCAT GCAAGCTATT CGATGCACAC TCACATTCTT CTCCTAATAC 540 GATAATAAAA CTTTCCATGA AAAATATGGA AAAATATATG AAAATTGAGA AATCCAAAAA 600 ACTGATAAAC GCTCTACTTA ATTAAAATAG ATAAATGGGA GCGGCAGGAA TGGCGGAGCA 660 TGGCCAAGTT CCTCCGCCAA TCAGTCGTAA. AACAGAAGTC GTGGAAAGCG GATAGAAAGA 720 ATGTTCGATT TGACGGGCAA GCATGTCTGC TATGTGGCGG ATTGCGGAGG AATTGCACTG 780 GAGACCAGCA AGGTTCTCAT GACCAAGAAT ATAGCGGTGA GTGAGCGGGA AGCTCGGTTT 840 CTGTCCAGAT CGAACTCAAA ACTAGTCCAG CCAGTCGCTG TCGAAACTAA TTAAGTtAAT 900 GAGTTTTTCA TGTTAGTTTC GCGCTGAGCA ACAATTAAGT TTATGTTTCA GTTCGGCTTA 960 GATTTCGCTG AAGGACTTGC CACTTTCAAT CAATACTTTA GAACAAAATC AAAACTCATT 1020 CTAATAGCTT GGTGTTCATC TTTTTTTTTA ATGATAAGCA TTTTGTCGTT TATACTTTTT 1080 ATATTTCGAT ATTAAACCAC CTATGAAGTc tATTTTAATC GCCAGATAAG CAATATATTG 1140 TGTAAATATT TGTATTCTTT ATCAGGAAAT TCAGGGAGAC G3GAAGTTAC TATCTACTAA 1200 AAGCCAAACA ATTTCTTACA GTTTTACTCT CTCTACTCTA GAGTAGCTTG GCACTGGCCG 1260 TCGTTTTACA ACGTCGTGAC TGGGAAAACC CTGGCGTTAC CCAACTTAAT CGCCTTGCAG 1320 CACATCCCCC TTTCGCCAGC TGGCGTAATA GCGAAGAGGC CCGCACCGAT CGCCCTTCCC 1380 AACAGTTGCG CAGCCTGAAT GGCGAATGGC GCCTGATGCG GTATTTTCTC CTTACGCATC 1440 TGTGCGGTAT TTCACACCGC ATATGGTGCA CTCTCAGTAC AATCTGCTCT GATGCCGCAT 1500 AGTTAAGCCA GCCCCGACAC CCGCCAACAC CCGCTGACGC GCCCTGACGG GCTTGTCTGC 1560 TCCCGGCATC CGCTTACAGA CAAGCTGTGA CCGTCTCCGG GAGCTGCATG TGTCAGAGGT 1620 TTTCACCGTC ATCACCGAAA CGCGCGAGAC GAAAGGGCCT CGTGATACGC CTATTTTTAT 1680 AGGTTAATGT CATGATAATA ATGGTTTCTT AGACGTCAGG TGGCACTTTT CGGGGAAATG 1740 TGCGCGGAAC CCCTATTTGT TTATTTTTCT AAATACATTC AAATATGTAT CCGCTCATGA 1800 GACAATAACC CTGATAAATG CTTCAATAAT ATTGAAAAAG GAAGAGTATG AGTATTCAAC 1860 ATTTCCGTGT CGCCCTTATT CCCTTTTTTG CGGCATTTTG CCTTCCTGTT TTTGCTCACC 1920 CAGAAACGCT GGTGAAAGTA AAAGATGCTG AAGATCAGTT GGGTGCACGA GTGGGTTACA 1980 TCGAACTGGA TCTCAACAGC GGTAAGATCC TTGAGAGTTT TCGCCCCGAA GAACGTTTTC 2040 CAATGATGAG CACTTTTAAA GTTCTGCTAT GTGGCGCGGT ATTATCCCGT ATTGACGCCG 2100 GGCAAGAGCA ACTCGGTCGC CGCATACACT ATTCTCAGAA TGACTTGGTT GAGTACTCAC 2160 CAGTCACAGA AAAGCATCTT ACGGATGGCA TGACAGTAAG AGAATTATGC AGTGCTGCCA 2220 TAACCATGAG TGATAACACT GCGGCCAACT TACTTCTGAC AACGATCGGA GGACCGAAGG 2280 AGCTAACCGC TTTTTTGCAC AACATGGGGG ATCATGTAAC TCGCCTTGAT CGTTGGGAAC 2340 CGGAGCTGAA TGAAGCCATA CCAAACGACG AGCGTGACAC CACGATGCCT GTAGCAATGG 2400 CAACAACGTT GCGCAAACTA TTAACTGGCG AACTACTTAC TCTAGCTTCC CGGCAACAAT 2460 TAATAGACTG GATGGAGGCG GATAAAGTTG CAGGACCACT TCTGCGCTCG GCCCTTCCGG 2520 CTGGCTGGTT TATTGCTGAT AAATCTGGAG CCGGTGAGCG TGGGTCTCGC GGTATCATTG 2580 CAGCACTGGG GCCAGATGGT AAGCCCTCCC GTATCGTAGT TATCTACACG ACGGGGAGTC 2640 AGGCAACTAT GGATGAACGA AATAGACAGA TCGCTGAGAT AGGTGCCTCA CTGATTAAGC 2700 ATTGGTAACT GTCAGACCAA GTTTACTCAT ATATACTTTA GATTGATTTA AAACTTCATT 2760 TTTAATTTAA AAGGATCTAG GTGAAGATCC TTTTTGATAA TCTCATGACC AAAATCCCTT 2820 AACGTGAGTT TTCGTTCCAC TGAGCGTCAG ACCCCGTAGA AAAGATCAAA GGATCTTCTT 2880 GAGATCCTTT TTTTCTGCGC GTAATCTGCT GCTTGCAAAC AAAAAAACCA CCGCTACCAG 2940 CGGTGGTTTG TTTGCCGGAT CAAGAGCTAC CAACTCTTTT TCCGAAGGTA ACTGGCTTCA 3000 GCAGAOCGCA GATACCAAAT ACTGTCCTTC TAGTGTAGCC GTAGTTAGGC CACCACTTCA 3060 AGAACTCTGT AGCACCGCCT ACATACCTCG CTCTGCTAAT CCTGTTACCA GTGGCTGCTG 3120 CCAGTGGCGA TAAGTCGTGT CTTACCGGGT TGGACTCAAG ACGATAGTTA CCGGATAAGG 3180 CGCAGCGGTC GGGCTGAACG GGGGGTTCGT GCACACAGCC CAGCTTGGAG CGAACGACCT 3240 ACACCGAACT GAGATACCTA CAGCGTGAGC ATTGAGAAA3 CGCCACGCTT CCCGAAGGGA 3300 GAAAGGCGGA CAGGTATCCG GTAAGCGGCA GGGTCOGAAC AGGAGAGCGC ACGAGGGAGC 3360 TTCCAGGGGG AAACGCCTGG TATCTTTATA GTCCTGTCGG GTTTCGCCAC CTCTGACTTG 3420 AGCGTCGATT TTTGTGATGC TCGTCAGGGG GGCGGAGCCT ATGGAAAAAC GCCAGCAACG 3480 CGGCCTTTTT ACGGTCCTGG CCTTTTGCTG GCCTTTTGCT CACATGTCTT TCCTGCGTTA 3540 TCCCCTGATT CTGTGGATAA CCGTATTACC GCCTTTGAGT GAGCTGATAC CGCTCGCCGC 3600 AGCCGAACcG ACCGAGCGCA GCGAGTCAGT GAGCGAGGAA GCGGAAGAGC GCCCAATACG 3660 CAAACCGCCT CTCCCCGCGC GTTGGCCGAT TCATTAATGC AGCTGGCACG ACAGGTTTCC 3720 CGACTGGAAA GCGGGCAGTG AGCGCAACGC AATTAATGTG AGTTAGCTCA CTCATTAGGC 3780 ACCCCAGGCT TTACACTTTA TGCTTCCGGC TCGTATGTTG TGTGGAATTG TGAGCGGATA 3840 ACAATTTCAC ACAGGAAACA GCTATGACAT GATTACCG 3878 (2) INFORMATION FOR SEQ ID NO:28: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 47 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SΞQ ID NO:28: GATCCTTATT AGATCTGCTT GGCGCCATCC TCAATTTGGG GGTTGAG 47 (2) INFORMATION FOR SEQ ID NO:29: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 47 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SΞQ ID NO:29: TCGACTCAAC CCCCAAATTG AGGATGGCGC CAAOCAGATC TAATAAG 47 (2) INFORMATION FOR SEQ ID NO: 30: ( ) SEQUENCE CHARACTERISTICS: (A) LENGTH: 3883 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO ( i) SEQUENCE DESCRIPTION: SΞQ ID NO:30: TTGCAGGACA GGATGTGGTG CCCGATGTGA CTAGCTCTTT GCTGCAGGCC GTCCTATCCT 60 TGGTTCCGAT AAGAGACCCA GAACTCCGGC CCCCCACCGC CCACCGCCAC CCCCATACAT 120 ATGTGGTACG CAAGTAAGAG TGCCTGCGCA TGCCCCATGT GCCCCACCAA GAGcTTTGCA 180 TCCCATACAA GTCCCCAAAG TGGAGAACCG AACCAATTCT TCGCGGGCAG AACAAAAGCT 240 TCTGCACACG TCTCCACTCG AATTTGGAGC CGGCCGGCGT GTGCAAAAGA GGTGAATCGA 300 ACGAAAGACC CGTGTGTAAA GCCGCGTTTC CAAAATGTAT AAAACCGAGA GCATCTGGCC 360 AATGTGCATC AGTTGTGGTC AGCAGCAAA. TCAAGTGAAT CATCTCAGTG CAACTAAAGG 420 GGAATTCGAG CTCGGTACCC GGGGATCCTT ATTAGATCTG CTTGGCGCCA TCCTCAATTT 480 GGGGGTTGAG TCGACCTGCA GGCATGCAAG CTATTCGATG CACACTCACA TTCTTCTCCT 540 AATACGATAA TAAAACTTTC CATGAAAAAT ATGGAAAAAT ATATGAAAAT TGAGAAATCC 600 AAAAAACTGA TAAACGCTCT ACTTAATTAA AATAGATAAA TGGGAGCGGC AGGAATGGCG 660 GAGCATGGCC AAGTTCCTCC GCCAATCAGT CGTAAAACAG AAGTCGTGGA AAGCGGATAG 720 AAAGAATGTT CGATTTGACG GGCAAGCATG TCTGCTATGT GGCGGATTGC GGAGGAATTG 780 CACTGGAGAC CAGCAAGGTT CTCATGACCA AGAATATAGC GGTGAGTGAG CGGGAAGCTC 840 GGTTTCTGTC CAGATCGAAC TCAAAACTAG TCCAGCCAGT CGCTGTCGAA ACTAATTAAG 900 TtAATGAGTT TTTCATGTTA GTTTCGCGCT GAGCAACAAT TAAGTTTATG TTTCAGTTCG 960 GCTTAGATTT CGCTGAAGGA CTTGCCACTT TCAATCAATA CTTTAGAACA AAATCAAAAC 1020 TCATTCTAAT AGCTTGGTGT TCATCTTTTT TTTTAATGAT AAGCATTTTG TCGTTTATAC 1080 TTTTTATATT TCGATATTAA ACCACCTATG AAGTctATTT TAATCGCCAG ATAAGCAATA 1140 TATTGTGTAA ATATTTGTAT TCTTTATCAG GAAATTCAGG GAGACGGGAA GTTACTATCT 1200 ACTAAAAGCC AAACAATTTC TTACAGTTTT ACTCTCTCTA CTCTAGAGTA GCTTGGCACT 1260 GGCCGTCGTT TTACAACGTC GTGACTGGGA AAACCCTGGC GTTACCCAAC TTAATCGCCT 1320 TGCAGCACAT CCCCCTTTCG CCAGCTGGCG TAATAGCGAA GAGGCCCGCA CCGATCGCCC 1380 TTCCCAACAG TTGCGCAGCC TGAATGGCGA ATGGCGCCTG ATGCGGTATT TTCTCCTTAC 1440 GCATCTGTGC GGTATTTCAC ACCGCATATG GTGCACTCTC A3TACAATCT GCTCTGATGC 1500 CGCATAGTTA AGCCAGCCCC GACACCCGCC AACACCCGCT GACGCGCCCT GACGGGCTTG 1560 TCTGCTCCCG GCATCCGCTT ACAGACAAGC TGTGACCGTC TCCGOGAGCT GCATGTGTCA 1620 GAGGTTTTCA CCGTCATCAC CGAAACGCGC GAGACGAAAG GGCCTCGTGA TACGCCTATT 1680 TTTATAGGTT AATGTCATGA TAATAATGGT TTCTTAGACG TCAGGTGGCA CTTTTCGGGG 1740 AAATGTGCGC GGAACCCCTA TTTGTTTATT TTTCTAAATA CATTCAAATA TGTATCCGCT 1800 CATGAGACAA TAACCCTGAT AAATGCTTCA ATAATATTGA AAAAGGAAGA GTATGAGTAT 1860 TCAACATTTC CGTGTCGCCC TTATTCCCTT TTTTGCGGCA TTTTGCCTTC CTGTTTTTGC 1920 TCACCCAGAA ACGCTGGTGA AAGTAAAAGA TGCTGAAGAT CAGTTGGGTG CACGAGTGGG 1980 TTACATCGAA CTGGATCTCA ACAGCGGTAA GATCCTTGAG AGTTTTCGCC CCGAAGAACG 2040 TTTTCCAATG ATGAGCACTT TTAAAGTTCT GCTATGTGGC GCGGTATTAT CCCGTATTGA 2100 CGCCGGGCAA GAGCAACTCG GTCGCCGCAT ACACTATTCT CAGAATGACT TGGTTGAGTA 2160 CTCACCAGTC ACAGAAAAGC ATCTTACGGA TGGCATGACA GTAAGAGAAT TATGCAGTGC 2220 TGCCATAACC ATGAGTGATA ACACTGCGGC CAACTTACTT CTGACAACGA TCGGAGGACC 2280 GAAGGAGCTA ACCGCTTTTT TGCACAACAT GGGGGATCAT GTAACTCGCC TTGATCGTTG 2340 GGAACCGGAG CTGAATGAAG CCATACCAAA CGACGAGCGT GACACCACGA TGCCTGTAGC 2400 AATGGCAACA ACGTTGCGCA AACTATTAAC TGGCGAACTA CTTACTCTAG CTTCCCGGCA 2460 ACAATTAATA GACTGGATGG AGGCGGATAA AGTTGCAGGA CCACTTCTGC GCTCGGCCCT 2520 TCCGGCTGGC TGGTTTATTG CTGATAAATC TGGAGCCGGT GAGCGTGGGT CTCGCGGTAT 2580 CATTGCAGCA CTGGGGCCAG ATGGTAAGCC CTCCCGTATC GTAGTTATCT ACACGACGGG 2640 GAGTCAGGCA ACTATGGATG AACGAAATAG ACAGATCGCT GAGATAGGTG CCTCACTGAT 2700 TAAGCATTGG TAACTGTCAG ACCAAGTTTA CTCATATATA CTTTAGATTG ATTTAAAACT 2760 TCATTTTTAA TTTAAAAGGA TCTAGGTGAA GATCCTTTTT GATAATCTCA TGACCAAAAT 2820 CCCTTAACGT GAGTTTTCGT TCCACTGAGC GTCAGACCCC GTAGAAAAGA TCAAAGGATC 2880 TTCTTGAGAT CCTTTTTTTC TGCGCGTAAT CTGCTGCTTG CAAACAAAAA AACCACCGCT 2940 ACCAGCGGTG GTTTGTTTGC CGGATCAAGA GCTACCAACT CTTTTTCCGA AGGTAACTGG 3000 CTTCAGCAGA GCGCAGATAC CAAATACTGT CCTTCTAGTG TAGCCGTAGT TAGGCCACCA 3060 CTTCAAGAAC TCTGTAGCAC CGCCTACATA CCTCGCTCTG CTAATCCTGT TACCAGTGGC 3120 TGCTGCCAGT GGCGATAAGT CGTGTCTTAC CGGGTTGGAC TCAAGACGAT AGTTACCGGA 3180 TAAGGCGCAG CGGTCGGGCT GAACGGGGGG TTCGTGCACA CAGCCCAGCT TGGAGCGAAC 3240 GACCTACACC GAACTGAGAT ACCTACAGCG TGAGCATTGA GAAAGCGCCA CGCTTCCCGA 3300 AGGGAGAAAG GCGGACAGGT ATCCGGTAAG CGGCAGGGTC GGAACAGGAG AGCGCACGAG 3360 GGAGCTTCCA GGGGGAAACG CCTGGTATCT TTATAGTCCT GTCGGGTTTC GCCACCTCTG 3420 ACTTGAGCGT CGATTTTTGT GATGCTCGTC AGGGGGGCGG AGCCTATGGA AAAACGCCAG 3480 CAACGCGGCC TTTTTACGGT CCTGGCCTTT TGCTGGCCTT TTGCTCACAT GTCTTTCCTG 3540 CGTTATCCCC TGATTCTGTG GATAACCGTA TTACCGCCTT TGAGTGAGCT GATACCGCTC 3600 GCCGCAGCCG AACCGACCGA GCGCAGCGAG TCAGTGAGCG AGGAAGCGGA AGAGCGCCCA 3660 ATACGCAAAC CGCCTCTCCC CGCGCGTTGG CCGATTCATT AATGCAGCTG GCACGACAGG 3720 TTTCCCGACT GGAAAGCGGG CAGTGAGCGC AACGCAATTA ATGTGAGTTA GCTCACTCAT 3780 TAGGCACCCC AGGCTTTACA CTTTATGCTT CCGGCTCGTA TGTTGTGTGG AATTGTGAGC 3840 GGATAACAAT TTCACACAGG AAACAGCTAT GACATGATTA CCG 3883 (2) INFORMATION FOR SEQ ID NO:31: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 879 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31: GAATTCATGG GCCACACACG GAGGCAGGGA ACATCACCAT CCAAGTGTCC ATACCTCAAT 60 TTCTTTCAGC TCTTGGTGCT GGCTGGTCTT TCTCACTTCT GTTCAGGTGT TATCCACGTG 120 ACCAAGGAAG TGAAAGAAGT GGCAACGCTG TCCTGTGGTC ACAATGTTTC TGTTGAAGAG 180 CTGGCACAAA CTCGCATCTA CTGGCAAAAG GAGAAGAAAA TGGTGCTGAC TATGATGTCT 240 GGGGACATGA ATATATGGCC CGAGTACAAG AACCGGACCA TCTTTGATAT CACTAATAAC 300 CTCTCCATTG TGATCCTGGC TCTGCGCCCA TCTGACGAGG GCACATACGA GTGTGTTGTT 360 CTGAAGTATG AAAAAGACGC TTTCAAGCGG GAACACCTGG CTGAAGTGAC GTTATCAGTC 420 AAAGCTGACT TCCCTACACC TAGTATATCT GACTTTGAAA TTCCAACTTC TAATATTAGA 480 AGGATAATTT GCTCAACCTC TGGAGGTTTT CCAGAGCCTC ACCTCTCCTG GTTGGAAAAT 540 GGAGAAGAAT TAAATGCCAT CAACACAACA GTTTCCCAAG ATCCTGAAAC TGAGCTCTAT 600 GCTGTTAGCA GCAAACTGGA TTTCAATATG ACAACCAACC ACAGCTTCAT GTGTCTCATC 660 AAGTATGGAC ATTTAAGAGT GAATCAGACC TTCAACTGGA ATACAACCAA GCAAGAGCAT 720 TTTCCTGATA ACCTGCTCCC ATCCTGGGCC ATTACCTTAA TCTCAGTAAA TGGAATTTTT 780 GTGATATGCT GCCTGACCTA CTGCTTTGCC CCAAGATGCA GA3A3A3AAG GAGGAATGAG 640 AGATTGAGAA GGGAAAGTGT ACGCCCTGTA TAAGGATTC 679 (2) INFORMATION FOR SEQ ID NO:32: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 738 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32: GAATTCATGG GCCACACACG GAGGCAGGGA ACATCACCAT CCAAGTGTCC ATACCTCAAT 60 TTCTTTCAGC TCTTGGTGCT GGCTGGTCTT TCTCACTTCT GTTCAGGTGT TATCCACGTG 120 ACCAAGGAAG TGAAAGAAGT GGCAACGCTG TCCTGTGGTC ACAATGTTTC TGTTGAAGAG 180 CTGGCACAAA CTCGCATCTA CTGGCAAAAG GAGAAGAAAA TGGTGCTGAC TATGATGTCT 240 GGGGACATGA ATATATGGCC CGAGTACAAG AACCGGACCA TCTTTGATAT CACTAATAAC 300 CTCTCCATTG TGATCCTGGC TCTGCGCCCA TCTGACGAGG GCACATACGA GTGTGTTGTT 360 CTGAAGTATG AAAAAGACGC TTTCAAGCGG GAACACCTGG CTGAAGTGAC GTTATCAGTC 420 AAAGCTGACT TCCCTACACC TAGTATATCT GACTTTGAAA TTCCAACTTC TAATATTAGA 480 AGGATAATTT GCTCAACCTC TGGAGGTTTT CCAGAGCCTC ACCTCTCCTG GTTGGAAAAT 540 GGAGAAGAAT TAAATGCCAT CAACACAACA GTTTCCCAAG ATCCTGAAAC TGAGCTCTAT 600 GCTGTTAGCA GCAAACTGGA TTTCAATATG ACAACCAACC ACAGCTTCAT GTGTCTCATC 660 AAGTATGGAC ATTTAAGAGT GAATCAGACC TTCAACTGGA ATACAACCAA GCAAGAGCAT 720 TTTCCTGATA ACGGATTC 738 (2) INFORMATION FOR SEQ ID NO:33: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1002 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: CDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33: GAGCTCATGG ATCCCCAGTG CACTATGGGA CTGAGTAACA TTCTCTTTGT GATGGCCTTC 60 CTGCTCTCTG GTGCTGCTCC TCTGAAGATT CAAGCTTATT TCAATGAGAC TGCAGACCTG 120 CCATGCCAAT TTGCAAACTC TCAAAACCAA AGCCTGAGTG AGCTAGTAGT ATTTTGGCAG 180 GACCAGGAAA ACTTGGTTCT GAATGAGGTA TACTTAGGCA AAGAGAAATT TGACAGTGTT 240 CATTCCAAGT ATATGGGCCG CACAAGTTTT GATTCGGACA GTTGGACCCT GAGACTTCAC 300 AATCTTCAGA TCAAGGACAA GGGCTTGTAT CAATGTATCA TCCATCACAA AAAGCCCACA 360 GGAATGATTC GCATCCACCA GATGAATTCT GAACTGTCAG TGCTTGCTAA CTTCAGTCAA 420 CCTGAAATAG TACCAATTTC TAATATAACA GAAAATGTGT ACATAAATTT GACCTGCTCA 460 TCTATACACG GTTACCCAGA ACCTAAGAAG ATGAGTGTTT TGCTAAGAAC CAAGAATTCA 540 ACTATCGAGT ATGATGGTAT TATGCAGAAA TCTCAAGATA ATGTCACAGA ACTGTACGAC 600 GTTTCCATCA GCTTGTCTGT TTCATTCCCT GATGTTACGA GCAATATGAC CATCTTCTGT 660 ATTCTGGAAA CTGACAAGAC GCGGCTTTTA TCTTCACCTT TCTCTATAGA GCTTGAGGAC 720 CCTCAGCCTC CCCCAGACCA CATTCCTTGG ATTACAGCTG TACTTCCAAC AGTTATTATA 780 TGTGTGATGG TTTTCTGTCT AATTCTATGG AAATGGAAGA AGAAGAAGCG GCCTCGCAAC 840 TCTTATAAAT GTGGAACCAA CACAATGGAG AGGGAAGAGA GTGAACAGAC CAAGAAAAGA 900 GAAAAAATCC ATATACCTGA AAGATCTGAT GAAGCCCAGC GTGTTTTTAA AAGTTCGAAG 960 ACATCTTCAT GCGACAAAAG TGATACATGT TTTTAAGGGC CC 1002 (2) INFORMATION FOR SEQ ID NO:34: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 751 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 34: GAGCTCATGG ATCCCCAGTG CACTATGGGA CTGAGTAACA TTCTCTTTGT GATGGCCTTC 60 CTGCTCTCTG GTGCTGCTCC TCTGAAGATT CAAGCTTATT TCAATGAGAC TGCAGACCTG 120 CCATGCCAAT TTGCAAACTC TCAAAACCAA AGCCTGAGTG AGCTAGTAGT ATTTTGGCAG 180 GACCAGGAAA ACTTGGTTCT GAATGAGGTA TACTTAGGCA AAGAGAAATT TGACAGTGTT 240 CATTCCAAGT ATATGGGCCG CACAAGTTTT GATTCGGACA GTTGGACCCT GAGACTTCAC 300 AATCTTCAGA TCAAGGACAA GGGCTTGTAT CAATGTATCA TCCATCACAA AAAGCCCACA 360 GGAATGATTC GCATCCACCA GATGAATTCT GAACTGTCAG TGCTTGCTAA CTTCAGTCAA 420 CCTGAAATAG TACCAATTTC TAATATAACA GAAAATGTGT ACATAAATTT GACCTGCTCA 480 TCTATACACG GTTACCCAGA ACCTAAGAAG ATGAGTGTTT TGCTAAGAAC CAAGAATTCA 540 ACTATCGAGT ATGATGGTAT TATGCAGAAA TCTCAAGATA ATGTCACAGA ACTGTACGAC 600 GTTTCCATCA GCTTGTCTGT TTCATTCCCT GATGTTACGA GCAATATGAC CATCTTCTGT 660 ATTCTGGAAA CTGACAAGAC GCGGCTTTTA TCTTCACCTT TCTCTATAGA GCTTGAGGAC 720 CCTCAGCCTC CCCCAGACCA CATTGGGGCC C 751 (2) INFORMATION FOR SEQ ID NO:35: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1611 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA 136 (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SΞQ ID NO*.35: GAATTCATGG CTCCCAGCAG CCCCCGGCCC GCGCTGCCCG CACTCCTGGT CCTGCTCGGG 60 GCTCTGTTCC CAGGACCTGG CAATGCCCAG ACATCTGTGT CCCCCTCAAA AGTCATCCTG 120 CCCCGGGGAG GCTCCGTGCT GGTGACATGC AGCACCTCCT GTGACCAGCC CAAGTTGTTG 180 GGCATAGAGA CCCCGTTGCC TAAAAAGGAG TTGCTCCTGC CTGGGAACAA CCGGAAGGTG 240 TATGAACTGA GCAATGTGCA AGAAGATAGC CAACCAATGT GCTATTCAAA CTGCCCTGAT 300 GGGCAGTCAA CAGCTAAAAC CTTCCTCACC GTGTACTGGA CTCCAGAACG GGTGGAACTG 360 GCACCCCTCC CCTCTTGGCA GCCAGTGGGC AAGAACCTTA CCCTACGCTG CCAGGTGGAG 420 GGTGGGGCAC CCCGGGCCAA CCTCACCGTG GTGCTGCTCC GTGGGGAGAA GGAGCTGAAA 480 CGGGAGCCAG CTGTGGGGGA GCCCGCTGAG GTCACGACCA CGGTGCTGGT GAGGAGAGAT 540 CACCATGGAG CCAATTTCTC GTGCCGCACT GAACTGGACC TGCGGCCCCA AGGGCTGGAG 600 CTGTTTGAGA ACACCTCGGC CCCCTACCAG CTCCAGACCT TTGTCCTGCC AGCGACTCCC 660 CCACAACTTG TCAGCCCCCG GGTCCTAGAG GTGGACACGC AGGGGACCGT GGTCTGTTCC 720 CTGGACGGGC TGTTCCCAGT CTCGGAGGCC CAGGTCCACC TGGCACTGGG GGACCAGAGG 780 TTGAACCCCA CAGTCACCTA TGGCAACGAC TCCTTCTCGG CCAAGGCCTC AGTCAGTGTG 840 ACCGCAGAGG ACGAGGGCAC CCAGCGGCTG ACGTGTGCAG TAATACTGGG GAACCAGAGC 900 CAGGAGACAC TGCAGACAGT GACCATCTAC AGCTTTCCGG CGCCCAACGT GATTCTGACG 960 AAGCCAGAGG TCTCAGAAGG GACCGAGGTG ACAGTGAAGT GTGAGGCCCA CCCTAGAGCC 1020 AAGGTGACGC TGAATGGGGT TCCAGCCCAG CCACTGGGCC CGAGGGCCCA GCTCCTGCTG 1080 AAGGCCACCC CAGAGGACAA CGGGCGCAGC TTCTCCTGCT CTGCAACCCT GGAGGTGGCC 1140 GGCCAGCTTA TACACAAGAA CCAGACCCG3 GAGCTTCGTG TCCTGTATGG CCCCCGACTG 1200 GACGAGAGGG ATTGTCCGGG AAACTGGACG TGGCCAGAAA ATTCCCAGCA GACTCCAATG 1260 TGCCAGGCTT GGGGGAACCC ATTGCCCGAG CTCAAGTGTC TAAAGGATGG CACTTTCCCA 1320 CTGCCCATCG GGGAATCAGT GACTGTCACT CGAGATCTTG AGGGCACCTA CCTCTGTCGG 1380 GCCAGGAGCA CTCAAGGGGA GGTCACCCGC GAGGTGACCG TGAATGTGCT CTCCCCCCGG 1440 TATGAGATTG TCATCATCAC TGTGGTAGCA GCCGCAGTCA TAATGGGCAC TGCAGGCCTC 1500 AGCACGTACC TCTATAACCG CCAGCGGAAG ATCAAGAAAT ACAGACTACA ACAGGCCCAA 1560 AAAGGGACCC CCATGAAACC GAACACACAA GCCACGCCTC CCTGAGGATC C 1611 (2) INFORMATION FOR SEQ ID NO:36: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1452 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36: GAATTCATGG CTCCCAGCAG CCCCCGGCCC GCGCTGCCCG CACTCCTGGT CCTGCTCGGG 60 GCTCTGTTCC CAGGACCTGG CAATGCCCAG ACATCTGTGT CCCCCTCAAA AGTCATCCTG 120 CCCCGGGGAG GCTCCGTGCT GGTGACATGC AGCACCTCCT GTGACCAGCC CAAGTTGTTG 180 GGCATAGAGA CCCCGTTGCC TAAAAAGGAG TTGCTCCTGC CTGGGAACAA CCGGAAGGTG 240 TATGAACTGA GCAATGTGCA AGAAGATAGC CAACCAATGT GCTATTCAAA CTGCCCTGAT 300 GGGCAGTCAA CAGCTAAAAC CTTCCTCACC GTGTACTGGA CTCCAGAACG GGTGGAACTG 360 GCACCCCTCC CCTCTTGGCA GCCAGTGGGC AAGAACCTTA CCCTACGCTG CCAGGTGGAG 420 GGTGGGGCAC CCCGGGCCAA CCTCACCGTG GTGCTGCTCC GTGGGGAGAA GGAGCTGAAA 480 CGGGAGCCAG CTGTGGGGGA GCCCGCTGAG GTCACGACCA CGGTGCTGGT GAGGAGAGAT 540 CACCATGGAG CCAATTTCTC GTGCCGCACT GAACTGGACC TGCGGCCCCA AGGGCTGGAG 600 CTGTTTGAGA ACACCTCGGC CCCCTACCAG CTCCAGACCT TTGTCCTGCC AGCGACTCCC 660 CCACAACTTG TCAGCCCCCG GGTCCTAGAG GTGGACACGC AGGGGACCGT GGTCTGTTCC 720 CTGGACGGGC TGTTCCCAGT CTCGGAGGCC CAGGTCCACC TGGCACTGGG GGACCAGAGG 780 TTGAACCCCA CAGTCACCTA TGGCAACGAC TCCTTCTCGG CCAAGGCCTC AGTCAGTGTG 840 ACCGCAGAGG ACGAGGGCAC CCAGCGGCTG ACGTGTGCAG TAATACTGGG GAACCAGAGC 900 CAGGAGACAC TGCAGACAGT GACCATCTAC AGCTTTCCGG CGCCCAACGT GATTCTGACG 960 AAGCCAGAGG TCTCAGAAGG GACCGAGGTG ACAGTGAAGT GTGAGGCCCA CCCTAGAGCC 1020 AAGGTGACGC TGAATGGGGT TCCAGCCCAG CCACTGGGCC CGAGGGCCCA GCTCCTGCTG 1080 AAGGCCACCC CAGAGGACAA CGGGCGCAGC TTCTCCTGCT CTGCAACCCT GGAGGTGGCC 1140 GGCCAGCTTA TACACAAGAA CCAGACCCGG GAGCTTCGTG TCCTGTATGG CCCCCGACTG 1200 GACGAGAGGG ATTGTCCGGG AAACTGGACG TGGCCAGAAA ATTCCCAGCA GACTCCAATG 1260 TGCCAGGCTT GGGGGAACCC ATTGCCCGAG CTCAAGTGTC TAAAGGATGG CACTTTCCCA 1320 CTGCCCATCG GGGAATCAGT GACTGTCACT CGAGATCTTG AGGGCACCTA CCTCTGTCGG 1380 GCCAGGAGCA CTCAAGGGGA GGTCACCCGC GAGGTGACCG TGAATGTGCT CTCCCCCCGG 1440 TATGAGGGAT CC 1452 (2) INFORMATION FOR SEQ ID NO:37: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 726 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37: GAGCTCATGG TTGCTGGGAG CGACGCGGGG CGGGCCCTGG GGGTCCTCAG CGTGGTCTGC 60 CTGCTGCACT GCTTTGGTTT CATCAGCTGT TTTTCCCAAC AAATATATGG TGTTGTGTAT 120 GGGAATGTAA CTTTCCATGT ACCAAGCAAT GTGCCTTTAA AAGAGGTCCT ATGGAAAAAA 180 CAAAAGGATA AAGTTGCAGA ACTGGAAAAT TCTGAATTCA GAGCTTTCTC ATCTTTTAAA 240 AATAGGGTTT ATTTAGACAC TGTGTCAGGT AGCCTCACTA TCTACAACTT AACATCATCA 300 GATGAAGATG AGTATGAAAT GGAATCGCCA AATATTACTG ATACCATGAA GTTCTTTCTT 360 TATGTGCTTG AGTCTCTTCC ATCTCCCACA CTAACTTGTG CATTGACTAA TGGAAGCATT 420 GAAGTCCAAT GCATGATACC AGAGCATTAC AACAGCCATC GAGGACTTAT AATGTACTCA 480 TGGGATTGTC CTATGGAGCA ATGTAAACGT AACTCAACCA GTATATATTT TAAGATGGAA 540 AATGATCTTC CACAAAAAAT ACAGTGTACT CTTAGCAATC CATTATTTAA TACAACATCA 600 TCAATCATTT TGACAACCTG TATCCCAAGC AGCGGTCATT CAAGACACAG ATATGCACTT 660 ATACCCATAC CATTAGCAGT AATTACAACA TGTATTGTGC TGTATATGAA TGTTCTTTAA 720 GGATCC 726 (2) INFORMATION FOR SEQ ID NO:38: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 657 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:38: GAGCTCATGG TTGCTGGGAG CGACGCGGGG CGGGCCCTGG GGGTCCTCAG CGTGGTCTGC 60 CTGCTGCACT GCTTTGGTTT CATCAGCTGT TTTTCCCAAC AAATATATGG TGTTGTGTAT 120 GGGAATGTAA CTTTCCATGT ACCAAGCAAT GTGCCTTTAA AAGAGGTCCT ATGGAAAAAA 180 CAAAAGGATA AAGTTGCAGA ACTGGAAAAT TCTGAATTCA GAGCTTTCTC ATCTTTTAAA 240 AATAGGGTTT ATTTAGACAC TGTGTCAGGT AGCCTCACTA TCTACAACTT AACATCATCA 300 GATGAAGATG AGTATGAAAT GGAATCGCCA AATATTACTG ATACCATGAA GTTCTTTCTT 360 TATGTGCTTG AGTCTCTTCC ATCTCCCACA CTAACTTGTG CATTGACTAA TGGAAGCATT 420 GAAGTCCAAT GCATGATACC AGAGCATTAC AACAGCCATC GAGGACTTAT AATGTACTCA 480 TGGGATTGTC CTATGGAGCA ATGTAAACGT AACTCAACCA GTATATATTT TAAGATGGAA 540 AATGATCTTC CACAAAAAAT ACAGTGTACT CTTAGCAATC CATTATTTAA TACAACATCA 600 TCAATCATTT TGACAACCTG TATCCCAAGC AGCGGTCATT CAAGACACAG AGGATCC 657 (2) INFORMATION FOR SEQ ID NO:39: <i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39: Glu Gin Leu Glu Ser lie lie Asn Phe Glu Lys Leu Thr Glu Trp Thr 1 5 10 15 Ser Ser Asn Val Met Glu Glu Arg 20 (2) INFORMATION FOR SEQ ID NO: 0: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 0: Asp Leu Arg Gly Tyr Val Tyr Gin Gly Leu 1 5 10 (2) INFORMATION FOR SEQ ID NO:41: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SΞQ ID NO:41 Phe Arg lie Gly Cys Arg His Ser Arg 1 5 (2) INFORMATION FOR SEQ ID NO:42: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 9 ammo acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SΞQ ID NO: 2 lie Leu Lys Glu Pro Val His Gly Val 1 5 (2) INFORMATION FOR SEQ ID NO:43: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 32 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43: ATATGGATCC TCACCATCTC AGGGTGAGGG GC 32 (2) INFORMATION FOR SEQ ID NO: 4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44: Arg Gly Tyr Val Tyr Gin Gly Leu Lys Ser 1 5 10 (2) INFORMATION FOR SEQ ID NO: 5: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 9 amino acids (B) TYPE: am o acid (C) STRANDEDNESS: Single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45: Phe Ala Pro Gly Asn Tyr Pro Ala Leu 1 5 (2) INFORMATION FOR SEQ ID NO:46: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (3) TYPΞ: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: internal ( i) SEQUENCE DESCRIPTION: SΞQ ID NO:46: Leu Ser Pro Phe Pro Phe Asp Leu 1 5 (2) INFORMATION FOR SEQ ID NO:47: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SΞQ ID NO: 7: Gin Leu Ser Pro Phe Pro Phe Asp Leu 1 5 (2) INFORMATION FOR SEQ ID NO: 8: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8: lie Leu Lys Glu Pro Val His Gly Val 1 5 (2) INFORMATION FOR SEQ ID NO: 9: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49: Tyr Met Asn Gly Thr Met Ser Gin Val 1 5 (2) INFORMATION FOR SEQ ID NO:50: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50: Gly He Leu Gly Phe Val Phe Thr Leu 1 5 (2) INFORMATION FOR SEQ ID NO:51: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: internal ( i) SEQUENCE DESCRIPTION: SEQ ID NO:51: Phe Leu Pro Ser Asp Phe Phe Pro Ser Val 1 5 10

Claims: WE CLAIM : 1. A synthetic antigen presenting cell line for use with T-cell lymphocytes comprising: a) a Class I MHC heavy chain gene operably linked to a first promoter and capable of expressing a Class I MHC heavy chain; b) a β- 2 microglobulin gene operably linked to a second promoter and capable of expressing a β- 2 microglobulin that with the MHC heavy chain forms MHC molecules; and c) a gene fcr an assisting molecule operably linked to a third promoter and capable of expressing an assisting molecule that interacts with a molecule on the T-cell lymphocytes; at least one of the MHC gene, β- 2 microglobulin gene and assisting molecule genes not being present in the cells from which the cell line is derived, such that the MHC molecules bind to a peptide, and the MHC molecules and assisting molecules are presented on the surface of the cell in sufficient numbers to activate a population of T-cell lymphocytes against the peptide when the peptide is bound to the MHC molecules. 2. The cell line of claim 1 wherein the assisting molecule is a costimulatory molecule. 3. The cell line of claim 2 wherein the costimulatory molecule is B7.1 or B7.2. 4. The cell line of claim 1 wherein the cell line is derived from a first species and the MHC heavy chain gene is from a second species. 5. The cell line of claim 4 wherein the first species is a poikilotherm and the second species is a homeotherm. 6. The cell line of claim 1 wherein the assisting molecule is an adhesion molecule. 7. The cell line of claim 6 wherein the adhesion molecule is ICAM-1, ICAM-2, ICAM-3 or LFA-3. 8. The cell line of claim 1 having a gene for a first assisting molecule and a gene for a second assisting molecule. 9. The cell line of claim 8 wherein the first assisting molecule is a costimulatory molecule and the εecond assisting molecule is an adhesion molecule. 10. The cell line of claim 1 wherein at least one promoter is inducible. 11. The cell line of claim 10 wherein the first promoter is inducible. 12. The cell line of claim 1 wherein the peptide is bound to the MHC molecules within the cell . 13. The cell line of claim 1 wherein the MHC molecules are presented empty on the surface of the cell. 14. A stable poikilotherm cell line used to stimulate human T-cell lymphocytes comprising: a) a Class I MHC gene operably linked to a first inducible promoter and capable of expresεing a Class I MHC heavy chain; b) a ,5-2 microglobulin gene operably linked to a second promoter and capable cf expresεing a β- 2 microglobulin that forms MHC molecules with the MHC heavy chain; and c) a gene for an assiεting molecule operably linked to a third promoter and capable of expreεεing an assisting molecule that interacts with a molecule on the T-cell lymphocytes; such that the cell assembles empty MHC molecules that are capable of binding to a peptide, and the MHC molecules and assisting molecules are presented on the surface of the cells in sufficient numbers to activate a population of T-cell lymphocytes against the peptide when the peptide is bound to the MHC molecules. 15. The cell line of claim 14 wherein the assisting molecule is a costimulatory molecule. 16. The cell line of claim 15 wherein the costimulatory molecule is B7.1 or B7.2. 17. The cell line of claim 14 wherein the assisting molecule is an adhesion molecule. 18. The cell line of claim 17 wherein the adhesion molecule is ICAM-1. 19. The cell line of claim 14 having a gene for a first assisting molecule and a gene for a second assisting molecule. 20. The cell line of claim 19 wherein the first assisting molecule is a costimulatory molecule and the second assisting molecule is an adhesion molecule. 21. A stable poikilotherm cell line used to stimulate human T-cell lymphocytes comprising: a) a Class I MHC gene operably linked to a first promoter and capable of expressing a Class I MHC heavy chain; b) a β- 2 microglobulin gene operably linked to a second promoter and capable of expreεεing a β- 2 microglobulin that forms MHC molecules with the MHC heavy chain; and c) a gene for a costimulatory molecule operably linked to a third promoter and capable of expressing a costimulatory molecule that interacts with a molecule on the T-cell lymphocytes,* d) a gene for an adhesion molecule operably linked to a fourth promoter and capable of expressing an adhesion molecule that interacts with a cooperative adhesion molecule on the T-cell lymphocytes; such that the cell is capable of assembling the MHC heavy chain and the β- 2 microglobulin into MHC molecules that bind to a peptide, and transporting MHC molecules, costimulatory molecules and adhesion molecules to the surface of the cell in sufficient numbers to activate a population of T-cell lymphocytes against the peptide when the peptide is bound to the MHC molecules. 22. The cell line of claim 21 wherein the costimulatory molecule is B7.1 or B7.2. 23. The cell line of claim 21 wherein the adhesion molecule is ICAM-1, ICAM-2, ICAM-3 or LFA-3. 24. The cell line of claim 21 wherein at least one of the promoters is inducible. 25. The cell line of claim 21 wherein the peptide is bound to the MHC molecules within the cell. 26. The cell line of claim 21 wherein the MHC molecules are presented empty on the surface of the cell. 27. Fragments of cells of claim 1 having MHC molecules and assisting molecules operably associated such that they activate the population of T-cell lymphocytes. 28. The cell fragment of claim 27 wherein the MHC molecules are empty. 29. The cell fragment of claim 27 wherein the peptide is bound to the MHC molecules. 30. Fragments of cells of claim 21 having MHC molecules, costimulatory molecules and adhesion molecules operably associated such that they activate the population of T-cell lymphocytes. 31. The cell fragment of claim 27 wherein the peptide is bound to the MHC molecules. 32. A method of producing a synthetic antigen presenting a cell line comprising: a) establishing of a culture of cells,* b) transfecting the culture with an expressible Class I MHC heavy chain gene operably linked to a first promoter; c) transfecting the culture with an expressible β-2 microglobulin gene operably linked to a second promoter; and d) transfecting the culture with an expressible assisting molecule gene operably linked to a third promoter. 33. The method of claim 32 wherein the asεisting molecule is a costimulatory molecule. 34. The method of claim 33 wherein the costimulatory molecule is B7.1 or B7.2. 35. The method of claim 32 wherein the cell line is derived from a first species and the MHC heavy chain gene is from a second species. 36. The method of claim 35 wherein the first species is a poikilotherm and the second species is a homeotherm. 37. The method of claim 32 wherein the assisting molecule is an adhesion molecule. 38. The method of claim 37 wherein the adhesion molecule is ICAM-1, ICAM-2, ICAM-3 or LFA-3. 39. The method of claim 32 including the step of transfecting the culture with a gene for a second assisting molecule. 40. The method of claim 39 wherein the first assiεting molecule is a costimulatory molecule and the second assisting molecule is an adhesion molecule. 41. The method of claim 32 wherein at least one of the promoters is inducible. 42. A method of producing a synthetic antigen presenting cell line comprising: a) establishing a culture of cells lacking a gene to at least one of: Class I MHC heavy chain, β- 2 microglobulin and assisting molecule; and b) transfecting the culture with an expressible gene for each of the genes of (a) lacking in the culture of cells, the gene being operably linked to a promoter. 43. A method of producing a synthetic antigen presenting a cell line comprising: a) establishing of a culture of cells,* b) transfecting the culture with an expressible Class I MHC heavy chain gene operably linked to a promoter; c) transfecting the culture with an expressible β-2 microglobulin gene operably linked to a second promoter; d) transfecting the culture with an expressible costimulatory molecule gene operably linked to a third promoter; and e) transfecting the culture with an expressible adhesion molecule gene operably linked to a third promoter. 44. A method of producing a synthetic antigen presenting cell line comprising: a) establishing a culture of cells lacking a gene to at least one of: Class I MHC heavy chain, β- 2 microglobulin and co-stimulatory molecule,* b) transfecting the culture with an expressible gene for each of the genes of (a) lacking in the culture of cells, the gene being linked to a first operable promoter; and c) transfecting the culture with an expressible adhesion molecule gene, the gene being linked to a second operable promoter. 45. A method for activating CD8 " T-cells against a selected peptide in vi tro, the method comprising: a) providing the cell line of claim 1; b) culturing the cell line under conditions such that MHC molecules are produced on the surface of the cell line that are bound to the selected peptide,* and c) contacting the cultured cells with the CD8 * T-cells against the selected peptide. 46. The method of claim 45 wherein the cell line is a poikilotherm. 47. The method of claim 45 further comprising the step of separating the activated CD8 * T-cells from the cell line. 48. The method of claim 47 further comprising the step of adding the activated CD8 * T-cells to an acceptable carrier or excipient to form a suspension. 49. The method of claim 48 further comprising the step of administering the suspension to a patient. 50. A synthetic antigen-presenting matrix comprising: a) a support; b) extracellular portion of MHC molecules capable of binding to a selected peptide and being operably linked to the support; and c) an assisting molecule operably linked to the support such that the extracellular, portion of the MHC and assisting molecules are present in sufficient numbers to activate a population of T-cell lymphocytes against the peptide when the peptide is bound to the extracellular portion of the MHC molecule. 51. The matrix of claim 50 wherein the εupport iε a cell fragment. 52. The matrix of claim 50 wherein the support is a cell. 53. The matrix of claim 52 wherein the extracellular portion of the MHC molecule is linked to the cell by a transmembrane domain of an MHC molecule. 53. The matrix of claim 53 wherein the extracellular portion of the MHC molecule is part of a complete MHC molecule. 54. The matrix of claim 50 wherein the support is liposome. 55. The matrix of claim 50 wherein the εupport is a solid surface. 56. The matrix of claim 50 wherein the extracellular portion of the MHC molecule is linked to an epitope which reacts with an antibody to link the portion to the support . 57. The matrix of claim 50 wherein the extracellular portion of the MHC molecule is linked to (His) 6 which reacts with nickel to link the portion to the support. 58. The matrix of claim 50 wherein the support is a porous material. 59. The matrix of claim 50 wherein the assisting molecule is a costimulatory molecule. 60. The matrix of claim 59 wherein the costimulatory molecule is B7.1 or B7.2. 61. The matrix of claim 59 wherein the costimulatory molecule is is at least a portion of an anti-CD80 antibody. 62. The matrix of claim 50 wherein the assisting molecule is an adhesion molecule. 63. The matrix of claim 62 wherein the adhesion molecule is ICAM-1, ICAM-2, ICAM-3 or LFA-3. 64. The matrix of claim 50 having two assisting molecules, the first asεiεting molecule being a coεtimulatory molecule and the second asεiεting molecule being an adhesion molecule. 65. The matrix of claim 50 wherein the extracellular portion of the MHC molecule is empty. 66. The matrix of claim 50 wherein the peptide is bound to the extracellular portion of the MHC molecule. 67. The matrix of claim 64 wherein the peptide is bound to the extracellular portion of the MHC molecule. 68. A synthetic antigen-presenting matrix comprising: a) a support; b) extracellular portion of MHC molecules capable of binding to a selected peptide and being operably linked to the εupport; c) B-7.1 or B-7.2 molecules or a combination thereof operably linked to the support; and d) ICAM-1 molecules operably linked to the support such that the mclecules are present in sufficient amount to activate a population of T-cell lymphocytes against the peptide when the peptide is bound to the extracellular portion of the MHC molecule. 69. A method of producing a synthetic T-cell lymphocyte antigen activating matrix comprising: a) establishing a first culture of cells; b) transfecting the culture with an expresεible Class I MHC heavy chain gene for at least the extracellular portion of the heavy chain operably linked to a promoter; c) transfecting the culture with an expressible β- 2 microglobulin gene operably linked to a second promoter; d) harvesting the produced MHC molecule portions; e) establishing a second culture of cells; f) transfecting the second culture with an expressible assisting molecule gene for at least the M I S S I N G P A G E b) contacting the matrix cells with the CD8 * T-cells. 79. A method for generating cytotoxic CD8* T-cells against a selected peptide, the method comprising: a) providing the matrix of claim 67; b) contacting naive CD8 * T-cells with the matrix in vi tro . 80. The matrix of claim 79 wherein the support is a cell. 81. The method of claim 79 wherein the costimulatory molecule is B7.1 or B7.2. 82. The method of claim 79 wherein the costimulatory molecule is at least a portion of an anti- CD28 antibody. 83. The method of claim 79 wherein the adhesion molecule is ICAM-1, ICAM-2, ICAM-3 or LFA-3. 84. A method of treating a tumor in a patient comprising: a) identifying a tumor specific antigen; b) collecting CD8 * T-cells from the patient; c) contacting the CD8 * T-cells with the matrix of claim 67 in vi tro in a sufficient amount and for a sufficient time to generate cytotoxic CD8 * T-cells, • and d) returning the cytotoxic CD8 * T-cells to the patient. 85. The method of claim 84 wherein the antigen is a self antigen of the patient. 86. The method of claim 85 wherein the asεisting molecule is B7.1 or B7.2 and the adhesion molecule is ICAM-1, ICAM-2 or ICAM-3. 87. The method of claim 84 wherein the assisting molecule is B7.1 and the adhesion molecule is ICAM-1.

Inventors: BRUNMARK ANDERS, CAI ZELING, JACKSON MICHAEL, LETURCQ DIDIER JEAN, LUXEMBOURG ALAIN M D PH D, MORIARTY ANN M, PETTERSON PER A, SPRENT JONATHAN, BRUNMARK, ANDERS, CAI, ZELING, JACKSON, MICHAEL, LETURCQ, DIDIER, JEAN, LUXEMBOURG, ALAIN, M., D., PH., D., MORIARTY, ANN, M., PETTERSON, PER, A., SPRENT, JONATHAN
Applicants: BRUNMARK ANDERS, CAI ZELING, JACKSON MICHAEL, LETURCQ DIDIER JEAN, LUXEMBOURG ALAIN M D PH D, MORIARTY ANN M, PETTERSON PER A, SCRIPPS RESEARCH INST, SPRENT JONATHAN, BRUNMARK, ANDERS, CAI, ZELING, JACKSON, MICHAEL, LETURCQ, DIDIER, JEAN, LUXEMBOURG, ALAIN, M., D., PH., D., MORIARTY, ANN, M., PETTERSON, PER, A., SPRENT, JONATHAN, THE SCRIPPS RESEARCH INSTITUTE
Application year: 1996
Main classification: A61K45
IPC codes: A61K35, A61K38, A61K48, A61P35, C07K14, C12N5, C12N15, C12P21