ICAM-1/LFA-1 SHORT-CHAIN PEPTIDES AND METHOD OF USING SAME

Abstract: Short-chain peptides replicating fragments of functional domains derived from LFA-1 and ICAM-1 parent protein sequences serve to modulate the ICAM/LFA binding interaction. In one aspect of the invention, this modulation serves to block interprotein binding reactivity, as a peptide of the invention binds to a target protein in a manner that precludes the normal binding reaction between ICAM-1 and LFA-1. In another aspect of the invention, this modulation enhances the reactivity of a first peptide, as a second peptide induces a conformational change in the target protein from a first conformation to a second, more reactive, conformation. The peptides are used according to a method including the steps of providing the proteins and applying them to a population of cells.

Description: ICAM-l/LFA-1 SHORT-CHAIN PEPTIDES AND METHOD OF USING SAME Sequence Listing A printed Sequence Listing accompanies this applica¬ tion, and has also been submitted with identical contents in the form of a computer-readable ASCII file on a floppy diskette. Background of the Invention 1. Field of the Invention The invention pertains to the field of protein chemistry and, more particularly, to short-chain peptides that serve to modulate interprotein interactions. More specifically, these short chain peptides have functional residue sequences that essentially replicate fragments of functional domain sequences in corresponding parent protein molecules, but without replicating the entire parent protein. A first such peptide may be mixed with a second peptide to enhance the functionality of the first peptide by inducing a conformational change in a target protein that is reactive with the first peptide. Simi¬ larly, the second peptide may induce a conformational change in the first peptide. Particularly preferred short-chain peptides are taken from parent protein mole¬ cule sequences selected from the family of integrins, proteins that react with integrins, and the superfamily of immunoglobulins. The peptides are utilized according to methods including the steps of providing the peptides and applying them to a population of cells. 2. Description of the Prior Art Certain immune system functions rely upon a critical interprotein binding interaction as a necessary element of immune response. One such binding interaction pertains to a heterophilic interaction between the glyco- proteins including Intercellular Adhesion Molecule ("ICAM- 1") and Leucocyte Function Associated antigen-1 ("LFA-1") . Examples of published sequences for LFA-1 and ICAM-1 are submitted as Sequence ID No.'s 1, 2, and 3. It is possible to derive immunological benefits by disrupting the intercellular ICAM/LFA binding interac¬ tion through the application of specific monoclonal antibodies ("mAbs"), i.e., anti-ICAM-1 or anti-LFA-1. As reported in Isobe et al . , Specific Acceptance of Cardiac Allograft After Treatment Wi th Antibodies to ICAM-1 and LFA-1 , 255 SCIENCE 1125-1127 (Feb. 1992), an induced immune tolerance enables the indefinite survival of cardiac allografts between fully incompatible mouse strains subsequent to a six day treatment including the simultaneous application of anti-ICAM-1 and anti-LFA-1 mAbs. Unfortunately, a major problem with the use of mAbs for the purpose of inducing immune-tolerance in a mammal is that these large (e.g., 80-150 kDa) molecules typically also induce an effectiveness-limiting immune response to the mAbs. LFA-1 has special significance in that it belongs to a family or class of proteins that are known as integrins, and is associated with leukocytes such as T and B cells. The integrin family includes similar glycopro- teins that combine two separate protein units as a func¬ tional member, i.e., a functional heterodimer formed of a β unit and an α. unit. These respective units each com¬ prise separate proteins that are anchored within interac- tive proximity to one another, and can extend outwardly from a cytoplasmic domain, across a transmembrane domain, and beyond the cell membrane. In LFA-1, binding avidity for ICAM-1 involves Mg 2+ and Ca 2* ions that, in part, govern the interaction between the β and α units. The integrin family may be subdivided into two groups wherein common group members share common β units. A first group of proteins share the common βl (CD29) unit, and typically function by binding to extracellular matrix proteins. This first group includes the VLA-1 (CD49a/CD29) , VLA-2 (CD49b/CD29) , VLA-3 (CD49c/CD29) , VLA- 4 (CD49d/CD29) , VLA-5 (CD49e/CD29) and VLA-6 (CD49f/CD29) proteins. A second group shares the β2 unit (CD18) , and typically functions in cell to cell interactions. This second group includes LFA-1 (CD18/CDlla) , MAC-1 (CD18/CDllb) , and pl50,95 (CD18/CDllc) . The LFA-lα. unit (CDlla protein, i.e., Sequence ID No. 2) is the specific heterodimer counterpart to the β2 unit in LFA-1. Li et al . , A Leukocyte Integrin Binding Peptide from Intercellular Adhesion Molecule-2 Stimulates T Cell Adhesion and Natural Killer Cell Activi ty, 268 J. Biol. Chem. 21474-21477 (July 20, 1993), report that β2 inte¬ grins, and particularly LFA-1, have a cytoplasmic domain which serves to alter protein configuration via a cellular phosphorylation pathway communicating the integrin and respective cell receptor areas. That is to say, phosphor¬ ylation may be activated at a remote receptor site with the result of indirectly inducing conformational changes in the integrin via cytoplasmic phosphorylation. On the other hand, ICAM-1 proteins are not known to incur phosphorylation induced conformational changes. ICAM-1 is found primarily upon monocytes and endothelial cells, and is widely inducible, or upregulated, on many cells including B and T lymphocytes, thymocytes, dendritic cells, endothelial cells, fibroblasts, keratinocytes, chondrocytes, and epithelial cells. This protein has a co-stimulatory effect upon cytotoxic T-cell interaction, and is utilized in a number of intercellular binding interactions. Recent developments in the field of protein chemistry confirm that short-chain peptides, which have amino acid residue sequences representing mere fragments of a corresponding parent protein molecule, may exhibit significant levels of biofunctionality. Short-chain peptides that derive from the ICAM-1 protein are known to have utility in blocking binding interactions between cells and viruses. Even so, only a very limited number of specific ICAM-1 based peptides have been shown to be useful. European Patent Publication No. EP 391,088 A2 indicates that functional derivatives of the intercellular adhesion molecule ("ICAM-1") may be used in anti-viral therapy. These functional derivatives may include func- tional domains and fragments of the ICAM-1 molecule. The method of using these fragments includes administering them in a manner that prevents viral infection of poten¬ tial viral host cells by impairing a binding interaction between the cells and rhinoviruses that may contact the cells. The binding interaction to be impaired includes one between ' an ICAM-1 cellular receptor and a correspond¬ ing viral adhesion site. The ICAM-1 fragments impair the cell-to-virus binding interaction by competing with normal cellular ICAM-1 molecules for adhesion to the correspond- ing viral binding site. Another publication, PCT/AU91/00205, describes the use of short-chain ICAM-1 based peptides for inhibit¬ ing intercellular adhesion in mammals. Summary of the Invention The present invention overcomes the problems outlined above by providing, for methods of use, a variety of short-chain ICAM-1 and LFA-1 based peptides having sufficiently low molecular weights to avoid the induction of a mammalian immune system response of a type that would attack the peptides. These peptides are useful for modulating intercellular binding interactions by blocking or enhancing interprotein reactivity. In some instances, a single peptide can function as both a binding blocker or a binding enhancer, depending upon the peptide concentra¬ tion. Another aspect of the invention pertains to the use of protein conformation modulator peptides that increase the avidity of these short-chain peptides for binding with naturally occurring ICAM-1 and LFA-1 based target pro- teins. Peptides of the invention may have further utility in immunosuppression applications. A specific immune tolerance to organ transplants may be induced by killing T-cells that recognize the transplanted tissues as foreign antigens by mimicking the thymic winnowing of developing T-cells. Cytotoxic T-cells require a two-stage activation, which includes a first signal that results from contacting the T-cell antigen receptor with a spe¬ cific antigen, and a second signal corresponding to for example the ICAM/LFA binding interaction. In the event that the second signal is blocked, the antigen-activated T-cell is eventually induced to die by apoptosis. Long- term immune tolerance is induced once the population of activatable antigen-specific T-cells dies. In a similar manner, these peptides may be used as an alternative treatment for various diseases where immunosuppression is a common therapy, e.g., Crohn's disease, ulcerative colitis, multiple sclerosis, rheumatoid arthritis, sys¬ temic lupus erythematosus, pemphigus vulgaris, pemphigoid, acquired epidermolysis bullous, allergic contact dermati¬ tis, pyoderma gangrenosum, psoriasis, asthma, and diabe¬ tes. Broadly speaking, one facet of the invention pertains to short-chain peptides that serve to promote the efficacy of other short-chain peptides in reacting with target proteins. It has been discovered that, in the case where a first short-chain peptide exhibits a specific type of biofunctionality, a second short-chain modulator peptide can induce conformational changes in a target protein, and these conformational changes can serve to increase the binding of the first protein for the target protein. The second peptide produces this increased level of binding by shifting the target protein from a first conformation to a second, more reactive conformation. Accordingly, a synergism is observed in that the first peptide has an increased level of reactivity with the target protein in the presence of the second peptide. A specific example of the use of modulator peptides includes the LFA-lβ based LJCRF-4 (Sequence ID No. 16) peptide, which synergistically increases binding of the LFA-lα. based LJCRF-6 (Sequence ID No. 18) peptide for binding with ICAM-1. Alternatively, these modulator peptides may induce a conformational change that decreases reactivity. Short-chain peptides of the invention, e.g., Sequence ID Nos. 4-18, each preferably replicate a frag¬ ment of a functional domain that is based upon or identi¬ fied from a parent protein molecule. These fragments have a residue sequence length less than the entire parent protein, and exhibit functionality as reactivity with naturally occurring proteins that are the reactive coun¬ terparts of the parent protein from which the peptides derive. By way of example, an ICAM-1 based peptide of the invention may consist essentially of the sequence from positions 28-37 of Sequence ID No. 3 (see Table 1) . ICAM- 1 based peptides compete with naturally occurring ICAM-1 proteins by targeting LFA-1 cellular proteins. Similarly, LFA-1 peptides target ICAM-1 cellular proteins. These short-chain peptides exhibit biofunc¬ tionality as a capacity for modulating the natural ICAM/LFA binding interaction by either acting to block or enhance the level of protein interaction. This modulation results from ICAM-1 or LFA-1 based short-chain peptides that bind to a corresponding natural ICAM-1 or LFA-1 target protein on a cell. By way of example, in this bound position the short-chain peptide can serve to block a T-cell from attaching to another cell by precluding the LFA-1 or ICAM-1 to which the peptide is bound on one cell from interacting in the normal manner with a counterpart binding protein on the other cell. A preferred form of the invention includes modulating the ICAM/LFA binding interaction by providing short-chain peptides that are based upon functional domain sub-sequences which are taken from the family of inte¬ grins. Sub-sequences that are taken from the LFA-1 and β units are particularly preferred. Short-chain peptides of the invention preferably have a molecular weight less than about 20 kDa, more preferably less than about 15 kDa, even more preferably less than about 10 kDa, and most preferably less than about 5 kDa, in order to reduce the chance for inducing an undesirable immune response, such as an allergic reaction or the production of antibodies for attacking the pep¬ tides, which might otherwise occur after the peptides are injected into a mammal or other organism. Alternatively, peptides of the invention preferably have an amino acid residue sequence of a length less than about 50 residues, more preferably have a length less than about 30 residues, and most preferably have a length less than about 25 residues . In a study of the ICAM/LFA interaction, the degree of peptide binding to a target protein can be acceptably quantified by using a flow cytometer to measure median fluorescence intensity peak shifts in terms of percentage change from .a control peak. In this type of device, a population of cells is individually subjected to a laser fluorescence study for purposes of identifying the presence of fluorescein labeled peptides that previously bound to a target protein on a living cell. The fluores¬ cence intensity results for each cell are plotted in the general shape of a bell curve, and a median intensity peak value is calculated for the test sample as well as a control sample. Median peak shifts greater than about 10% with respect to the control usually indicate a level of functionality beyond that which might be induced by mere laboratory error, and shifts greater than about 20% are particularly good indicators of modulation in the ICAM/LFA binding interaction. The peptides are used by introducing them into a population of cells. A blocking peptide can be mixed with a modulator peptide for increased in vivo efficacy of the blocking peptide. Peptides of the invention may be produced by any available method including, by way of example, automated protein synthesis and recombinant host expression. Brief Description of the Drawings Figure 1 schematically depicts the LFA-1 and ICAM-1 proteins, including special regions of interest such as signal sequences, sequence repeat regions, and suspected functional domains from which short-chain peptides were selected for synthetic replication; Fig. 2 depicts a cartesian plot of flow cytome- ter results that were obtained from one of several fluorescence runs that were conducted to quantify the capacity of the ICAM-1 based SC-3-111 (Sequence ID No. 10) peptide for blocking the specific antibody binding of anti-LFA-1 mAb; Fig. 3 is a plot similar to Fig. 2, but derived from a sample of the LJCRF-1 (Sequence ID No. 6) peptide; Fig. 4 depicts a plot of flow cytometer results for the LFA-1 based LJCRF-4 (Sequence ID No. 16) peptide, which functions to promote the specific antibody binding of anti-ICAM-1 mAb; Fig. 5 is a photograph obtained from a control sample in a T-cell intercellular adhesion assay, wherein the cells demonstrated a natural lack of intercellular clumping; Fig. 6 is a photograph of a sample similar to that of Fig. 5, but in which a chemical additive has induced substantially complete intercellular clumping by way of the ICAM/LFA binding interaction; Fig. 7 is a photograph of a sample similar to that of Fig. 6, wherein a peptide of the invention has overcome the effect of the chemical additive to restore the sample to a state wherein substantially no intercellu¬ lar clumping is observed; Fig. 8 depicts a plot of flow cytometer results that demonstrate substantially no T-cell binding enhance- ment of fluorecescein labeled LJCRF-6 (Sequence ID No. 18) peptide in the presence of SC-3-111 (Sequence ID No. 10) peptide; Fig. 9 depicts a plot of flow cytometer results that demonstrate significant T-cell binding enhancement of fluorecescein labeled LJCRF-6 (Sequence ID No. 18) peptide in the presence of LJCRF-4 (Sequence ID No. 16) peptide; and Fig. 10 is a graph illustrating homotypic adhesion properties of LFA-1 and ICAM-1 derived enhancer peptides, as compared with a control peptide, at varying concentrations. Detailed Description of the Preferred Embodiment The following examples provide, by way of example, specific methodology that may be utilized to practice preferred forms of the invention. Example 1 Short Chain Polypeptide Identification and Synthesis Table 1 below identifies synthetically produced amino acid residue sequences replicating functional domain polypeptide fragments from the parent proteins (LFA-1 and ICAM-1) upon which they are based. Parent protein amino acid residue sequences of the integrin LFA-1 include the β or CD18 subunit (Sequence ID NO. 1) and the α. or CDlla subunit (Sequence ID NO. 2) . The ICAM-1 or CD54 (Sequence ID NO. 3) sequence was used as another parent protein. These functional domains were selected to include suspected ICAM/LFA interprotein binding sites. The fragments were arbitrarily selected in sub-regions over the domains of interest as short-chain sequences for synthetic replication, with each sequence having no more than about 30 amino acid residues, in order to reduce the molecular weight and reduce the chance of provoking an im¬ mune system response in mammals injected with the pep¬ tides. These selected sequences are identified as Se- quence ID Nos. 4-15 in Table 1. In particular, the synthetic peptides reproduced sub-regions of Sequence ID No. 3 within positions 25-95, and these respective sub- regions randomly overlapped. Other parent protein regions included positions 130-160 of Sequence ID No. 1, as well as Sequence ID No. 2 positions 260-290 and 465-495. Table 1 provides a specific positional cross- reference between the synthetic peptide sequences and the parent protein sequences according to the Sequence List¬ ing. Those synthetic peptide sequences having a designa- tion including a "LJCRF" prefix were synthesized on commercial order by La Jolla Cancer Research Foundation of La Jolla, California using a Model 430A peptide synthe¬ sizer (manufactured by ABI of Foster City, California) . This entire peptide synthesis was conducted according to standard solid phase protocols employing t-butyloxy- carbamate amino acid chemistry; i.e., standard techniques described in Atherton and Sheppard, Solid Phase Peptide Synthesis, which generally follow the protein synthesis techniques as identified in Merrifield, Automated Synthe- sis of Peptides, 150 Science 178-185 (Oct. 1965) . The remaining sequences were synthesized according to the same protocols at the University of Kansas, Department of Pharmaceutical Chemistry, in Lawrence, Kansas. Table 1 SHORT CHAIN SYNTHETIC PEPTIDES AND SEQUENCE LISTING COMPARISON TO NATURAL PROTEINS NOTES : a Xaa is coumarin . b Cyclic peptide having a disulfide bond between Cys and/or Pen residues formed by dropwise addition of 2% (aqueous ) potassium ferricyanide solution to a dilute solution of their respective linear peptides at pH 7 . 8 . c Xaa is penicillamine . d Xaa is fluorescein isothiocyanate . Example 2 Antibody Binding Assay An antibody binding assay was performed to demonstrate that peptides of the invention can change the ability of anti-LFA-1 and anti- ICAM- 1 mAbs to bind with to human T-cells . The T-cells were selected as Molt3 cells , a leukemia-derived human T-cell line that was purchased from American Type Culture Collection of Rockville , Maryland, and which had a catalog number CRL-1552 . A 1 ml quantity of the Molt3 inoculum was warmed to 37°C and transferred by pipette into about 25 ml of a growth culture medium including RPMI 1640 lymphocyte growth medium (Fisher Scientific of St. Louis, Missouri) mixed with 10% by volume fetal calf serum, 100 units/ml penicillin, 100 μg/ml streptomycin and 2mM glutamine. The inoculated culture was incubated for three to four days at 37°C under a normal atmosphere having about 5% C0 2 added thereto. This culture was capable of being maintained indefinitely by siphoning away the top (non-cell contain¬ ing) layer and adding 50 ml (or more) of growth culture medium at intervals of three to four days. An aqueous buffer solution including Dulbecco's phosphate buffered saline and bovine serum albumin ("- PBS/BSA solution") was prepared for use, and included a mixture of 200 mg KH 2 P0 4 , 1.15 g Na 2 HP0 4 , 100 mg CaCl 2 , 200 mg KCl, 100 mg MgCl 2 -6H 2 0, 8 g NaCl, with distilled water added to a final 1 liter volume at room temperature, thus, forming a "PBS" solution of pH 7.4 to which was added 0.5% by weight bovine serum albumin. An aliquot of about 7 ml was removed from the incubated culture medium and subjected to washing with the PBS/BSA solution. The washing procedure was initiated by placing the aliquot in a centrifuge (Model GT422 from Jouan, distributed Piruscio & Assoc. of Manchester, Missouri) at 1400 rpm for 6.5 minutes to separate the cells from the growth medium. The supernatant fluid was decanted, a 5 ml portion of PBS/BSA solution was trans¬ ferred by pipette into the pellet, and the mixture was again centrifuged as before. The resulting supernatant fluid was decanted and the cells from the pellet were resuspended in PBS/BSA solution at a titer of about 1 x 10 6 cells/ml. A 500 μl portion of the solution containing the resuspended cells was placed in a microcentrifuge (Mara¬ thon 13K/M from Fisher Scientific of St. Louis, Missouri) at 3500 rpm for six minutes, to provide a washed pellet. The washed pellet was resuspended in a 25 μl peptide cocktail including PBS solution mixed with 455 nmol of the LJCRF-1 peptide described in Table 1 above. The cells were incubated with the peptide cocktail at 37°C for 45 minutes. After incubation with the peptide cocktail, several antibodies were successively mixed into the incubated solution. A 75 μl portion of anti-IgM solution (a mixture of 10 μl of goat anti-human IgM stock purchased from Jackson Immuno Research of estgrove, Pennsylvania; and 65 μl of PBS/BSA solution) was added to the peptide cocktail for blocking of nonspecific binding of the secondary antibody. The resulting mixture was incubated for 10 minutes on ice. About 0.1 μg of a primary antibody including mouse IgG anti-human CDlla (purified according to conventional protocols at the University of Kansas Department of Pharmacology and Toxicology from an American Type Culture hybridoma cell line having Catalogue Number HB-202) was next stirred into the mixture, which was then incubated for 30 minutes at room temperature. The incu¬ bated mixture was placed in a microcentrifuge at 3500 rpm for six minutes to remove the cells from the mixture, and the cell pellet was washed two times by adding 500 μl aliquots of PBS/BSA solution and microfuging to remove non-bound antibodies from the cells as before. The resulting washed pellet was resuspended in 100 μl of a secondary antibody cocktail including 5 μl of fluorescein isothiocyanate ("FITC") labeled goat anti-mouse IgG (from CALTAG of South San Francisco, California) and 95 μl of PBS/BSA solution. The resulting solution was incubated for 30 minutes at room temperature in the dark. The incubated solution including the secondary antibody cocktail was microfuged at 3500 rpm for six minutes to remove the cells from the solution, and the cell pellet was washed two times with 500 μl portions of PBS/BSA solution, as before, to remove unbound antibodies. The washed pellet was resuspended in 500 μl of PBS/BSA solution, and the mixture was assayed in a flow cytometer (FACscan from Becton-Dickinson of San Jose, California) to obtain a fluorescein fluorescence intensity analysis at 520 nm following laser excitation of the individual cells at 488 nm. Identical peptide assays to that described above were conducted for several of the synthetic peptides listed in Table 1, using respective 455 nmol amounts of each peptide in the corresponding peptide cocktails. In these additional assays, mouse IgG anti-human CD54 (pur¬ chased from Cambridge Research Biochemicals of Wilmington, Delaware) was substituted for the primary antibody in each instance where the peptide was based on an LFA-1 posi¬ tional reference. The FACscan computer was used to calculate the relative amount of antibody bound per cell using the computer generated median of the peak. Figs. 2 and 3 provide sample curves produced by this instrumentation, and each includes a two dimensional plot of fluorescence intensity (x axis) versus relative number of cells (y axis) for the synthetic peptides SC-3-111 (Sequence ID No. 10 corresponding to an above average 51% blocking effi¬ ciency at a 4550 μM concentration) and LJCRF-1 (Sequence ID No. 6 corresponding to an above average 65% blocking efficiency at 4550 μM) . In Fig. 2, by way of example, a first SC-3-111 (Sequence ID No. 10) peptide sample median peak value at A has an intensity value of 128.74. A control sample was obtained from an identical procedure to that of the SC-3- 111 (Sequence ID No. 10) sample, but one without the step of adding a synthetic peptide cocktail. A control sample median peak value at B has an intensity value of 264.41. Accordingly, shifting of Peak A away from Peak B in the direction of arrow 20 indicates, by a reduction of cell fluorescence intensity values, that less mouse IgG anti- human CDlla is specifically bound to the T-cells. The degree of shifting was quantified by applying a calcula¬ tion for percentage change: % change = [(observed - control/ (control) ] * 100%; i.e., %change = [ (128.74-264.41) / (264.41) ] * 100% = -51%. The shifting of Peak A in the negative direction of arrow 20 indicates successful antibody blocking, i.e., the peptides modulated the antibody to T-cell binding interac- tion by inhibiting or blocking the antibody from attaching to its complimentary LFA or ICAM target. Similarly, a shifting of Peak A in the opposite (positive) direction of arrow 22 would indicate an enhanced level of binding as compared to the control . Fig. 4 (for sample LJCRF-4 at 4550 μM) is similar to Figs. 2 and 3, but the +447% curve shift occurs in a positive direction, which indicates enhancement of the anti-ICAM antibody binding activity, not inhibition thereof . Table 2, in part, provides a summary of the FACscan results for each of the peptides from which results were obtained. Most samples received multiple measurements, and the results varied appreciably for repeat tests within a given concentration. Accordingly, the Table 2 FACscan results include an average value for all of the median intensity peak values thus obtained; e.g., Fig. 4 depicts a +447% median peak enhancement for LJCRF-4 at 4550 μM, which was one enhancement value out of five such values that contributed to the overall average of +275 ± 33 in Table 2. Standard error was calculated for each peptide as the standard deviation in the popula¬ tion of median peak intensity values divided by the square root of the number of intensity values. The results of Table 2 indicate that several ICAM-1 or LFA-1 peptides having lengths from about 10-25 amino acid residues serve to regulate the binding of antibodies which are specific for the ligands of these peptides. For example, an ICAM-1 derived peptide such as SC-3-111 can bind with the ligand LFA-1 and inhibit the binding of a monoclonal antibody that is specific for LFA- 1. The LJCRF-2 peptide exhibited a capacity to block antibody binding at a 4550 μM concentration, but func- tioned to enhance such binding at a 2275 μM concentration. Example 3 Homotypic Adhesion Assay An assay was performed to demonstrate that short-chain peptides can inhibit or enhance the intercellular adhesion between human T-cells by modulating the LFA/ICAM binding interaction between respective cells. As before, the T-cells were Molt3 cells prepared in RPMI 1640 growth medium according to Example 2. An approximate 7 ml aliquot of the growth medium was centrifuged (Jouan Model GT422 at 1400 rpm for 6.5 minutes) , and the superna¬ tant fluid was decanted. The pellet was resuspended in the supernatant fluid at titer of about 1 x 10 6 cells/ml, with the excess supernatant fluid being retained for future resuspension of the peptides. A 228 nmol quantity of the LJCRF-1 (Sequence ID No. 6) peptide described in Table 1 was diluted with the supernatant fluid to a 500 μl final volume, and filtered in microcentrifuge filter tubes (0.22 micron filter; 3500 rpm for 6 minutes) . Serial dilution of the filtered peptide with supernatant fluid was conducted to yield five 125 μl peptide solutions to give respective concentrations of about 115 μM, 230 μM, 460 μM, 910 μM, and 1.82 mM. At each concentration, a 125 μl portion of the cell suspen¬ sion was pipetted into each 125 μl portion of the peptide solutions to yield combined samples having a 250 μl volume. The 250 μl samples were seeded into individual wells in a 48-well tissue culture grade plate (from Costar of Cambridge, Maryland) and incubated at 37°C for 30 minutes. Each sample received 5 μl of 2,5-methyl dihy- droxy cinnamate ("MDHC") for a 10 μM final concentration, and was subsequently incubated for about 10 hours at 37°C. MDHC induces intercellular clumping of human T-cells (Tibbetts, Butler, and Benedict, submitted) by activating the LFA/ICAM binding interaction between respective cells. Pictures were taken of five random fields per sample using an Olympus photomicroscope having a 0.8 mm field at 400X magnification, and a 3.2 mm field of view at 100X magnification. The amount of clumping observed was quantified by determining the mean number of clumps per field ± a standard error. The assay was repeatedly conducted for most of the short-chain peptides of Table 1, and these results are summarized below in Table 2. Figs. 5, 6, and 7 replicate photographs taken from exemplary peptide assays. Fig. 5 is a photograph of the control sample having no peptides and no MDHC added, from which essentially no clumping of cells is observed. Fig. 6 depicts a photograph taken from a sample having MDHC added with no synthetic peptides, and demonstrates substantial clumping of cells. Fig. 7 depicts a photograph taken from the LJCRF-6 (Sequence ID No. 18) peptide assay at 910 μM, and demonstrates an almost total inhibition of MDHC induced clumping activity. Any sample having a score of "+++" or "++++" is considered to have a significant ability to modulate T-cell binding interaction. This is similar to results obtained by incubating similarly treated cells with antibody against either LFA or ICAM. The homotypic adhesion assay assesses the actual ability of a candidate peptide to modulate the ability of T-cells to adhere with one another. At high concentra¬ tions, some of these peptides can function as blockers of intercellular adhesion, while having a capacity for functioning as binding enhancers at lower concentrations in growth medium; e.g., ICAM-derived LJCRF-2 at respective 910 and 460 μM concentrations, and LFA-derived LJCRF-4 at respective 230 and 460 μM concentrations. Example 4 Cell Metabolic Assay After taking the photographs in Example 3 (and in some instances no photographs were taken) , cell meta¬ bolic activity was assessed by an MTT assay. A 37.5 μl portion of MTT dye (purchased from SIGMA of St. Louis, Missouri) , i.e., 3- [4, 5-dimethyl thiazole-2yl] -2, 5- diphenyl tetrazolium bromide; thiazolyl blue) was added to each well, and incubated with the cells at 37°C for about six hours. After incubation, a 250 μl quantity of a MTT solubilization solution (of a type that may be purchased from either Fischer Scientific of St. Louis, Missouri or BCD Labs of Lawrence, Kansas) was added to each well, and incubated with the cells at room temperature overnight. In the morning, the spectrophotometric absorption of each plate was determined at 570 nm in a microplate ELISA reader manufactured by Cambridge Technologies of Water- town, Massachusetts, in order to assess cell viability as an indicator of potential peptide toxicity. The MTT assay is an index of the metabolic activity of the cells; any decrease in MTT absorbance reflects a decrease in metabolic activity., or indicates cell death. Some samples indicated a decrease in meta¬ bolic activity, especially at the higher concentrations, e.g., LJCRF-5 at 1820 μM. In other samples, cellular respiration was enhanced, e.g., MC-1-80 at 910 μM. The precise reasons for the observed decrease in metabolic activity are unknown, because this decrease may be caused by a number of factors. The possible causes of a decrease in metabolic activity include peptide toxicity, peptide toxicity in combination with MDHC, a simple reduction in the cellular metabolic rate. Another possible cause of metabolic decline is that the peptides may cause the bound T-cells to die by apoptosis in a process similar to the thymic winnowing of immature T-cells. T-cells have very specific antigen receptors that serve to recognize foreign or non-self antigen complexes, and receive a first signal when they contact their particular antigen. After this initial contact, T-cells are activated to become cytotoxic T-cells by a second signal, i.e., the LFA/ICAM binding interac- tion. If the second signal never arrives, the cell is never activated, and is eventually induced to die by apoptosis. The peptides may induce this type of cellular death in these leukemic T-cells which are already acti- vated by blocking the second signal (cytotoxic activation signal) . Accordingly, the peptides may be utilized to induce specific immune tolerance by causing the deaths of T-cells that recognize antigens that are present at a specific time of treatment. Example 5 Use of Conformation Modulator Peptides The LJCRF-2 and LJCRF-4 peptides ("modulator peptides") of Table 1 demonstrated a significant positive intensity shift in the corresponding FACS analysis of Table 2, i.e., a positive intensity shift indicating a capacity for enhancing the LFA/ICAM interaction between the T-cells and the antibody binding interaction. Follow- ing these results, a further assay was performed to ascertain the effect of sequentially combining short-chain blocker peptides with the short-chain modulator peptides of the invention. It was discovered that a combination of specific blocker and modulator peptides significantly improves the level of observed binding enhancement as quantified by a flow cytometric assay using fluorescinated peptides. The antibody binding assay of Example 2 was twice repeated at a peptide concentration of 4550 μM, each time substituting a 455 nmol quantity of synthetic peptide selected from the group consisting of ICAM-based peptide SC-3-111 (Sequence ID No. 10) and LFA-based peptide LJCRF- 4 (Sequence ID No. 16) in respective assays. Following incubation for 30 minutes at 37°C after introduction of the first peptides, both incubated culture were supple¬ mented by an additional quantity of FITC-labeled RB-1-88 (Sequence ID No. 19) peptide in a quantity sufficient to provide a 1.90 μM concentration in the cell/PBS solution. Following addition of the RB-1-88 (Sequence ID No. 19) peptide, the mixture was incubated for 30 minutes at 4°C, washed three times with 500μl PBS/BSA, resuspended in 500μl PBS/BSA and immediately subjected to a flow cytometer analysis as before. Table 3 presents the results of this analysis, which are also depicted in Figs 8 (SC-3-111) and 9 (LJCRF-4) . Fig. 9 depicts binding of RB-1-88 to T cells and the ability of LJCRF-4 to enhance the binding. Con- versely, Fig. 8 demonstrates that the SC-3-111 peptide had no effect upon the ability of cell surface ICAM-1 to bind RB-1-88. The results of Table 3 and Fig. 9 indicate that the LJCRF-4 peptide significantly enhanced the binding of RB-1-88 peptide to ICAM-1. Similar enhancement of the RB- 1-88 peptide binding avidity was not observed in the presence of SC-3-111 peptide, inasmuch as the negative 8% blocking shift (Fig. 8) is probably not significant in terms of laboratory error. The peptides that serve to promote interprotein binding capacities do so according to the steps of: (1) binding to the protein ligand at a first site, and (2) changing the conformation of the protein ligand to make a second site more available for binding with the corre- sponding monoclonal antibody or peptide counterpart to the protein ligand. By way of example, the LFA-derived LJCRF- 4 modulator peptide inherently targets a corresponding ICAM protein. The resultant enhanced ICAM/LFA-type of reactive binding must result from a conformational change, but cannot result from phosphorylation-induced conforma¬ tional changes because ICAM configuration is generally not influenced by cellular phosphorylation pathways. TABLE 2 PEPTIDE RESULTS COMPARISON CO CO C :O rτrι 5 rrt Co CD CO CO I t z I m en change from control of the medium fluorescence intensity TABLE 3 SEQUENTIALLY COMBINED BLOCKER AND MODULATOR PEPTIDES CO cz CO CO — J I rυ I ι* CD Example 6 Blocking of Homotypic Adhesion by LFA-1 and ICAM-1 Enhancer and Blocker Peptides In this example, the ability of LFA-1 and ICAM-1 enhancer peptides were tested for their abilities to block homotypic adhesion of Molt-3 cells. In particular, the assay procedure of Example 3 was followed wherein homo¬ typic adhesion of Molt-3 cells was induced with tyrosine kinase inhibitor MDHC (10 μM) . These values were compared to MDHC-treated controls which were deemed to have 100% adhesion. The ICAM-1 derived peptides are Sequence ID #6 (IB) and Sequence ID #11 (IE) . The LFA-1 derived protein is Sequence ID #16 (LBE) . The control peptide was a cyclic form of Sequence ID #10 and prepared as set forth in footnote b of Table 1. Fig. 10 is a graph illustrating these homotypic adhesion results. As can be seen, all of the test pep¬ tides inhibited homotypic adhesion, demonstrating that the enhancer peptides also have an additional functionality. Example 7 Antibody Binding of Cyclic and Linear Peptides In this example, the antibody binding assay of Example 2 was performed using peptides, namely Sequence ID Nos. 6, 15 (cyclic form) and 16. The cyclic peptide was prepared as recited in footnote b of Table 1. The data derived from this assay is set forth in the following Table 4, and demonstrates that the cyclic peptides show activities similar to their linear counterparts. Table 4 SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: Benedict, Stephen H. Siahaan, Teruna Chan, Marcia Tibbetts, Scott (ii) TITLE OF INVENTION: ICAM-l/LFA-l PEPTIDES FOR INDUCING IMMUNE TOLERANCE (iii) NUMBER OF SEQUENCES: 19 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: HOVEY, WILLIAMS, TIMMONS & COLLINS (B) STREET: 1101 Walnut St. (C) CITY: Kansas City (D) STATE: MO (E) COUNTRY: USA (F) ZIP: 64106 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentln Release #1.0, Version #1.25 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: (B) FILING DATE: (C) CLASSIFICATION: (viii) ATTORNEY/AGENT INFORMATION: (A) NAME: Collins, John M. (B) REGISTRATION NUMBER: 26262 (C) REFERENCE/DOCKET NUMBER: 22833 (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: (816)474-9050 (B) TELEFAX: 816)474-9057 (C) TELEX: 434-363 (2) INFORMATION FOR SEQ ID NO:1: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 769 amino acids (B) TYPE: amino acid (D) TOPOLOGY: both (ii) MOLECULE TYPE: protein (iii) HYPOTHETICAL: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (F) TISSUE TYPE: Tonsil (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 1..22 (D) OTHER INFORMATION: /label= signal /note= "signal sequence" (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 449..496 (D) OTHER INFORMATION: /label= repeat /note= "cysteine rich repeat" (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 497..540 (D) OTHER INFORMATION: /label= repeat /note= "cysteine rich repeat" (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 541..581 (D) OTHER INFORMATION: /label= repeat /note= "cysteine rich repeat" (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 582..617 (D) OTHER INFORMATION: /label= repeat /note= "cysteine rich repeat" (ix) FEATURE: (A) NAME/KEY: Domain (B) LOCATION: 701..723 (D) OTHER INFORMATION: /label= trans /note= "transmembrane domain" (ix) FEATURE: (A) NAME/KEY: Domain (B) LOCATION: 724..769 (D) OTHER INFORMATION: /label= cyto /note= "cytoplasmic domain" (x) PUBLICATION INFORMATION: (A) AUTHORS: Pigott, Power, (B) TITLE: LFA-1 Amino acid sequence (B2) (from human tonsil) (C) JOURNAL: The Adhesion Molecule Facts Book (F) PAGES: 96-96 (G) DATE: 1993 (K) RELEVANT RESIDUES IN SEQ ID NO:1 : FROM 1 TO 769 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l: Met Leu Gly Leu Arg Pro Pro Leu Leu Ala Leu Val Gly Leu Leu Ser 1 5 10 15 Leu Gly Cys Val Leu Ser Gin Glu Cys Thr Lys Phe Lys Val Ser Ser 20 25 30 Cys Arg Glu Cys lie Glu Ser Gly Pro Gly Cys Thr Trp Cys Gin Lys 35 40 45 Leu Asn Phe Thr Gly Pro Gly Asp Pro Asp Ser lie Arg Cys Asp Thr 50 55 60 Arg Pro Gin Leu Leu Met Arg Gly Cys Ala Ala Asp Asp lie Met Asp 65 70 75 80 Pro Thr Ser Leu Ala Glu Thr Gin Glu Asp His Asn Gly Gly Gin Lys 85 90 95 SUBSTITUTE SHEET (RU!£ 26) Gln Leu Ser Pro Gin Lys Val Thr Leu Tyr Leu Arg Pro Gly Gin Ala 100 105 ' 110 Ala Ala Phe Asn Val Thr Phe Arg Arg Ala Lys Gly Tyr Pro lie Asp 115 120 125 Leu Tyr Tyr Leu Met Asp Leu Ser Tyr Ser Met Leu Asp Asp Leu Arg 130 135 140 Asn Val Lys Lys Leu Gly Gly Asp Leu Leu Arg Ala Leu Asn Glu lie 145 150 155 160 Thr Glu Ser Gly Arg lie Gly Phe Gly Ser Phe Val Asp Lys Thr Val 165 170 175 Leu Pro Phe Val Asn Thr His Pro Asp Lys Leu Arg Asn Pro Cys Pro 180 185 190 Asn Lys Glu Lys Glu Cys Gin Pro Pro Phe Ala Phe Arg His Val Leu 195 200 205 Lys Leu Thr Asn Asn Ser Asn Gin Phe Gin Thr Glu Val Gly Lys Gin 210 215 220 Leu lie Ser Gly Asn Leu Asp Ala Pro Glu Gly Gly Leu Asp Ala Met 225 230 235 240 Met Gin Val Ala Ala Cys Pro Glu Glu lie Gly Trp Arg Asn Val Thr 245 250 255 Arg Leu Leu Val Phe Ala Thr Asp Asp Gly Phe His Phe Ala Gly Asp 260 265 270 Gly Lys Leu Gly Ala lie Leu Thr Pro Asn Asp Gly Arg Cys His Leu 275 280 285 Glu Asp Asn Leu Tyr Lys Arg Ser Asn Glu Phe Asp Tyr Pro Ser Val 290 295 300 Gly Gin Leu Ala His Lys Leu Ala Glu Asn Asn lie Gin Pro lie Phe 305 310 315 320 Ala Val Thr Ser Arg Met Val Lys Thr Tyr Glu Lys Leu Thr Glu lie 325 330 335 lie Pro Lys Ser Ala Val Gly Glu Leu Ser Glu Asp Ser Ser Asn Val 340 345 350 Val His Leu lie Lys Asn Ala Tyr Asn Lys Leu Ser Ser Arg Val Phe 355 360 365 Leu Asp His Asn Ala Leu Pro Asp Thr Leu Lys Val Thr Tyr Asp Ser 370 375 380 Phe Cys Ser Asn Gly Val Thr His Arg Asn Gin Pro Arg Gly Asp Cys 385 390 395 400 Asp Gly Val Gin lie Asn Val Pro lie Thr Phe Gin Val Lys Val Thr 405 410 415 Ala Thr Glu Cys lie Gin Glu Gin Ser Phe Val lie Arg Ala Leu Gly 420 425 430 Phe Thr Asp lie Val Thr Val Gin Val Leu Pro Gin Cys Glu Cys Arg 435 440 445 Cys Arg Asp Gin Ser Arg Asp Arg Ser Leu Cys His Gly Lys Gly Phe 450 455 460 Leu Glu Cys Gly lie Cys Arg Cys Asp Thr Gly Tyr lie Gly Lys Asn 465 470 475 480 Cys Glu Cys Gin Thr Gin Gly Arg Ser Ser Gin Glu Leu Glu Gly Ser 485 490 495 Cys Arg Lys Asp Asn Asn Ser lie lie Cys Ser Gly Leu Gly Asp Cys 500 505 510 Val Cys Gly Gin Cys Leu Cys His Thr Ser Asp Val Pro Gly Lys Leu 515 520 525 lie Tyr Gly Gin Tyr Cys Glu Cys Asp Thr lie Asn Cys Glu Arg Tyr 530 535 540 Asn Gly Gin Val Cys Gly Gly Pro Gly Arg Gly Leu Cys Phe Cys Gly 545 550 555 560 Lys Cys Arg Cys His Pro Gly Phe Glu Gly Ser Ala Cys Gin Cys Glu 565 570 575 Arg Thr Thr Glu Gly Cys Leu Asn Pro Arg Arg Val Glu Cys Ser Gly 580 585 590 Arg Gly Arg Cys Arg Cys Asn Val Cys Glu Cys His Ser Gly Tyr Gin 595 600 605 Leu Pro Leu Cys Gin Glu Cys Pro Gly Cys Pro Ser Pro Cys Gly Lys 610 615 620 Tyr lie Ser Cys Ala Glu Cys Leu Lys Phe Glu Lys Gly Pro Phe Gly 625 630 635 640 Lys Asn Cys Ser Ala Ala Cys Pro Gly Leu Gin Leu Ser Asn Asn Pro 645 650 655 Val Lys Gly Arg Thr Cys Lys Glu Arg Asp Ser Glu Gly Cys Trp Val 660 665 670 Ala Tyr Thr Leu Glu Gin Gin Asp Gly Met Asp Arg Tyr Leu He Tyr 675 680 685 Val Asp Glu Ser Arg Glu Cys Val Ala Gly Pro Asn He Ala Ala He 690 695 700 Val Gly Gly Thr Val Ala Gly He Val Leu He Gly He Leu Leu Leu 705 710 715 720 Val He Trp Lys Ala Leu He His Leu Ser Asp Leu Arg Glu Tyr Arg 725 730 735 Arg Phe Glu Lys Glu Lys Leu Lys Ser Gin Trp Asn Asn Asp Asn Pro 740 745 750 Leu Phe Lys Ser Ala Thr Thr Thr Val Met Asn Pro Lys Phe Ala Glu 755 760 765 Ser (2) INFORMATION FOR SEQ ID NO:2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1170 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (iii) HYPOTHETICAL: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 1..25 (D) OTHER INFORMATION: /label= signal /note= "Signal sequence" (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 32..79 (D) OTHER INFORMATION: /label= Repeat /note= "Repeat I" (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 82..132 (D) OTHER INFORMATION: /label= Repeat /note= "Repeat II" (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 339..391 (D) OTHER INFORMATION: /label= Repeat /note= "Repeat III" (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 392..446 (D) OTHER INFORMATION: /label= Repeat /note= "Repeat IV" (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 447..508 (D) OTHER INFORMATION: /label= Repeat /note= "Repeat V" (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 509..567 (D) OTHER INFORMATION: /label= Repeat /note= "Repeat VI" (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 568..629 (D) OTHER INFORMATION: /label= Repeat /note= "Repeat VII" (ix) FEATURE: (A) NAME/KEY: Domain (B) LOCATION: 170..349 (D) OTHER INFORMATION: /label= IDomain /note= "I-Domain" (ix) FEATURE: (A) NAME/KEY: Domain (B) LOCATION: 1089..1112 (D) OTHER INFORMATION: /label= Trans /note= "Transmembrane Domain" (ix) FEATURE: (A) NAME/KEY: Domain (B) LOCATION: 1113..1170 (D) OTHER INFORMATION: /label= Cyto /note= "Cytoplasmic domain" (x) PUBLICATION INFORMATION: (A) AUTHORS: Pigott, Power, (B) TITLE: LFA-1 Amino acid sequence (alphaL) (from PMA-stimulated HL-60 cells) (C) JOURNAL: The Adhesion Molecule Facts Book (F) PAGES: 94-95 (G) DATE: 1993 (K) RELEVANT RESIDUES IN SEQ ID NO:2 : FROM 1 TO 1170 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2 : Met Lys Asp Ser Cys He Thr Val Met Ala Met Ala Leu Leu Ser Gly 1 5 10 15 Phe Phe Phe Phe Ala Pro Ala Ser Ser Tyr Asn Leu Asp Val Arg Gly 20 25 30 Ala Arg Ser Phe Ser Pro Pro Arg Ala Gly Arg His Phe Gly Tyr Arg 35 40 45 Val Leu Gin Val Gly Asn Gly Val He Val Gly Ala Pro Gly Glu Gly 50 55 60 Asn Ser Thr Gly Ser Leu Tyr Gin Cys Gin Ser Gly Thr Gly His Cys 65 70 75 80 Leu Pro Val Thr Leu Arg Gly Ser Asn Tyr Thr Ser Lys Tyr Leu Gly 85 90 95 Met Thr Leu Ala Thr Asp Pro Thr Asp Gly Ser He Leu Ala Cys Asp 100 105 110 Pro Gly Leu Ser Arg Thr Cys Asp Gin Asn Thr Tyr Leu Ser Gly Leu 115 120 125 Cys Tyr Leu Phe Arg Gin Asn Leu Gin Gly Pro Met Leu Gin Gly Arg 130 135 140 Pro Gly Phe Gin Glu Cys He Lys Gly Asn Val Asp Leu Val Phe Leu 145 150 155 160 Phe Asp Gly Ser Met Ser Leu Gin Pro Asp Glu Phe Gin Lys He Leu 165 170 175 Asp Phe Met Lys Asp Val Met Lys Lys Leu Ser Asn Thr Ser Tyr Gin 180 185 190 Phe Ala Ala Val Gin Phe Ser Thr Ser Tyr Lys Thr Glu Phe Asp Phe 195 200 205 Ser Asp Tyr Val Lys Trp Lys Asp Pro Asp Ala Leu Leu Lys His Val 210 215 220 Lys His Met Leu Leu Leu Thr Asn Thr Phe Gly Ala He Asn Tyr Val 225 230 235 240 Ala Thr Glu Val Phe Arg Glu Glu Leu Gly Ala Arg Pro Asp Ala Thr 245 250 255 Lys Val Leu He He He Thr Asp Gly Glu Ala Thr Asp Ser Gly Asn 260 265 270 He Asp Ala Ala Lys Asp He He Arg Tyr He He Gly He Gly Lys 275 280 285 His Phe Gin Thr Lys Glu Ser Gin Glu Thr Leu His Lys Phe Ala Ser 290 295 300 Lys Pro Ala Ser Glu Phe Val Lys He Leu Asp Thr Phe Glu Lys Leu 305 310 315 320 Lys Asp Leu Phe Thr Glu Leu Gin Lys Lys He Tyr Val He Glu Gly 325 330 335 Thr Ser Lys Gin Asp Leu Thr Ser Phe Asn Met Glu Leu Ser Ser Ser 340 345 350 Gly He Ser Ala Asp Leu Ser Arg Gly His Ala Val Val Gly Ala Val 355 360 365 Gly Ala Lys Asp Trp Ala Gly Gly Phe Leu Asp Leu Lys Ala Asp Leu 370 375 380 Gin Asp Asp Thr Phe He Gly Asn Glu Pro Leu Thr Pro Glu Val Arg 385 390 395 400 Ala Gly Tyr Leu Gly Tyr Thr Val Thr Trp Leu Pro Ser Arg Gin Lys 405 410 415 Thr Ser Leu Leu Ala Ser Gly Ala Pro Arg Tyr Gin His Met Gly Arg 420 425 430 Val Leu Leu Phe Gin Glu Pro Gin Gly Gly Gly His Trp Ser Gin Val 435 440 445 Gin Thr He His Gly Thr Gin He Gly Ser Tyr Phe Gly Gly Glu Leu 450 455 460 Cys Gly Val Asp Val Asp Gin Asp Gly Glu Thr Glu Leu Leu Leu He 465 470 475 480 Gly Ala Pro Leu Phe Tyr Gly Glu Gin Arg Gly Gly Arg Val Phe He 485 490 495 Tyr Gin Arg Arg Gin Leu Gly Phe Glu Glu Val Ser Glu Leu Gin Gly 500 505 510 Asp Pro Gly Tyr Pro Leu Gly Arg Phe Gly Glu Ala He Thr Ala Leu 515 520 525 Thr Asp He Asn Gly Asp Gly Leu Val Asp Val Ala Val Gly Ala Pro 530 535 540 Leu Glu Glu Gin Gly Ala Val Tyr He Phe Asn Gly Arg His Gly Gly 545 550 555 560 Leu Ser Pro Gin Pro Ser Gin Arg He Glu Gly Thr Gin Val Leu Ser 565 570 575 Gly He Gin Trp Phe Gly Arg Ser He His Gly Val Lys Asp Leu Glu 580 585 590 Gly Asp Gly Leu Ala Asp Val Ala Val Gly Ala Glu Ser Gin Met He 595 600 605 Val Leu Ser Ser Arg Pro Val Val Asp Met Val Thr Leu Met Ser Phe 610 615 620 Ser Pro Ala Glu He Pro Val His Glu Val Glu Cys Ser Tyr Ser Thr 625 630 635 640 Ser Asn Lys Met Lys Glu Gly Val Asn He Thr He Cys Phe Gin He 645 650 655 Lys Ser Leu Tyr Pro Gin Phe Gin Gly Arg Leu Val Ala Asn Leu Thr 660 665 670 Tyr Thr Leu Gin Leu Asp Gly His Arg Thr Arg Arg Arg Gly Leu Phe 675 680 685 Pro Gly Gly Arg His Glu Leu Arg Arg Asn He Ala Val Thr Thr Ser 690 695 700 Met Ser Cys Thr Asp Phe Ser Phe His Phe Pro Val Cys Val Gin Asp 705 710 715 720 Leu He Ser Pro He Asn Val Ser Leu Asn Phe Ser Leu Trp Glu Glu 725 730 735 Glu Gly Thr Pro Arg Asp Gin Arg Ala Gin Gly Lys Asp He Pro Pro 740 745 750 He Leu Arg Pro Ser Leu His Ser Glu Thr Trp Glu He Pro Phe Glu 755 760 765 Lys Asn Cys Gly Glu Asp Lys Lys Cys Glu Ala Asn Leu Arg Val Ser 770 775 780 Phe Ser Pro Ala Arg Ser Arg Ala Leu Arg Leu Thr Ala Phe Ala Ser 785 790 795 800 Leu Ser Val Glu Leu Ser Leu Ser Asn Leu Glu Glu Asp Ala Tyr Trp 805 810 815 Val Gin Leu Asp Leu His Phe Pro Pro Gly Leu Ser Phe Arg Lys Val 820 825 830 Glu Met Leu Lys Pro His Ser Gin He Pro Val Ser Cys Glu Glu Leu 835 840 845 Pro Glu Glu Ser Arg Leu Leu Ser Arg Ala Leu Ser Cys Asn Val Ser 850 855 860 Ser Pro He Phe Lys Ala Gly His Ser Val Ala Leu Gin Met Met Phe 865 870 875 880 Asn Thr Leu Val Asn Ser Ser Trp Gly Asp Ser Val Glu Leu His Ala 885 890 895 Asn Val Thr Cys Asn Asn Glu Asp Ser Asp Leu Leu Glu Asp Asn Ser 900 905 910 Ala Thr Thr He He Pro He Leu Tyr Pro He Asn He Leu He Gin 915 920 925 Asp Gin Glu Asp Ser Thr Leu Tyr Val Ser Phe Thr Pro Lys Gly Pro 930 935 940 Lys He His Gin Val Lys His Met Tyr Gin Val Arg He Gin Pro Ser 945 950 955 960 He His Asp His Asn He Pro Thr Leu Glu Ala Val Val Gly Val Pro 965 970 975 Gin Pro Pro Ser Glu Gly Pro He Thr His Gin Trp Ser Val Gin Met 980 985 990 Glu Pro Pro Val Pro Cys His Tyr Glu Asp Leu Glu Arg Leu Pro Asp 995 1000 1005 Ala Ala Glu Pro Cys Leu Pro Gly Ala Leu Phe Arg Cys Pro Val Val 1010 1015 1020 Phe Arg Gin Glu He Leu Val Gin Val He Gly Thr Leu Glu Leu Val 1025 1030 1035 ■ 1040 Gly Glu He Glu Ala Ser Ser Met Phe Ser Leu Cys Ser Ser Leu Ser 1045 1050 1055 He Ser Phe Asn Ser Ser Lys His Phe His Leu Tyr Gly Ser Asn Ala 1060 1065 1070 Ser Leu Ala Gin Val Val Met Lys Val Asp Val Val Tyr Glu Lys Gin 1075 1080 1085 Met Leu Tyr Leu Tyr Val Leu Ser Gly He Gly Gly Leu Leu Leu Leu 1090 1095 1100 Leu Leu He Phe He Val Leu Tyr Lys Val Gly Phe Phe Lys Arg Asn 1105 1110 1115 1120 Leu Lys Glu Lys Met Glu Ala Gly Arg Gly Val Pro Asn Gly He Pro 1125 1130 1135 Ala Glu Asp Ser Glu Gin Leu Ala Ser Gly Gin Glu Ala Gly Asp Pro 1140 1145 1150 Gly Cys Leu Lys Pro Leu His Glu Lys Asp Ser Glu Ser Gly Gly Gly 1155 1160 1165 Lys Asp 1170 (2) INFORMATION FOR SEQ ID NO:3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 531 amino acids (B) TYPE: amino acid (D) TOPOLOGY: both (ii) MOLECULE TYPE: protein (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 1..25 (D) OTHER INFORMATION: /label= Signal /note= "Signal sequence" (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 41..100 (D) OTHER INFORMATION: /label= Igl (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 128..190 (D) OTHER INFORMATION: /label= Ig2 (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 230..294 (D) OTHER INFORMATION: /label= Ig3 (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 325..375 (D) OTHER INFORMATION: /label= Ig4 (ix) FEATURE: (A) NAME/KEY: Region (B) LOCATION: 413..461 (D) OTHER INFORMATION: /label= Ig5 (ix) FEATURE: (A) NAME/KEY: Duplication (B) LOCATION: 481..503 (D) OTHER INFORMATION: /label= Trans /note= "Transmembrane domain" (ix) FEATURE: (A) NAME/KEY: Binding-site (B) LOCATION: 152..154 (D) OTHER INFORMATION: /label= Attachment /note= "Cell attachment site" (x) PUBLICATION INFORMATION: (A) AUTHORS: Pigott, Power, (B) TITLE: ICAM-1 Amino acid sequence (from HL-60) (C) JOURNAL: The Adhesion Molecule Facts Book (F) PAGES: 75-75 (G) DATE: 1993 (K) RELEVANT RESIDUES IN SEQ ID NO:3: FROM 1 TO 531 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: Met Ala Pro Ser Ser Pro Arg Pro Ala Leu Pro Ala Leu Leu Val Leu 1 5 10 15 Leu Gly Ala Leu Phe Pro Gly Pro Gly Asn Ala Gin Thr Ser Val Ser 20 25 30 Pro Ser Lys Val He Leu Pro Arg Gly Gly Ser Val Leu Val Thr Cys 35 40 45 Ser Thr Ser Cys Asp Gin Pro Lys Leu Leu Gly He Glu Thr Pro Leu 50 55 60 Pro Lys Lys Glu Leu Leu Leu Pro Gly Asn Asn Arg Lys Val Tyr Glu 65 70 75 80 Leu Ser Asn Val Gin Glu Asp Ser Gin Pro Met Cys Tyr Ser Asn Cys 85 90 95 Pro Asp Gly Ser Thr Ala Lys Thr Phe Leu Thr Val Tyr Trp Thr Pro 100 105 110 Glu Arg Val Glu Leu Ala Pro Leu Pro Ser Trp Gin Pro Val Gly Lys 115 120 125 Asn Leu Thr Leu Arg Cys Gin Val Glu Gly Gly Ala Pro Arg Ala Asn 130 135 140 Leu Thr Val Val Leu Leu Arg Gly Glu Lys Glu Leu Lys Arg Glu Pro 145 150 155 160 Ala Val Gly Glu Pro Ala Glu Val Thr Thr Thr Val Leu Val Arg Arg 165 170 175 Asp His His Gly Ala Asn Phe Ser Cys Arg Thr Glu Leu Asp Leu Arg 180 185 190 Pro Gin Gly Leu Glu Leu Phe Glu Asn Thr Ser Ala Pro Tyr Gin Leu 195 200 205 Gin Thr Phe Val Leu Pro Ala Thr Pro Pro Gin Leu Val Ser Pro Arg 210 215 220 Val Leu Glu Val Asp Thr Gin Gly Thr Val Val Cys Ser Leu Asp Gly 225 230 235 240 Leu Phe Pro Val Ser Glu Ala Gin Val His Leu Ala Leu Gly Asp Gin 245 250 255 Arg Leu Asn Pro Thr Val Thr Tyr Gly Asn Asp Ser Phe Ser Ala Lys 260 265 270 Ala Ser Val Ser Val Thr Ala Glu Asp Glu Gly Thr Gin Arg Leu Thr 275 280 285 Cys Ala Val He He Gly Asn Gin Ser Gin Glu Thr Leu Gin Thr Val 290 295 300 Thr He Tyr Ser Phe Pro Ala Pro Asn Val He Leu Thr Lys Pro Glu 305 310 315 320 Val Ser Glu Gly Thr Glu Val Thr Val Lys Cys Glu Ala His Pro Arg 325 330 335 Ala Lys Val Thr Leu Asn Gly Val Pro Ala Gin Pro Leu Gly Pro Arg 340 345 350 Ala Gin Leu Leu Leu Lys Ala Thr Pro Glu Asp Asn Gly Arg Ser Phe 355 360 365 Ser Cys Ser Ala Thr Leu Glu Val Ala Gly Gin Leu He His Lys Asn 370 375 380 Gin Thr Arg Glu Leu Arg Val Leu Tyr Gly Pro Arg Leu Asp Glu Arg 385 390 395 400 Asp Cys Pro Gly Asn Trp Thr Trp Pro Glu Asn Ser Gin Gin Thr Pro 405 410 415 Met Cys Gin Ala Trp Gly Asn Pro Leu Pro Glu Leu Lys Cys Leu Lys 420 425 430 Asp Gly Thr Phe Pro Leu Pro He Gly Glv Ser Val Thr Val Thr Arg 435 440 445 Asp Leu Glu Gly Thr Tyr Leu Cys Arg Ala Arg Ser Thr Gin Gly Glu 450 455 460 Val Thr Arg Glu Val Thr Val Asn Val Leu Ser Pro Arg Tyr Glu He 465 470 475 480 Val He He Thr Val Val Ala Ala Ala Val He Met Gly Thr Ala Gly 485 490 495 Leu Ser Thr Tyr Leu Tyr Asn Arg Gin Arg Lys He Lys Lys Tyr Arg 500 505 510 Leu Gin Gin Ala Gin Lys Gly Thr Pro Met Lys Pro Asn Thr Gin Ala 515 520 525 Thr Pro Pro 530 (2) INFORMATION FOR SEQ ID NO:4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: Gin Thr Ser Val Ser Pro Ser Lys Val He Leu Pro Arg Gly Gly Ser 1 5 10 15 Val Leu Val Thr Gly 20 (2) INFORMATION FOR SEQ ID NO:5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5 : Xaa Gin Thr Ser Val Ser Pro Ser Lys Val He Leu Pro Arg Gly Gly 1 5 10 15 Ser Val Leu Val Thr Gly 20 (2) INFORMATION FOR SEQ ID NO:6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: Gin Thr Ser Val Ser Pro Ser Lys Val He Leu Pro Arg Gly Gly Ser 1 5 10 15 Val Leu Val Thr Gly 20 (2) INFORMATION FOR SEQ ID NO:7 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 12 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7 : Cys Leu Pro Arg Gly Gly Ser Val Leu Val Thr Cys 1 5 10 (2) INFORMATION FOR SEQ ID NO:8: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 12 amino acids (B) TYPE: amino acid (D) TOPOLOGY: cyclic (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8 : Cys Leu Pro Arg Gly Gly Ser Val Leu Val Thr Cys 1 5 10 (2) INFORMATION FOR SEQ ID NO:9: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 11 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: Leu Pro Arg Gly Gly Ser Val Leu Val Thr Gly 1 5 10 (2) INFORMATION FOR SEQ ID NO:10: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10: Gin Thr Ser Val Ser Pro Ser Lys Val He 1 5 10 (2) INFORMATION FOR SEQ ID NO:11: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 25 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11: Asp Gin Pro Lys Leu Leu Gly He Glu Thr Pro Leu Pro Lys Lys Glu 1 5 10 15 Leu Leu Leu Pro Gly Asn Asn Arg Lys 20 25 (2) INFORMATION FOR SEQ ID NO:12 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (F) TISSUE TYPE: tonsil (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12: Lys Glu Leu Leu Leu Pro Gly Asn Asn Arg Lys Val Tyr Glu Leu Ser 1 5 10 15 Asn Val Gin Glu Asp Ser Gin Pro 20 (2) INFORMATION FOR SEQ ID NO:13: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13: Pro Ser Lys Val He Leu Pro Arg Gly Gly 1 5 10 (2) INFORMATION FOR SEQ ID NO:14: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 12 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14: Xaa Pro Ser Lys Val He Leu Pro Arg Gly Gly Cys 1 5 10 (2) INFORMATION FOR SEQ ID NO:15: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 12 amino acids (B) TYPE: amino acid (D) TOPOLOGY: cyclic (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15: Xaa Pro Ser Lys Val He Leu Pro Arg Gly Gly Cys 1 5 10 (2) INFORMATION FOR SEQ ID NO:16: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 25 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (F) TISSUE TYPE: tonsil (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: Asp Leu Ser Tyr Ser Leu Asp Asp Leu Arg Asn Val Lys Lys Leu Gly 1 5 10 15 Gly Asp Leu Leu Arg Ala Leu Asn Glu 20 25 (2) INFORMATION FOR SEQ ID NO:17: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17: Gly Val Asp Val Asp Gin Asp Gly Glu Thr Glu Leu He Gly Ala Pro 1 5 10 15 Leu Phe Tyr Gly Glu Gin Arg Gly 20 (2) INFORMATION FOR SEQ ID NO:18: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18: He Thr Asp Gly Glu Ala Thr Asp Ser Gly Asn He Asp Ala Ala Lys 1 5 10 15 Asp He He Tyr He He Gly He 20 (2) INFORMATION FOR SEQ ID NO:19: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (v) FRAGMENT TYPE: internal (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (G) CELL TYPE: T-cell (H) CELL LINE: HL-60 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: He Thr Asp Gly Glu Ala Thr Asp Ser Gly Asn He Asp Ala Ala Lys 1 5 10 15 Asp He He Tyr He He Gly He 20

Claims: Claims : 1. A peptide composition, comprising: a first peptide having an amino acid residue sequence replicating a portion of a functional domain ' sequence from a first parent protein molecule, said first peptide exhibiting functionality as a capacity for reacting with a target protein at a first level of reactivity, but having a length less than that of said parent protein molecule; and a second peptide providing means for modulating said functionality by inducing a conformational change from a first conformation to a second conformation, said first conformation being associated with said first level of reactivity, said second conformation being associated with a second level of reactivity. 2. The composition as set forth in Claim 1, said first level of reactivity being less than said second level of reactivity. 3. The composition as set forth in Claim 1, said first level of reactivity being greater than said second level of reactivity. 4. The composition as set forth in Claim 1, said first level of reactivity being greater than said second level of reactivity at a first concentration of said second peptide, and less than said second level of reactivity at a second concentration of said second peptide. 5. The composition as set forth in Claim 1, said first and second peptides having molecular weights sufficiently low to fall beneath a threshold for avoiding an immune system antibody response when administered to a mammal. 6. The composition as set forth in Claim 5, said threshold being about 20 kDa. 7. The composition as set forth in Claim 6, said threshold being about 15 kDa. 8. The composition as set forth in Claim 7, said threshold being about 10 kDa. 9. The composition as set forth in Claim 1, at least one of said first and second peptides including an amino acid sub-sequence selected from a group consisting essentially of sub-sequences taken from integrins and sub- sequences from immunoglobulin superfamily members. 10. The composition as set forth in Claim 1, said parent protein molecule being selected from a group consisting essentially of LFA-1 and ICAM-1 molecules. 11. The composition as set forth in Claim 10, said parent protein molecule including an LFA-1 integrin having sequences from CDlla (Sequence ID No. 2) and CD18 (Sequence ID No. 1) . 12. The composition as set forth in Claim 1, said first peptide consisting essentially of Sequence ID No. 16. 14. The composition as set forth in Claim 1, said parent protein molecule including an ICAM-1 molecule having a sequence from CD54 (Sequence ID No. 3) . 15. The composition as set forth in Claim 1, said first peptide functionality being the capacity for inducing peptide binding to target protein-containing cells, said binding being induced in an amount sufficient to produce a percentage shift exceeding ± 10% in median peak intensity values obtained from a flow cytometer, said shift being calculated as a comparison between a primary intensity value determined for a control sample median intensity peak and a secondary intensity value determined for a test sample containing a fluorescein labeled compo- sition. 16. The composition as set forth in Claim 15, said cells including Molt3 T-cells. 17. The composition as set forth in Claim 15, one of said first and second peptides having a concentra¬ tion from about 115 to 4550 μM in an aqueous buffer solution. 18. The composition as set forth in Claim 1, including means for stabilizing at least one of said first and second peptides for protection against bodily trans¬ portation environments. 19. The composition as set forth in Claim 18, said stabilizing means resulting from the formation of cyclic peptides and peptide mimetics. 20. The composition as set forth in Claim 1, including said first and second peptides being in the form of separate molecules. 21. A short-chain peptide for use in modulating interprotein protein reactions between respective target proteins, said peptide consisting essentially of an amino acid residue sequence selected from a group consisting of functional domain sub-sequences for integrin family members, said peptide having a capacity for modulating said reactions by contacting one of said target proteins. 22. The peptide as set forth in Claim 21, at least one of said target proteins selected from a group consisting of integrin family members and reactive coun¬ terparts to said members. 23. The peptide as set forth in Claim 21, said sequence including said sub-sequence selected from LFA-1 (Sequence ID Nos. 1 and 2) . 24. The peptide as set forth in Claim 21, said sub-sequence consisting essentially of LJCRF-4 (Sequence ID No. 16) . 25. The peptide as set forth in Claim 21, said sub-sequence consisting essentially of LJCRF-5 (Sequence ID No. 17) . 26. The peptide as set forth in Claim 21, said sub-sequence consisting essentially of LJCRF-6 (Sequence ID No. 18) . 27. The composition as set forth in Claim 21, including means for stabilizing said peptide for protec tion against bodily transportation environments . 28. The composition as set forth in Claim 27, said stabilizing means resulting from the formation of cyclic peptides and peptide mimetics. 29. The peptide as set forth in Claim 21, including said peptide having a capacity for inducing peptide binding to target protein-containing cells, said binding being induced in an amount sufficient to produce a percentage shift exceeding + 10% in median peak inten¬ sity values obtained from a flow cytometer, said shift being calculated as a comparison between a primary inten¬ sity value determined for a control sample median inten¬ sity peak and a secondary intensity value determined for a test sample containing a fluorescein labeled composi¬ tion. 30. A short-chain peptide for use in modulating interprotein protein reactions between respective target proteins, said peptide consisting essentially of an ICAM-1 based amino acid residue sequence selected from a group consisting of Sequence ID Nos. 4-15. 31. The peptide as set forth in Claim 30, including said sequence selected from a group consisting of Sequence ID Nos. 6, 10, and 11. 32. The peptide as set forth in Claim 30, said sequence consisting essentially of Sequence ID No. 11. 33. The composition as set forth in Claim 30, including means for stabilizing said peptide for protec tion against bodily transportation environments. 34. The composition as set forth in Claim 30, said stabilizing means resulting from the formation of cyclic peptides and peptide mimetics. 36. A method for modulating an interprotein reaction of target proteins, comprising the steps of: providing a short-chain peptide having an amino acid residue sequence selected from a group consist¬ ing of sub-sequences derived from integrin family member functional domains and Sequence ID Nos. 4-15; and contacting a population of target proteins with said peptide. 37. The method as set forth in Claim 36, said peptide consisting essentially of a sub-sequence selected a group consisting of CDlla (Sequence ID No. 2) and CD18 (Sequence ID No. 1) functional domains. 38. The method as set forth in Claim 36, said peptide consisting essentially of a sub-sequence selected from the group consisting of Sequence ID Nos. 16, 17, and 18. 39. The method as set forth in Claim 36, said peptide consisting essentially of a sub-sequence selected from the group consisting of Sequence ID Nos. 6, 10, and 11. 40. The method as set forth in Claim 36, said target proteins including ICAM-1 and LFA-1. 41. A method for enhancing the functionality of a first short-chain peptide, comprising the steps of: providing a first peptide having an amino acid resi¬ due sequence replicating a portion of a functional domain sequence from a first parent protein molecule, said first peptide having a length sufficient to provide a first level of functionality, but having a length less than that of said parent protein molecule; and mixing, with said first peptide, a second peptide having an amino acid residue sequence sufficient to exhibit synergism by increasing said func¬ tionality beyond said first level when combined with said first peptide; and reacting said first and said second peptides with a protein in a reaction wherein said second pep- tide induces a conformational change in said protein from a first conformation to a second conformation, said protein in said second con¬ formation being reactive with said first, said first peptide having greater avidity for said protein in said second conformation than for said protein in said first conformation. 42. The method as set forth in Claim 41, at least one of said first and second peptides consisting essentially of an amino acid sub-sequence selected from a group consisting of functional domain sub-sequences taken from the family of integrins and Sequence ID Nos 4-15.

Inventors: BENEDICT STEPHEN, CHAN MARCIA A, SIAHAAN TERUNA J, TIBBETTS SCOTT A, BENEDICT, STEPHEN, CHAN, MARCIA, A., SIAHAAN, TERUNA, J., TIBBETTS, SCOTT, A.
Applicants: BENEDICT STEPHEN, CHAN MARCIA A, SIAHAAN TERUNA J, TIBBETTS SCOTT A, UNIV KANSAS, BENEDICT, STEPHEN, CHAN, MARCIA, A., SIAHAAN, TERUNA, J., THE UNIVERSITY OF KANSAS, TIBBETTS, SCOTT, A.
Application year: 1995
Main classification: A61K38
IPC codes: A61K38, A61P1, A61P3, A61P11, A61P37, A61P43, C07K7, C07K14