bluuebunny/crossref_metadata_2025 huggingface dataset:
ANTIGEN RECOGNITION AND THE IMMUNE RESPONSE
Authors: Bush Maurice E. , Alkan Sefik S. , Nitecki Danute E. , Goodman Joel W.
Published: June 2004

L-Tyrosine-p-azobenzenearsonate (RAT) induces cellular immunity without humoral antibody in guinea pigs. Asymmetric bifunctional antigens composed of one RAT moiety and one dinitrophenyl (DNP) group separated by flexible spacers induce anti-RAT cellular immunity and an anti-DNP humoral response. Symmetrical bifunctional antigens of similar design but comprised of two RAT determinants induce cellular immunity without demonstrable anti-RAT antibody. However, when the flexible spacer is replaced by a rigid decaproline chain, humoral anti-RAT responses are provoked.

Since RAT contains both electropositive (azo) and electronegative (arsonate) centers, the failure of bifunctional RAT compounds with flexible spacers to induce humoral immunity might be ascribed either to intramolecular stacking, which compromises their bifunctional character, or to interaction of both determinants with receptors on the same cell surface, which would fail to satisfy the requirement for cooperation. In order to distinguish between these alternatives, symmetrical bifunctional antigens composed of two L-tyrosine-p-azophenyltrimethylammonium (TAT) determinants separated by flexible or rigid spacers were synthesized. TAT is immunogenic and does not cross-react with RAT. Furthermore, it contains only electropositive centers and consequently bifunctional molecules do not undergo intramolecular stacking. Immunization with either flexibly or rigidly spaced bifunctional TAT antigens raised anti-TAT antibody. These results conclusively demonstrate that "self-help," cooperation between bone marrow-derived and thymus-derived lymphocytes of identical or similar specificity, can occur, provided the determinants on the antigen are prevented from associating with each other.
A NEUTROPHIL-IMMOBILIZING FACTOR DERIVED FROM HUMAN LEUKOCYTES
Authors: Goetzi Edward J. , Austen K. Frank
Published: June 2004

A factor has been derived from human leukocytes which irreversibly inhibits the response of human neutrophils to diverse chemotactic stimuli without impairing their viability. It is released by both polymorphonuclear and mononuclear leukocytes during incubation in acidic medium, after endotoxin exposure and subsequent incubation in low potassium medium, and during phagocytosis of particles. It is extractable from both leukocyte types and therefore must be preformed. This chemotactic inhibitor is completely separable from contaminating chemotactic activity in the crude supernatants, has a mol wt of 5000, and is inactivated by digestion with trypsin or chymotrypsin. It has been termed a neutrophil-immobilizing factor because it inhibits neutrophils directly and independently of the chemotactic stimulus, and has relatively little effect on human monocyte chemotaxis.
REJECTION OF ASCITES TUMOR ALLOGRAFTS
Authors: Berke Gideon , Sullivan Karen A. , Amos Bernard
Published: June 2004

A pathway for cell-mediated tumor destruction in vitro by immune peritoneal exudate lymphoid cells has been proposed. The union of lymphocytes and tumor cells precedes the formation of an intermediate phase leading to lysis. The initial interaction is only partially temperature dependent. The cytolytic process per se is highly temperature dependent, being negligible at 25°C but proceeding rapidly at 37°C. 51Cr release from tumor cells is demonstrable within 10 min at 37°C and can be reversibly arrested by cooling. Once initiated, lysis is largely independent of additional interactions and continues at almost full rate for 30 min. The effector cells are not lysed and appear to be free to enter into further effector cycles.
CELLULAR EVENTS DURING THE INDUCTION OF EXPERIMENTAL THYROIDITIS IN THE RABBIT
Authors: Clinton Bruce A. , Weigle William O.
Published: June 2004

With a modification of the Jerne plaque technique to enumerate plaque-forming cells (PFC) to bovine and rabbit thyroglobulin, the cellular kinetics of the antibody response were followed during two 5-day series of injections of an aqueous preparation of bovine thyroglobulin. The results support the suggestion that thyroiditis in the rabbit is mediated by antibody. The peak PFC appear in the spleen at the end of the second series of injections and are considerably greater for bovine than for rabbit thyroglobulin. PFC also appear in the thyroid gland; however, the numbers of PFC for bovine and rabbit thyroglobulin were similar, and they did not reach a peak until 7 days after the peak PFC in the spleen. There was an excellent correlation between the appearance of PFC in the thyroid gland and the appearance of thyroid lesions. The disappearance of antibody to rabbit thyroglobulin from the serum also correlated with the appearance of lesions. Migration inhibition factor (MIF) activity was not produced at any time throughout the study when rabbit thyroglobulin was added to peritoneal exudates of immunized rabbits containing circulating antibody to rabbit thyroglobulin. MIF activity was observed when bovine thyroglobulin was added to similar cells in the later stages of the study after lesions were present.
IMMUNE RESPONSES AGAINST NATIVE AND CHEMICALLY MODIFIED ALBUMINS IN MICE
Authors: Schirrmacher Volker , Wigzell Hans
Published: June 2004

Immune cells induced by bovine serum albumin (BSA) and its methylated derivative (MBSA) have been compared in a cooperative cell transfer system for their content of BSA-specific antibody-forming cell precursors (AFCP, B) and BSA-specific helper (T) cells. When MBSA immune cells were transferred together with hapten-primed cells into recipient mice which were stimulated by a hapten-BSA conjugate, their cooperative secondary anti-hapten response was as good as in case of transferred BSA immune cells. Their secondary anti-BSA response, however, was markedly reduced (reduction factor > 30). Hapten-MBSA conjugates had the same capacity to react with BSA-specific helper cells in the cooperative secondary anti-hapten response as hapten-BSA conjugates but had a reduced ability to react with BSA-specific AFCP cells. In spite of the pronounced reduction of the B cell response, MBSA had the same threshold dose as BSA for activating BSA-specific T cells.

These data suggest that B and T cells recognize different epitopes on the BSA molecule, only those recognized by B cells being affected by the methylation procedure.
THE EFFECT OF GLUCOCORTICOSTEROIDS ON THE PROLIFERATION AND KINETICS OF PROMONOCYTES AND MONOCYTES OF THE BONE MARROW
Authors: Thompson Jan , van Furth Ralph
Published: June 2004

To elucidate mechanisms underlying the prolonged monocytopenia induced in the peripheral blood of mice by injection of a subcutaneous depot of hydrocortisone acetate, the effect of this compound on the production of monocytes and their release from the bone marrow was studied.

Hydrocortisone was found to cause a rapid reduction of the bone marrow promonocytes to about 65% of their initial number. The number of monocytes in the bone marrow decreased gradually, over a period of 96 h, to 75% of the initial value.

The mitotic activity of the promonocytes was not diminished, as judged from the labeling in vitro with [3H]thymidine and the DNA-synthesis and cell-cycle times of these cells. The production of monocytes was only moderately diminished, i.e., to about 80% of the normal amount.

The release of monocytes from the bone marrow was found to be influenced by hydrocortisone. After in vivo labeling with [3H]thymidine the monocyte-labeling indices were initially significantly higher in hydrocortisone-treated than in normal mice.

It is concluded that a decreased production of monocytes in the bone marrow cannot account for the prolonged monocytopenia in the peripheral blood after hydrocortisone administration. However, hydrocortisone interferes with the release of newly formed monocytes from the bone marrow, resulting in a prolonged sojourn of these cells in this compartment.
PRODUCTS OF ACTIVATED LYMPHOCYTES
Authors: Sorg Clemens , Bloom Barry R.
Published: June 2004

General methods were developed and applied to the biosynthesis and purification of products of activated lymphocytes available in minute quantities. The activity studied here was the migration inhibitory factor (MIF) produced by purified protein derivative (PPD)- or concanavalin A (Con A)-stimulated lymphocytes obtained from one guinea pig or less. The methods selected yielded results in terms of two chemical parameters characteristic of the molecules involved, namely Kd on Sephadex G-75 and isoionic point, pI, on isoelectric focusing.

When supernatants were fractionated on G-75 columns, there were several areas even in control supernatants which produced migration inhibition relative to medium controls. However, in PPD- and Con A-stimulated supernatants, at least one peak of MIF activity was found solely in the stimulated cultures, with a Kd of 0.15. A double-labeling technique was used to characterize the proteins of this peak. Control, unstimulated cultures were labeled with [14C]leucine and stimulated cultures were labeled with [3H]leucine. After mixing the supernatants and G-75 filtration, a major "ratiolabeled" broad peak. i.e. one with increased 3H/14C ratio, was found. When a narrow portion of this peak about Kd 0.15, containing most of the MIF activity, was subjected to analytical isoelectric focusing, all of the label was associated with proteins of lower net charge than albumin. A unique ratiolabeled peak was found in PPD- and Con A-stimulated fractions with a pI of approx. 5.3. A micropreparative isoelectric focusing technique was developed and yielded MIF activity in the same region as the major ratiolabeled peak. Further study will be required to ascertain whether the ratiolabeled protein is MIF. By following the Kd, pI, and 3H/14C labeling ratio, at least 14 products of activated lymphocytes, synthesized either de novo or in increased amounts, could be distinguished.
IMMUNOLOGICAL TOLERANCE TO A HAPTEN
Authors: Fidler John M. , Golub Edward S.
Published: June 2004

Treatment of mice with a nonimmunogenic preparation of free reactive hapten, trinitrobenzene sulfonic acid (TNBS), leads to the induction of a state of tolerance to the hapten, 2,4,6-trinitrophenyl (TNP). This is determined by the lack of response to the haptenic moiety in an immunogenic hapten-carrier conjugate (TNP-SRBC) as assayed both by serum antibody titrations and the hemolytic plaque assay. The tolerance produced is specific for the hapten, since the anticarrier responses are essentially unaltered compared with the control values. The unresponsiveness induced by TNBS treatment is a dose-dependent phenomenon, becoming less complete at lower doses of TNBS. The tolerance is of a definite length, both in its induction phase and in the duration of the established unresponsive state. Tolerance can be maintained and extended, and may also be reentered once escape has been initiated.
DIRECT DEMONSTRATION OF THETA-POSITIVE ANTIGEN-BINDING CELLS, WITH ANTIGEN-INDUCED MOVEMENT OF THYMUS-DEPENDENT CELL RECEPTORS
Authors: Ashman Robert F. , Raff Martin C.
Published: June 2004

Anti-θAKR antibody conjugated to fluorescein has been used in direct immunofluorescence tests to identify spleen θ+ (T) sheep erythrocyte rosette-forming cells in AKR mice. Specificity studies involving A and cogenic A/θAKR mice clearly demonstrated that the cell surface fluorescence and cytotoxicity produced by the antiserum is directed solely toward the θAKR alloantigen. Approximately ⅜ of rosette-forming and non-rosette-forming spleen cells were found to be θ+. The tendency for T cells to bind less antigen and the tendency for antigen-binding T cells to bear less θ than other spleen T cells, first suggested by other studies involving rosette-elimination by anti-θC3H plus complement, were confirmed by direct immunofluorescence. All AKR rosettes are specifically inhibitable by anti-immunoglobulin, including T rosettes. Antigen-induced redistribution of T cell receptors, analogous to that previously described for B cell receptors (16), occurs as readily in θ+RFC as in θ- RFC, without altering the symmetrical ring distribution of θAKR antigen.
BIOLOGICAL EXPRESSIONS OF LYMPHOCYTE ACTIVATION
Authors: Rich Robert R. , Pierce Carl W.
Published: June 2004

The effects of nonspecific phytomitogens on primary plaque-forming cell (PFC) responses of mouse spleen cells to heterologous erythrocytes in vitro were studied. Spleen cell cultures treated with concanavalin A or phytohemagglutinin in vitro or established with spleen cells derived from mice injected with concanavalin A 24 h previously were similarly affected. In both cases, submitogenic doses resulted in substantial enhancement of PFC responses, whereas 10-fold larger doses were profoundly inhibitory. In contrast to the suppressive effects of mitogenic doses of phytomitogens added at culture initiation, addition of these same doses to cultures 48 h later resulted in increased PFC responses. This enhancement could be observed within 1 h after treatment and consequently could not be ascribed only to mitotic expansion of the antibody-synthesizing clone. Activation of spleen cells with specific antigen before mitogen treatment was not required for expression of the enhancing or suppressing effects on PFC responses. IgM and IgG PFC responses were similarly affected. Studies of cell interactions revealed that as few as 105 spleen cells obtained from mice treated with concanavalin A in vivo synergistically enhanced the PFC responses of 107 normal spleen cells. This enhancement was mediated by mitogen-activated T lymphocytes which were resistant to 2000 R irradiation 24 h after activation. The relevance of these observations to emerging concepts of helper and suppressor T cell activity is discussed.
THYMUS CELL MATURATION
Authors: Weissman Irving L.
Published: June 2004

Outer thymic cortical large lymphocytes were labeled by transcapsular administration of tritiated thymidine. By 2–4 days after labeling, small and medium labeled lymphocyte descendants were found throughout the cortex and in the medulla. The labeled cortical lymphocytes undergo pycnosis after parenteral administration of hydrocortisone 24 h previously, but their (labeled) medullary descendants do not. In addition, parenteral administration of hydrocortisone at the time of surface labeling results in the absence of the appearance of labeled medullary descendants.
THE MEDIATOR OF CELLULAR IMMUNITY
Authors: McGregor D. D. , Logie Pamela S.
Published: June 2004

The antimitotic drug vinblastine (Vbl) has a profound impact upon the specifically sensitized lymphocytes that transfer cellular resistance to Listeria monocytogenes. A 12-h pulse of the drug given to prospective donors during the first week of an immunizing Listeria infection inhibits the delivery of protective lymphocytes to the thoracic duct and their subsequent movement into an inflammatory exudate induced in the peritoneal cavity. The effect of Vbl is clearly related to its antimitotic activity, not to an effect on lymphocytes regardless of their position in the division cycle. This conclusion was drawn from an autoradiographic analysis of cells in the lymph of Vbl-treated rats and from failure of the drug to abrogate a known function of small lymphocytes, namely, their ability to initiate a graft-vs.-host reaction. The results imply that large lymphocytes, the rapidly proliferating cells in central lymph, are the principal effector cells responsible for transmitting resistance to L. monocytogenes and provide a plausible explanation for their rapid turnover and short circulating life-span.
PREVENTION OF CELL-TO-CELL SPREAD OF HERPES SIMPLEX VIRUS BY LEUKOCYTES
Authors: Lodmell Donald Louis , Niwa Akira , Hayashi Kozaburo , Notkins Abner Louis
Published: June 2004

Antibody to herpes simplex virus (HSV) plus complement destroyed HSV-infected cells but did not stop the spread of the infection. Studies on the relationship between the time of appearance of viral antigens on the cell surface, immunological destruction of the cells by antiviral antibody and complement, and transfer of the virus to adjacent cells showed that the virus spread from infected to uninfected cells before the infected cells were susceptible to immunological destruction. Incubation of infected monolayers with leukocytes, however, stopped the spread of the virus by nonspecifically damaging both infected and uninfected cells and by presumably breaking intercellular bridges. When leukocytes were removed from infected monolayers, viral plaques developed. If, however, antiviral antibody and complement were added to monolayers before the leukocytes were removed, the development of plaques was prevented. These findings suggest that both antibody and leukocytes are needed to cure HSV infections.
THE CYTOTOXIC EFFECT OF MOUSE MACROPHAGES ON SYNGENEIC AND ALLOGENEIC ERYTHROCYTES
Authors: Melsom H. , Seljelid R.
Published: June 2004

A cytotoxic effect of mouse peritoneal macrophages against syngeneic and allogeneic erythrocytes was demonstrated by isotope release and release of hemoglobin. The cytotoxic effect was dependent on the contact between viable, activated macrophages and target cells. Activation was accomplished by prolonged cultivation of macrophages and by the presence of Zn++ and Con-A. Immunization did not prove necessary. Morphological observations as well as experiments with various salt concentrations indicate that the cytotoxic reaction may involve some kind of osmotic effect upon the target cells.
GLUCOCORTICOID SUPPRESSION OF MACROPHAGE MIGRATION INHIBITORY FACTOR
Authors: Balow James E. , Rosenthal Alan S.
Published: June 2004

The ability of hydrocortisone to modify antigen-mediated inhibition of macrophage migration, an in vitro correlate of cellular immunity in the guinea pig, was investigated. Only the glucocorticoids, hydrocortisone and dexamethasone, significantly blocked migration inhibitory factor (MIF) activity in pharmacologic concentrations. Hydrocortisone had no effect on antigen "processing" by macrophages, nor on the ability of antigen-stimulated peritoneal exudate lymphocytes to produce MIF. Rather, hydrocortisone antagonized directly the inhibitory effect of MIF on the macrophage.
THE PRODUCTION OF VESICULAR STOMATITIS VIRUS BY ANTIGEN- OR MITOGEN-STIMULATED LYMPHOCYTES AND CONTINUOUS LYMPHOBLASTOID LINES
Authors: Nowakowski Maja , Feldman Joseph D. , Kano Shogo , Bloom Barry R.
Published: June 2004

A variety of lymphoid cell populations were examined in terms of their ability to replicate vesicular stomatitis virus (VSV), a lytic, RNA-containing virus maturing at the cell surface. The number of cells capable of producing VSV was estimated in terms of infectious centers by the virus plaque assay (VPA), and morphologically by electron microscopy (EM). The lymphoid cells examined in this study included: (a) lymph node cells from delayed hypersensitive guinea pigs stimulated by specific antigen, (b) mouse spleen cells activated by selective bone marrow-derived (B) cell and thymus derived (T) cell mitogens, and (c) cells of human and murine continuous lymphoblastoid or lymphoma lines.

In unstimulated cultures of guinea pig lymph node cells there is a background of approximately 1 in 1,000 cells which produces VSV; in purified protein derivative (PPD)-stimulated cultures the number of cells producing virus was 1.6% in the VPA and 1.9% by EM. These cells were large lymphocytes with some morphological features of transformed lymphocytes but were not typical blast cells. A few macrophages were associated with virus in both stimulated and control cultures. These observations indicate that (a) cells responsive to antigens, as detected by a marker virus, were lymphocytes; (b) cells other than lymphocytes (macrophages) were capable of replicating VSV even without antigenic stimulation; and (c) the correlation of results obtained by VPA and morphologic examination was usually quite good.

Of the total number of mouse spleen cells stimulated with concanavalin (Con A), a T cell mitogen, 4.5 (EM)–5.7% (VPA) were associated with VSV. These were characteristic transformed lymphocytes, similar to phytohemagglutinin (PHA)-stimulated human lymphocytes. In contrast Escherichia coli lipopolysaccharide (LPS)-treated mouse spleen cultures contained lower numbers of virus plaque-forming cells. The majority of such cells associated with virus displayed extensive rough endoplasmic reticulum.

Two cultured murine lymphomas containing lymphocytes with the θ surface marker (L5178Y and EL-4) showed a 15–100-fold higher incidence of virus-producing cells than leukemias (L1210 and C57Bl/6) which did not carry this marker. Similarly, the L2C guinea pig leukemia, a known B cell leukemia, yielded a low percent of virus plaque-forming cells (<2%). However, MOPC-104, a plasma cell tumor presumed to be of B cell origin, was found to be an efficient virus producer. There was a wide variation in the efficiency of VSV replication among human lymphoblastoid lines. One line, Wil-2, produced 80% infectious centers after 24 h of exposure to VSV, and all cells were associated with virus at the EM level. The relationship between the virus-producing cells and different lymphocyte subpopulations as well as the efficiency of the two assays for studying virus-producing lymphocytes is discussed.
BLOCKING AND REDISTRIBUTION ("CAPPING") OF ANTIGEN RECEPTORS ON T AND B LYMPHOCYTES BY ANTI-IMMUNOGLOBULIN ANTIBODY
Authors: Roelants Georges , Forni Luciana , Pernis Benvenuto
Published: June 2004

Antigen-binding T and B lymphocytes were studied by combined autoradiography and immunofluorescence; mouse spleen lymphocytes binding the antigens, [125I]MSH or [125I]TIGAL, were incubated with rhodamine-labeled anti-Ig reagents or with a rhodamine-labeled IgG fraction of anti-θ serum. B cells were identified as Ig+ or θ-, T cells as Ig- or θ+.

It was found that: (a) 20% (1–2 mo after priming) to 30% (3.5–4 mo after priming) of the antigen-binding cells were T cells. (b) The range of antigen molecules bound by B and T cells was similar. (c) Binding of antigen to B and T cells was inhibited by polyvalent anti-Ig, anti-µ, or anti-L reagents. Binding to T cells was more readily inhibited than to B cells. Normal rabbit serum, antimouse lymphocyte serum, or anti-θ did not inhibit antigen binding. (d) When Ig at the surface of B cells was induced, by noninhibiting concentrations of anti-Ig reagents, to redistribute into polar caps and the cells subsequently exposed to [125I)antigen under noncapping conditions, the [125I]antigen silver grains were distributed in caps superimposed on the Ig fluorescent cap. Of crucial importance, antigen was found in cap in the same proportion of T cells as B cells. Significant capping of antigen receptors was not induced in B or T cells with normal rabbit serum or by anti-Ig reagents absorbed with mouse Ig.

The main conclusions of this series of experiments using direct visualization of antigen-binding B and T lymphocytes is that T cells have antigen-specific receptors, probably of IgM nature, and that the number of these receptors appears to range in the order of thousands.
INDUCTION OF ARGINASE ACTIVITY WITH THE SHOPE PAPILLOMA VIRUS IN TISSUE CULTURE CELLS FROM AN ARGININEMIC PATIENT
Authors: Rogers Stanfield , Lowenthal A. , Terheggen H. G. , Columbo J. P.
Published: June 2004

Inoculation of the Shope virus in tissue cultures of human fibroblasts from a patient with a deficiency of the enzyme arginase results in an induction of arginase activity, apparently virus coded.
RELATION OF CHROMOSOME 4 (LINKAGE GROUP VIII) TO MURINE LEUKEMIA VIRUS-ASSOCIATED ANTIGENS OF AKR MICE
Authors: Ikeda H. , Stockert E. , Rowe W. P. , Boyse E. A. , Lilly F. , Sato H. , Jacobs S. , Old L. J.
Published: June 2004

Genes specifying or controlling the expression of GIX (cell surface), GCSA (cell surface), and gs (internal viral) antigens are located in chromosome 4 (linkage group [LG] VIII) of the AKR mouse.

All three antigens may exhibit mendelian inheritance, mice being antigen positive or antigen negative, but each may also appear in leukemic cells of mice whose inherited genotype was antigen negative. The GIX-determining gene in LG VIII of AKR mice apparently is equivalent to Gv-1, which determines expression of the same antigen in 129 strain mice, but which in the latter strain is located in LG IX. As the estimated distance of Gv-1 from H-2 in 129 mice is considerable (37 units) further tests are now indicated to assess the possibility of pseudolinkage in this case.

The Fv-1 locus, also located in LG VIII, influences the mouse's titer of MuLV, and might thereby be thought to regulate the GIX and gs phenotypes of AKR backcross segregants. But the data indicate a discrete LG VIII locus for GIX, since expression of this antigen is mendelian and independent of infectious virus titer. Since the GIX and GCSA phenotypes of AKR backcross segregants were invariably concordant, these two antigens must be specified or controlled by closely linked genes, and the latter also is presumably independent of virus titer. The question as to what extent expression of gs antigen in the segregants is secondary to virus production is undecided.
CRITICAL ROLE OF DETERMINANT PRESENTATION IN THE INDUCTION OF SPECIFIC RESPONSES IN IMMUNO-COMPETENT LYMPHOCYTES
Authors: Katz David H. , Unanue Emil R.
Published: June 2004

A detailed analysis of the role of determinant presentation in the process of triggering immunocompetent lymphocytes has been made utilizing cell-bound hapten-carrier conjugates to elicit secondary antihapten antibody responses, primarily in vitro. The results of these experiments demonstrate that: (a) hapten-protein conjugates will attach to the surface membranes of macrophages directly, in the absence of specific antibodies, in a highly immunogenic form; (b) such macrophage-bound conjugates serve as remarkedly efficient stimuli to trigger both thymus-derived (T) and bone marrow-derived (B) cells in a specific manner, lowering the optimal threshold antigen dose (in molar terms) by several logs as compared with soluble antigen; (c) the macrophage is not unique in this regard, since fibroblasts are essentially comparable in the capacity to present antigen in highly immunogenic form; (d) cell surface-bound antigen clearly favors secondary in vitro responses of the IgG as compared with the IgM antibody class; (e) in terms of triggering B or T cells, antigen bound to macrophages in the form of immune complexes does not appear to possess any appreciable advantage over equimolar quantities of directly attached antigen; (f) the increased immunogenicity of cell-bound antigen appears to reflect certain crucial, and undefined, features of cell surface membranes and not merely the stabilization of determinants on a relatively immobile surface; and (g) although the efficiency of lymphocyte triggering is markedly enhanced by cell-bound antigen, the presence of macrophages is apparently not an absolute requirement for eliciting secondary in vitro antibody responses to soluble hapten-protein conjugates.

The relevance of these observations to the nature of the signal induced upon antigen interaction by specific lymphocytes and the sequential cellular events involved in the regulatory influence of activated T cells on B cell responses to antigen is discussed. We postulate that T lymphocytes are best triggered by cell-bound antigen and that after this step the activated T lymphocytes regulate the triggering of B cells with antigen.
GENETIC CONTROL OF THE IMMUNE RESPONSE
Authors: Hämmerling Günter J. , Masuda Tohru , McDevitt Hugh O.
Published: June 2004

The influence of immunization with (T,G)-A--L on the frequency and characteristics of [125I] (T,G)-A--L-binding cells (ABC) was investigated in high and low responder mice, whose ability to respond to (T,G)-A--L is under control of an H-2-linked immune response gene, Ir-1. Unimmunized high and low responder mice have about the same number of ABC in spleen and lymph nodes (6–12 ABC/104). However, after immunization with (T,G)-A--L in aqueous solution, ABC in high responders increase to a much greater extent than they do in low responders. By inhibition of ABC with class-specific anti-Ig sera, it was demonstrated that in nonimmune and primed mice antigen is bound to IgM receptors, which is in agreement with the exclusive production of 19S anti-(T,G)-A--L antibody in primed animals. In contrast, after secondary challenge with antigen, ABC in high and low responder mice have mainly IgG receptors, although under the conditions used for immunization, low responders are not able to produce detectable amounts of 7S anti-(T,G)-A--L antibody. From these results and from the evidence that low responders very probably have a T cell defect, it is suggested that the switchover from IgM to IgG precursor cells can be induced by antigen itself, without the action of specific T cells. Furthermore, the failure of marked proliferation of ABC in low responders after antigenic stimulation is explained by the lack of stimulation by specific T cells. By independent methods it has been shown that all ABC detected in this study are B cells. Preliminary experiments indicate that purified peripheral T cells bind antigen, but much less per cell than do B cells.
SYNTHESIS OF ENDOGENOUS PYROGEN BY RABBIT LEUKOCYTES
Authors: Moore Douglas M. , Murphy Patrick A. , Chesney P. Joan , Wood W. B.
Published: June 2004

Rabbit ieukocytes from peritoneal exudates and from blood were stimulated to form leukocyte pyrogen in the presence of radiolabeled amino acids. The stimuli used were endotoxin, phagocytosis, and tuberculin. The crude leukocyte pyrogen samples were purified; pyrogen from exudate cells was rendered homogeneous; pyrogen from blood cells was still contaminated with other proteins. All the purified pyrogens were radioactive; and for all it was shown that radioactivity and pyrogenic activity coincided on electrophoresis at pH 3.5 and pH 9 in acrylamide and on isoelectric focusing in acrylamide. Furthermore, pyrogens obtained from exudate cells stimulated in different ways, or from blood cells and exudate cells stimulated with endotoxin, appeared to be identical. These results suggest that leukocyte pyrogen was synthesized de novo from amino acid precursors and that leukocytes made the same pyrogen whatever the stimulus used to activate them.
BIOLOGICAL CHARACTERISTICS OF T AND B MEMORY LYMPHOCYTES IN THE RAT
Authors: Strober Samuel , Dilley Jeanette
Published: June 2004

The adoptive secondary antibody response of rats to the hapten-protein conjugate dinitrophenyl-diphtheria toxoid (DNP-DT) was used to investigate the migratory properties and rate of formation of T and B memory cells in the spleen. The experimental findings show that hapten (DNP-BSA)- and carrier (DT)-primed spleen cells act synergistically in the restoration of the adoptive anti-DNP response. Passage of both hapten- and carrier-primed spleen cells through an intermediate host (intravenous injection and subsequent collection in the thoracic duct lymph) showed that both cell types are able to recirculate from the blood to the lymph. In addition, memory to the hapten or carrier could be withdrawn from the spleen by prolonged thoracic duct drainage.

The rate of formation of hapten- and carrier-primed spleen cells was studied by treating donors with [3H]thymidine for 48 h before cell transfer in an attempt to "suicide" rapidly dividing cells. Only a slight reduction in the adoptive response to the hapten or carrier was noted upon transfer of treated cells to irradiated hosts. In further experiments, the cell lineage of hapten- and carrier-primed cells was determined by treating each cell type in vitro with rabbit antirat B cell serum (RARBS) and complement. Although treatment with RARBS did not affect the adoptive response restored by carrier-primed cells, the same treatment abolished the response restored by hapten-primed cells.

These findings indicate that T and B memory cells in the spleen of the rat are relatively long-lived, recirculating lymphocytes. The contribution of fixed or rapidly turning over cells to immunological memory is small or negligible as compared with the latter cells.
ACTIVE SUPPRESSION OF IMMUNOGLOBULIN ALLOTYPE SYNTHESIS
Authors: Herzenberg Leonore A. , Chan Eva L. , Ravitch Myrnice M. , Riblet Roy J. , Herzenberg Leonard A.
Published: June 2004

Thymus-derived cells (T cells) that actively suppress production of IgG2a immunoglobulins carrying the Ig-1b allotype have been found in adult (SJL x BALB/c)F1 mice exposed to anti-Ig-1b early in life. The suppression is specific for Ig-1b. The allelic product, Ig-1a, is unaffected.

Spleen, lymph node, bone marrow, or thymus cells from suppressed mice suppress production of Ig-1b by syngeneic spleen cells from normal F1 mice. When a mixture of suppressed and normal cells is transferred into lethally irradiated BALB/c mice, there is a short burst of Ig-1b production after which Ig-1b levels in the recipient fall rapidly below detectability. Pretreatment of the cells from the suppressed mice with antiserum specific for T cells (anti-Thy-1b) plus complement before mixture destroys the suppressing activity.

Similar results with suppressor cells were obtained in vitro using Mishell-Dutton cultures. Mixture of spleen cells from suppressed animals with sheep erythrocyte (SRBC)-primed syngeneic normal spleen before culture suppresses Ig-1b plaque-forming cell (PFC) formation while leaving Ig-1a PFC unaffected. Treatment of the suppressed spleen with anti-Thy-1b before transfer removes the suppressing activity.
REGULATION OF THE SECONDARY ANTIBODY RESPONSE IN VITRO
Authors: Ambrose Charles T.
Published: June 2004

A material inhibiting antibody synthesis in vitro is produced during the productive phase by rabbit lymph node organ cultures undergoing a secondary response. This antibody inhibitory material (AIM) has been isolated from serum-free medium taken from the cultures and also extracted from lymph node fragments as late as their 4th wk in vitro.

AIM inhibits most strikingly the early productive phase of the secondary response in vitro (i.e, during the 2nd wk). AIM isolated from cultures undergoing a given immune response inhibits the same as well as different responses, thus indicating an immunologically nonspecific effect.

Ultrafiltration and related studies reveal that the molecular size of AIM is 10,000–50,000 daltons and that it is not antibody. AIM can readily be separated from 7S globulin by use of CM-cellulose. The inhibitory activity of AIM is lost by digestion with ribonuclease. Thus the avoidance of serum with its high levels of ribonucleases may be crucial in the study of this material.

The presence in eukaryotic cells of metabolic regulators, governors, etc. has been postulated largely by analogy with microbial systems (27). There is little direct evidence about the chemical nature of these presumed regulators. Our data on the RNase sensitivity of AIM raises the possibility in this lymphoid system of regulation by a species of RNA.
MECHANISMS IN THE SUPPRESSION OF DELAYED HYPERSENSITIVITY IN THE GUINEA PIG BY 6-MERCAPTOPURINE
Authors: Phillips S. Michael , Zweiman Burton
Published: June 2004

The mechanism of suppression, of delayed hypersensitivity to tuberculoprotein by 6-mercaptopurine (6-MP) was studied in guinea pigs. Under the conditions of the protocol, suppression of tuberculin delayed skin test reactivity was not associated with a significantly altered end-organ response to mediators of permeability. No significant alteration of in vivo lymphoid activity, as measured by reconstitution studies, was found. In addition, lymphoid cells from 6-MP-treated animals reacted in a fashion similar to those of placebo-treated animals with respect to (a) antigen-induced lymphocyte proliferation, (b) antigen-induced liberation of macrophage inhibitory factor activity, (c) direct inhibition by antigen of peritoneal exudate cell migration.

Conversely, suppression was seen in levels of blood monocytes and in vitro function of macrophages from 6-MP-treated animals in several respects: (a) adherence to glass, (b) migratory rate, (c) phagocytic capacity. Therefore, it would appear that a ma]or mechanism of 6-MP-induced suppression of delayed hypersensitivity is through its action on effector cells.
STUDIES ON CELL-MEDIATED IMMUNITY TO LYMPHOCYTIC CHORIOMENINGITIS VIRUS IN MICE
Authors: Marker Ole , Volkert Mogens
Published: June 2004

A large amount of experimental evidence has already been presented indicating the great importance of the cell-mediated immunity in the pathogenesis of the LCM virus infection in mice. In this laboratory a method which makes it possible to measure this cellular immunity quantitatively in vitro has been developed. The method is based on the determination of the radioisotope released after the interaction between specifically sensitized lymphocytes and syngeneic 1Cr-labeled LCM virus-infected target cells. By using this technique the time-course of the cell-mediated immunity has been established in acutely infected mice and in virus carriers adoptively immunized with syngeneic sensitized lymphocytes. Lymphocytes from acutely infected mice showed a strong lysing effect on the target cells, with a sharp maximum at about the 9th day after infection. The cell-mediated immunity in adoptively immunized virus carrier mice showed the same time-course, but in these animals the lytic effect of the lymphoid cells was considerably less pronounced. Lymphocytes from untreated virus carriers did not, however, have any effect on the target cells, and in these animals it was not possible to demonstrate any evidence of enhancing antibodies, In experiments employing serial dilutions of sensitized lymphocytes in normal cells a direct linear relationship between the number of sensitized lymphocytes and target cell destruction was found. These experiments seem to indicate that the underlying mechanism in the cytotoxic reaction is a direct cell-to-cell interaction.
IN VITRO INDUCTION OF TUMOR-SPECIFIC IMMUNITY
Authors: Wagner Hermann , Röllinghoff Martin
Published: June 2004

Induction of tumor-specific immunity in vitro was accomplished by cocultivation of cortisone-resistant murine thymocytes or spleen cells with irradiated syngeneic plasma cell tumors (PCT). The cytotoxic activity generated could be detected in a short-term 51Cr-release assay. Optimal cytotoxic activity against PCT-associated transplantation antigens (TATA) was generated after 7 days in culture. Unlike cytotoxic responses to tumor allografts in which the cytotoxic activity was directed against allogeneic transplantation antigens, the cytotoxic activity obtained in the syngeneic tumor system was specific to the immunizing syngeneic PCT.

Similar parameters of induction of cytotoxic responses in in vitro tumor allograft responses and in the syngeneic tumor system suggested that both reactions are cell-mediated cytotoxic immune responses. With regard to the magnitude of cytotoxic responses obtained, allogeneic transplantation antigens induced about a 30-fold higher cytotoxic immune response than plasma cell TATA. The results are consistent with the concept that in vitro tumor allograft responses and in vitro responses against TATA of PCT are similar in quality, but differ in the magnitude of the cytotoxic response provoked.
SUPPRESSION OR AUGMENTATION OF THE ANTIHAPTEN RESPONSE IN MICE BY ANTIBODIES OF DIFFERENT SPECIFICITIES
Authors: Haughton G. , Mäkelä O.
Published: June 2004

We have measured the production by (C57 x CBA)F1 mice of hapten-binding antibody in response to a standard dose of 50 µg of alum-precipitated NIP12-CG and the influence on this response of the prior administration of hyperimmune antisera raised against the homologous conjugate, the carrier globulin alone, the hapten conjugated to a non-cross-reactive carrier (NIP4-OA), or a related hapten (NP) coupled to CG.

The homologous antiserum was strongly immunosuppressive; a dose capable of binding about 1% of the administered hapten caused significant suppression. High doses of anticarrier serum caused significant but modest suppression (about 50%); low doses had no effect. High doses of the serum prepared against NIP4-OA suppressed the 19 day response by more than 97%, while 100–1,000 times lower doses caused the response to be elevated to about double the control level. The antibodies responsible for immunosuppression could be removed from this serum, as could the NIP-binding antibodies, by absorption with NIP coupled through ethylenediamine to insoluble Sepharose. The ability of this serum to augment the response was not reduced by such absorption. Augmenting antibodies could be removed by absorption with HOP-BSA-Sepharose. Thus, immunosuppression and augmentation are functions of two different populations of antibody. The former are specific hapten-binding antibodies, the latter seem to be directed against new antigenic determinants created by coupling any of the family of haptens through lysine to protein carriers. In support of this contention, it was observed that rabbit antiserum to NP-CG, after absorption with CG-Sepharose, augmented the response of mice to standard immunization with NIP12-CG.

Female mice produced significantly more NIP-binding antibody than did males.
TEMPERATURE-DEPENDENT VARIATION IN THE SYNTHESIS OF GROUP-SPECIFIC CARBOHYDRATE BY STREPTOCOCCAL VARIANT STRAINS
Authors: Ayoub Elia M. , Dudding Burton A.
Published: June 2004

A temperature-dependent alteration in the synthesis of the group-specific polysaccharide was found to occur in two "variant" streptococcal strains, A-486-Var and C 121/46/4. These strains synthesize a polysaccharide with variant immunochemical characteristics when grown at 37°C. However, when these organisms are grown at lower temperatures, 22°C, an enhanced synthesis of Group A carbohydrate occurs. Other variant strains show no appreciable alteration of the cell wall carbohydrate composition when grown at lower temperatures. Studies on an intermediate strain show that this organism has a propensity for the synthesis of a polysaccharide with higher glucosamine content and enhanced Group A serological reactivity when grown at 22°C.

Immunochemical studies performed on the carbohydrates produced by the A-486-Var at various temperatures revealed that the appearance of Group A serological reactivity at lower temperatures is due to the additional synthesis of a polysaccharide with Group A specificity along with the continued synthesis of a variant carbohydrate. This finding contrasts with data obtained on the carbohydrate produced by the intermediate organisms that appears to consist predominently of one molecule bearing dual A and variant antigenic determinants.
MEMBRANE ANTIGENS SPECIFIC FOR HUMAN LYMPHOID CELLS IN THE DIVIDING PHASE
Authors: Thomas D. B. , Phillips B.
Published: June 2004

Heteroantisera were raised in rabbits against human thymocytes or Burkitt's lymphoma EBI cells. After suitable absorption, the sera were specific for thymocytes and T lymphoblasts, or B lymphoblasts and several lymphoblastoid cell lines. The specificity of the antisera was confirmed by preparing purified populations of T or B lymphocytes on immunoabsorbent columns, followed by mitogenic stimulation with phytohemagglutinin. The antisera did not react with normal resting lymphocytes.
MUTANTS OF NONPRODUCER CELL LINES TRANSFORMED BY MURINE SARCOMA VIRUS
Authors: Hatanaka M. , Klein R. , Long C. W. , Gilden R.
Published: June 2004

Tumorigenic and nontumorigenic mutants induced by a single 5'-bromodeoxyuridine (BrdU) treatment of a nonproducer (NP) tumorigenic cell line were isolated and characterized. Among the cloned derivatives were examples of virus-free and sarcoma virus-producing cell lines. Oncogenicity did not correlate with production of virus or ease of rescue of the sarcoma genome. All lines, including nononcogenic derivatives, retained the sarcoma genome.

Phenotypic reversion of some cell mutants was observed after in vivo inoculation or long term in vitro cultivation. The M-50T cell line, obtained from a tumor induced by M-50 cells, had a sarcoma genome rescuable by direct superinfection; this was only achieved with parental M-50 cells by a cell fusion rescue technique. The M-43-2T cell, obtained from a single small static tumor induced by otherwise nononcogenic M-43-2 cells, shed sarcoma virus and became tumorigenic. M-58-4-48 became tumorigenic after passage 48 of the M-58-4 line, which was originally nontumorigenic. These observations of phenotypic reversion demonstrate that the presence of the sarcoma gene in cells is an essential but not sufficient condition of tumorigenesis.
THE MODULATING INFLUENCE OF CYCLIC NUCLEOTIDES UPON LYMPHOCYTE-MEDIATED CYTOTOXICITY
Authors: Strom Terry B. , Carpenter Charles B. , Garovoy Marvin R. , Austen K. Frank , Merrill John P. , Kaliner Michael
Published: June 2004

The capacity of allosensitized thymus-derived lymphocytes to destroy target cells bearing donor alloantigens is modulated by the cellular levels of cyclic AMP and cyclic GMP. Increases in the cyclic AMP levels of attacking lymphocytes by stimulation with prostaglandin E1, isoproterenol, and cholera toxin inhibit lymphocyte-mediated cytotoxicity; whereas, depletion of cyclic AMP with imidazole enhances cytotoxicity. The augmentation of cytotoxicity produced by cholinergic stimulation with carbamylcholine is not associated with alterations in cyclic AMP levels and is duplicated by 8-bromo-cyclic GMP. The effects of activators of adenylate cyclase, cholinomimetic agents, and 8-bromocyclic GMP are upon the attacking and not the target cells and occur at the time of initial interaction of attacking and target cells. Indeed, the level of cyclic nucleotide (cyclic AMP and cyclic GMP) at the time of initial cell-to-cell interaction determines the extent of cytotoxicity.
THE MEMBRANE ATTACK MECHANISM OF COMPLEMENT
Authors: Kolb William P. , Müller-Eberhard Hans J.
Published: June 2004

The membrane attack mechanism of complement, C5 to C9, has previously been postulated to associate on the target cell surface to a stable decamolecular complex with a calculated mol wt of 995,000.

A soluble and stable complex consisting of C5, C6, C7, C8, and C9 has now been demonstrated to arise as a consequence of complement activation by the classical or alternate pathway. It has a sedimentation coefficient of 22.5S and a mol wt of 1 million daltons, and it migrates on electrophoresis at pH 8.6 as an α-globulin.

The stable and soluble C5b-9 complex cannot bind to erythrocytes and has no demonstrable cytolytic activity. However, due to partially unsaturated binding sites for C9, it can bind additional C9 and thus function as an inhibitor of lysis of EAC1-8 by C9.

These results support the concept according to which the membrane-bound attack system of complement represents a stable, decamolecular assembly of C5b-9. Unlike its analogue in free solution, the membrane-bound complex is cytolytically active.