bluuebunny/crossref_metadata_2025 huggingface dataset:
Specific, Transient Suppression of the Immune Response by HGG Tolerant Spleen Cells
Authors: Doyle Michael V. , Parks Elliot , Weigle William O.
Published: December 2022

In a previous report, it was shown that spleen cells from mice made tolerant to human γ-globulin (HGG)5 could specifically inhibit the immune response of normal spleen cells after adoptive transfer to lethally irradiated recipients. However, that report also showed that the suppressive activity was only transiently associated with tolerant spleen cell populations. It was concluded from those experiments that while suppressive activity could be demonstrated in tolerant spleen cells under certain conditions, such activity was not obligatory for the maintenance of the tolerant state. The experiments presented here were performed to determine the nature of the effector cell(s) and the target cell(s) involved in this system of suppression of the immune response. Treatment of cells from tolerant animals with anti-thymocyte serum and complement to remove thymus-derived (T) cells completely abrogated suppressive activity. Removal of adherent cells from tolerant spleen cells by passage over glass wool columns resulted in partial loss of the suppression. The inhibitory activity of the suppressor cells was resistant to 900 R irradiation regardless of whether the tolerant spleen cells were irradiated before or after adoptive transfer.

The cellular target(s) for the suppressor cells was examined by using lipopolysaccharide (LPS) as an alternative source of helper activity for the response to HGG. LPS, injected at the time of the initial antigenic challenge of mice that had been reconstituted with tolerant and normal spleen cells, prevented the expression of suppression against bone marrow-derived (B) cells. However, when LPS was presented only at the time of the secondary antigenic challenge, it was unable to overcome suppression of the immune response of reconstituted recipients. Thus, LPS could produce a state where the B cells were resistant to suppression, but LPS could not rescue the responsiveness of B cells once the cells in the reconstituted recipient had been suppressed. In addition, the immune responses to both the hapten dinitrophenol (DNP) and the carrier (HGG) were suppressed when recipients of tolerant and normal spleen cells were challenged with DNP 6HGG. This indicates that T helper cells are also a target for suppression.

The results presented in this paper are discussed in relation to a possible mechanism of suppression which proposes that suppressive activity represents the induction of tolerance in immunologically competent cells by HGG which is closely associated with the tolerant spleen cells.
Early and Late Contact Sensitivity Reactions in Guinea Pigs Sensitized to Oxazolone
Authors: Baer Harold , Stone Sanford H. , Malik Frank
Published: December 2022

Inbred strain 2 and random-bred guinea pigs injected with oxazolone in incomplete or complete Freund's adjuvant showed contact reactions within an hour after topical application when tested 3 weeks post-sensitization. The time for appearance of skin reactions at 5 days post-sensitization was 8 to 24 hr, more along classic delayed-type hypersensitivity lines. Immune serum harvested several weeks after sensitization was effective for passive transfer of early skin reactivity, while serum at 5 to 10 days was not. Lymph node cells taken 5 days after sensitization of donors could transfer only the late skin reactivity to histocompatible recipients; lymph nodes taken at 15 days did not contain cells capable of such transfers. The heterogeneity of mechanisms involved in the production of contact dermatitis suggested by these results provides new approaches to studies in this area of allergy research.
Reaginic Antibody Formation in the Mouse
Authors: Takatsu Kiyoshi , Ishizaka Kimishige
Published: December 2022

The ongoing IgE antibody formation against ovalbumin (OA) in high responder mice was depressed by i.v. injections of either native or urea-denatured ovalbumin (UD-OA). Adoptive transfer experiments to determinethe helper function of spleen cells from the treated animals showed that helper function for both IgE and IgG antibody responses diminished after treatment. Evidence was obtained that treatment suppressed the expansion of IgE-B memory cells. When the same treatment with OA or UD-OA was given to OA-primed mice before the appearance of IgE antibody in their serum, OA-specific splenic suppressor T cells were demonstrable. Thus, the transfer of splenic T cells from treated mice into normal mice suppressed the primary IgE and IgG antibody responses of the recipeints to DNP-OA. It was also found that the transfer of the splenic T cells from UD-OA-treated mice into OA-primed mice depressed ongoing IgE antibody formation in the recipients. The results suggested strongly that the decrease of helper function and the depression of ongoing IgE antibody formation by repeated injections of UD-OA was caused by generation of antigen (OA)-specific suppressor T cells.
Proliferation of Murine Thymic Lymphocytes in Vitro Is Mediated by the Concanavalin A-Induced Release of a Lymphokine (Costimulator)
Authors: Paetkau Verner , Mills Gordon , Gerhart Stanley , Monticone Vicky
Published: December 2022

Mitogen-induced proliferation of lymphocytes may in theory result directly from the interaction of mitogen with the cells, or indirectly as a result of the mitogen-stimulated release of lymphokines. In the case of murine thymic lymphocytes exposed to concanavalin A (Con A) in tissue culture, we have determined that mitogenesis depends upon a lymphokine. Interaction of the thymic lymphocytes with lectin is necessary, but not sufficient, for mitogenesis. A lymphokine, or costimulator for mitogenesis, is released by normal spleen or thymus cells during the first 16 hr of their exposure to Con A, and in the presence of a phytomitogen it stimulates thymic mitogenesis. Under conditions of low costimulator levels, no mitogenesis follows the interaction of Con A with cells. The response of adult CBA/J mouse thymocytes to phytohemmaglutinin (PHA) is very low, compared to their response to Con A. When costimulator is added to PHA, the cells respond as well as they do to Con A. Costimulator does not act through Con A-binding sites on thymus cells. Its production is dependent on both cells carrying θ surface antigen (T lymphocytes) and adherent cells of the macrophage-monocyte series. The adherent population, but not the T cells, may be heavily irradiated without affecting production of costimulator. Costimulator is not a mitogen on its own.
The Role of Macrophages in Thymocyte Mitogenesis
Authors: Mills Gordon , Monticone Vicky , Paetkau Verner
Published: December 2022

The thymic lymphocytes of CBA/J mice respond to the mitogen Concanavalin A (Con A) only in the presence of adherent cells of the monocyte-macrophage series. Depletion of adherent cells abrogates the response, and macrophage-rich populations of cells restore it. The need for macrophages and mitogen is completely provided by irradiated splenic macrophages which have been exposed to Con A and washed free of the soluble mitogen. The mitogenmacrophage effect in this case is apparently not due to soluble factors.

Even more striking than the effect of macrophages on fresh cultures of thymic lymphocytes is their ability to restimulate quiescent cells 72 hr after their first stimulation with Con A. The quiescent cells respond immediately and quantitatively to Con A in the presence of fresh macrophages. This stimulation, like that of fresh thymocytes, is also controlled by a lymphokine (“costimulator”) produced by mixing macrophages, mitogen, and T lymphocytes.

Our data suggest a model in which two signals are required for mitogenesis. First, the interaction of macrophage, T cell, and mitogen elicits a soluble costimulator, which is itself not mitogenic. Secondly, in the presence of costimulator, the mitogen (either soluble, or, more efficiently, bound to macrophages) induces a proliferative response in the T cell.
Lysis of Tumor Cells by Antibody and Complement
Authors: Boyle Michael D. P. , Ohanian Sarkis H. , Borsos Tibor
Published: December 2022

The guinea pig hepatoma (line-1) treated with anti-Forssman antibody (TA) and GPC sequentially released 86Rb, 14C from 14C aminoisobutyric acid and failed to exclude trypan blue. Incubation of TA with fluid phase GPC for 1 min caused maximal 86Rb release; however, if the GPC was removed at this time, the cells were not subsequently killed. Using a number of naturally occurring human sera deficient in a complement component we have shown 86Rb release requires the binding of the complement components 1 through 8, but there was no absolute requirement for C9. Irreversible damage to the cell as measured by 14C AIB release or uptake of trypan blue required the complete sequence of complement components. These observations indicate that 86Rb release is not a reliable indicator of cytotoxicity.
Characterization of Ia Antigens in Mouse Serum
Authors: Callahan Gerald N. , Ferrone Soldano , Poulik M. David , Reisfeld Ralph A. , Klein Jan
Published: December 2022

Sera obtained from normal B10.BR mice were shown to inhibit selectively a specific anti-Ia alloantiserum. Partial purification of the Ia antigenic activity was accomplished by isolation of the high density lipoproteins from these sera by fractional precipitation with sodium phosphotungstate and MgCl2. Both H-2.23 and Iak antigens present in this high density lipoprotein fraction were completely adsorbed by rabbit anti-rat β2-microglobulin immunoadsorbents, whereas specific anti-H-2.23 immunoadsorbents removed only the H-2 activity. These data demonstrate that Ia antigens, like H-2 antigens in the sera of B10.BR mice are associated with high density lipoproteins and further suggest that both H-2 and Ia antigens are associated with a β2-microglobulin-like molecule.
Activation of the Complement Attack Mechanism in the Fluid Phase and its Control by C5̄6̄7̄-INH: Lysis of Normal Erythrocytes Initiated by Zymosan, Endotoxin, and Immune Complexes
Authors: Lint Thomas F. , Behrends Carole L. , Baker Patricia J. , Gewurz Henry
Published: December 2022

Addition of zymosan-serum complexes to guinea pig erythrocytes in guinea pig complement-EDTA was found to result in substantial lysis of the bystander cells in the presence of polycations such as poly-L-lysine of 178,000 daltons. Involvement of the alternative C pathway was shown, and the optimum time, temperature, and eruthrocyte and polycation concentrations were defined; a surprising efficiency was observed at low temperatures and high cell concentrations. Several lines of evidence indicated that this hemolysis was mediated via the complex of the C system and modulated by serum inhibitors of (-INH): lysis was observed only with zymosan-serum complexes possessing C-consuming activity; it was not observed in C5-depleted guinea pig serum but was restored upon addition of purified C5; the addition of partially purified -INH substantially depressed hemolysis; and poly-L-lysine which is known to neutralize -INH in solution resulted in substantial enhancement of hemolysis. We also sought to determine whether the addition of complement activators directly to erythrocyte-serum mixtures could result in the hemolysis of bystander erythrocytes. It was found that zymosan, endotoxin, antigen-antibody complexes, and aggregated human γ-globulin each could initiate such bystander lysis under appropriate conditions. Lysis again was favored by increased erythrocyte concentrations, low temperatures, and the presence of polycations such as poly-L-lysine, and was found to be mediated via the C system. -INH blocked cytolysis whereas poly-L-lysine potentiated hemolysis by neutralization of -INH. These experiments emphasize the propensity for formation and lysis of bystander erythrocytes during C activation generally, the role of -INH in the control of this lysis, and the susceptibility of these interactions to modulation by highly charged macromolecules.
Investigations of the Complement-Fixing Sites of Immunoglobulins
Authors: Johnson Brian J. , Thames Kenneth E.
Published: December 2022

Indirect evidence for the complement-fixing site of human IgG1 has been found by using chemical modification of the immunoglobulin. These results implicate the amino acid residues, tryptophan, tryosine, and arginine. Furthermore, by the use of synthetic peptides it has been shown that the area surrounding the tryptophan and tyrosine at positions 277 and 278 probably constitutes the locus of the complement-fixing site of human IgG1. By anology, the tryptophan at position 488 of human IgM is implicated in the complement-fixing site of this immunoglobulin. Evidence for this is presented by a peptide which mimics the sequence 487–491 of human IgM and possesses the ability to consume complement. The relative complement-fixing ability of this peptide is 200 times less effective than human IgG1.
H-2 Compatibility Requirement for Virus-Specific T Cell-Mediated Effector Functions in Vivo
Authors: Zinkernagel Rolf M. , Welsh Raymond M.
Published: December 2022

Adoptive immunization of recipient mice preinfected with lymphocytic choriomeningitis virus (LCMV) is mediated exclusively by virus-specific thymus-derived lymphocytes, when assayed in a short-term transfer model. Protection, measured as reduction of LCMV plaque-forming units in spleens, is conferred only if donors of immune spleen cells and recipients share the K or the D region of the H-2 gene complex. I region compatibility is neither necessary nor sufficient. The F1 → Parent combination is as effective as a syngeneic system. Admixture of a 6-fold excess of immune allogeneic cells did not impair the protective effect exerted by syngeneic immune spleen cells in vivo. Furthermore, allogeneic spleen cells or target cells added in syngeneic systems in vitro did not allogeneicly inhibit or suppress cytolytic activity. H-2 mutant mice B6.H-2bf did not protect wild type H-2Kb B10.A(5r) or vice versa. Therefore, these mice define the gene(s) coding for the relevant cell-surface structure involved.

These results are consistent with the idea that immune T cells, which are specific for virally altered cell-surface self structures impair virus growth in vivo either by lysing target cells, probably before infectious virus is released, or alternatively via activities exerted by lymphokines.
Relationship between β2-Microglobulin and Cell-Surface Alloantigens of the Mouse
Authors: Geib Roy , Poulik Miroslav D. , Vitetta Ellen S. , Kearney J. F. , Klein Jan
Published: December 2022

Molecular relationships between β2m and other cell surface antigens (H-2, Tla, Ia, and Thy-1) were studied with the double immunofluorescence method. Cells were incubated with an antiserum against one antigen, capped, and then tested with an antiserum against a second antigen. Capping of β2m on thymocytes led to simultaneous capping of H-2 and Tla but not Thy-1 antigens; capping of H-2 and Tla (but not Thy-1) antigens resulted in capping of all β2m detectable by the immunofluorescence method. Similarly, capping of β2m on B or T lymphocytes resulted in capping of H-2 and vice versa. Ia antigens on B lymphocytes were not capped after the redistribution of β2m. We conclude from these data that, in the cell membrane of thymocytes, virtually all the β2m molecules are associated with H-2 and Tla, but not with Thy-1, and that on the cell surface of T or B lymphocytes, virtually all β2m is associated with H-2 but not with Ia. We found no evidence of any significant free β2m on either thymocytes or splenocytes.
Serotonin-Induced Production of a Monocyte Chemotactic Factor by Human Peripheral Blood Leukocytes
Authors: Foon Kenneth A. , Wahl Sharon M. , Oppenheim Joost J. , Rosenstreich David L.
Published: December 2022

Serotonin and histamine are released during the early phase of inflammatory reactions. To determine if these vasoactive amines might play a role in the cell-mediated phase of inflammation, studies were undertaken to assess their influence on some immunologic functions of human peripheral blood leukocytes (PBL). PBL were cultured with serotonin or histamine, and assayed for a proliferative response and their cell-free supernatants were assayed for the presence of a mediator with chemotactic activity for monocytes. Neither serotonin nor histamine were directly chemotactic for monocytes. When serotonin was cultured with PBL factors chemotactic for monocytes as well as polymorphonuclear leukocytes were produced in the absence of a proliferative response. Production of the factor chemotactic for monocytes was specifically blocked by methysergide, a serotonin antagonist and nonspecifically blocked by cycloheximide which interferes with leukocyte protein synthesis. Some decarboxylated precursors of serotonin also induced the monocyte chemotactic factor. This serotonin-induced chemotactic factor was demonstrated to be a product of mononuclear cells, and the majority of chemotactic activity was physicochemically similar to the previously characterized lymphocyte-derived chemotactic factor, suggesting that this factor is a lymphocyte product.

In contrast to serotonin, histamine did not stimulate the production of monocyte chemotactic factor, nor did it induce proliferation. However, when 10-5 M histamine was added to a substimulatory concentration of serotonin (10-5 M), chemotactic activity comparable to or greater than the peak activity of serotonin alone was measured. This synergistic effect between serotonin and histamine suggests that the combination of these two amines may contribute to the induction of monocyte exudation in inflammatory reactions.
Analysis of the Cross-Reactive Immune Suppression Induced by the Random Copolymers L-Glutamic Acid50-L-Tyrosine50 (GT), L-Glutamic Acid60-L-Alanine40 (GA), and L-Glutamic Acid60-L-Alanine30-L-Tyrosine10 (GAT)
Authors: Waltenbaugh Carl , Debré Patrice , Benacerraf Baruj
Published: December 2022

Cross-reactivity in the induction of immune suppression by the structurally related polymers of L-glutamic acid, L-alanine, and/or L-tyrosine (GAT, GA, GT) was studied. SJL (H-2s) and DBA/1 (H-2q) mice are nonresponders to GAT and GT, whereas SJL, but not DBA/1 responds to GA. Both strains respond to these polymers when complexed to the immunogenic carrier methylated bovine serum albumin (MBSA). Preimmunization with GA suppresses GA-MBSA as well as GAT-MBSA and GT-MBSA plaque-forming cell (PFC) responses in DBA/1 mice. Likewise, GAT suppresses GAT-MBSA and GA (SJL) and GA-MBSA (DBA/1) PFC responses in both strains while being totally ineffective on GT-MBSA responses. GT preimmunization suppressed GA, GAT-MBSA, and GT-MBSA PFC responses in SJL but not DBA/1 responses. This study demonstrates that cross-reactive immune suppression does not follow predictable patterns based upon cross-reactivity of specific antibodies and that cross-suppression is not reciprocal.
Synergism between Immune Complexes and Serum from Tumor-Bearing Mice in the Suppression of Mitogen Responses
Authors: Kilburn Douglas G. , Fairhurst Maureen , Levy Julia G. , Whitney Richard B.
Published: December 2022

Immune complexes either adhering to the cells, or added to cultures, synergized with an inhibitor present in serum to suppress the proliferative response of lymphocytes to specific antigens or to B cell or T cell mitogens. The suppressive effect was dependent on the concentration of both the complex and the serum and was substantially higher when the serum was taken from tumor-bearing, rather than normal mice. Immune complexes alone were not suppressive, but appeared to activate or increase the effectiveness of an inhibitor present in serum. Complexes of syngeneic mouse serum or specifically purified rabbit or sheep antibody, as well as heat aggregated mouse or rabbit Ig, were all effective, whereas heat-aggregated F(ab′)2 fragments of rabbit Ig were not, suggesting that any aggregate bearing exposed Fc fragments could mediate synergy. Purified T lymphocytes were equally as sensitive to inhibition as the whole spleen cell population, implying that the suppression acted directly on responding cells rather than via an accessory cell.
Guinea Pig Lymphocyte-Derived Macrophage Aggregation Factor: Its Separation from Macrophage Migration Inhibitory Factor
Authors: Postlethwaite Arnold E. , Kang Andrew H.
Published: December 2022

Lymphocytes from guinea pigs having delayed hypersensitivity to horse-radish peroxidase (HRPO) when cultured in vitro with HRPO produce a large m.w. factor (τ; 100,000 daltons) that causes peritoneal macrophages from nonimmune animals to aggregate. The macrophage aggregation factor (MAF) can be separated from macrophage migration inhibitory factor (MIF) by gel filtration of active lymphocyte supernatants on Sephadex G-150. MAF is heat stable (56°C for 30 min) but inactivated by trypsin.

These data suggest that aggregation of macrophages in vitro by lymphokine-rich culture supernatants is nót due to MIF but is caused by a separate large m.w. factor.
IgA Half Molecules
Authors: Mushinski J. Frederic , Dungan Miranda M.
Published: December 2022

Six two chain IgA myeloma proteins are found to have very similar molecular weights and to share immunochemical characteristics suggesting that they represent examples of an unusually small-sized variant of normal mouse IgA. With the use of homologous antiserums in 2-dimensional double diffusion and hemagglutination inhibition, small amounts of this unusual IgA were detected in normal BALB/c intestinal contents. Gel filtration of the intestinal contents suggests that these gut immunoglobulins A exist in a size consistent with a single light chain plus a single heavy chain and can be regarded as IgA “half molecules” or IgAh. We feel that IgAh represents a normal variant of mouse IgA, but the possibility of its being a second mouse IgA subclass is discussed.
Partial Characterization of Human C5a Anaphylatoxin
Authors: Fernandez Horacio N. , Hugli Tony E.
Published: December 2022

Human C5a was isolated from complement-activated serum and was characterized for protein and carbohydrate content. The purified C5a was judged to be homogeneous by both polyacrylamide gel electrophoresis and immunologic techniques. The polypeptide moiety of C5a contains 73 amino acid residues which represent a m.w. of 8,200. Analysis of the carbohydrate moiety in C5a indicated 4 moles of glucosamine, 3 to 4 moles of sialic acid, 4 moles of mannose and 2 moles of galactose. The total carbohydrate content in C5a, therefore, amounts to approximately 25% of the apparent m.w. of the anaphylatoxin molecule. The protein and carbohydrate portions of C5a together equal a m.w. of approximately 11,000 which is considerably less than the 15 to 16,000 indicated by physical measurements. Human C5a contains a COOH-terminal arginine which is essential for anaphylatoxin activity and a sequence of Gln-Leu-Gly-Arg-COOH at the COOH-terminus which compares favorably with that of human C3a (Gly-Leu-Ala-Arg-COOH). Additional similarities between the C3a and C5a molecules include length of the polypeptide chain, number of disulfide bonds and an absence of tryptophan residues. A major chemical difference does exist between these two human anaphylatoxins, namely that carbohydrate is associated with C5a but is absent in the C3a molecule. The partial NH2-terminal sequence of C5a was determined as NH2-Thr-Leu-Glx-Lys-Lys-Ile-Glx-Glx-Ile-Ala- and direct comparison with the known sequence of human C3a shows little homology.
Replication of Herpes Simplex Virus in Mouse Spleen Cell Cultures Stimulated by Lipopolysaccharide
Authors: Kirchner Holger , Hirt Hans-Martin , Kleinicke Christa , Munk Klaus
Published: December 2022

Replication of HSV was demonstrated in spleen cell cultures of D2 and several other strains of mice after prestimulation with mitogenic doses of LPS for 2 days. No viral replication occurred in unstimulated cultures or in cultures prestimulated with PHA and Con A, whereas there was some viral replication in spleen cell cultures of D2 mice after prestimulation with Poly I-C. Spleen cells of B6 mice did not support replication of HSV under any of the conditions we have tested thus far. The reasons for this defect are not clear, but it was obviously not caused by a defective lymphoproliferative response to LPS or by an active anti-viral principle elaborated by B6 spleen cells. F1 hybrids between B6 and D2 mice were capable of HSV replication to the same extent as were spleen cells of D2 mice. Several strains of both HSV-1 and HSV-2 could be replicated in D2 spleen cell cultures. Nylon column treatment of D2 spleen cells removed the ability to replicate HSV, whereas macrophage removal from the spleens by plastic adherence was without effect. Purified peritoneal exudate cells from D2 mice did not support replication of HSV. Together these data suggest that B cells activated by LPS represent the target cell of HSV replication in mouse spleen cell cultures.
Membrane Expression of Thy-1.2 and GM1 Ganglioside on Differentiating T Lymphocytes
Authors: Milewicz Christine , Miller Harold C. , Esselman Walter J.
Published: December 2022

C3H mouse bone marrow cells were separated by discontinuous BSA gradient centrifugation. Marrow cells from the 17 to 19%, 19 to 21%, 21 to 23%, 23 to 25%, 25 to 27% interfaces and the cell pellet were treated with thymic factor (TF) or with Vibrio cholerae neuraminidase (VCN), followed by anti-Thy-1.2 and anti-GM1 ganglioside antisera. Antigens determined by anti-Thy-1.2 or anti-GM1 were expressed either with TF or VCN within a 30-min incubation. Cells expressing these antigens after VCN or TF treatment were concentrated in the 19 to 21% layer and the 21 to 23% layer whereas there was little or no antiserum cytotoxicity in the other layers. A small amount of Thy-1.2 or GM1 could be detected by cytotoxicity on 19 to 21% layer marrow cells within 15 min of either TF or VCN treatment. Treatment of 19 to 21 % layer cells with TF or VCN had no effect on anti-H-2 cytotoxicity. Pretreatment of TF-treated marrow cells with cholera toxin or choleragenoid (which bind cell surface GM1) abrogated the cytotoxicity of anti-Thy-1.2 or anti-GM1 without affecting anti-H-2 cytotoxicity. Pretreatment of Thy-1.2 positive C3H thymocytes with cholera toxin or choleragenoid greatly reduced the cytotoxicity of anti-Thy-1.2 antiser a without affecting the cytotoxicity of anti-H-2 antisera. Nude mouse splenocytes, after treatment with VCN or TF, were susceptible to the lytic action of anti-Thy-1.2 and anti-GM1. The possibility that nonspecific autologous antibodies were responsible for anti-Thy-1.2 cytotoxicity toward VCN-treated marrow cells was eliminated because anti-Thy-1.2 was not cytotoxic for VCN treated AKR (Thy-1.1) marrow cells.
Mouse Factor B of the Alternative Pathway of Complement Activation
Authors: Bitter-Suermann Dieter , Burger Reinhard , Brade Volker , Hadding Ulrich
Published: December 2022

Mouse factor B was purified and a monospecific anti-serum was raised. The physicochemical data of this protein (108,000 m.w., 5.9 S, 5.9 to 6.05 isoelectric point) were determined. The functional behavior resembles that of human and guinea pig factor B and it operates efficiently in the C3 feedback cycle of the alternative pathway of complement activation. A provisional scheme is given for the operation of the C3bB̄ enzyme on the cellular (macrophage) level.
Complement Receptor Lymphocytes in the Rabbit
Authors: Green William R. , Fanger Michael W.
Published: December 2022

In an attempt to determine the effect of immunization on the size and nature of the complement receptor lymphocyte (CRL) and surface immunoglobulin (SIg)-bearing lymphocyte (B cell) populations, rabbits were injected with 1 mg alum-precipitated keyhole limpet hemocyanin and the cells of the draining popliteal lymph nodes were examined at various intervals for these populations. Comparison to unimmunized control lymph nodes revealed that immunization resulted in an approximate doubling of the percentages of both CRL and B cell populations by the peak of the response, while the cell number increased about 7- to 8-fold. Kinetic analysis of the response indicated that the percentages of CRL and B cells increased essentially in parallel beginning within 40 hr after immunization. Simultaneous assay of complement receptor (CR) and SIg disclosed that CRL remained a B cell subpopulation throughout the response. To determine whether cell division could account for the increases in total cell number and per cent CRL and B cells, immunized rabbits were given 3H-thymidine in vivo, and the fraction of labeled (dividing) cells was determined by autoradiography. Results of such experiments were consistent with the possibility that the early increase in cell number and the increase in CRL and B cell percentages were a result of trapping and/or homing of lymphocytes migrating from the peripheral blood. A significant amount of lymphocyte division occurred during later stages of the response.
Induction or Suppression of B Cell Proliferation and Differentiation by Phytohemagglutinin or Concanavalin A in Mouse Spleen Cell Cultures
Authors: Piguet Pierre-F. , Dewey Henry K. , Vassalli Pierre
Published: December 2022

Cultures of mouse spleen cells or various mixtures of mouse T and B cells were stimulated with PHA or Con A, and the T, B, or plasmablast nature of the transformed cells, was determined by immunofluorescence 2 to 4 days later. The lectins enhanced B cell proliferation and plasmablast differentiation (“helper” effect) provided one of the following conditions was fulfilled: a) suboptimal doses of lectin were used, b) cultures were performed at low cell concentration, c) cultures were made of spleen cells containing a small percentage of T cells, d) the cultures contained a mixture of T-depleted spleen cells and T cells rendered unable to proliferate by irradiation. In contrast, cultures performed with 1.5 106 or more spleen cells/ml and optimal doses of lectin contained almost exclusively T blasts, as did cultures stimulated in the same conditions with both PHA and LPS. This last observation indicates the existence of a lectin-induced “suppressor” effect, since LPS, a B cell mitogen, induces, in the absence of PHA, a marked B cell proliferation and differentiation into plasmablasts. These helper and suppressor effects were entirely mediated by T cells, since they were not observed in spleen cell cultures depleted in T cells by anti-θ + C.

Analysis of the cultures by immunofluorescence and radioautography after pulses of 3H-thymidine showed that these antagonistic effects could be related to the number of T blasts present in the culture and to their proliferative behavior. Helper effect is observed in cultures containing a relatively low number of T blasts (or none in cultures made with irradiated T cells), whereas suppressive effect is observed in cultures containing a high number of T blasts, a large proportion of them having left the proliferative cell cycle. It is proposed that when a critical concentration of T blasts is reached (“saturation density”), further proliferation and differentiation is prevented, resulting in a suppressive effect on the generation of plasmablasts.

The helper effect of lectin-activated T cells seems to be exerted on a subpopulaion of B cells which was, at least in part, already proliferating in vivo, and to result in a polyclonal IgM plasmablast differentiation.
Complement-Induced Ultrastructural Membrane Lesions: Requirement for Terminal Components
Authors: Packman Charles H. , Rosenfeld Stephen I. , Weed Robert I. , Leddy John P.
Published: December 2022

The step in the complement (C) sequence at which 8- to 11-nm ring-shaped lesions are formed on antibody-coated erythrocytes (EA) has remained controversial. Some workers have concluded that these lesions appear at the C5 step and are not ultrastructural correlates of lysis; others hold that these lesions are formed only after the action of C8 and C9 in association with lysis. We have re-examined this problem by using sheep EA and human sera genetically lacking C5, C6, C7, or C8. Electron micrographs of negatively stained membranes (× 220,000) were read in blind fashion and the results correlated with 125I-C5 binding.

Rare structures resembling C-induced ring lesions were found on EA exposed to C5-deficient (C5D), C6D, C7D and C8D sera or to heated normal serum, with no significant differences among these sera (lesion density 0 to 0.26/µm2). Fresh normal serum (NHS) produced 140 to 220 ring lesions/µm2. C5 binding to EA in C8D serum was 60% of that observed in an NHS control; in C6D and C7D sera C5 binding was 4 to 11% of the normal value. Iodine treatment of sera (to enhance C5 uptake by C2 oxidation) increased C5 binding in C6D serum to 40 to 65% of that seen in native NHS; in iodine-treated C7D and C8D sera C5 binding was 250 and 440%, respectively, of the native NHS value. No increase in ring lesions was observed, however, except in the iodine-treated NHS.

Thus, in whole serum, C5 binding is not sufficient to produce ultrastructural membrane rings in the absence of later-acting C components, at least through C8. The formation of ring lesions appears to have C requirements similar to those necessary for lysis.
Comparative Immunogenic Properties of N-Substituted Phosphatidylethanolamine Derivatives and Liposomal Model Membranes
Authors: Nicolotti Robert A. , Kochibe Naohisa , Kinsky Stephen C.
Published: December 2022

This study describes some of the parameters that quantitatively or qualitatively influence the immunogenicity in guinea pigs of synthetic lipid antigens: phosphatidylethanolamine (PE) derivatives in which the amino (N) group has been substituted with either dinitrophenyl (DNP), dinitrophenylaminocaproyl (DNP-Cap), fluoresceinthiocarbamyl (Fl), or mono (p-azobenzenearsonic acid) tyrosyl (ABA-Tyr) residues. Previous experiments have shown that the non-covalent insertion of DNP-Cap-PE and ABA-Tyr-PE into the same lipid bilayers of sphingomyelin-cholesterol-dicetylphosphate liposomes markedly enhanced anti-DNP -Cap antibody formation over that produced by liposomes sensitized with only DNP-Cap-PE. The humoral response to Fl-PE and DNP-PE-sensitized liposomes is also augmented by the simultaneous incorporation of ABA-Tyr-PE. Moreover, micelles containing both DNP-Cap-PE and ABA-Tyr-PE induce more antibodies to the DNP-Cap determinant than do micelles of DNP-Cap-PE alone, or a mixture of DNP-Cap-PE and ABA-Tyr-PE micelles. Nevertheless, in regard to a humoral response, liposomes were more potent immunogens than were their micellar counterparts.

Of all the N-substituted derivatives examined so far, ABA-Tyr-PE is unique in that it can elicit cell-mediated immunity in addition to antibodies. The cellular response to ABA-Tyr-PE is not, however, stimulated by incorporation into liposomal bilayers and requires administration of either micelles or liposomes in complete Freund's adjuvant. In contrast, the ability of ABA-Tyr-PE to enhance a humoral response to another N-substituted PE derivative present in the same immunogen is also observed when the latter are given with incomplete Freund's adjuvant. The relationship of these findings to the immunogenicity of naturally occurring lipid antigens, as well as conventional immunogens having at least one determinant covalently attached to a protein carrier, is discussed.
The L1210 Leukemia Cell Bears a B Lymphocyte Specific, non-H-2 Linked Alloantigen
Authors: Freund John G. , Ahmed Aftab , Budd Richard E. , Dorf Martin E. , Sell Kenneth W. , Vannier Wilton E. , Humphreys Robert E.
Published: December 2022

The L1210 murine lymphoblast cell line possessed a B lymphocyte-specific alloantigen which was detected with C57BL/Ks anti-L1210 serum. The antigen was found on splenic B lymphocytes but not on thymocytes, T lymphocytes, or erythrocytes. It was present on only 7% of bone marrow cells. The reactivity of C57BL/Ks anti-DBA/2-spleen serum was indistinguishable from that of the anti-L1210 serum, confirming that the antigen was a normal component of B lymphocytes of the DBA/2 mouse. The strain distribution pattern of the antigen detected by the C57BL/Ks anti-L1210 serum indicated that this alloantigen was not an allele of the H-2K, H-2D, Mls, or Ig loci. Genetic analysis indicated that the antigen was inherited as a single, Mendelian dominant trait. Segregation analysis of (B10.BR × DBA/2)F1 × AKR/J offspring indicated that this B cell marker was was not linked to genes coding for H-2.31 or Ly 4.2. The C57BL/Ks anti-L1210 serum identified a new polymorphic genetic locus, the product of which was B lymphocyte specific.
Studies on Non-H-2-Linked Lymphocyte-Activating Determinants
Authors: Ahmed Aftab , Scher Irwin
Published: December 2022

Minor lymphocyte-stimulating (Mls) coded non-H-2-linked lymphocyte-activating determinants (LADs) are present on a late developing subpopulation of murine B lymphocytes. Spleen cells derived from CBA/N mice, a strain with an X-linked defect in B cell function, failed to stimulate a response in Mls-defined mixed lymphocyte reactions (MLRs) with H-2 identical CBA/J or C3H/N responder spleen cells. However, both CBA/J and C3H/N spleen cells induced significant responses in CBA/N responder spleen cells. The inability of CBA/N cells to induce an Mls-defined response was not due to the presence of the nonstimulatory Mlsb genotype, since spleen cells of immunologically normal F1 mice derived from crosses of CBA/N and C3H/N mice were stimulatory in MLRs for C3H/N responder cells. Immunologically abnormal CBA/N ♀ × C3H/N ♂) F1 male mice, which are hemizygous for the CBA/N X chromosome, were also unable to induce a response in Mls-defined MLRs. Although the Mls-coded LADs are not X linked in immunologically normal mouse strains, these data indicate that the failure of CBA/N mice to functionally express the Mls-coded LADs is X linked. This characteristic was not secondary to the presence of suppressor cells and is most likely related to the absence of a late developing (mature) subpopulation of B lymphocytes in these mice.
Preparation and Subfractionation of Isologous and Heterologous Anti-Idiotypes, with FV Fragments
Authors: Helman Miriam , Shreier Irit , Givol David
Published: December 2022

Fv fragments prepared from the BALB/c anti-dinitrophenyl (DNP) myeloma proteins, produced by plasmacytoma MOPC 315 and XRPC 25, were used to raise anti-idiotypic antibodies in rabbits. Anti-idiotypes to these proteins were also prepared in the syngeneic BALB/c mice by injecting the intact proteins, whereas Fv fragment failed to induce antibodies in these mice. Rabbit anti-Fv of late bleedings did not show the presence of antibodies to “framework determinants” (determinants common to all V regions of mouse immunoglobulins), and they contained precipitating anti-idiotypes whereas mouse antisera contained non-precipitating anti-idiotypes. No cross-reaction was observed between anti-idiotypes to these myeloma proteins and induced anti-DNP antibodies from BALB/c mice, when either rabbit or mouse antisera were analyzed. The antisera were fractionated into anti-binding site and anti-non-binding site by adsorption onto the immunogen-Sepharose column and elution by DNP lysine, followed by elution with acetic acid. Only the anti-binding sites were inhibited by hapten in their interaction with the homologous myeloma protein. The rabbit anti-Fv antibodies were predominantly (over 90%) anti-non-binding site whereas the mouse anti-idiotypes were predominantly (60 to 70%) anti-binding site.
Antigen-Specific and Nonspecific Mediators of T Cell/B Cell Cooperation
Authors: Hoffeld J. Terrell , Marrack Philippa , Kappler John W.
Published: December 2022

Previous work in this laboratory has defined two functionally distinct T cell subpopulations (viz., antigen-specific helper and antigen-nonspecific helper) and characterized some of the parameters of these subpopulations derived from the keyhole limpet hemocyanin (KLH)-primed mouse spleen. Characterization of further parameters such as relative susceptibility to tolerance induction and relative degree of specificity was not possible with the use of KLH as the antigen. In order to overcome this impediment, an experimental protocol was developed, as herewith described, to permit the study of these two T cell subpopulations in response to the antigen human γ-globulin (HGG). This protocol qualitatively duplicates the parameters defined for the KLH system and provides an extremely useful model for the study of the response to serum proteins in the Mishell-Dutton culture system.
In Vitro Synthesis of Immunoglobulin and Anti-Viral Antibody in Aleutian Disease Viral Infection of Mink
Authors: Perryman Lance E. , McGuire Travis C. , Banks Keith L.
Published: December 2022

Lymphoid cells from Aleutian disease virus (ADV)-infected and noninfected mink were cultured in vitro and the synthesis of IgM, IgG, and viral-specific antibody measured. Compared to noninfected mink, lymph node cells from mink with Aleutian disease (AD) contained a higher percentage of plasma cells and immunoglobulin-bearing lymphocytes, synthesized significantly more IgG, and produced detectable quantities of ADV-specific antibody. No significant difference in IgM synthesis was observed.
Natural Immunity in Cats to Feline Leukemia Viral Antigens
Authors: Ruscetti Sandra K. , Parks Wade P.
Published: December 2022

An isotopic antiglobulin assay was developed to measure antibodies in “normal” cats to FOCMA, a cell surface antigen associated with immunity to FeLV-induced tumors. The assay was shown to be specific for detecting FOCMA antibody by several criteria including a correlation with cell membrane immunofluorescence and absorption tests. Antibody to FOCMA developed following FeLV infection and was expressed in heterologous species. In combination with an assay for antibody to FeLV p30, the major virion structural protein, 200 cat sera from three different geographic areas were examined for evidence of FeLV type-C virus infection. Approximately 17% of all sera examined contained antibody to FOCMA. Antibodies directed to the FeLV p30 were detected in 8 of the sera and only in cats with FOCMA antibody. The correlation in cat sera between antibody to FOCMA and FeLV p30 antigen suggests that both are associated with the same process, infection with FeLV type-C virus. The relationship of the observed pattern of antibody response is discussed in relation to other horizontally transmitted RNA-containing viruses.
Blocking of HLA and B Lymphocyte Alloantigens with F(ab′)2 Fragments of Rabbit Antibodies
Authors: Billing Ronald J. , Honig Ronald , Peterson Patricia , Safani Madjid , Terasaki Paul I.
Published: December 2022

F(ab′)2 fragments of rabbit antibodies specific for human B lymphocytes will block human anti-B lymphocyte alloantibodies. F(ab′)2 of anti-β2m will block HLA alloantibodies. The F(ab′)2 fragments can be used to improve the typing of B cell lines and certain leukemia cells which express both HLA and B lymphocyte alloantigens. Pretreatment of these cells with anti-β2m F(ab′)2 preparations allows the observation of B cell alloantigen reactions independently of HLA reactions. Similar use of rabbit anti-B cell F(ab′)2 allows HLA reactions to be observed free from the “extra reactions” caused by B alloantibodies that are present in many HLA typing sera.
Lack of Antibody Activity Directed against the Most Common Human Bacteria in Human Myeloma Protein
Authors: Pillot Jacques , Creau-Goldberg Nicole , Gonzales Yolanda
Published: December 2022

The fact that sera of germ-free animals contain very little Ig suggests that most Ig probably correspond to antibacterial antibodies. We have detected an antibacterial activity against 40 bacterial strains, pathogenic and nonpathogenic, in 85 M-components of human origin. This work has been conducted by indirect immunofluorescence withipure labeled antibodies. The reasons for which indirect immunofluorescence seems to be the best technique for this kind of research are discussed.

No activity whatsoever could be detected. The reasons of this lack of activity remain to be explained.
Antibody-Induced Modulation of Leishmania Donovani Surface Membrane Antigens
Authors: Dwyer Dennis M.
Published: December 2022

The effects of specific antibodies on the surface membrane antigens of Leishmania donovani (strain 1S, clone 2D) amastigote and promastigote developmental stages was detected in vitro by using several direct and indirect immunofluorescence methods. Stage specific anti-L. donovani sera and sera from L. donovani-infected hamsters induced parasite surface membrane antigens to aggregate, move along the longitudinal cell axis, form polar cell caps, and subsequently disappear. No membrane fluorescence was observed in cells treated identically with normal sera. In amastigotes a single anterior cell pole cap was formed after antibody treatment. However, antibody-treated promastigotes showed tripartite membrane antigen capping consisting of a major anterior cell pole cap with minor caps at the posterior cell pole and flagellar tip regions. Loss of surface fluorescence from the two latter capped cell regions was frequently concurrent with the formation of extracellular fluorescent plasmanemes. The cell-capping process was antibody concentration, temperature, time, and energy dependent. No cell caps were formed at low temperature or in the presence of several metabolic inhibitors. Cell cap formation was also selectively inhibited by certain types of antibody-induced intercellular agglutination. Parasite membrane antigens removed by capping were regenerated and detectable at the cell surface only after a 3.5 to 4-hr period. Antibody-induced membrane antigen movement in L. donovani is apparently similar to phenomena observed with mammalian cells. Results of direct cross-staining and cross-absorption cell-capping experiments showed that the two parasite developmental forms shared some common or at least cross-reacting membrane antigens and that each form also possessed some unique stage-specific surface antigens. Cell-capping results with sera from L. donovani infected hamsters showed that this host produces some IgG which is directed specifically against certain parasite surface membrane antigens. The results suggest that host antibody-induced parasite membrane antigen modulation might have a role in the infectious process of this human pathogen.
The Genetics of Cell-Mediated Lympholysis
Authors: Long Mary Ann , Handwerger Barry S. , Amos D. Bernard , Yunis Edmond J.
Published: December 2022

The role of HLA antigens in the generation of cytotoxic cells in CML has been investigated. Cytotoxic effector cells were generated in MLC among HLA-A or HLA-A and HLA-B disparate, HLA-D identical siblings, and among HLA-A and HLA-B disparate, MLC identical (%RR ≦2 3.6) unrelated individuals. The data indicate that HLA-D differences and poliferative MLC responses as measured by 3H-thymidine incorporation are not requisite for the in vitro generation of cytotoxic cells and suggest the existence of a CML-S locus (loci) distinct from HLA-A, HLA-B and HLA-D. The degree of cytotoxicity generated in a proliferative versus a “nonproliferative” MLC was comparable.

In addition, these studies demonstrate that antigens other than the currently definable HLA-A, HLA-B, HLA-C, and HLA-D can serve as target determinants in cell-mediated lympholysis.
Analysis of Mononuclear Cell Surfaces with Fluoresceinated Staphylococcal Protein A Complexed with IgG Antibody or Heat-Aggregated γ-Globulin
Authors: Ades Edwin W. , Phillips Don J. , Shore Steven L. , Gordon David S. , LaVia Mariano F. , Black Charlotte M. , Reimer Charles B.
Published: December 2022

Fluorescein-conjugated staphylococcal protein A (SPA) was complexed with either: 1) heat-aggregated IgG, 2) B cell specific antibody, or 3) T cell specific antibody and then used for an immunofluorescent analysis of mononuclear cell surfaces. Cellular Fc receptors failed to recognize the Fc region of aggregated IgG that had been blocked by SPA. Moreover, fluoresceinated SPA that had been complexed either with anti-Fab (B-cell specific) or T cell-specific antisera prevented the nonspecific binding of these reagents to the IgG-Fc receptors on mononuclear cells, thereby permitting the latter to be properly identified as B or T lymphocytes. In addition, when unconjugated SPA was added to presensitized target cells in a test for antibody-dependent cell-mediated cytotoxicity, cytolysis was abrogated.