bluuebunny/crossref_metadata_2025 huggingface dataset:
Antibody Synthesis in Synchronized Mouse Spleen Cells during the Secondary Response to Sheep Erythrocytes in Vitro
Authors: Thomas David W. , Roberts Walden K. , Talmage David W.
Published: December 2022

Immunoglobulin synthesis by mouse spleen cells during the secondary response to sheep erythrocytes in vitro was examined in cells synchronized by methotrexate and adenosine treatment. Antibody synthesis by the synchronized cells was discontinuous through the cell cycle and highest in the early S phase. Previous results from this laboratory indicated that antibody secretion, as measured by plaque production to sheep erythrocytes, was also greatest in early S, and taken together these results suggest little difference in the expression of antibody synthesis and secretion through the cell cycle. Actinomycin D was employed in several experiments in an attempt to determine if the variation in antibody synthesis was regulated by transcription of messenger RNA for immunoglobulin. Lower actinomycin concentrations (0.05 µg/ml) reportedly permitting messenger, but not ribosomal, RNA synthesis as well as higher concentrations (1 µg/ml) rapidly reduced immunoglobulin and total protein synthesis. These results indicated that actinomycin was not suitable for determining the stability of messenger RNA in mouse spleen cells. Therefore, the reasons for fluctuation in antibody synthesis during the cell cycle could not be determined, but several possibilities are suggested.
Effects of Concanavalin A on the in Vitro Responses of Mouse Spleen Cells to T-Dependent and T-Independent Antigens
Authors: Jacobs Diane M.
Published: December 2022

The stimulatory and inhibitory effects of concanavalin A (Con A) on the in vitro primary immune responses to a T-dependent antigen, sheep erythrocytes (SRBC) and a T-independent antigen, TNP-lipopolysaccharide (TNP-LPS) have been studied. Inhibition of the response to both antigens was optimal when 2 µg Con A were added at the initiation of the culture period. The response to SRBC was considerably enhanced by the addition of Con A 24 hr later. In contrast, this late addition did not stimulate the TNP-LPS response and often inhibited it. Inhibition of the TNP-LPS response required the participation of T cells since it was not observed in cells from adult thymectomized irradiated bone marrow-reconstituted (ATxBM) mice. The response to TNP-LPS was somewhat enhanced in ATxBM cells, but the degree of enhancement was strikingly less than that observed for SRBC. LPS per se did not block the stimulatory effect of Con A on the SRBC response, and was observed to act synergistically with this lectin. None of the Con A effects observed required the participation of adherent cells. These observations are consistent with a model in which different subpopulations of T cells are responsible for the inhibitory and stimulatory effects. They further suggest that the Con A inhibitory activity acts via a T cell to inhibit directly the B cell response to antigen.
Role of the Clotting System in Cell-Mediated Hypersensitivity
Authors: Colvin Robert B. , Dvorak Harold F.
Published: December 2022

Radioactive tracers and immunofluorescence were employed to detect and quantitate fibrinogen/fibrin deposition in two types of cell-mediated hypersensitivity reactions in the guinea pig. Classic delayed hypersensitivity (DH) reactions to Old Tuberculin and to the azobenzenearsonate hapten were characterized by a progressive increase in the fibrinogen (125I-HF) content which exceeded that of the albumin tracer (131I-HSA) and paralleled the development of induration and erythema. Accumulation of 125I-HF could be related both to increased vascular permeability to 125I-HF and, more specifically, to retarded efflux of extravascular 125I-HF from tuberculin reaction sites. Warfarin inhibited 125I-HF accumulation and the formation of urea-insoluble 125I-HF (cross-linked fibrin) as well as induration in tuberculin reactions. Immunofluorescence studies revealed the site of Fib deposition to be extravascular, among the connective tissue fibers of the dermis, similar to that in DH reactions in man.

In contrast, little 125I-HF accumulated in cell-mediated reactions rich in basophils—cutaneous basophil hypersensitivity (CBH) reactions to keyhole limpet hemocyanin, ovalbumin, and dinitrochlorobenzene—due in part to less vascular leakage of macromolecules and to decreased formation of urea-insoluble fibrin. By immunofluorescence Fib deposits were found in CBH reactions in a pattern similar to that in DH reactions, but the intensity of staining was appreciably less. Thus, fibrin accumulation further distinguishes DH from CBH reactions and is very likely responsible for the induration characteristic of DH reactions.
Activation of Guinea Pig Macrophages by Bacterial Lipopolysaccharide Requires Bone Marrow-Derived Lymphocytes
Authors: Wilton J. Mark , Rosenstreich David L. , Oppenheim Joost J.
Published: December 2022

The activation of guinea pig peritoneal macrophages by Salmonella typhimurium lipopolysaccharide (LPS) was studied by using 14C-glucosamine uptake. Peritoneal exudate cells incorporated significant amounts of 14C-glucosamine when stimulated with LPS but neither purified macrophages nor nonadherent lymphocytes by themselves incorporated glucosamine. The activation of macrophages could be restored by adding nonadherent peritoneal lymphocytes, spleen cells, and lymph node cells but not thymocytes. Removal of B lymphocytes abolished the restorative capacity from active lymphoid cell populations. In contrast, B lymphocytes would restore glucosamine incorporation by macrophages stimulated with LPS but T lymphocytes did not. In addition, cell-free supernatants from LPS stimulated B lymphocytes but not from T lymphocytes could restore glucosamine incorporation by macrophages. These experiments demonstrate that LPS does not directly activate macrophages as measured by glucosamine incorporation but stimulates B lymphocytes which in turn activate macrophages.
The Role of Antibody Feedback Inhibition in the Regulation of the Secondary Antibody Response after High and Low Dose Priming
Authors: Grantham William G. , Fitch Frank W.
Published: December 2022

Spleen cells from mice immunized with a low dose of SRBC produce a greater secondary response to subsequent antigenic challenge in vivo than spleen cells from mice initially immunized with a larger dose. However, when the spleen cells are removed from the mice and given a secondary challenge in tissue culture, a greater response is produced by cells from animals primed with the larger antigen dose. Serum from the mice receiving the high dose of antigen contains low, but significant levels of specific antibody, and can inhibit further antibody production by lymphoid cells in vivo and in vitro. The characteristics of this suppression are similar to those known for passively administered specific antibody. Thus, antibody produced as a result of primary immunization may act through an inhibitory feedback mechanism in specifically limiting the magnitude of the secondary response to subsequent antigenic challenge in the normal animal.
Inhibition of the Primary in Vitro Antibody Response by Interferon Preparations
Authors: Johnson Howard M. , Smith Bennett G. , Baron Samuel
Published: December 2022

The primary in vitro plaque forming cell (PFC) response of mouse (C57BL/6J) spleen cells to sheep red blood cells (SRBC) was inhibited by two sources of crude and two sources of partially purified mouse interferon. The substance(s) in the interferon preparations which induced PFC and antiviral inhibitory activity have the following common characteristics: a) The interferon preparations were of different potencies and specific activities, but they inhibited the PFC response in proportion to their activities; 20 to 60 units were the minimal amounts required for τ;90% inhibition of the PFC response. b) Both the antiviral activity and the PFC inhibitory activity of the interferons were neutralized by antibody specific for mouse interferon. c) Both activities were partially or completely inactivated by heating at 60°C for 1 h. d) Human interferons had neither antiviral activity nor PFC inhibitory activity in mouse cells. e) Limited exposure (4 h) of cells to interferon significantly inhibits both viral infection and the PFC response. f) Both the antiviral activity and the PFC inhibitory activity of the interferon preparations are acid stable. It is concluded, therefore, that the inhibition of the primary in vitro PFC response is due to interferon in the preparations.

Maximum inhibition of the PFC response occurred when interferon was added to cultures at the same time as SRBC. Significant inhibition was also observed when interferon was added 1 day later, and slight enhancement was observed when added at days 2 and 3 of SRBC addition. Kinetic data showed that the greater the concentration of interferon added to the cultures, the earlier the effect on the PFC response. The presence of interferon in cultures for the first 4 h is sufficient to inhibit the PFC response. Interferon, then, appears to affect some early event(s), which leads to inhibition of the PFC response.

Since mature T cells can be one of the sources of interferon, we have demonstrated a suppressor effect of a T cell lymphokine. The precise relationship of interferon to various expressions of suppressor T cell effects remains to be determined.
Micro-Radioimmunoassay for Antibodies to Newcastle Disease Virus in the Chicken
Authors: Cleland Glen B. , Perey Daniel Y. E. , Dent Peter B.
Published: December 2022

An indirect micro-radioimmunoassay is described in which chicken anti-Newcastle disease virus antibody was detected, with radioactively labeled rabbit anti-chicken Fab, on virus-infected microcultures of chick embryo fibroblasts. Newcastle disease virus-infected microcultures were formalin fixed and stored at 4°C for up to 4 months without affecting the sensitivity of the test. The micro-technique was found to be a highly sensitive and specific assay of anti-viral antibody and may allow detection of immunoglobulin class of anti-Newcastle disease virus antibody.
Spleen Cell-Mediated Cytotoxicity of Hamster Cells Transformed by Herpes Simplex Virus: Evidence for Virus-Specific Membrane Antigen
Authors: Lausch Robert N. , Jones Constance , Christie Diane , Hay Kathleen A. , Rapp Fred
Published: December 2022

The nature of the host's immune response to isografts of hamster embryo fibroblasts (HEF) transformed by herpes simplex virus type 1 (HSV-1) was investigated by the microcytotoxicity assay. It was found that spleen cells from tumor-bearing hamsters killed homologous tumor cells but not HEF transformed by cytomegalovirus or PARA-(defective SV40)-adenovirus 7 (PARA-7). Cytotoxicity was lost as the tumor increased in size. Spleen cells from animals bearing isografts of HSV type 2 (HSV-2) transformed cells also killed HSV-1 target cells whereas spleen cells from PARA-7 tumor bearers did not. Further studies showed that animals immunized with HSV-1 or HSV-1-infected rabbit kidney cells produced spleen cells specifically cytotoxic for HSV-transformed cells. On the other hand, sera from virus-immunized hosts or tumor bearers had no effect on the cells in the presence of guinea pig complement. However, in blocking experiments such sera could significantly reduce spleen cell cytotoxicity.

These experiments established that cells transformed by HSV could elicit a cellular immune response in the syngeneic host, and provided evidence that the immunity was directed against virus-specific antigens on the cell surface.
Characterization of Human B Lymphocyte Specific Antigens
Authors: Ishii Yoshifumi , Koshiba Hirofumi , Ueno Hiroo , Maeyama Iwaki , Takami Tsuyoshi , Ishibashi Fumitoshi , Kikuchi Kokichi
Published: December 2022

Anti-human B cell serum (ABS) was developed by sequentially absorbing a rabbit anti-human tonsil serum (AHTS) with human red cells, liver, serum, and thymocytes. AHTS was also absorbed quantitatively with tissues and cells from different sources. ABS was nontoxic for thymocytes but lysed the majority of chronic lymphocytic leukemia (CLL) cells. This also killed a number of tonsil and blood lymphocytes which bound erythrocyte-antibody-complement complexes but not sheep erythrocytes. In further studies, it was shown that phytohemagglutin-responsibility of peripheral blood lymphocytes was not affected by treating them with ABS and complement. Anti-B cell activity of AHTS was not changed by absorption with thymus, liver, kidney, and brain tissues but absorbed with tonsil and CLL cells. These data confirm the B cell specificity of ABS.

Indirect immunofluorescence was carried out on tissue sections of human tonsils and lymph nodes, which indicated that ABS-reactive cells were concentrated in lymphoid follicles including germinal centers, while plasma cells and cells located in the thymus-dependent area were essentially devoid of immunofluorescence.
Immunoglobulin D-Bearing Lymphocytes in Primary Immunodeficiencies
Authors: Preud'Homme Jean-Louis , Clauvel Jean-Pierre , Seligmann Maxime
Published: December 2022

Surface IgD on blood lymphocytes was studied in 10 normal adults and 24 patients with primary immunodeficiencies by direct immunofluorescence, together with surface immunoglobulins of the other classes and with spontaneous rosette formation with sheep erythrocytes. In the normal adults, 8% of the lymphocytes bore δ chains (the figures for µ chains being 11%) and, among the cells positive for µ and/or δ, 70% were mixed stained, 22% and 8% being single stained for µ and δ respectively. In 10 patients with sex-linked agammaglobulinemia or variable immunodeficiency, practically no cells bearing surface immunoglobulins, including IgD, were detectable. A normal distribution of surface immunoglobulins, including the results of double labeling for µ and δ, was found in five other immunodeficiency patients in whom there was a block of the terminal differentiation of B lymphocytes into plasma cells. A new kind of block in the differentiation of the B cell line was observed in two patients affected with sex-linked severe combined immunodeficiency and variable immuno-deficiency respectively. They showed high figures for IgD-bearing lymphocytes, some of which carried simultaneously µ chains, contrasting with the absence of lymphocytes carrying IgM without IgD and of IgG- or IgA-bearing cells. The data obtained in several other patients with low figures for IgG- and IgA-bearing lymphocytes and a predominance of IgD-carrying cells with an excess of single producers for δ chains over single producers for µ chains suggest an analogous but incomplete maturation arrest.
Evidence for the Absence of Noncovalent Bonds in the Fcµ Region of IgM
Authors: Hester Raymond B. , Mole John E. , Schrohenloher Ralph E.
Published: December 2022

IgM isolated from the sera of five patients with Waldenström's macroglobulinemia was subjected to tryptic digestion at 60°C. The Fc5µ fragments recovered from the digests were reduced by 0.05 M cysteine and alkylated by iodoacetamide, producing large quantities of an Fcµ fragment having a sedimentation velocity (so20,w) of 2.9S and a molecular weight of 33,500 by sedimentation equilibrium in neutral buffer. Further studies on the Fcµ fragment from one of the proteins demonstrated that it was not dissociated into smaller fragments by 5 M guanidine-HCl, even after reduction with 0.1 M 2-mercaptoethanol in 5 M guanidine at pH 7.5. The number of sulfhydryl groups released by the latter treatment indicated the presence of two intrachain disulfide bonds. These observations provide evidence that this portion of the µ-chain demonstrates minimal noncovalent interactions.

Tryptic digestion of the Fcµ fragment at 37°C resulted in the production of several lower molecular weight fractions. The three major fractions demonstrated apparent molecular weights of 21,000, 13,800 and 6800 by sedimentation equilibrium in 5 M guanidine-HCl. The latter fraction (fraction C) had no detectable carbohydrate and consisted of two disulfide-bonded peptides having molecular weights of approximately 3800 and 2200. Studies on the amino acid composition and amino-terminal sequences indicated that fraction C was derived from the Cµ4 homology region and consisted of residues 468 through 546 of the µ-chain with the tryptic peptides encompassing residues 492 through 514 missing.
Prevention of Calcium Ionophore-Induced Release of Histamine in Rat Mast Cells by Disodium Cromoglycate
Authors: Johnson Herbert G. , Bach Michael K.
Published: December 2022

Discussion and Summary

Our results confirm the observations of Foreman et al. (10) that calcium-specific ionophores can initiate histamine release. In an accompanying communication (17) the production of a substance resembling SRS-A under the influence of A23, 187 is also reported. In this study we addressed ourselves to the question: “Can these ionophores be used as a tool to study DSCG's mechanism of inhibition of secretory histamine release?” DSCG does not chelate divalent cations very well (2, 12), but this conclusion is based on measurements of Ca in the free medium. Under similar conditions in which the EDTA shows significant evidence of complex formation with divalent cations the pH titration curve of DSCG is unaffected. The calcium involved in histamine release may be cell bound in a specific spatial arrangement that lends itself to “tying” up by DSCG at the cell surface. Thus, it is possible that one of the ways that DSCG inhibits is by preventing the increased uptake of calcium by cells under the influence of histamine release initiators such as compound 48/80 or antigen-antibody reactions, and thereby inhibiting mediator release. Physicochemical studies on binding to cell-bound ions by DSCG will be required to answer this point. DSCG can exist in a water lattice form that may have a specific binding capacity when divalent cations like calcium are present (18). It may thus exert its effect by existing in juxtaposition to the cell surface and binding ions, rather than actually binding to the mast cells. This juxtaposition could result in decreased calcium movement and diminished release of histamine.

These studies do not answer the questions of peculiar self-inhibition effects seen with DSCG, nor do they explain the biphasic nature of the drug's dose response in most in vitro cells systems. They are consistent with DSCG being involved in Ca economy of the cells, but it may also inhibit some unidentified enzyme needed for release. Self-inhibition effects of DSCG in ionophore-induced release are being studied as well as desensitization as a consequence of calcium deprivation to the cell and will be reported at a later date.
Complement Receptor in Synchronized Cultures of Human Hematopoietic Cell Lines
Authors: Papenhausen Peter , Papageorgiou Photini , Hirschhorn Kurt
Published: December 2022

Three previously established human lymphoid cell lines (HLV-1, SWB-16A, and B-1) were synchronized by double thymidine block and the lymphocyte surface receptor for complement was studied by the EAC-rosetting technique during the cell cycle. It was found that the proportion of cells with EAC rosettes remained unchanged throughout the various stages, indicating that the presence of the complement receptor in cultured lymphoid cells is independent of the cell cycle.
Serum IgE Levels during the Potentiated Reagin Response to Egg Albumin in Rats Infected with Nippostrongylus Brasiliensis
Authors: Carson Dennis , Metzger Henry , Bloch Kurt J.
Published: December 2022

Rats were immunized with 10 µg EA and alum; a low titer IgE anti-EA response was observed; subsequent infection with N. brasiliensis greatly potentiated this response. Serum from these rats was tested for its IgE content by a recently developed radioimmunoassay for rat IgE. The average serum level of IgE was 1.15 µg/ml; this level was not altered by immunization, but infection with N. brasiliensis was accompanied by an 80-fold increase in concentration of serum IgE. This increase could not be accounted for by IgE anti-EA or anti-worm antibodies. Infection with N. brasiliensis appears to have an adjuvant effect on IgE antibody responses to diverse environmental antigens as well as well as the experimentally induced IgE anti-EA response.
Canine Marrow Transplantation: Are Serum Blocking Factors Necessary to Maintain the Stable Chimeric State?
Authors: Tsoi Mang-So , Storb Rainer , Weiden Paul L. , Graham Theodore C. , Schroeder Maria-Louise , Thomas E. Donnall , Jacinto Pacita
Published: December 2022

Nine long-term canine radiation chimeras and their canine histocompatibility locus (DL-A) compatible, mixed leukocyte culture negative littermate marrow donors were studied between 545 and 1282 days after 1,200 R total body irradiation and marrow grafting. Before the time of testing, marrow donors were immunized against their chimeras by repeated skin grafts which they rejected. Skin fibroblasts from chimeras and their donors were tested for cell inhibition (CI) in the microcytotoxicity assay by exposure to lymphocytes from chimeras, donors and normal dogs and the effects of various sera on CI were evaluated. Lymphocytes from sensitized marrow donors consistently inhibited fibroblasts from the chimeras (eight of nine dogs); CI was abrogated by chimeric serum in only three of eight cases. Only two chimeras showed consistent CI of their “own” fibroblasts; CI was blocked by chimeric serum in one of the two. The remaining seven chimeras did not show consistent CI.

Sequential studies in 16 additional recipients of DL-A compatible littermate marrow were carried out from 45 to 439 days after marrow grafting. Seven of the 16 did not show CI of chimeric fibroblasts by chimeric lymphocytes at any time. Nine showed CI on one or several occasions. Serum blocking factors were seen on one occasion in each of two chimeras.

In conclusion, the CI assay is able to detect immunity against “minor” histocompatibility systems in dogs. Both long- and short-term chimeras occasionally demonstrated CI of chimeric fibroblasts but serum blocking factors did not appear to be necessary for maintaining stable grafthost “tolerance.”.
Immune Precipitation of Immunoglobulin Producing Polysomes from Mouse Myeloma Cells
Authors: Laskov Reuven , Mitelman Sara
Published: December 2022

MPC-11 myeloma cells synthesizing IgG were labeled in vitro with 3H-uridine and the cell lysates, or purified polysomes treated with rabbit antiserum against the purified MPC-11 protein. The specificity of the immune precipitation was tested by rabbit anti-egg albumin and by precipitation of polysomes from “non-producer” myeloma and HeLa cells. The specific precipitation was 10 to 15% of the total cytoplasmic RNA for the IgG-producing clone of MPC-11 and about 2% for the non-producer clone or HeLa cells. Up to 20% of the total polysomal and ribosomal RNA were precipitated from the IgG producing clone. This value correlated with the IgG synthesis of these cells which is also about 20% of the total protein produced. The results suggest that myeloma cells produce a relatively large quantity of IgG due to the presence of a large amount of specific m-RNA molecules.
Broad Antigenic Relationships among Rhinovirus Serotypes Revealed by Cross-Immunization of Rabbits with Different Serotypes
Authors: Cooney Marion K. , Wise John A. , Kenny George E. , Fox John P.
Published: December 2022

The rhinoviruses are a large group of at least 89 serotypes and extensive cross-relationships have been demonstrated among these serotypes using both rabbit and human antisera. The extent of these cross-relationships was further investigated by sequential intravenous immunization of rabbits with different virus serotypes, known to be both related and unrelated. In all cases in which a relationship had previously been demonstrated with monovalent antiserum, after heterotypic stimulation with the related virus, the antibody titer to the virus used as primary immunogen showed an anamnestic response, whereas the antibody response to the secondary immunogen was good, but resembled a primary response. In some instances when immunized rabbits were injected with an unrelated antigen, previously unsuspected relationships were revealed. A number of clusters of related rhinoviruses were demonstrated: 5, 17, 42; 3, 4, 6, 14; 9, 32, 67; and 13, 14, 41. Heterotypic immunization proved to be a powerful biologic probe for detecting relationships among the rhinoviruses, and together with newly available epidemiologic data it offers a possible approach for design of a vaccine for the common cold.
Inhibition of Lymphocyte Trapping by Cell-Free Ascitic Fluids Cultivated in Syngeneic Mice
Authors: Mongini Patricia K. A. , Rosenberg Leon T.
Published: December 2022

Several cellfree ascitic fluids were tested for their influence on the trapping of lymphocytes in peripheral lymph nodes after subcutaneous antigen administration. Fluid from three ascitic tumors cultivated in syngeneic mice diminished the trapping response significantly. Fluid from one of these ascitic tumors cultivated in allogeneic mice showed no inhibition of trapping. Syngeneic normal mouse serum did not significantly affect the trapping phenomenon. It was concluded that syngeneic ascitic fluids contain a substance capable of inhibiting lymphocyte trapping.
Differential Regulatory Effects of Accessory Cells on the Generation of Cell-Mediated Immune Reactions
Authors: Miller Carol L. , Mishell Robert I.
Published: December 2022

Conditions for the in vitro generation of cell-mediated immunity to H-2 and non-H-2 alloantigens by mouse spleen cells were compared to conditions for generation of in vitro humoral immunity to sheep red cells. Normal spleen cells grown in standard or allogeneic cell-conditioned media developed strong cellular responses to H-2 antigens but little or no response to non-H-2 antigens. Subpopulations of spleen cells from which the adherent cells had been removed generated strong cellular responses to both H-2 and non-H-2 antigens, provided the cells were cultured in conditioned media. Both cellular responses were shown to be mediated by T cells. Accessory cells from the peritoneum and/or spleen differentially inhibited the non H-2 cellular responses at doses which did not inhibit humoral responses.
Thymus-Independent Antigens: The Preparation of Covalent, Hapten-Ficoll Conjugates
Authors: Inman John K.
Published: December 2022

A general approach is presented for covalently attaching carboxyl and primary amino groups to Ficoll and other high molecular weight polysaccharides in a readily controlled manner. The polysaccharides are o-carboxymethylated with chloroacetate in NaOH solutions. Amino groups are next introduced by monoamide formation with ethylenediamine using a water-soluble carbodiimide. Reactions are carried out in aqueous solutions; excess reactants and by-products are removed by dialysis. Three different ways for coupling the 2,4-dinitrophenyl hapten to the amino derivative of Ficoll are described. Preliminary evidence indicates that 2,4-dinitrophenyl-Ficolls, prepared by procedures developed in this study, are potent thymus-independent antigens which can specifically stimulate B cells both in vivo and in vitro. Hapten-Ficoll conjugates should be exceptionally useful for studies of B lymphocyte activation because of the ready availability of the starting material and the ease with which a wide variety of hapten derivatives may be prepared.
Characterization of the Lymphocyte Surface Receptors for Con A and PHA
Authors: Henkart Pierre A. , Fisher Richard I.
Published: December 2022

Human peripheral blood lymphocytes were iodinated by the lactoperoxidase technique, and the surface receptors for the mitogens concanavalin A (Con A) and phytohemagglutinin (PHA) were analyzed by solubilizing the membrane protein and incubating with mitogen and antimitogen antibody. The resulting precipitates were washed and analyzed by gel electrophoresis. Con A was shown to bind to about twice as much labeled protein as PHA. Sodium dodecyl sulfate gels showed that multiple surface membrane proteins bind to these mitogens and that the pattern of peaks of Con A receptors is generally similar to the pattern of PHA receptors. Long-term lymphoid tissue culture cell lines also show multiple receptor peaks for mitogens, indicating that the heterogeneity of receptors is not caused entirely by lymphocyte heterogeneity.
The Stimulation of Immunoglobulin Production in Murine Spleen Cells by the Pokeweed Mitogens
Authors: Basham T. Y. , Waxdal M. J.
Published: December 2022

We have shown that of the five mitogens purified from the salt extracts of pokeweed, only Pa-1, the B cell mitogen, has the ability to stimulate immunoglobulin production in murine B cells in the absence of classical T cells. Furthermore, the T cell mitogens (Pa-2-Pa-5) not only are unable to stimulate DNA or immunoglobulin synthesis by B cells in the absence of T cells, but cannot stimulate immunoglobulin production by B cells even in the presence of T cells. This suggests that Pa-2-Pa-5 do not stimulate adequate helper function in T cells or that if such a helper function is stimulated, an additional stimulatory factor is required for immunoglobulin production, which is not provided for by Pa-2-Pa-5.
Anti-Receptor Antibody
Authors: Strayer David S. , Vitetta Ellen S. , Köhler Heinz
Published: December 2022

We have demonstrated a phosphorylcholine-binding protein in lysates from radioiodinated splenocytes of immunized mice. Immunoprecipitation with anti-receptor antibody of lysates from splenocytes obtained from animals undergoing a primary response or from mice immunized 4 to 6 months earlier with antigen demonstrated that this protein contained H and L chains.

Therefore, we have isolated an antigen-specific receptor from the surface of spleen cells from both immunized and “memory” animals. This receptor Ig comprises approximately 10% of total cell surface Ig.
Anti-Receptor Antibody
Authors: Strayer David S. , Lee William M. F. , Rowley Donald A. , Köhler Heinz
Published: December 2022

We have examined the ability of anti-receptor antibody (ARA) to induce specific unresponsiveness to the hapten, phosphorylcholine (PC), in neonatal and adult mice. When ARA is given to adult mice, suppression is of short duration. Cells from such mice are responsive in vitro, indicating that suppression in vivo is probably due to blockade of receptors by persisting ARA. ARA given to neonatal mice induces long-term unresponsiveness. These mice apparently have decreased numbers of PC-responsive cells, since cells from such mice are unresponsive both in vitro and in adoptive transfer. Furthermore, cells from neonatally suppressed animals do not suppress the response of normal cells either in vitro or in adoptive transfer, indicating that unresponsiveness is most likely not due to active suppression. We therefore conclude that ARA given to neonates depletes the clone of receptor-bearing cells at the time ARA is given. Clonal depletion may result from antibody-dependent cell mediated cytotoxicity.
Comparison of the Ocular Effects of Circulating Endotoxin and Immune Complexes: Role of Vasoactive Amines
Authors: Howes Edward L. , McKay Donald G.
Published: December 2022

Both bacterial endotoxin and soluble immune complexes (BGG-anti BGG) injected i.v. in rabbits produce an alteration in ocular vascular permeability confined primarily to the vessels of the iridial portion of the ciliary processes. These effects have been measured by the ocular accumulation of 125I-albumin relative to cardiac plasma. Ten micrograms per kilogram of Escherichia coli bacterial endotoxin (055:B5) produce a consistent alteration in ocular vascular permeability over a 90-min period after injection. Fifty to 100 µg/kg of endotoxin result in a prolonged effect that is maximum 4 hr after injection.

Large quantities of immune complexes (BGG-antiBGG) prepared in 20 to 25 times antigen excess produce an anaphylaxis and approximately a 70% reduction in platelets and CH50. The alteration in ocular vascular permeability is half as great as that produced by 10 µg/kg of endotoxin over the 90-min period after their injection. A combined treatment with antihistamine, pyrilamine maleate, and antiserotonin, methysergide maleate, results in a significant reduction in ocular 125I-albumin in normal animals, virtually prevents the ocular effect of immune complexes, and reduces the average effect of endotoxin by 30%. Increasing the quantities of injected antihistamine and antiserotonin does not have a further effect on the response to 10 µg of endotoxin.
Functional Heterogeneity Among the T-Derived Lymphocytes of the Mouse
Authors: Araneo Barbara A. , Marrack Philippa C. (Hunter) , Kappler John W.
Published: December 2022

A number of T cell functions were examined in mice after treatment with rabbit anti-mouse thymocyte serum (ATS). Two populations of T cells were distinguished; the first, sensitive, and a second, resistant, to elimination by ATS. The helper cell involved in the primary humoral response to sheep red blood cells (SRBC) was found within the sensitive population, whereas the T-cells required for the priming of mice with SRBC for helper activity or delayed hypersensitivity were found within both populations. Previous work and experiments utilizing both adult thymectomy and ATS treatment indicated that the ATS-sensitive population is long-lived, whereas the ATS-resistant population is short-lived.
The Effect of Fluorescein Conjugation on Fc-Dependent Properties of Rabbit Antibody
Authors: Thrasher Susan G. , Bigazzi Pierluigi E. , Yoshida Takeshi , Cohen Stanley
Published: December 2022

Conjugation of rabbit antibody with fluorescein isothiocyanate results in loss of the ability of that antibody to fix complement, mediate passive cutaneous anaphylaxis, or participate in cytophilic binding to macrophage surfaces. Fluorescein, perhaps because of molecular size or configuration, is a more potent suppressor of Fc function than are other reagents previously studied in this regard.
Rat Cell Surface Antigens
Authors: Callahan Gerald N. , Dewitt Charles W.
Published: December 2022

Rat major histocompatibility or Ag-B antigen was isolated from soluble extracts of lymphoid cells by affinity chromatography. The purified antigen was shown in SDS gel electrophoresis to have a molecular weight of 30,000 to 35,000 daltons. In addition, amino acid composition was found to resemble, generally, that of HL-A and H-2 antigens. It thus appears that the demonstrated homology between H-2 and HL-A extends also to Ag-B.
The Effect of Plasmacytomas on Serum Immunoglobulin Levels of BALB/c Mice
Authors: Fenton Marilyn R. , Havas H. Francis
Published: December 2022

The effect of tumor growth on serum immunoglobulin levels and on the immune response to sheep erythrocytes (SRBC) was studied in BALB/c mice bearing MOPC-315 (IgA), MOPC-460 (IgA), MOPC-173 (IgG2a) and MOPC-104E (IgM) to gain insight into the immunologic competence of the plasmacytoma-bearing host.

The initial increase of myeloma protein coincided with the first appearance of the tumor and increased as the tumor progressed. However, at the time of death there was little correlation between spleen weights, tumor size, and myeloma-protein levels. The mean serum concentration of the myeloma proteins reached a higher level in the mice bearing tumors transplanted i.p. compared to those with tumors transferred subcutaneously (s.c.). Non-myeloma immunoglobulin levels in the serum were reduced: IgM was significantly lowered in the presence of MOPC-315 injected i.p. and MOPC-460 injected s.c. and the IgG2 levels were depressed in mice injected i.p. with MOPC-315 and MOPC-104E. The only significant reduction of IgA levels was seen when MOPC-173 was transplanted i.p. The decreases observed in immunoglobulin levels correlated with plasmacytoma growth. They were specific for myeloma and were not due to tumor growth per se since the levels of all immunoglobulins tested increased in the presence of Sarcoma 37, a pleomorphic neoplasm.

The primary plaque-forming cell (PFC) response of tumor-bearing animals after the injection of 0.5 ml of 10% SRBC was either similar or enhanced compared to the controls. However, with a lower SRBC dosage (0.5 ml of 2% SRBC) the indirect PFC were reduced with mice bearing MOPC-104E and MOPC-173. Tumor sizes did not seem to correlate with the reduction of the PFC responses. MOPC-460-bearing mice had a comparable number of PFC per spleen to those of the controls, but reduced numbers when calculated per 106 spleen cells. Consistently, hemagglutination titers were reduced in all tumor-bearing animals. The number of direct and indirect PFC per spleen was increased in mice bearing Sarcoma 37, compared to the controls. The possible implications of these findings are discussed.
Two Distinct Chemotactic Factor Inactivators in Human Serum
Authors: Till Gerd , Ward Peter A.
Published: December 2022

The chemotactic factor inactivator (CFI) has been isolated from whole human serum by a combination of techniques including salt precipitation, anionic exchange, and gel filtration chromatography. Two inactivators have been obtained, a β-globulin with a sedimentation velocity of approximately 7S and an α-globulin with a sedimentation velocity of approximately 4S. The former has a specificity for inactivation of the chemotactic activity associated with the C3 fragments, whereas the C5 chemotactic fragment is specifically inactivated by the α-globulin CFI. CFI in crude fractions of human serum is heat labile, time and temperature dependent for its activity, and pH dependent, expressing optimal activity at a pH range of 7.2 to 7.4.
Anti-Dinitrophenyl Antibody Production in Strain 13 Guinea Pigs Fed or Sensitized with Dinitrochlorobenzene
Authors: Reese Robert T. , Cebra John J.
Published: December 2022

Guinea pigs fed dinitrochlorobenzene (DNCB) in corn oil become unresponsive to sensitization by later percutaneous applications of DNCB in olive oil. Sera taken from animals after feeding DNCB or after feeding and intradermal stimulation with DNCB contained no antibody detectable with 2,4-dinitrophenyl (DNP)-labeled T4 bacteriophage; if any antibody was present, its concentration was below 2 µg/ml. Guinea pigs fed DNCB and then immunized with DNP on a foreign carrier, keyhole limpet hemocyanin (KLH), produced as much antibody 16 to 18 days later as did animals which had only been sensitized by intradermal injection of DNCB in phosphate-buffered saline and then immunized with DNP-KLH. The amount of antibody produced to DNP-KLH was measured at intervals. In the fed animals, the titer either remained approximately the same or increased with time. In contrast, the sera of sensitized guinea pigs contained a decreasing antibody titer when the first and last bleedings were compared. Isoelectric focusing was carried out with sera from DNP-KLH-immunized animals after the sera were first mixed with a radiolabeled hapten. The antibody-hapten profiles revealed that the DNCB-fed animals contained predominantly IgG2 in their serum by the time of their initial bleeding, whereas sensitized animals still contained a considerable proportion of more acidic antibodies having marked charge heterogeneity. Equilibrium dialysis measurements on purified antibody from fed animals indicated an average binding affinity for hapten equivalent to that of purified antibody from sensitized animals. Feeding of DNCB did not cause any obvious decrease in the number of lymphoid cells able to bind 125I-labeled guinea pig IgG. Lack of reduction of the average binding affinity coupled with the retention of antigen-binding lymphoid cells, permit the conclusion that feeding DNCB to guinea pigs under the conditions used did not induce an unresponsiveness or a deletion of those cells which have the potential to produce anti-DNP antibody. Thus, feeding of guinea pigs with DNCB in oil seems to induce a central tolerance at the T cell level without adversely affecting their B lymphocytes. The antibody response obtained by immunization with DNP conjugated to a foreign carrier is not only quantitatively equivalent to that obtained in control animals but qualitatively it appeared to be more mature with respect to charge at a relatively early time (16 to 18 days). Since the antibody levels in serum from fed animals increased or remained approximately constant with time instead of waning as occurred in the DNCB-sensitized animals, feeding or sensitization with DNCB may also have affected some cell or cell product important in the regulation of antibody production.
Identification of Macrophage-Like Characteristics in a Cultured Murine Tumor Line
Authors: Koren Hillel S. , Handwerger Barry S. , Wunderlich John R.
Published: December 2022

A mouse tumor line P388D1 passaged in tissue culture for many years has been characterized morphologically and functionally as a macrophage like cell. P388D1 cells phagocytize latex particles and firmly adhere to glass and plastics. In addition they have been shown to carry cell-bound receptors for immunoglobulin (Fc) and complement (C3). They fail to stain with fluorescent anti-mouse Ig or heterologous anti-mouse. Functionally, these cells exhibited high effector activity in an antibody-dependent, cell-mediated cytotoxic system. The accessibility of well characterized and pure macrophage-like cell lines, such as P388D1, will facilitate studies where cell purity is essential.
Phenotypic Correction of Low Reagin Production: A Genetic Defect in the SJL Mouse
Authors: Gollapudi V. S. Sastry , Kind Leon S.
Published: December 2022

Levine and Vas (1) have reported a system of genetic control of the immune response in mice which is not correlated with H-2 and does not appear to be correlated with allotype. In contrast to most other inbred strains, SJL mice were shown to be poor producers of serum reagins to high or low doses of eight different protein or hapten-protein antigens; breeding studies indicated high serum reagin production to be the dominant trait (2). In a recent communication (3) we confirmed the low serum reagin production by SJL mice and showed by the method of heterologous adoptive cutaneous anaphylaxis (HACA)2 that in contrast to a good serum reagin producing strain (SWR) the SJL mice showed: 1) a decreased production of reagins by spleen cells and 2) an absence of reaginic antibody-forming cells (RAFC) in the bone marrow.
Sensitive Detection of Immunoglobulin Light and Heavy Chains on Rabbit Lymphocytes by Antibody-Coated Erythrocytes
Authors: Molinaro Giuseppe A. , Bessinger Beverly A. , Gilman-Sachs Alice , Dray Sheldon
Published: December 2022

Rabbit bone marrow-derived (B)2 lymphocytes have immunoglobulin (Ig) on their surface (1–12). Oddly, various workers have reported for the last decade that in lymphocyte populations, the cells bearing the genetic markers of the κ-light chain (b locus allotypes) outnumbered 2 to 4 times the cells bearing the a locus allotypes of the heavy chain (1–5, 10). By using a more sensitive immunocytoadhesion method, we found in rabbit blood that the cells bearing heavy chains numerically equaled the cells bearing light chains. We also directly demonstrated that most B lymphocytes bore the light chains together with the heavy chains.

We have recently (13–15) shown that erythrocytes artificially coated with an antibody (Ab-E) agglutinated and lysed, when reacted with the complementary soluble antigen. When the antigen was cell bound, Ab-E adhered to the cell membrane (rosetting of antigen-bearing cells with Ab-E). Thus, rabbit lymphocytes rosetted with Ab-E specific for the b allotypes.
The Cytotoxic Effector Potential of Some Common Non-Lymphoid Tumors and Cultured Cell Lines with Phytohemagglutinin and Heterologous Anti-Erythrocyte Antisera
Authors: Muchmore A. V. , Nelson D. L. , Blaese R. M.
Published: December 2022

We have found that a) phytohemagglutinin can induce red blood cell cytolysis by non-lymphoid tumors and cell lines, b) rabbit anti-chicken red blood cell antibody but not turkey anti-chicken red blood cell antibody can mediate an antibody-directed cellular cytotoxicity type of cytotoxicity utilizing P815Y mastocytoma cells or rat embryo fibroblasts as effectors, and c) none of these cell line effectors was capable of killing metabolically active cultured cell line targets in the presence of phytohemagglutinin.
T Mitogens Trigger LPS Responsiveness in Mouse Thymus Cells
Authors: Forbes James T. , Nakao Yoshinobu , Smith Richard T.
Published: December 2022

Mouse thymus cells are essentially unresponsive to lipopolysaccharide (LPS) stimulation. However, when cultured with minimally mitogenic levels of concanavalin A or submitogenic ratios of mitomycin-treated allogeneic spleen cells, in combination with LPS, they demonstrate levels of DNA synthesis de novo or greater than those induced by the T cell mitogen alone. Dose-response kinetics were characteristic of LPS. The subpopulation containing the LPS responsive cells was of low net buoyant density. Neither phytohemagglutinin nor pokeweed mitogen acted synergistically with LPS in this model to trigger thymus cells. The data suggest that LPS triggering may involve interaction with a T cell subpopulation.
IgG and IgA Diphtheria Antitoxin Responses from Human Tonsil Lymphocytes
Authors: Platts-Mills Thomas A. E. , Ishizaka Kimishige
Published: December 2022

Human tonsil lymphocytes were stimulated with diphtheria toxoid and then cultured in a Marbrook culture system so that antibodies could be measured in the culture supernatant. Specific antibodies were measured with excess radiolabeled antigen and antisera specific for each immunoglobulin class. Good IgG and IgA diphtheria antitoxin responses have been obtained and responding culture supernatants were shown to neutralize toxin. The relationship between antitoxin response in vitro and immunization of donors with toxoid was investigated. It was found that at least two immunizations after the age of 6 months were necessary to prime the tonsils for an in vitro antibody response.

The IgG and IgA in culture supernatants were demonstrated by immunodiffusion and were measured by radioimmunoassay. By sucrose density gradient ultracentrifugation, it was shown that 40% of the IgA produced in the cultures was greater than 7S. Evidence was obtained that neither the IgA nor the specific IgA antitoxin bears secretory piece. It appears that human lymphocytes from tonsils produce polymer IgA in vitro without secretory piece.
Characterization of Measles Virus-Specific Cytotoxic Antibodies by Use of a Chronically Infected Cell Line
Authors: Ehrnst Anneka C.
Published: December 2022

The complement-dependent cytotoxic effects of measles virus specific antibodies on a chronically infected cell line were studied by a microcytotoxicity assay. Cell damage was determined by trypan blue staining. Monospecific antisera against the hemagglutinin (HA) and the hemolysin (HL) of the virus were both cytotoxic. Rabbits, immunized with measles virus, developed cytotoxic antibodies in parallel with the appearance of antibodies against the measles virus HA and HL. Human sera with high antibody titers against HA and HL also gave a specific cytotoxic reaction. The main part of the activity was carried by IgG in rabbit sera tested within 2 weeks after primary immunization and only IgM. The serum reaction was characterized by a prozone. The maximal degree of cytotoxicity varied between tests performed on different occasions. Complement, at higher concentrations than were used for analysis of antibody-mediated cytolysis, caused a significant lysis of infected cells.

Cerebrospinal fluid (CSF) samples containing measles virus-specific antibodies were investigated. CSF from patients with subacute sclerosing panencephalitis displayed a pronounced cytotoxicity without a prozone. CSF samples from four patients with multiple sclerosis were identified by a specific cytotoxic reaction despite low titers of antibodies against HA and HL.