bluuebunny/crossref_metadata_2025 huggingface dataset:
Studies on the Effect of Specific Antisera on the Metabolism of Cellular Antigens
Authors: Yu Alice , Cohen Edward P.
Published: December 2022

Thymus leukemia antigens and H-2a antigens were released in soluble form from ASL-1 leukemia cells (H-2a; TL 1,2,3) during extraction with the nonionic detergent, NP-40. They were isolated by immunoprecipitation with specific antisera and rabbit anti-mouse IgG. Specific antigens were obtained by the procedure because: 1) increasing radioactivity in immunoprecipitates of the NP-40 extracts of 3H-fucose or 125I-labeled ASL-1 cells was obtained with increasing amounts of H-2a or TL 1,3 antisera, but no significant amounts of radioactivity above control precipitates were detected in extracts incubated with H-2b antiserum; 2) the complete removal of TL antigens from the NP-40 extract of ASL-1 cells by immunoprecipitation with excess TL antisera had little effect upon the subsequent immunoprecipitation of H-2a antisera; 3) immunoprecipitates of NP-40 extracts of ASL-1 cells with H-2a antisera yielded one radioactive peak on acrylamide gel electrophoresis whereas the control immunoprecipitates with normal mouse serum or H-2b antiserum failed to yield any measurable peaks of radioactivity. The immunoprecipitates of NP-40 extracts of ASL-1 cells with TL 1,3 antiserum obtained from (BALB/c × C3H) F1 mice yielded two major peaks and one minor peak of radioactivity. Further analysis suggested that they represented TL, tumor antigens associated with ASL-1 leukemia cells, and an antigen of strain A mice which was foreign to (BALB/c × C3H) F1 mice, respectively.
The Isolation and Characterization of an Anti-Sheep Red Blood Cell Antibody Having Limited Heterogeneity
Authors: Bratcher Raymond L. , Chong Christine A. , Dray Sheldon
Published: December 2022

A rabbit (RB1) was induced to produce anti-sheep red blood cell antibodies of limited heterogeneity in abnormally high concentrations. These antibodies were isolated by a new procedure which involves the cross-linking of stroma with glutaraldehyde to produce a reusable immunosorbent. This immunosorbent permitted the isolation of a pure antibody preparation, primarily of the IgG class. The small amount of IgM antibodies present was removed by Sephadex chromatography to yield a pure IgG product (99.7%) which gave a discrete dense band by cellulose acetate electrophoresis and three or four bands by isoelectric focusing. Also, by isoelectric focusing, one major and one minor light chain band and two major heavy chain bands were observed. Moreover, all of the antibody molecules were of the same allotype and had a characteristic idiotypic specificity.
Requirement for Bone Marrow-Derived Cells in Resistance to Listeria
Authors: Campbell Priscilla A. , Martens Barbara L. , Cooper H. Rebecca , McClatchy J. Kenneth
Published: December 2022

Experiments were designed to study the participation of bone marrow-derived cells in mediating resistance to Listeria monocytogenes. Lethally irradiated normal mice were injected with various combinations of bone marrow, spleen, thymus, and peritoneal exudate cells. Eight to 10 days later, the mice were challenged with Listeria, and assayed for resistance by determining the number of Listeria in their spleens, livers, and blood 48 hr after challenge. In some experiments, survival was used as an indicator of resistance. Since the injected cells were from normal animals, and since antigen was not knowingly administered to the recipient mice, the resistance studied here is probably due primarily to nonspecific rather than specific mechanisms. Mice injected with bone marrow with or without the addition of thymus cells were resistant to challenge; whereas mice injected with thymus cells, with peritoneal exudate cells, with both these cell types, with spleen cells, or with no cells were not resistant to Listeria.

Since neither peritoneal macrophages nor spleen cells would substitute for bone marrow cells, and since anti-ϑ-treated bone marrow cells were effective, these data suggest that a bone marrow-derived cell in addition to the macrophage is required. In addition, it was found that these irradiated, bone marrow-injected mice appear to be more resistant to Listeria than are normal mice, and can resist supra-lethal doses of the bacterium as do previously immunized mice.
Crossed Immunoelectrophoresis and Electroimmunoassay of IgM
Authors: Grubb Anders O.
Published: December 2022

Analysis of IgM by crossed immunoelectrophoresis is possible if the original technique is modified by the use of: agarose gel with little, or preferably no, electroendosmosis; a high pH of 10.3; and an antibody population essentially isoelectric at pH 10.3. Such an antibody population can easily be produced by a chemical modification, under mild conditions, of about one-third of the carboxylic acid groups of the antibodies. This analytical system also permits rapid quantitation of IgM by the electroimmunoassay.
Early Detection of Lymphocyte Stimulation and Mixed Lymphocyte Interaction in Man with a Semimicro Protein Synthesis Assay
Authors: Adkinson N. Franklin , Rosenberg Steven A. , Terry William D.
Published: December 2022

A rapid semimicro assay for lymphocyte activation in man is described. It is based upon the stimulation of protein synthesis (SPS) in peripheral blood lymphocytes cultured in serum-free medium with soluble antigens, phytomitogens or allogeneic lymphocytes. Cellular response to PHA is discernible in 4 hr and response to antigen in 16 to 24 hr. Stimulation in mixed lymphocyte cultures can be detected as early as 8 to 10 hr and invariably by 42 hr. The assay permits detection of lymphocyte activation earlier and with greater precision than the widely employed thymidine incorporation blast transformation system.
Immunocyte Response to a Purified Bacterial Toxin (Choleragen) and Toxoid: Cytokinetics, Immunoglobulin Class, and Specificity
Authors: Kateley John R. , Lyons Susan F. , Friedman Herman
Published: December 2022

A unique indirect plaque-forming cell (PFC) assay has been developed to assess the immune response to choleragen, a highly purified exo-enterotoxin derived from Vibrio cholerae cultures, at the cellular level. Spleen cells releasing anti-choleragen antibody were readily detectable with choleragen-sensitized sheep erythrocytes as “target” cells in a variation of the localized hemolysin in agar technique. Injection of 1-µg doses of the toxin into BALB/c mice was sufficient to stimulate large numbers of specific antibody PFC. Heat-treated choleragen preparations, i.e., procholeragenoid and choleragenoid, also induced a detectable immune response at 1-µg doses; however, the results indicate that the PFC response varied as a function of the dose and form of the cholera antigen used for immunization. Choleragen and procholeragenoid induced essentially similar immune responses and were more immunogenic than choleragenoid, although all immunogens stimulated IgM (19S) and IgG (7S) PFC. Peak IgM PFC responses were detected on days 6 and 7 whereas peak IgG PFC responses occurred 1 day later in all cases. A booster injection of choleragen (1 µg) increased the numbers of IgM and IgG PFC approximately 4- to 5-fold. IgA PFC were also detected after a booster injection. Specificity experiments revealed that the toxin and toxoid, but not a vibrio whole cell vaccine or lipopolysaccharide preparation, could inhibit the anti-choleragen PFC response.
Products of Activated Lymphocytes: Leukocyte Inhibitory Factor (LIF) Distinct from Migration Inhibitory Factor (MIF)
Authors: Rocklin Ross E.
Published: December 2022

The mechanism of inhibition in the leukocyte migration system was investigated. The migration of human buffy coat cells was inhibited by a soluble product of activated lymphocytes. This factor was found to have an elution pattern in Sephadex G-100 gels similar to that of albumin (69,000 daltons), thus differentiating it from human migratory inhibitory factor (MIF) (23,000 daltons). Leukocyte inhibitory factor (LIF), like MIF, was shown to be elaborated by sensitized lymphocytes following stimulation by specific antigen. LIF selectively inhibited the migration of polymorphonuclear (PMN) leukocytes but did not affect the migration of guinea pig macrophages or human monocytes. Thus, the leukocyte migration system has been shown to be mediated by a separate factor (LIF) and different indicator cells (PMN leukocytes) than the MIF-mediated system of migration inhibition which utilizes guinea pig macrophages or human monocytes as indicator cells.
Effect of Prior Blood Transfusions on Marrow Grafts: Abrogation of Sensitization by Procarbazine and Antithymocyte Serum
Authors: Storb Rainer , Floersheim Georg L. , Weiden Paul L. , Graham Theodore C. , Kolb Hans-Jochem , Lerner Kenneth G. , Schroeder Maria-Louise , Thomas E. Donnall
Published: December 2022

Previous studies have shown that a single blood transfusion can immunize a dog and lead to rejection of a subsequent marrow graft even when donor and recipient are littermates matched at the major histocompatibility complex (DL-A). The present study was designed to evaluate whether immunization of a marrow graft recipient by prior blood transfusions could be abrogated by procarbazine and/or antithymocyte serum. On day 0, recipient dogs were given 1200 R total body irradiation followed by marrow from DL-A incompatible unrelated donors of different breed. All recipients were transfused with blood from the marrow donor on days -20 and -13 before irradiation. Group 1: seven of 10 dogs given no further therapy rejected the marrow graft. Group 2: four of eight dogs given procarbazine 12.5 mg/kg i.v. on days -8, -6 and -4 rejected. Group 3: five of 10 dogs given anti-dog thymocyte serum (ATS), 0.6 ml/kg subcutaneously on days -7, -5, and -3 rejected. Group 4: one of 10 dogs given procarbazine, 12.5 mg/kg i.v. on days -8, -6, and -4 and ATS, 0.6 ml/kg subcutaneously on days -7, -5, and -3 rejected the graft. Nine had sustained engraftment. Group 5: six of six dogs given the combination of procarbazine and ATS and further donor transfusions on days -6, -4, and -2 rejected.

It was concluded that immunization by preceding blood transfusions can be successfully abrogated by a combination of procarbazine and ATS (p < 0.01). ATS or procarbazine alone were not sufficient. Continuation of donor blood transfusions during the procarbazine-ATS regimen precluded successful engraftment.
Structural Studies on a Human IgAl Myeloma Protein with a Carboxy-Terminal Deletion
Authors: Despont Jean-Pierre J. , Abel Carlos A. , Grey Howard M. , Penn Gerald M.
Published: December 2022

A human IgA1 λ monomeric myeloma protein (Vo) was found to be antigenically deficient to other IgA proteins. The molecular weight of the heavy chains was 42,000 daltons, compared with a figure of 58,000 daltons found for normal α chains. In contrast, both the light (L) chains and the Fd′ fragment had molecular weights identical to those of other IgAl proteins. Two cyanogen bromide fragments were isolated from the heavy (H) chains: one fragment consisted of the first 85 amino acid residues of the variable region, whereas the second one contained the remainder of the foreshortened α chain. Neither the carboxy-terminal octapeptide of normal α chains nor the cyanogen bromide fragment containing 30 amino acid residues and located in normal α chains in a position preceding that of the octapeptide were found. The structural moiety missing in Vo and responsible for its antigenic deficiency was found to be a peptide consisting of more than 100 amino acid residues from the C-terminal end of the α chain, probably involving the entire CH3 domain.
Development of Reaginic Antibody-Forming Cells in the Spleen and Bone Marrow of Immunized Mice
Authors: Kind Leon S. , Malloy William F.
Published: December 2022

In a recent communication (1) we described a method, heterologous adoptive cutaneous anaphylaxis (HACA),2 for the detection of reaginic antibody-forming cells (RAFC). Spleen, bone marrow, or lymph node cells taken from immunized CBA mice 28 and 56 days after immunization were transferred to the skin of rats. Twenty-four hours later the rats were challenged with antigen and Evans blue dye and a blue area appeared within minutes at the site of deposition of mouse cells. Since mouse IgG1 antibody will fix to mouse but not to rat mast cells (2), one can feel confident that HACA detects only reaginic antibody formation (1).

We now report the optimal conditions for HACA as well as the development of RAFC in the spleen and bone marrow of a moderate (SWR) and a poor (SJL) reagin-producing strain of mice from 3 to 61 days post-immunization. Levine (3) has proposed two kinds of genetic control of reagin production in mice.
Glycopeptides of Heavy Chains from Human IgA Myeloma Proteins
Authors: Despont Jean-Pierre J. , Abel Carlos A.
Published: December 2022

Glucosamine-containing glycopeptides were isolated from the heavy chains of four human IgA1 and three IgA2 myeloma proteins. The amino acid sequence of two α1 glycopeptides (1a and 4a) and of three α2 glycopeptides (1b, 2b, and 3b) isolated from the constant region of α chains was determined. Peptides 1a and 1 b were localized to the N-terminal half of the Fc fragment, whereas peptides 4a and 3b were localized to the C-terminal half of this fragment.
Immunogenicity and Partial Purification of Soluble H-2 Antigens Extracted with Hypertonic Salt
Authors: Götze Dietrich , Reisfeld Ralph A.
Published: December 2022

H-2d antigen was solubilized by 3 M KCl extraction from cultured murine lymphoid cells L1210, derived from DBA/2 mice. Yields of protein and specific inhibitory activity were found to be optimal when the cells were harvested during log growth. A 5- to 7-fold increase in specific H-2d antigen activity was achieved by Sephadex gel filtration and cellulose ion exchange chromatography. Antibodies against H-2Kd and H-2Dd determinants were produced by immunization experiments within the allogeneic system. Immunization with soluble H-2d antigen led to specific sensitization with respect to cellular immunity, indicating that the soluble material maintained its integrity as a serologically defined histocompatibility antigen.
A Solid Phase Radioimmunoassay for the Diagnosis of Human Hydatidosis
Authors: Musiani Piero , Piantelli Mauro , Arru Efisio , Pozzuoli Roberto
Published: December 2022

This report describes the application of solid phase radioimmunoassay in the diagnosis of human hydatidosis. Using plastic tubes or plastic cups coated with pure Echinococcus antigens (EA) and incubating with 125I-radioiodinated human anti-EA IgG and appropriate amounts of hydatidosis sera, we have demonstrated a competitive inhibition of the 125I anti-EA-binding reaction. The sensitivity and the specificity of this test was compared with the radioimmunoelectrophoretic, passive hemagglutination, and radioimmunoprecipitation tests. The solid state competitive binding radioassay offers clear advantages as regards the other tests so far utilized. Sensitivity as well as specificity is very high so that this method can be considered the test of choice in the serologic diagnosis of human hydatidosis.
Regulation of Antibody Response in Vitro
Authors: Kishimoto Tadamitsu , Ishizaka Kimishige
Published: December 2022

Mesenteric lymph node cells were obtained from rabbits that were primed with dinitrophenylated ragweed antigen (DNP-Rag) and supplemented with free Ascaris antigen (Asc); in vitro anti-hapten antibody response of the primed cells was studied. It was found that stimulation of Rag-sensitized cells with free Rag enhanced anti-DNP antibody response to DNP-unrelated carrier conjugate (DNP-KLH) but suppressed the response of the same cell suspension to DNP-Asc. It was also found that both IgG and IgE antibody responses to DNP-Rag were suppressed by the addition of free Asc with the homologous antigen. Mechanisms involved in the enhancement and suppression of antibody response by free carrier were explored with cell-free supernatant (CFS) obtained from the culture of primed lymph node cells with free carrier. The results showed that essentially all CFS preparations studied enhanced antibody response when they were added to primed lymph node cells which had been treated with a hapten-protein conjugate. Some of the CFS preparations, however, suppressed antibody response of the same primed cells, if they were added to the cells together with the antigen. Evidence was obtained which indicated that suppression of the antibody response by CFS is not due to an excess amount of enhancing soluble factor. Several preparations of CFS were incubated with the same primed cell suspension, either before or after incubation with antigen, to determine their suppressive effect and enhancing effect. The results showed that suppressive effect of CFS on the unstimulated cells did not correlate with their enhancing effect on the antigen-stimulated cells. The experiment eliminated the possibility that the same T cell factor expresses two opposite effects depending on the stage of B cell differentiation, and suggested the presence of both suppressive and enhancing soluble factors in the preparations of cell free supernatant.

The experiment also showed that enhancing soluble factor for IgE antibody response was obtained from carrier-specific cells primed with alum-precipitated antigen but the cells proliferated by the immunization with the carrier included in complete Freund's adjuvant did not release the enhancing soluble factor for IgE. It was also found that enhancing soluble factor for IgG antibody neither enhanced nor suppressed IgE antibody response.
Effect of Anti-µ-Specific Immunosuppression on Friend Virus Leukemia
Authors: Manning Dean D. , Kouttab Nicola M. , Jutila John W.
Published: December 2022

Evidence is presented that neonatally initiated immunosuppression of BALB/c mice with rabbit anti-µ antiserum renders them refractory to Friend virus leukemia. Mice so treated failed to develop typical leukemic leukocytosis, splenomegaly, or splenic foci upon injection of 2.2 LD50 doses of Friend leukemia virus. A brief regimen of anti-µ treatment in young adult mice was shown to depress moderately the immune response against sheep red blood cells but to have no discernible effect upon the development of Friend virus leukemia. Several possible explanations for these observations are discussed.
Histamine Release: In Vitro Studies of the Inhibitory Region of the Dose-Response Curve
Authors: Magro Albert M. , Alexander Alice
Published: December 2022

Histamine release from the leukocytes of ragweed sensitive donors, which also release well to Fc-specific anti-IgE, was studied. In one type of experiment the effect of increasing concentrations of monomer anti-IgE (Fab) upon the release induced by anti-IgE was observed. for a suboptimum concentration of anti-IgE, increasing concentrations of the monomer inhibited the histamine release. However, for a supraoptimum concentration of anti-IgE, increasing concentrations of the monomer enhanced the histamine release. Evidence was also obtained which shows that supraoptimum concentrations of anti-IgE render the cells unresponsive to a near optimum concentration of antigen E. As the anti-IgE concentrations were increased into the inhibitory region the release due to antigen E was not maintained but dropped in parallel to the decrease due to the anti-IgE. High concentrations of monomer anti-IgE caused no inhibition of the antigen E induced release. The data imply, that for anti-IgE-induced release, the descending portion of the dose-response curve may be the result of a turn-off mechanism caused by an excess of bridging.
Rosette-Plaque Assay: Fine Specificity Differences among Individual Hapten-Primed Lymph Node Cells
Authors: Bankert Richard B. , Wolf Benjamin
Published: December 2022

A combination rosette-plaque assay was used to demonstrate simultaneously hapten binding and antibody secretion by hapten-primed rabbit lymph node cells. A family of cross-reacting haptens was employed to investigate fine specificity differences of hapten-binding among early antibody-forming cells. The compounds used included 3-nitro-4-hydroxy-5-iodophenyl acetic acid (NIP), 3,5-dinitro-4-hydroxyphenyl acetic acid (NNP), 3,5 diiodo-4-hydroxyphenyl acetic acid (DIP), and 3-nitro-4-hydroxyphenyl acetic acid (NP). The hapten-primed lymph node cells were first rosetted with two different rosette target cells simultaneously; the priming (homologous) hapten coupled to a sheep red blood cell stabilized with pyruvic aldehyde (PASRBC). One of the cross-reacting haptens was coupled to the other target PASRBC. The two rosette target PASRBC were distinguished by labeling one with a fluorescent marker. After rosette formation via centrifugation at 4°C, the cells were plaqued on fresh SRBC coupled with one of the several haptens. The simultaneous expression of the specificity of hapten-binding and antibody secretion was manifested as a rosette in the center of a plaque.

NIP-primed lymph node cells produced three different types of rosettes within plaques. The majority bound only the homologous hapten-coupled rosette target PASRBC (NIP-PASRBC). A smaller number of cells bound only the cross-reacting hapten-coupled target cells, i.e., NNP-PASRBC, DIP-PASRBC, or NP-PASRBC. The last group of cells bound both the homologous and cross-reactive target PASRBC simultaneously. In animals immunized to either NNP or DIP, these being relatively non-cross-reactive, very little to no cross-reactive binding was observed when the rosetted cells were plaqued on the homologous hapten-conjugated plaque target SRBC. for example NNP-primed lymph node cells rosetted by employing NNP-PASRBC plus DIP-PASRBC, and then plaqued on NNP-SRBC, were observed to bind the homologous NNP-PASRBC exclusively. However, when the same hapten-primed cells were rosetted as before, but now plaqued on the relatively non-cross-reactive plaque target cells (DIP-SRBC), a significant increase was noticed in the cross-reactive binding.

It would appear that the use of the heterologous plaque target SRBC has prejudiced the expression of heterologous binding which was not manifested when plaqued on the homologous plaque target SRBC. Moreover, the data considered collectively suggest a heterogeneity in the cross-reactivity of hapten-binding by plaque-forming cells.
Immunity to Murine Sarcoma Virus-Induced Tumors
Authors: Gorczynski Reginald M.
Published: December 2022

BALB/c mice immunized with murine sarcoma virus (MSV) have been studied at various times after injection for their immunity to either extracts of MSV-induced sarcomas or preparations of disrupted whole MSV virus. The assays used were the inhibition of normal macrophage migration, in the presence of immunizing antigen and sensitized cells, or the stimulation of protein or DNA synthesis in immune cells themselves, caused by the addition of the immunizing antigen.

Each assay system is shown to detect specific anti-MSV-induced immunity in both the spleen and lymph node of infected mice. The immune cells causing the effects observed in the systems tested seem to be T lymphocytes, as judged by the effect of anti-θ antiserum. Evidence is presented that two physically distinct populations of cells exist which can respond to MSV and show a net increase in total DNA or protein synthesis. However, only one of these cells seems able to release factors able to cause macrophage migration inhibition.
Immunologic Tolerance to a Hapten
Authors: Fidler John M. , Golub Edward S.
Published: December 2022

The induction of tolerance to the TNP hapten in populations of B cells in various states of differentiation was studied with the free, reactive hapten trinitrobenzene-sulfonic acid (TNBS). An adoptive transfer system was used in which tolerance was induced in an adoptively transferred B cell source before the addition of reconstituting thymus cells and immunization with hapten-carrier. Bone marrow cells and cells from fetal liver were rendered tolerant to TNP in this manner, showing that tolerance can be induced in cells from the primary B cell precursor and stem cell source of both the adult and fetal mouse. Tolerance was induced in normal spleen cells after adoptive transfer and in spleen cells from ATXBM mice (B cell spleens) after transfer to ATX hosts. These experiments show the induction of tolerance to TNP in B cells in another state of differentiation (peripheral B cells). Tolerance could be induced in B cell sources depleted of T cells indicating that immunologic tolerance to TNP may be induced in B cells in the absence of T cells. The possibility of residual TNBS affecting the thymus cells used for reconstitution was ruled out by transferring fetal liver cells to a secondary host after tolerance induction in a primary adoptive host, and also by rendering ATXBM mice tolerant before the transfer of their spleen cells to an adoptive host. These maneuvers remove the B cells from the environment used for tolerance induction, and indicate that TNBS did not act upon reconstituting thymus cells. The data indicate that tolerance to TNP can be induced in early differentiation states of the B cell.
Reversal of Tumor-Mediated Suppression of Immune Reactivity by in Vitro Incubation of Spleen Cells
Authors: Laux David , Lausch Robert N.
Published: December 2022

The cellular immune response of hamsters to PARA-7 tumor isografts was investigated with the 51Cr-release assay. It was found that spleen cells obtained from tumor-bearing animals were not cytotoxic for PARA-7 target cells on direct testing. However, after overnight incubation in vitro the cells became specifically reactive. Activation was temperature dependent occurring at 37°C but not at 4°C.

Spleen cells from SV40-immunized animals behaved differently. They produced significant cytotoxicity without preincubation. if the virus-immunized hamsters were given a tumor graft then incubation was required. Conversely, if the graft was excised from tumor-bearing animals 3 days before spleen cell harvest, an immune response was obtained on direct testing. Spleen cells pretreated with sera from PARA-7 tumor bearers were significantly less cytotoxic than cells exposed to normal or heterologous tumor bearer sera.

These results indicate that the tumor, or more likely its by-products, can inhibit the activity of sensitized spleen cells, and that this phenomenon can be reversed by incubation of the cells in a tumor-free environment.
Isolation of IgE from Reaginic Rat Serum
Authors: Isersky Chaviva , Kulczycki Anthony , Metzger Henry
Published: December 2022

IgE from the serum of rats immunized with Nippostrongylus brasiliensis was substantially purified. As measured by passive cutaneous anaphylaxis (PCA) activity an overall yield of 45% was reproducibly achieved by a combination of ammonium sulfate precipitation, gel and ion-exchange chromatography, and immunoadsorbents. The iodinated final product was 80% precipitable by specific antisera to IgE, and in radio-immunoelectrophoresis with antiserum to light chains gave only a single line corresponding to the line given by antisera to IgE. No lines could be detected with an antiserum to normal serum proteins devoid of antibodies to light chains. About 80% of the IgE related counts were shown to bind specifically to neoplastic rat basophils. On the basis of these results it was calculated that 18 ng of rat IgE are adequate to give a detectable PCA response.
Evidence for T Lymphocyte Subpopulations in the Rabbit
Authors: Fanger Michael W. , Reese Andy C. , Schoenberg Melvin D. , Stavitsky Abram B. , Reese Alice L.
Published: December 2022

A goat antibody to rabbit thymus cells (ATG) was used to study T lymphocyte distributions and subpopulations in the rabbit. ATG inhibited transformation of peripheral blood lymphocytes stimulated by phytohemagglutinin (PHA) and concanavalin A (Con A) but not by goat anti-rabbit Fab. The distribution of T lymphocytes was determined by the ability of lymphocytes from the various tissues to absorb this inhibitory activity. Lymph node, peripheral blood, and thymus lymphocytes, in that order, were most capable of absorbing the ATG activity. Fluorescein-conjugated ATG caused fluorescence of 100% of thymus cells but lower percentages of lymphocytes from other organs. In addition, the intensity of staining by fluorescent antibody varied among lymphocytes even from the same organ. The results indicate the presence of rabbit T cell subpopulations which differ in the quantity of T antigen per cell. The effect of ATG on mitogen stimulation of lymphocytes from the various lymphoid organs indicated that the T cell subsets differed in their response to PHA and Con A, and in their susceptibility to the inhibitory effect of ATG. Evidence is also presented which suggests the presence of lymphocytes which possess both surface T antigen and Ig determinants.
In Vitro Demonstration of Complement-Dependent Cytotoxic Antibodies to Moloney Sarcoma Cells
Authors: Tamerius John D. , Hellström Ingegerd
Published: December 2022

An in vitro microcytotoxicity assay was used to demonstrate complement-dependent cytotoxic antibodies to Moloney sarcoma virus-induced cells throughout the stages of tumor progression and regression in BALB/c mice. Complement-dependent cytotoxic antibodies were present in the sera of mice whose tumors were in the late stages of tumor growth, and most consistently in the sera of mice whose tumors had completely regressed (regressors). Regressor sera were strongly cytotoxic for at least 125 days after the onset of tumor regression. Preliminary evidence indicates that the complement-dependent cytotoxic antibody in regressor sera is a 7S immunoglobulin.
Regulation of the Antibody Response to Type III Pneumococcal Polysaccharide
Authors: Baker Phillip J. , Stashak Philip W. , Amsbaugh Diana F. , Prescott Benjamin
Published: December 2022

Prior treatment with a marginally immunogenic dose of Type III pneumococcal polysaccharide (SSS-III) resulted in the development of an unresponsive state. Such low-dose paralysis, which is antigen-specific, could not be induced in athymic nude mice and could be abrogated by treatment with anti-lymphocyte serum (ALS), a procedure reported to inactivate thymic-derived suppressor cells (suppressor T cells). These findings provide support for the view that low-dose paralysis to SSS-III is a T cell-dependent phenomenon in which suppressor T cells, activated by low doses of antigen, play a major role.
Cellular Basis of Immunodepression in Mice with Plasmacytomas
Authors: Zolla Susan , Naor David , Tanapatchaiyapong Prasert
Published: December 2022

The primary immune response to sheep red cells (SRC) in plasmacytoma (PC)-bearing mice is consistently depressed; the degree of depression is dependent on tumor size and the dose of antigen administered. Mice bearing the two PC which most severely depress the primary response also have an impaired ability to mount a secondary response to SRC. The primary response of PC-bearing mice to a thymus-independent antigen is also depressed, indicating that B cells are at least one of the cell populations adversely affected in tumor-bearing hosts. However, the B cell precursors of antibody-forming cells, antigen-binding cells, are found in normal quantities and appear capable of binding antigen. The data also suggest that the PC which cause the most severe immunodepression affect the maturation of the precursors of antibody-forming cells whereas those tumors that have a significant but less marked effect on antibody production do not appear to alter the process of differentiation in antigen-triggered cells.
Interaction of Rheumatoid Factor with Infectious Vaccinia Virus-Antibody Complexes
Authors: Gipson Thomas G. , Daniels Charles A. , Notkins Abner L.
Published: December 2022

Rheumatoid factor (RF), isolated from sera of patients with rheumatoid arthritis, attached to infectious vaccinia virus-IgG (Vac-IgG) complexes. Neutralization did not occur until the Vac-IgG-RF complexes were incubated with either goat antihuman IgM or complement. The amount of RF that attached was proportional to the concentration of IgG incubated with the virus. The enhancement by RF of complement-mediated neutralization of Vac-IgG was not dependent upon the order of addition of the immunoreactants. Complement addition before, with, or after RF resulted in the same amount of neutralization. These observations suggest that under certain conditions RF may play a role in the defense against viral infections.
Synergistic Cooperation between Isoantiserum and Immune Lymphoid Cells: In Vitro Studies with a Syngeneic Rat Lymphoma
Authors: de Landazuri M. Ortiz , Kedar E. , Fahey J. L.
Published: December 2022

The immune response of W/Fu rats to a syngeneic tumor was notable for the absence of serum-blocking activity and the presence of serum potentiation of cellular cytotoxicity (synergistic effect).

Heat-inactivated isoimmune serum potentiated the in vitro cell-mediated cytotoxicity (CMC) of lymphoid cells immune to a syngeneic Gross virus-induced lymphoma (C58NT)D in W/Fu rats. This isoimmune serum contains antibodies capable of mediating antibody-dependent cellular cytotoxicity (ADCC) in the presence of normal rat lymphoid cells. The synergistic effect observed demonstrated greater cytotoxicity than the sum of CMC and ADCC measured independently.

Inoculation of W/Fu rats with (C58NT)D tumor cells resulted in a 2- to 3-fold increase (compared to normal lymphoid cells) in the ADCC activity of immune spleen and peripheral blood cells. This was true for both the specific target cell (C58NT)D antibody complex and an unrelated target cell antibody complex. The kinetics of this increase in ADCC activity parallels the development of CMC activity as measured in vitro.

The synergistic effect is primarily attributable to an increased capacity of immune lymphoid cells to lyse antibody-coated target cells. The concomitant increase in Fc receptor-bearing cells (non-T cells) may contribute to this effect.
Basophil Activation by Concanavalin a: Characteristics of the Reaction
Authors: Siraganian Patricia A. , Siraganian Reuben P.
Published: December 2022

Concanavalin A (Con A) induced histamine release from the basophils or mast cells of several mammalian species including human, rabbit guinea pig, mouse, and rat. The mechanism was investigated in detail in humans. Wide variation in the response of the basophil of different individuals was found. The optimum release was produced with 1 to 3 µg/ml of Con A. The response is characterized by inhibition by excess amounts of Con A and has great similarity to antigen activation of basophils. The release of histamine by Con A requires Ca2+, is modulated by the level of cyclic AMP, is inhibited by chlorphenesin and diethylcarbamazine and enhanced by deuterium oxide. The activation of the basophil depends on the binding of Con A to pyranosyl sugar residues on the cell membrane.
Fluorescent Antibody Responses to Chlamydial Infection in Patients with Lymphogranuloma Venereum and Urethritis
Authors: Philip Robert N. , Casper Elizabeth A. , Gordon Francis B. , Quan Alice L.
Published: December 2022

Indirect immunofluorescence (the micro IF test) was used to study trachomainclusion conjunctivitis and lymphogranuloma venereum (LGV) serum antibody patterns in a venereal disease clinic population. Antibodies undifferentiated as to immunoglobulin class, but probably IgG, were detected in 80% of the study participants and titers were particularly high in LGV patients. Two kinds of antibody were observed. One was broadly reactive to all test antigens, and one was specifically directed toward strains of a particular antigenic type. Although most sera were broadly reactive, 40% also showed some type specificity. Specific antibody patterns in persons from whom chlamydiae were isolated usually corresponded to the immunotypes of isolates, and LGV reaction patterns were particularly associated with LGV patients. IgM antibodies were observed in 20% of the study participants. These were usually type specific and perhaps indicated current infection. The sensitive micro IF test is considered useful in the epidemiologic study of chlamydial infections of genital origin and can provide information about the nature of infecting strains.
The Allogeneic Effect: Increased Cellular Immune and Inflammatory Responses
Authors: Elfenbein Gerald J. , Green Ira , Paul William E.
Published: December 2022

It has been previously shown that infusion of allogeneic lymphoid cells into primed inbred guinea pigs produces an augmentation of humoral immune responses. In this communication we have investigated the effects of such treatment on host cellular immune and inflammatory responses. Dinitrophenyl (DNP) ovalbumin- or DNP-bovine serum albumin-primed strain 2 guinea pigs received infusions of strain 13 guinea pig lymphoid cells. Six days later the recipients were challenged with the priming antigen intradermally. These “allogeneic” guinea pigs displayed larger delayed hypersensitivity skin reactions than did “control” guinea pigs which had been primed but had not received allogeneic lymphoid cells. The in vitro migration of peritoneal exudate cells from allogeneic guinea pigs was inhibited by antigen to a greater extent than was the migration of control peritoneal exudate cells. In addition, allogeneic guinea pigs displayed larger Arthus reactions than did control guinea pigs at a time when both the total serum and the IgG2 immunoglobulin DNP-lysine binding capacity were virtually identical for allogeneic and control guinea pigs. Also, both passive Arthus reactions and croton oil skin responses were greater in allogeneic guinea pigs. Thus, the transient graft-vs-host reaction induces both enhanced host thymus-derived lymphocyte function and inflammatory responses.
Release of 51Cr-Endotoxin from Bacteria as an Assay of Serum Bactericidal Activity
Authors: Fierer Joshua , Finley Fred , Braude Abraham I.
Published: December 2022

In order to facilitate the study of normal serum on Gram-negative bacteria we have developed a bactericidal assay based on the release of 51Cr from labeled bacteria. Release of radioactivity was shown to be mediated by complement and paralleled the bactericidal action of sera measured by change in colony-forming units. Serum from the C6 deficient rabbits and serum that had been treated to inactivate complement did not release 51Cr. The rate of release of 51 Cr was decreased by absorption of serum with Bentonite but the maximum percentage released was not decreased. 51Cr was not released from serum resistant bacteria.

The 51Cr appears to be associated with the lipopolysaccharide fraction of the bacteria. Since this structure is the target for the bactericidal reaction, the release of 51Cr is a direct measure of the effect of complement on the bacteria. Under most conditions this reaction leads to cell death so that the release of 51Cr can be used as an assay of bactericidal activity. It was shown that the assay was feasible with Escherichia coli, Salmonella and Hemophilus influenza B.
Cytotoxic Immune Cells with Specificity for Defined Soluble Antigens
Authors: Schirrmacher Volker , Rubin Bent , Pross Hugh
Published: December 2022

Antigen-coated target cells were specifically lysed (a) by spleen cells from mice that had been immunized against the antigen presented in a soluble form and (b) by non-immune mouse spleen cells in the presence of specific antibodies. In the immune cell cytotoxic system, which consists of antibody-producing cells (APC) and cytotoxic effector cells (CEC), limiting dilution assays showed that the APC were diluted out before the CEC so that the amount of antibody being produced in vitro was limiting the cytotoxicity. In the presence of optimal concentrations of antibody, CEC from immune and non-immune cells were equally effective, regardless of whether the immune cells and the antibody were of the same or of different specificity.

In vitro cytotoxicity in the immune spleen cell system started after a lag period of about 3 hr and was depressed to about 50% after the cells had been treated with trypsin. The cytotoxic activity of non-immune spleen cells in the presence of antibody started without a lag period and was not affected by trypsin treatment of the cells.

Both the immune and the non-immune cell cytotoxic system were blocked by unrelated antigen-antibody complexes. Antibody-complexed target cells were much more potent inhibitors than were soluble complexes. Small numbers of target cells heavily coated with antibody were as efficient in blocking as were larger numbers of target cells lightly coated with antibody. The inhibition by antibodycomplexed antigen-coated target cells was neutralized by the addition of soluble antigen. This “unblocking” was thought to be due to a competition between soluble and insoluble antigen for antibody and the creation of non-inhibitory soluble complexes. The relevance of these findings for anti-tumor immune systems is discussed.
The IgM Antibody Site
Authors: Oriol Rafael , Rousset Monique
Published: December 2022

Two different populations of sites on IgM have been described. The high affinity population of 19S antibody sites is independent of antibody concentration, as can be expected for the interaction of a single antibody site with a monovalent hapten. The low affinity population of 19S antibody sites reacts with the hapten in a different way. The measurements made at low antibody concentration give higher apparent binding affinities than those made at high antibody concentration. A positive correlation can be established between the amount of free hapten needed to saturate the low affinity population of sites and the total amount of antibody sites available.

Based on these data and taking into account other authors results, we propose a model which may explain the controversial facts reported concerning the IgM valence and the maturation of its binding affinity.
Abnormalities in the Mixed Leukocyte Reaction During Infectious Mononucleosis
Authors: Twomey Jeremiah J.
Published: December 2022

Lymphocytes obtained from patients during the lymphoproliferative phase of infectious mononucleosis were hyporesponsive in the mixed leukocyte reaction. Lymphocyte hypore-sponsiveness was only demonstrated on patients with significant serum titers of heterophil antibody. Blood from two granulocytopenic patients obtained shortly after the onset of symptoms contained reduced concentrations of monocytes; this monocytopenic blood provided insufficient macrophage precursors to mediate a mixed leukocyte reaction. Both in vitro abnormalities were of transient duration.
IgM-Forming Cells as the Immediate Precursor of IgA-Producing Cells during Ontogeny of the Immunoglobulin-Producing System of the Chicken
Authors: Martin Louis N. , Leslie Gerrie A.
Published: December 2022

Chickens bursećtomized at hatch and treated with either anti-α or anti-µ had depressed serum levels of both IgM and IgA, whereas IgY production was unaffected. Chickens bursectomized at hatch and treated with anti-α + anti-µ combined had more severely depressed levels of serum IgM and IgA than bursectomized chickens treated with either antibody alone. The suppressive effects of anti-µ on IgA, anti-α on IgM, and the the additive suppressive effect of combined antibody treatment on both IgM and IgA suggest that IgA-producing cells are derived from IgM-forming precursor cells through a doubly-producing intermediate. The lack of suppressive effect on IgY production in birds having severe depression of IgM and IgA indicates that few if any IgY-forming cells serve as precursors for IgA-producing cells.
The Splenic Suppressor Cell
Authors: Folch Hogo , Waksman Byron H.
Published: December 2022

3H-thymidine incorporation was measured in mixed lymphocyte cultures of allogeneic rat spleen cells, the stimulating (donor) cells having been treated with mitomycin C (MC). Removal of adherent cells from either stimulating or reactive cells, or both, by passage through a glass wool column gave a substantial elevation of the response. The effect of such treatment of the stimulating cells could be duplicated by treating them with cycloheximide or by use of cells from doubly thymus-deprived donors. MC treatment, on the other hand, did not affect their supporessor activity. It was inferred that a subpopulation of splenic T cells adherent to glass wool act as suppressors in this immunologic system and that their action requires protein synthesis but not cell division. A close similarity was noted between these suppressor cells and those active in earlier experiments with PHA and concanavalin A and with suppressor cells studied in intact animals subjected to the graft-vs-host reaction.

A general classification of suppressor cell systems is proposed.
Dissociation and Recombination of the Subunits of the Cholera Enterotoxin (Choleragen)
Authors: Finkelstein Richard A. , Boesman Mary , Neoh Sim Hee , LaRue Michael K. , Delaney Robert
Published: December 2022

By gel filtration under dissociating conditions in acid urea, the cholera enterotoxin (choleragen) is separated into two fractions, A and B. Fraction A, which elutes first from columns of Bio-Gel P-60, is unique to the toxin and is not found in the spontaneously formed enterotoxoid (choleragenoid) after similar treatment. Neither A nor B alone has significant toxicity in either the infant rabbit model or adult rabbit skin tests. However, after neutralization and dialysis of mixtures of A and B (regardless of whether the latter is derived from choleragen or choleragenoid) the toxin activity in both systems is restored as is the characteristic electrophoretic mobility of the original toxin. Antiserum against the holotoxin precipitates both fragments but A and B give reactions of non-identity. Antiserum against choleragenoid does not precipitate A. Peptide mapping indicates that choleragen contains a number of peptides not present in choleragenoid. The A fragment can further be dissociated into two parts by treatment with thiol reagents. It now seems evident that the cholera enterotoxin consists of two major kinds of non-covalently associated subunits. Of these, the B type, comprising choleragenoid, is likely to be responsible for interaction with GM1 ganglioside-like host cell receptors and the A fragment for biologic activity, i.e., the activation of adenyl cyclase.
The Correlation of Serum IgD Concentration with Gm Allotype
Authors: Walzer Peter D. , Kunkel Henry G.
Published: December 2022

Serum IgD concentrations were measured in normal Caucasian blood donors homozygous for either the Gm(a) Gm(g) or the Gm(f) Gm(b) linkage groups. IgD levels were measured by hemagglutination inhibition, single radial diffusion, and solid phase radioimmunoassay. The Gm(f) Gm(b) group had a significantly lower serum IgD level than did the Gm(a) Gm(g) group by all three methods of quantitation. The difference between the two groups was due primarily to the high frequency of low serum IgD levels among Gm(f) Gm(b) homozygotes. Although no genetic markers are available in IgD, the correlation with allotype markers of the IgG class indicates close linkage between IgD and IgG heavy chain C-region genes.