bluuebunny/crossref_metadata_2025 huggingface dataset:
Role of Intracellular and Extracellular Cationic Composition in Immune Lysis of Mammalian Erythrocytes
Authors: Dalmasso Agustin P. , Giavedoni Elda B. , Lelchuk Rosalia , de Bracco Maria M. E.
Published: December 2022

Immune lysis of human, rat, and sheep erythrocytes (E) containing high K and low Na (HK) was markedly influenced by the alkali metal ions present in the reaction mixture. The degree of lysis in the various cations conforms to the series K > Rb > Li > Cs > Na or to a slightly different one. Immune lysis of dog, cat, and sheep E containing low K and high Na (LK) was unaffected or only slightly affected by changes in the alkali metal ions present in the reaction system.

Human E which were converted artificially into LK E had a larger degree of immune lysis in Na buffer than the original HK E, approaching the degree of lysis obtained in K buffer. This suggests that the intracellular cationic composition of naturally occurring HK E is essential for their different reactivity with complement (C) in media with various alkali metal cations. In contrast, experimental conversion of sheep LK E into HK E caused no modification of their initial reactivity with C in Na and K media. This indicates that the membranes of naturally occurring LK E react optimally with C independently of the intra- and extracellular cationic composition.

Previous work demonstrated that immune lysis in Na buffer could be enhanced by treatment of the cells with K before the C reaction. The degree of C lysis in Na buffer of the sheep HK E examined in the present study, however, was not enhanced by preliminary treatment with K buffer. Therefore, it appears that this assay permits differentiation of two types of sheep HK E. Human E failed to respond with enhancement of immune lysis in Na buffer after pretreatment with K buffer. Ouabain treatment of HK sheep E caused no modification in the degree of C lysis obtained in either Na or K buffer.
Immunogenicity and Arthritogenicity in the Rat of an Antigen from Mycobacterium Tuberculosis Wax D
Authors: Koga Toshitaka , Pearson Carl M.
Published: December 2022

Most of the conventional wax D preparations from human strain Mycobacterium tuberculosis induced adjuvant arthritis (AA) and typical delayed skin reaction to purified protein derivative (PPD) in the rat. However, the tuberculin-sensitizing ability of these waxes D as judged by skin testing with PPD was variable from preparation to preparation. “Pure” wax D containing trace amounts of tuberculin-sensitizing materials has recently been extracted from a human strain M. tuberculosis H37Ra, cultivated on Sauton medium for 4 weeks. The water-soluble portion (WSP) subfractions designated as F-IA and F-IB were obtained from this pure wax D. These subfractions, which are homogeneous as revealed by Ouchterlony tests and immunoelectrophoresis, were capable of inducing AA and a delayed hypersensitivity elicited by themselves, but not by PPD. PPD and π, which were less immunogenic than F-IA and F-IB, failed to produce AA. Arthritogenicity of the parent wax D correlated more closely with the degree of skin reaction to the WSP subfractions than with the PPD skin reaction. However, after development of AA the skin reaction to F-IB decreased significantly, especially in severely diseased rats, whereas PPD positivity remained constant and strong.

Although Ouchterlony double diffusion tests with F-IA, F-IB, PPD and π revealed that an identical or partially identical component is present in all, F-IA and F-IB are peptidopolysaccharides and differ in their immunologic behavior from the tuberculoproteins, PPD and π. Evidence of two different delayed-type antigens in conventional wax D as opposed to one in pure wax D was revealed in Ouchterlony tests.
Subpopulations of Human Lymphocytes Defined by β2-Microglobulin
Authors: Fanger Michael W. , Bernier George M.
Published: December 2022

A human serum protein with structural homology to immunoglobulins, β2-microglobulin, was synthesized by normal lymphocytes in vitro. Production of microglobulin was increased when lymphocytes were stimulated with phytohemagglutinin (PHA) or concanavalin A (Con A) but was not increased by stimulation with pokeweed mitogen, suggesting that microglobulin was elaborated by thymus-dependent lymphocytes. Maximal β2-microglobulin production occurred at a concentration of PHA (2 µg/106 cells) which was far lower than that which led to maximal DNA and protein synthesis. Treatment of normal lymphocytes in culture with anti-β2-microglobulin and complement removed 20% of the cells and abolished β2-microglobulin production while stimulation of DNA synthesis by PHA was only slightly decreased. Fluorescein coupled antibody to β2-microglobulin stained virtually all normal lymphocytes, but 15 to 20% stained brightly. Although 15 to 25% of normal lymphocytes stained with anti-light chain antibody, none of these cells stained brightly with antibody to microglobulin when double-staining experiments were performed with two fluorochromes. Lymphocytes from a patient with acquired agammaglobulinemia synthesized microglobulin at a normal rate although the thymus independent (B) cell population as determined by staining for immunoglobulin was less than 3%. On the other hand, synthesis of microglobulin by lymphocytes from four patients with stage IV Hodgkin's disease was minimal. Production of β2-microglobulin by peripheral lymphocytes of patients with chronic lymphatic leukemia (CLL) was low when the lymphocytes were identified as B-cells; synthesis was normal, however, by lymphocytes lacking surface immunoglobulin in one case of CLL. These results suggest that in man β2-microglobulin is in large part the product of a thymic dependent lymphocyte subpopulation.
In Vitro Assessment of Cellular Immunity to Rauscher Leukemia Virus
Authors: Mortensen Richard F. , Ceglowski Walter S. , Friedman Herman
Published: December 2022

Cell-mediated immunity to Rauscher leukemia virus (RLV) was studied utilizing the macrophage migration inhibition assay. Cell-mediated immunity to purified RLV antigen was detected in splenic lymphocytes of the leukemia resistant C57BL/6 mouse strain inoculated with infectious RLV. In contrast, no cellular immunity to RLV could be detected in splenic lymphocytes of the leukemia susceptible BALB/c mouse strain upon infection with RLV. Both strains of mice exhibited an active cell-mediated immune response following inoculation with heat-inactivated RLV. Appropropriate controls ruled out a contribution of either antibody or antibody-antigen complexes as mediators of the observed inhibition of macrophage migration. These data support the view that an inhibition of cell-mediated immune function may be a prerequisite to the induction of viral leukemia.
Purification of the Human Complement Protein Clq by Affinity Chromatography
Authors: Sledge Carlos R. , Bing David H.
Published: December 2022

A subunit of the first component of human complement C1q, was purified by the technique of affinity chromatography. The chromatographic resin was cyanogen bromide-activated Sepharose covalently linked to human IgG. For the removal of traces of IgM it was necessary to subject further the C1q obtained from the chromatographic step to ultracentrifugation in sucrose gradients. The highly purified C1q was characterized immunochemically and according to its electrophoretic mobility in polyacrylamide gel. The purified material was capable of combining with C1r and C1s to reconstitute fully active macromolecular C1.
Alterations in Lymphocyte Populations in Tumorigenesis
Authors: Zatz Marion M. , White Abraham , Goldstein Allan L.
Published: December 2022

Studies have been conducted of the migration of 51Cr-labeled lymph node cells into the lymphoid tissues of mice inoculated with the Moloney sarcoma virus. Significant numbers of labeled cells were trapped in the draining lymph nodes and spleens of tumor-bearing mice. The kinetics of lymphocyte trapping correlated with the cycle of growth and spontaneous regression which is characteristic of this tumor in healthy mice. Trapping first occurred after injection of the virus and maximal retention of lymphocytes corresponded to the time of tumor regression.
Cellular Events in Tolerance
Authors: Scott David W.
Published: December 2022

Lymph node cultures from Lewis rats rendered unresponsive to ultracentrifuged sheep immunoglobulins (soluble ShIg or ShIgSo) were examined at various times after exposure to tolerogen in vivo to determine if activation of macromolecular synthesis occurred during tolerance induction. Cells from ShIgSo treated animals could not be stimulated by antigen to incorporate precursors of DNA, RNA, or protein. “Tolerant” cells could not be stimulated over a 10,000-fold dose range in vitro and did not show early or delayed DNA synthesis in vitro. In contrast, cells from rats injected with ShIg in adjuvant showed dose-dependent stimulation at all times examined. In addition, thoracic duct lymphocytes from tolerant rats were not “activated” in terms of their ability to support the replication of vesicular stomatitis virus. These results suggest that tolerance induction to this soluble protein does not proceed through the active steps in the immune response which are readily detected after the injection of an immunogen.

Furthermore, the in vitro proliferation of ShIg-immune lymph node cells, stimulated with ShIg, occurred whether these cells were cultured with normal rat serum or serum from rats tolerant to ShIg. In addition, co-cultivation of lymph node cells from tolerant animals with immune lymphocytes did not lead to suppression of the responsiveness of the latter cells in vitro. These results suggest that the induction and maintenance of tolerance to ShIg occur through mechanisms involving neither blocking factors nor suppressor cells. Multiple pathways of tolerance to different types of antigens are proposed.
Antigenicity of Formaldehyde-Inactivated Staphylococcal Enterotoxin B
Authors: Warren John R. , Spero Leonard , Metzger Joseph F.
Published: December 2022

The reactivity toward or ability to stimulate production of antibodies specific for native staphylococcal enterotoxin B (nSEB) demonstrated by formaldehyde-inactivated toxin (fSEB) is directly related to the extent to which SEB is polymerized by aqueous formaldehyde. Combined use of polyacrylamide gel electrophoresis in sodium dodecyl sulfate and agarose gel chromatography revealed that: 1) 30-days incubation of SEB in 1% formaldehyde solution buffered to pH 5.0 or 7.5 resulted in a series of small to very large SEB polymers, while SEB incubated at pH 9.5 formed heterogeneous monomers and a small amount of dimer; 2) SEB was polymerized to maximal extent in pH 7.5 formaldehyde. Polymeric fSEB obtained with neutral or acid formaldehyde retained the capacity to precipitate quantitatively nSEB antibodies from rabbit antiserum and to induce the formation of nSEB hemagglutinins in rhesus monkeys. In contrast, the nonpolymerized fSEB failed to react with, or stimulate production of nSEB antibodies. Furthermore, the relative in vitro antibody-combining activities of the two polymeric fSEB products were inversely related to in vivo antibody production. Thus, although quantitative precipitation experiments revealed that 50% of the native antigenic determinants were retained after polymerization in acid aqueous formaldehyde (5.0-fSEB), while only 15% were preserved upon polymerization in neutral formaldehyde solution (7.5-fSEB), hemagglutinin responses of rhesus monkeys were generally several fold greater after parenteral injection of 7.5-fSEB than of 5.0-fSEB. The greater antibody response to 7.5-fSEB may be explained by its more extensive polymerization; the greater average molecular size of the 7.5-fSEB antigenic complex may permit more efficient processing by macrophages or provide a more effective presentation of reactive sites for attachment with, or activation of, immunocompetent cells.
Graft-vs-Host Reaction with an H-2 Mutant
Authors: Klein J. , Egorov I. K.
Published: December 2022

GVHR can be induced across the H-2D locus, presumably uncontaminated by any other genetic differences (Ir); it can also be induced across differences that do not have any histocompatibility effect on skin grafts. This is interpreted as evidence for two types of genes in the H-2 complex, H-genes involved in both GVHR and skin graft rejection, and Gvh-genes involved in GVHR but not in skin graft rejection.
Thymus Function in Spontaneous Lymphoid Leukemia
Authors: Nagaya Hiroshi
Published: December 2022

Lymphoid cells from normal, “preleukemic” and leukemic AKR mice were cultured in vitro to evaluate their response to mitogens, concanavalin A (Con A) and phytohemagglutinin (PHA).

Normal thymus cells were stimulated by Con A to increase their 3H-thymidine uptake 455-fold (median) whereas the thymidine uptake by autologous leukemic thymus cells was increased only 5-fold by Con A, the latter response resembling that of normal bone marrow cells. The median increases in thymidine uptake by “preleukemic” 4- and 5-month-old thymus cells stimulated by Con A were 280-fold and 269-fold, respectively. Failure in the thymic function to convert Con A-unresponsive bone marrow precursor cells to Con A-responsive normal thymus cells appears to precede leukemogenesis in the thymus of AKR mice.
Factors Involved in the Cytotoxicity of Normal Guinea Pig Serum for Cells of Murine Tumor TA3 Sublines Treated with Neuraminidase
Authors: Hughes R. Colin , Palmer Peter D. , Sanford Barbara H.
Published: December 2022

Treatment of two ascites sublines of the TA3 tumor cell with neuraminidase under conditions that removed 70 to 75% of the total cell surface sialic acid exposes receptors on the cell surface that react with cytotoxins in normal serum from unsensitized guinea pigs. Removal of comparable amounts of cell surface sialic acid from TA3 cells by trypsin in the form of sialoglycopeptides did not render the cells susceptible to lysis by the guinea pig serum. Treatment of neuraminidase-treated cells with trypsin under identical conditions reduced the susceptibility of the cells to lysis by only 35 to 40%. The requirement for neuraminidase appears to lie in the hydrolysis of glycoside linkages between sialic acid residues and penultimate sugars in cell surface glycoproteins and possibly glycolipids, with exposure of new terminal oligosaccharide sequences. Simple changes in the net surface charge or membrane fluidity of TA3 cells brought about by neuraminidase treatment were considered less likely to be responsible for the increased susceptibility of the treated cells to lysis. Hapten inhibition studies on the cytotoxic effects of normal guinea pig serum on neuraminidase-treated TA3 cells using glycoproteins with defined terminal oligosaccharide sequences have in general been unconvincing. However, glycoproteins containing terminal β-galactosyl residues showed weak but definite inhibitory activity. The presence of such residues at the surface of both sublines of the TA3 tumor cell was demonstrated by agglutination of the cells with a galactose-binding lectin from Ricinus communis.

The cytotoxicity of the normal guinea pig serum against neuraminidase-treated TA3 cells was concentrated in the 19S IgM fractions of guinea pig immunoglobulins and was labile to mild heat treatment and during storage at 4°C. No agglutination of neuraminidase-treated TA3 was found with guinea pig serum either at 37° or 4°C. The role of similar cytotoxins in C3H mouse serum in the transplantation properties of the strain-specific TA3-St and the nonspecific subline TA3-Ha and derivatives derived from them by neuraminidase treatment are briefly discussed.
Stimulation of γ-Globulin Synthesis and Specific Antibody Production by a Factor Released by Activated Lymphocytes
Authors: Rosenthal Moshe , Stastny Peter , Ziff Morris
Published: December 2022

By using a radioactive immune coprecipitation technique, supernatants of lymph node cells from immunized rabbits incubated with keyhole limpet hemocyanin (KLH) were shown to enhance γ-globulin synthesis and anti-KLH antibody production of KLH-primed and nonprimed lymphoid cells from the peripheral blood and lymph nodes in the presence of KLH in vitro. Similarly, when KLH-stimulated supernatants were added both to BSA-primed and nonprimed lymphocyte cultures in the presence of BSA, increases in γ-globulin and anti-BSA synthesis were observed. The data presented provide evidence that a soluble factor is released by antigen-stimulated lymphocytes which enhances immunoglobulin and specific antibody production in a nonspecific manner. The evidence obtained suggests that the factor may be a lymphokine which either directly or indirectly stimulates bone marrow-derived lymphocyte function in both the primary and secondary responses.
Immunoglobulin Biosynthesis by Normal and Leukemic Human Peripheral Blood Lymphocytes
Authors: Nies Kenneth M. , Oberlin M. Anne , Brown John C. , Halpern Michael S.
Published: December 2022

Short-term tissue cultures of peripheral blood lymphocytes isolated from normal individuals and patients with chronic lymphocytic leukemia (CLL) were endogenously labeled with radioactive amino acid and the intracellular and extracellular immunoglobulin isolated by immune precipitation. Even though individual cells from occasional CLL cultures expressed both γ- and µ-chains on their surface as determined by immunofluorescence, each culture in all cases studied synthesized either IgG or IgM but not both. By contrast, cultures of normal peripheral-blood lymphocytes synthesized both IgG and IgM. All normal cultures and certain CLL cultures released labeled immunoglobulin into supernatant fluids.
Spleen Cell Cytotoxicity in New Zealand Black Mice (NZB) with Autoimmune Disease
Authors: Liburd E. M. , Russell A. S. , Dossetor J. B.
Published: December 2022

New Zealand black mice (NZB)2 develop disease spontaneously which is similar in many ways to human systemic lupus erythematosis (SLE) and is characterized by autoantibodies, nephritis, (1, 2) and perivascular cellular infiltration. These mice have also shown a decreased capacity with ageing to reject tumors induced by Moloney sarcoma virus (3). Cantor and co-workers (4) have demonstrated that the capacity of spleen cells from NZB mice to mount a graft-vs-host reaction also decreases with age. Both of these observations indicate reduced efficacy of the thymus-dependent immune system or thymus-dependent lymphocytes (T cells), but do not show whether programmed T cells are responsible for some of the cellular autoimmunity in this disease.

We have investigated the ability of spleen cells from diseased NZB mice to elicit a cytotoxic effect against fetal target cells, in vitro, by a modified form of the colony inhibition, or lymphocyte cytotoxicity system (5, 6).
Specific Heterologous Enhancement of Immune Responses
Authors: Rubin Arnold S. , MacDonald A. Bruce , Coons Albert H.
Published: December 2022

In cultures of thymocytes or spleen cells from tetanus toxoid-primed mice, the addition of 1 ng of toxoid generated the release of a soluble factor which was capable of nonspecifically enhancing the in vitro immune response of normal mouse spleen cells against sheep erythrocytes. When active supernatants were fractionated by gel filtration on Sephadex G-150 and Bio-Gel P-150, the active material eluted with molecules of approximately 75,000 m.w. Column fractions from similarly-treated supernatants from cultures of normal thymocytes or spleen cells, maintained with or without tetanus toxoid, were inactive. The enhancing activity recovered from the column fractions was degraded by treatment with the enzyme protease. A factor present in the supernatant fluid of cultures containing equal numbers of histoincompatible (allogeneic) spleen cells had similar properties. The enhancing factor was further purified by a combination of ammonium sulfate fractionation, Sephadex gel filtration, and polyacrylamide gel electrophoresis.
Differentiation of Lymphoid Cells: Effect of Serum and Other Mitogenic Agents on the Selective Induction of Immunoglobulin M Secreting Lymph Node Cells in Tissue Culture
Authors: Zimmerman Daniel H. , Kern Milton
Published: December 2022

Rabbit lymph node cells incubated in a serum-free medium exhibit a relatively low level of incorporation of [3H]thymidine into DNA. Fetal calf serum stimulated this incorporation as much as 65-fold. On the other hand, rabbit serum from the same animal that was the source of the cells did not effectively support [3H]thymidine incorporation (only a 0- to 2.7-fold greater incorporation than serum-free medium) and such autologous serum was inhibitory when added in conjunction with fetal calf serum. This same behavior for both fetal calf serum and autologous serum was observed when the emergence of enhanced immunoglobulin M production was assessed rather than [3H]thymidine incorporation.

The emergence of enhanced immunoglobulin M production was markedly inhibited when either concanavalin A, phytohemagglutinin, or pokeweed mitogen was incubated with such cells in tissue culture. In contrast, bacterial endotoxin increased immunoglobulin M production about 2- to 4-fold. Concanavalin A did not inhibit the secretion of immunoglobulin M when added to cells which already exhibit enhanced immunoglobulin M production, i.e., at 48 hr in tissue culture. However, if concanavalin A was added at 24 hr or earlier its full inhibitory activity was manifested.
Immunoglobulin (Ig) Allotype Markers on Rabbit Lymphocytes: Separation of Cells Bearing Different Allotypes and Demonstration of the Binding of Ig to Lymphoid Cell Membranes
Authors: Jones Patricia P. , Cebra John J. , Herzenberg Leonard A.
Published: December 2022

Lymphocytes from peripheral blood and Peyer's patches of b5b9 and b4b5 rabbits were membrane stained with rhodamine- and fluorescein-labeled anti-allotype reagents. Up to 63% of peripheral blood lymphocytes and 15% of Peyer's patch lymphocytes were found to double stain for both allotypes on their membranes. With a fluorescence-activated electronic cell sorter, cell populations were fractionated according to membrane allotype. A double pass separation procedure after staining yielded four distinct cell populations: one each of cells single-stained for one of the two allotypes, one containing cells double-stained for both allotypes, and one of unstained cells. Properties of the cells bearing two allotypes were examined. Cells put into culture after they were stripped of their membrane immunoglobulin (Ig) with Pronase regenerated their membrane Ig; however, the proportion of cells bearing both allotypes was greatly reduced. This suggested that the lymphocytes were restricted to the synthesis of membrane Ig molecules of a single allotype at a given time. It was found that a high proportion of rabbit lymphocytes could bind exogenous Ig from serum in vitro and probably also in vivo, explaining the presence of molecules of both allotypes on individual cells.
Studies of Antibodies to Non-Type-Specific Antigens Associated with Streptococcal M Protein in the Sera of Patients with Rheumatic Fever
Authors: Beachey Edwin H. , Ofek Itzhak , Bisno Alan L.
Published: December 2022

Antibodies against the non-type-specific (NTSM) antigens in purified M protein preparations were found to be widely distributed in human sera but were consistently present, and in highest titer, in the sera of patients with acute rheumatic fever. These NTSM antibodies were measured by complement-fixation (CF) and passive hemagglutination (PHA) using as antigen purified M30, a serotype now rarely encountered. The sensitization of human blood group O erythrocytes for the PHA test afforded the opportunity to identify antigens in M protein preparations that have, like teichoic acid, an affinity for red cell membranes. Red blood cells were sensitized with M protein and with teichoic acid, respectively, and were compared as absorbents of serum NTSM antibodies. PHA antibodies were removed in reciprocal fashion by both sensitized erythrocyte preparations suggesting that the absorbed M protein contains teichoic acid. Either teichoic acid or M protein sensitized erythrocytes absorbed only a portion of the CF antibodies to M30 protein present in rheumatic fever sera. In contrast, streptococcal protoplast membranes absorbed virtually all of the CF antibody. These results show that purified M proteins of group A streptococci contain at least two NTSM antigens which are also present in streptococcal protoplast membranes and to which rheumatic fever patients are hyperimmune. One of these is teichoic acid or a closely related antigen.
Induction of Cell-Mediated Immunity and Tolerance to Homologous Collagen in Guinea Pigs: Demonstration of Antigen-Reactive Cells for a Self-Antigen
Authors: Senyk George , Michaeli Dov
Published: December 2022

Although normal guinea pigs exhibit no immunity against heterologous (human, mouse) and homologous collagens, cell-mediated immunity to these collagens can be induced in these animals after immunization with a heterologous (human) or the homologous (guinea pig) collagen in Freund's complete adjuvant. Furthermore, guinea pigs may be made specifically tolerant of their own collagen, and such tolerance is not broken by the subsequent induction of cellular immunity to a cross-reacting antigen, the human collagen. These results demonstrate the presence in guinea pigs of antigen-reactive cells capable of recognizing a self-antigen, with the possibility of subsequent induction of either immunity or tolerance, and argue against the assumption that self-tolerance is due to the absence of specific antigen-recognition cells.
Analysis of Antibody Populations to Oligo-Alanine Determinants
Authors: Kolb Hubert , Weiler Eberhardt
Published: December 2022

In order to characterize the variable (v-) regions of anti-oligo-D-alanine antibodies, we determined the velocity constant of the antibody-antigen-association reaction. This was done in an indirect hemolysis system by measuring the rate of the binding reaction between antibodies to oligo-D-alanine and their corresponding antigenic determinants coupled to sheep red cells. A strong correlation was found between the forward rate constant of IgG1- and IgG2a-anti-D-ala antibodies. In “fast” sera, antibodies of both subclasses were fast, in “slow” sera both were slow. It was concluded that in a given individual the v-regions of IgG1- and IgG2a-anti-D-ala antibodies are similar.
Role of Humoral and Cell-Mediated Immunity on the Recovery of Chickens from Fowlpox Virus Infection
Authors: Morita Chiharu
Published: December 2022

The influence of thymectomy and bursectomy in chickens on the recovery from fowlpox virus infection was examined. It was found that the virus titer in the lesions the 2nd week after infection was well correlated with the migration inhibition test of splenic cells. Healing time after the infection also was correlated with the migration inhibition test of splenic cells. These findings suggest that cell-mediated immunity plays a more important role in the recovery than does humoral immunity. However, the mechanism of humoral immunity was not excluded since the mortality was significantly higher in bursectomized-thymectomized chickens than in thymectomized chickens.
Cellular Site of Action of Various Adjuvants in Antibody Responses to Hapten-Carrier Conjugates
Authors: Hamaoka Toshiyuki , Katz David H.
Published: December 2022

The adjuvant action of concanavalin A (Con A), Escherichia coli lipopolysaccharide (LPS), beryllium sulphate (Be salt) and Bordetella pertussis vaccine have been analyzed in a system measuring humoral immune responses to hapten-carrier conjugates in mice. For the definition of the cellular locus of their action, the effect of these adjuvants on adoptive secondary anti-hapten antibody responses was studied. Spleen cells from 2,4-dinitrophenyl (DNP)-keyhole limpet hemocyanin (KLH) primed donors, which normally fail to develop adoptive secondary anti-DNP responses to a heterologous conjugate such as DNP-bovine γ-globulin (BGG), can be stimulated to do so when an appropriate dose of adjuvant is administered with DNP-BGG. The capacity for the adjuvants employed to exert this effect requires the presence of thymus-derived (T) lymphocytes, since depletion of such cells by treatment of the donor cell inoculum with anti-θ serum and complement in vitro before adoptive transfer abrogates the response to DNP-BGG plus adjuvant. Moreover, evidence is presented which demonstrates that these substances exert their adjuvant action through the small number of unimmunized BGG-specific T lymphocytes in the donor cell inoculum. This conclusion derives from the failure of adjuvants to augment adoptive secondary anti-DNP responses to the DNP derivative of a non-immunogenic copolymer of d-glutamic acid and d-lysine for which there are few or no specific, functional T cells.
Alternate Complement Pathway: Elevation of Cobra Venom-Induced Hemolytic Activity in Sera of Tumor-Bearing Mice
Authors: Brai Melchiorre , Osler Abraham G.
Published: December 2022

We have recently reported that sera of cancer patients possess greater lytic capacity for unsensitized erythrocytes after interaction with a cobra venom protein, CVF, (1). This antibody independent lytic process results from utilization of the terminal complement components (C3 to C9) through an alternate pathway that can be initiated by an enzymatically active complex formed by CVF, C3 proactivator (properdin factor B) and other serum constituents (2–4). A hemolytic assay for this reaction sequence has been described in (5).

For minimizing the number of variables inherent in studies of this type with sera of hospitalized patients, an experiment was designed to evaluate levels of CVF-mediated lytic activity in the sera of tumor-bearing mice. Toward this end, mice of both sexes (25 ± 2g), were inoculated subcutaneously with 0.5 mg of 3-methylcholanthrene (3MC) dissolved in sterile corn oil. The largest experimental group contained mice of the Swiss-Webster strain maintained as an outbred colony in this Institute.
Mapping the H-2-Linked Genes Governing, Respectively, the Immune Responses to a Glutamic Acid-Alanine-Tyrosine Copolymer and to Limiting Doses of Ovalbumin
Authors: Dunham Edward K. , Dorf Martin E. , Shreffler Donald C. , Benacerraf Baruj
Published: December 2022

Mice of inbred strains having known crossovers within the H-2 gene complex have been used to map the two H-2-linked immune response genes governing, respectively, responsiveness to a synthetic glutamic acid-alanine-tyrosine terpolymer and to limiting doses of ovalbumin. Both genes have been found to be within the H-2 complex between the K and D loci and have been further localized to the segment between the K and Ss-Slp loci.
Macrophage-Mediated Resistance to Intracellular Infection Induced by Poly-L-Glutamic Acid
Authors: Axline Stanton G. , Mendenhall Joy W. , Remington Jack S.
Published: December 2022

The ability of poly-L-glutamic acid (PLGA) to induce resistance to infection with an intracellular bacteria was examined. PLGA conferred significant protection in mice against subsequent challenge with an ordinarily lethal inoculum of Listeria monocytogenes. Enhanced resistance, as manifest by a decrease in overall mortality and prolongation of time to death, could be demonstrated for time periods up to 44 days after PLGA treatment. Peritoneal macrophages from PLGA-treated animals were resistant to in vitro challenge with Listeria. Such resistance could be demonstrated for time periods up to 60 days after PLGA treatment. These findings suggest that “activated” macrophages are the effector cells in the PLGA-induced resistance observed.
Effect of Concanavalin A on the Homing of Labeled T Lymphocytes
Authors: Gillette Ronald W. , McKenzie Gail O. , Swanson Margaret H.
Published: December 2022

Treatment in vitro with concanavalin A was found to alter markedly the distribution of 51Cr-labeled lymph node cells to the lymphoid organs of syngeneic recipient mice. The accumulation of labeled cells in recipient spleens was greatly increased at the expense of cell migration to the lymph nodes. This effect was abrogated by addition of a competitive inhibitor of concanavalin A. The data suggest that pyranoside groups on lymphocyte membranes are associated with the homing and recirculating properties of thymus derived cells.
Characterization of a Second Normal Antigen that Cross-Reacts with CEA
Authors: Burtin P. , Chavanel G. , Hirsch-Marie H.
Published: December 2022

The recent discovery of an antigen present in normal tissues that cross-reacts with the carcinoembryonic antigen of gastrointestinal tumors (CEA)3 was made in several laboratories (1, 2). This antigen received different names, including that of nonspecific cross-reacting antigen (NCA) given by von Kleist, Chavanel, and Burtin (2). A study of the perchlorosoluble antigens present on the stools of non-cancerous and cancerous patients (3) led us to identify another antigen that cross-reacts with CEA which later on was found in normal tissues. We propose to name it NCA 2.

Materials and Methods. The different categories of samples of stools and meconium were described in another paper (3). These samples were extracted volume/volume by 1 M perchloric acid, then precipitated with ammonium sulfate at 100% saturation, dialyzed and lyophilized.

These perchloric extracts, dissolved in deionized water, were precipitated by an equal volume of absolute ethanol.
Rate of Hemolytic Antibody Production by Single Cells in Vivo in Rabbits
Authors: Conrad Robert E. , Ingraham Joseph S.
Published: December 2022

Published estimates of the rate of synthesis and secretion of antibody molecules by antibody-forming tissues in vitro range from 0.5 to 5000 molecules per cell per second. In the present paper we describe an experimental approach to the estimation of the absolute rate of antibody secretion per cell by undisturbed antibody-forming cells in vivo.

There is evidence that most of the serum antibody appearing during the rapid rise to peak titer after a single intravenous (i. v.) injection of sheep erythrocytes into rabbits is made in the spleen. Plots of serum antibody titers typically exhibit an initial exponential rise followed by an arithmetic rise during which most of the antibody is produced and from which a constant rate of antibody production can be calculated. Rabbits immunized with a single i. v. injection of boiled sheep erythrocyte stromata were bled frequently, then sacrificed during the period of constant arithmetic antibody production and their spleen cells assayed by the liquid plaque assay technique to detect plaque-forming cells (PFC) making IgM hemolysin. Total splenic PFC were calculated based on spleen weights. The number of extrasplenic PFC was also taken into consideration and an allowance was made for PFC in the liver and blood. The rate of antibody production per milliliter in each animal was obtained from computer analysis of data for photometric 50% hemolysin units vs time and was converted to the rate of total hemolysin production based on total serum volume for each animal. This data, together with a literature value for molecules per RBC for 50% hemolysis, led to values for molecules of hemolytic antibody per PFC per second ranging from 8000 to 20,000 with a mean of 12,000 for seven rabbits. Our estimation of the protein-synthesizing machinery required for a cell to manufacture IgM antibody at this rate leads us to conclude that an average-sized plasma cell could do this if the messenger RNA is relatively stable or if gene expansion occurs to provide rapid transcription to maintain the level of messenger required.
Stimulation of Phagocytic Release of Neutral Protease from Human Neutrophils by Cholinergic Amines and Cyclic 3′,5′-Guanosine Monophosphate
Authors: Ignarro Louis J.
Published: December 2022

The purpose of this study was to elucidate the actions of cholinergic agents and cyclic 3′,5′-guanosine monophosphate (cyclic GMP) on phagocytic release of a lysosomal neutral protease from human neutrophils in the presence of aggregated IgG-rheumatoid factor complex. Neutrophils (5 × 106) and complex were incubated in a balanced salt solution, pH 7.4, at 37°C for 15 min in the absence and presence of various cholinergic agents and cyclic GMP analogs. Acetylcholine, acetyl β-methylcholine, carbamylcholine, and pilocarpine, but not choline, provoked a 74% to 94% increase in enzyme release at 10-7 M. Atropine, but not hexamethonium, blocked the accelerating action of acetylcholine on enzyme release. Cyclic GMP as well as three of its analogs also enhanced release of neutral protease from phagocytosing human neutrophils (71% to 136% increase at 10-7 M). Guanosine 5′-monophosphate or triphosphate did not provoke enzyme release and atropine did not block the effect of cyclic GMP. Neither the cholinergic agents nor cyclic GMP affected release of cytoplasmic lactate dehydrogenase, thus indicating maintenance of cell viability during lysosomal enzyme release. The data suggest that human neutrophils have muscarinic receptors that respond to cholinergic agents by acceleration of lysosomal enzyme release, perhaps via a mechanism involving intracellular cyclic GMP.
Properties of Rabbit Anti-Rat Epidermal Cell Sera. Demonstration of Tissue-Specific Differentiation Antigens
Authors: Lloyd Kenneth O. , Darnule Tukaram V.
Published: December 2022

Immunization of rabbits with rat tail epidermal cell suspensions produced antisera which, after absorption with rat red and spleen cells, showed specific reactivity with cell surface antigens of stratified squamous epithelia and their derived cells. Indirect immunofluorescence on tissue sections and a 51Cr-release cytotoxicity method on cell suspensions showed that the sera contained antibodies which were rat epidermal cell-specific and antibodies which cross-reacted with the corresponding cells and tissues of other species (mouse, guinea pig, dog, cat, and man). Using the indirect immunofluorescence technique, we showed individual sera to be specific for the basal layer or for all the cell layers of the epidermal epithelium. This specificity difference was not detected when trypsinized cell suspensions were examined. In epidermal cells isolated by a method entailing minimal trypsinization and by using EDTA to suspend the cells, the antigens were more mobile than in those prepared by the standard trypsinization method. The antiserum which reacted with all the epidermal layers showed many similarities to sera from patients with the autoimmune disease, pemphigus. In general, the specificity of these antisera may be considered as being directed toward cell surface, tissue-specific differentiation antigens of epidermal cells.
Autoantibodies in NZB Mice Detected by Lysis of DBA/2 Lymphoma Cells
Authors: Auer Ignaz O. , Tomasi Thomas B. , Milgrom Felix
Published: December 2022

Natural antibodies lytic for DBA/2 lymphoma cells, L5178Y, were shown in NZB mice by means of plaque assay demonstrating antibody-forming cells and spot test detecting serum antibodies. The antibodies were of the IgM class, resisted heating to 56°C for 30 min, and showed optimal binding activity at 4°C. They were efficiently removed by murine kidney and spleen cells and less so by thymocytes, brain, and liver cells. Since syngenic cells removed them as readily as allogenic cells, it could be concluded that the antibodies under investigation are autoantibodies. On the basis of physicochemical properties and because of distribution of their reactive antigens, the antibodies under study could be distinguished from previously described autoantibodies of NZB mice.

Lysins for L5178Y cells were also shown in BALB/c, DBA/2, and C3H mice. However, in these strains the optimal binding activity was at 37°C rather than at 4°C. The relation of these antibodies to those in NZB mice was not established by this study.
Preformed Eosinophil Chemotactic Factor of Anaphylaxis (ECF-A)
Authors: Wasserman Stephen I. , Goetzl Edward J. , Austen K. Frank
Published: December 2022

The eosinophil chemotactic factor of anaphylaxis (ECF-A), previously recognized as being released from sensitized human or guinea pig lung tissue by antigen challenge, has been demonstrated to exist preformed in these tissues and in isolated rat mast cells. The presence of performed ECF-A in the mast cell of the rat was established by the full recovery of ECF-A with mast cells purified from a mixed peritoneal cell population and cellular subfractionation which revealed the preformed ECF-A to be largely associated with the mast cell granules. The sum of the ECF-A released by antigen and that remaining in the sensitized challenged human or guinea pig lung tissue or isolated rat mast cells equalled that extractable from the equivalent unchallenged lung tissue or rat mast cells, indicating that the ECF-A available exists entirely in a preformed state.
Effect of Concanavalin a in Vivo in Suppressing the Antibody Response in Mice
Authors: Egan Harwood S. , Reeder Walter J. , Ekstedt Richard D.
Published: December 2022

Adult mice injected intraperitoneally with concanavalin A 2 days prior to primary immunization with a thymus-dependent, sheep erythrocyte antigen by the same route, showed a marked suppression of their immunoglobulin response as measured by a plaque-forming spleen cell assay. The effect was less pronounced or not demonstrable if the concanavalin A was injected at longer intervals before, at the same time as, or after antigen. A comparable immunosuppression was seen in the secondary response to sheep erythrocytes injected 10 days apart, provided the concanavalin A was injected 2 days prior to the second injection of antigen. Treatment with concanavalin A 2 days prior to the first injection of antigen had no effect upon the secondary response. It was also shown that concanavalin A treatment did not suppress the immunoglobulin response to a thymus-independent, Escherichia coli lipopolysaccharide antigen. The most striking immunosuppression was observed when both concanavalin A and antigen were injected by the same route, either intraperitoneally or intravenously.
Thymus-Dependent (T) Lymphocytes in the Rat
Authors: Balch Charles M. , Feldman Joseph D.
Published: December 2022

Thymus-dependent (T) lymphocytes in the rat may be recognized by their reactions with fluorescein-labeled heteroantibodies and by their functional activities. The Ig fractions of horse and rabbit antisera to rat thymocytes, when labeled with fluorescein, stained more than 95% thymocytes, less than 2% bone marrow cells, and 30 to 60% of the lymphocytes in peripheral blood, lymph nodes and spleen, and did not stain IgG-bearing lymphocytes. Cell-mediated cytotoxicity and stimulation by PHA and Con A were eliminated by these heteroantisera and C in vitro; survival of skin allografts was prolonged in vivo. T lymphocytes in the rat resembled, by analogy, T lymphocytes in the mouse.
The Effect of Anti-Plasma Cell Serum on B Cells
Authors: Nariuchi Hideo , Matuhasi Tyoku
Published: December 2022

Anti-plasma cell serum prepared against plasmacytoma 5563 was found to contain antibodies which are specific for surface antigens of lymph node cells bearing a receptor for C3. When lymph node cells were exposed to the antiserum and guinea pig complement prior to assay, a large number of the cells lost their capacity to bind the sheep red blood cells coated with IgM antibody and complement. This antiserum did not affect the cells bearing the receptor which requires Mg++ for the attachment of C3.
Cyclic AMP Content and Histamine Release in Rat Mast Cells
Authors: Johnson Alice R. , Moran Neil C. , Mayer Steven E.
Published: December 2022

The relationship between the content of cyclic AMP (cAMP) in rat mast cells in vitro and release of histamine was studied with compound 48/80 and a variety of hormones and drugs that affect cAMP in tissues. Papaverine (0.1 mM) increased cAMP in mast cells 3-fold. No increases in cAMP were observed in response to other drugs unless papaverine was in the medium. Isoproterenol, epinephrine, prostaglandins E1, E2, or F2α produced further increases of cAMP in the presence of papaverine. Norepinephrine only slightly increased cAMP. Angiotensin II, vasopressin, bradykinin, and glucagon did not increase cAMP even when the drugs were used in concentrations as high as 0.1 mM. Compound 48/80 did not affect cAMP content in mast cells. Release of histamine induced by 48/80 was partially inhibited by PGE2, PGF2α, vasopressin, and angiotensin II in the presence of papaverine and by angiotensin II alone. Neither papaverine alone nor the combination of PGE1 with papaverine inhibited release of histamine. These data suggest, therefore, that changes in cAMP in rat mast cells are not correlated with inhibition of 48/80-induced release of histamine.
Evidence for in Vivo Protection against Murine-Sarcoma Virus-Induced Tumors by T Lymphocytes from Immune Animals
Authors: Gorczynski Reginald M.
Published: December 2022

Normal 3- to 4-week old BALB/c mice inoculated with murine sarcoma virus (MSV) develop sarcomas within 7 to 10 days, which generally regress by day 15 to 20. Sublethal irradiation of the recipients, prior to inoculation of MSV, leads to the development of progressively growing tumors and ultimate death of the host within 30 days. Small numbers (1 × 107) of spleen cells from mice whose tumor has regressed, but not similar numbers of cells from age-matched control mice, protect these irradiated mice from developing progressively growing tumors. Pretreatment of the protecting cells with anti-θ antibody and complement, but not with anti-immunoglobulin antibody and complement, abolishes the protective effect.
Allelic Selection of Light Chain Allotypes by Antibodies Raised in Heterozygous Rabbits
Authors: Kindt Thomas J.
Published: December 2022

A marked selection for allotype b4 over b9 was observed for anti-streptococcal antibodies produced in rabbits heterozygous for allotypes of groups a and b. Less than 2% of the streptococcal antibodies was of allotype b9 while 20% of the IgG in preimmune sera from the same rabbits was of allotype b9. No such effect was observed with respect to the group a allotypes. In spite of this marked selectivity in b4,b9 heterozygotes for the production of antistreptococcal antibodies of allotype b4, no evidence for linkage between the gene controlling this allotype and immune responsiveness to streptococcal antigens could be detected. Equal amounts of antibody were produced by all immunized offspring (b4,b4; b4,b9; or b9,b9) of heterozygous rabbits.
Human T Lymphocyte Receptors for Sheep Erythrocytes: Conditions for Binding including Inhibition by Cytochalasin B
Authors: Kersey John H. , Hom Denis J. , Buttrick Peter
Published: December 2022

Requirements for binding of sheep erythrocytes (E) to human thymus-derived (T) lymphocytes were studied. The drug cytochalasin B (CB) was found to inhibit reversibly activity of the lymphocyte receptor for E. Similarly, binding was inhibited by preincubation of E with CB; the effects of CB on lymphocytes and E may be related to inhibition of microfilament function. Colchicine and vinblastine did not inhibit binding. Binding was temperature dependent but temperature dependence was reversed by Vibrio cholera neuraminidase. T lymphocyte binding of E is a physiologically active process requiring energy and intact membrane functions.