bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunoglobulins in Intact, Immunized, and Contaminated Axenic Mice: Study of Serum IgA
Authors: Benveniste J. , Lespinats G. , Adam C. , Salomon J.-C.
Published: December 2022

The study of immunoglobulin levels of intact, immunized and/or contaminated axenic mice showed that: 1) axenic mice had lower levels of serum IgG2 and IgG1 than did holoxenic mice, but both immunoglobulin levels rose sharply after immunization or contamination of the former. In contrast, serum IgA remained undetectable in axenic mice after immunization and at 21 days after contamination; 2) serum IgA appeared about 2 months after massive contamination of axenic mice or 6 days after administration of prednisolone to them. The effect of prednisolone on serum IgA seemed to be related to passage of IgA from the intestinal system into the blood without true activation of the serum system.
Isolation of the Anticomplementary Protein from Cobra Venom and its Mode of Action on C3
Authors: Müller-Eberhard Hans J. , Fjellström Karl-Erik
Published: December 2022

Cobra factor, a protein of the venom of Naja naja, was isolated. It was characterized as a glycoprotein with a molecular weight of 144,000 and an electrophoretic mobility of a β globulin at pH 8.6. Added to whole human serum it converted and inactivated C3. The effect on C3 was found to be dependent upon a non-complement plasma protein and on bivalent cations. To generate C3-directed enzymatic activity, cobra factor had to enter into a firm complex with the plasma protein, a 5S thermolabile β pseudoglobulin.
Antibody Against Neoplastic Plasma Cells
Authors: Watanabe Takeshi , Yagi Yasuo , Pressman David
Published: December 2022

Rabbit antisera against cellular components of BALB/c mouse myeloma were found to contain antibodies which are specific for surface antigens present on both normal and neoplastic plasma cells. When spleen cells from mice previously immunized with sheep erythrocytes were exposed to the antisera and rabbit complement prior to assay, the formation of hemolytic plaques was strongly suppressed, even though the viability of spleen cells in general was not affected. The suppressive effects of anti-myeloma sera on direct and indirect plaque-forming cells (PFC) were similar in both primary and secondary responses. The suppressive effect was seen with PFC of all the inbred strains of mice tested. The surface antigens which react with the PFC-suppressive antibodies are also present on myeloma cells, but do not appear to be present in any significant amounts on lymphocytes of thymus, lymph node and spleen, or on cells of liver and kidney.
Immune Adherence Reactions of Human Erythrocytes Sensitized with Complement In Vitro and In Vivo
Authors: Petz Lawrence , Fudenberg H. Hugh , Fink Diane
Published: December 2022

Human erythrocytes and erythrocyte stroma sensitized in vitro with antibody and complement are reactive in immune adherence (I.A.). This was demonstrated by using isoantibodies (anti-A, anti-Lea) and with autoantibodies obtained from patients with the cold hemagglutinin syndrome or paroxysmal cold hemoglobinuria (P.C.H.). Although trypsin destroys the I.A. receptor site on human erythrocytes used as indicator particles, it did not diminish the ability of the cells to fix complement and participate in I.A. as the antigen-antibody-complement complex. Erythrocytes from patients with warm antibody autoimmune hemolytic anemia (A.I.H.A.), the cold hemagglutinin syndrome, or P.C.H., which were strongly sensitized in vivo with C3 as judged by agglutination tests (and stroma prepared from such erythrocytes), were uniformly negative in I.A., indicating a lack of biologic reactivity of cell-bound C3 in these disease states. Paroxysmal nocturnal hemoglobinuria (P.N.H.) erythrocytes lysed by fresh acidified normal human serum were negative in I.A., in striking contrast to positive results obtained with the same concentration of stroma of erythrocytes sensitized with complement by isoantibodies or autoantibodies. Erythrocytes that had fixed C3 by mechanisms possibly of a nonimmunologic nature were negative in I.A. Although quantitative factors may be the explanation for negative immune adherence in some of these test systems, qualitatively different C3 fixation may also account for such findings. The I.A. receptor site on human erythrocytes was shown to be present on P.N.H. erythrocytes and umbilical cord erythrocytes and was not inhibited by sensitization with a variety of isoantibodies or penicillin. However, the erythrocytes of one of 13 patients with warm antibody A.I.H.A. gave negative results when used as indicator particles, and the autoantibody in this patient's serum inhibited the I.A. receptor site of normal erythrocytes.
Potentiation of Cultured Mouse Thymocyte Responses by Factors Released by Peripheral Leucocytes
Authors: Gery I. , Gershon R. K. , Waksman B. H.
Published: December 2022

Lymphoid cell interactions in the immune response, both in vivo and in vitro, have been the subject of much interest recently (1, 2). In this regard, substances have been found in supernatants of cell cultures which can affect the response of other lymphoid cells (3, 4). The following report describes a new effect wherein mouse thymocytes are highly stimulated by a combination of nonspecific stimulants and substances released by human leucocytes.

Five × 106 thymus or spleen cells from male CBA/H or CBA/J mice, aged 6 to 12 weeks, were cultured in duplicate in plastic tubes (Falcon #2001) in 1.0 ml MEM-S (Microbiological Associates, Bethesda, Md., with added antibiotics and l-glutamine), supplemented with 8% normal human serum. These cultures were stimulated by the following, added singly or in combination: a) 1.0 µl phytohemagglutinin-P (PHA, Difco Laboratories, Detroit, Mich.); b) different concentrations of human white blood cells (obtained by gravitational sedimentation of the erythrocytes); c) supernatants (SUP) from cultures of human leucocytes.
Amino Terminal Sequences of Heavy and Light Chains of Chicken Anti-Dinitrophenyl Antibody
Authors: Kubo Ralph T. , Rosenblum Irwin Y. , Benedict Albert A.
Published: December 2022

Based on their amino terminal sequences human κ and λ light (L) chains form several subgroups (1–3). The L chains of the immunoglobulins of other animal species have been classified as homologues of the κ and/or λ classes (4). The κ class has been characterized as having a free (unblocked) amino terminus and a carboxyl terminal half cystine residue, whereas the λ class has been considered generally to have a blocked amino terminus, usually pyrrolidone carboxylic acid, and a penultimate half cystine residue at the carboxyl terminus (4, 5). Based on these criteria, Hood et al. (4) suggested that the chicken L chain was a κ type. Although the L chain from normal chicken IgG has an unblocked N-terminal amino acid (4, 6), we reported that the first 17 residues appeared to fit better in the N-terminal sequence of human λ rather than κ chains (6).
Functional Heterogeneity of Murine Lymphoid Cells
Authors: Stobo John D. , Rosenthal Alan S. , Paul William E.
Published: December 2022

Concanavalin A is a potent stimulant of in vitro blast transformation and of DNA synthesis by murine lymphoid cells. In contradistinction to phytohemagglutin which stimulates thymocytes minimally, concanavalin A causes marked responses on the part of both thymus cells and spleen cells. Both of these stimulants lead to meager responses on the part of bone marrow cells.

In the spleen and the bone marrow, the cells responsive to both agents appear to be thymus derived or thymus dependent. Thus, treatment of bone marrow cells with complement (C) and iso-antisera directed against the thymus dependent differentiation antigen ϑ abolishes the Con A and phytohemagglutinin (PHA) responsiveness of these cells. Spleen cells from adult mice which have been thymectomized, irradiated and reconstituted with bone marrow cells exhibit depressed responses to Con A and are unresponsive to PHA. Treatment of these cells with anti-ϑ antisera and C abolishes their residual Con A responsiveness.

Despite differences in the reactivity in various lymphoid cell populations to Con A and to PHA, similar amounts of radioiodinated mitogen can be bound to the surface of these cells. Thus, the number of Con A molecules which can bind to the highly responsive spleen and thymus cell populations is similar (∼10 × 106) to that which binds to the weakly reactive bone marrow cells. Spleen cells from thymus-deprived mice bind similar amounts of Con A and PHA as do spleen cells from intact mice. Among given lymphoid cell populations, a heterogeneity in Con A binding can be demonstrated either by fractionation of the population on Con A-polyacrylamide affinity columns or by direct light and electron microscopic visualization using covalent complexes of Con A with horseradish peroxidase or with ferritin.

An estimate of the number of Con A molecules which are required to bind to a cell surface to yield maximal stimulation was obtained by presensitizing thymus and spleen cells at 4°C with varying concentrations of Con A. Binding of approximately 1.5 to 2.5 × 106 molecules to these cells yielded optimal stimulation.

These results are discussed in terms of heterogeneity of mouse lymphoid cell populations and of the complex requirements for cellular stimulation by mitogen.
A Study of Tumor Allograft Sensitized Lymph Nodes in Mice
Authors: Irvin George L. , Eustace John C. , McAleer James A. , Levi Donald F.
Published: December 2022

The immunoglobulin synthesis by immune lymph node cells, which in vivo have been shown to transfer adoptively accelerated tumor allograft rejection and enhancement, was studied by in vitro techniques. Autoradiographs of tissue culture supernatants showed that lymph node cells that were sensitized for 5 days and were capable of mediating rejection immunity were producing peak amounts of IgM and increased amounts of 7Sγ2, but there was no increase in 7Sγ1 immunoglobulin when compared to unsensitized nodes. The autoradiographs of lymph node cells sensitized for 14 days, which in vivo transferred enhancement activity, showed that these cells synthesized maximal amounts of 7Sγ2 and 7Sγ1 immunoglobulins, but decreased amounts of IgM when compared to 5-day immune cells. There were no consistent differences in the synthesis of IgA by normal or immune lymph node cells. Fluorescent antibody studies confirmed the results of the tissue culture experiments and extended these findings by localizing the site of immunoglobulin production within the node to the hilar and medullary cord areas. While antisera specific for all immunoglobulin classes stained the mature germinal centers of 14-day nodes, the quality of the staining reaction suggested entrapment of immunoglobulin in these cortical areas rather than active synthesis. Concomitant with the development of transferable biologic activities and immunoglobulin synthesis by the allograft sensitized lymph node cells was the appearance of many large, immature blastoid cells within the node. A correlation of immunoglobulin synthesis by immune lymph node cells in vitro with accelerated rejection immunity and enhancement activity mediated in vivo is presented.
Naturally-Occurring Antibodies to Bacillary Teichoic Acids
Authors: Decker Gene P. , Chorpenning Frank W. , Frederick G. Thomas
Published: December 2022

Extracts from two Bacillus species were purified and characterized chemically as glycerol teichoic acids. Although neither alanine nor other amino acids were present, the purified substances adsorbed spontaneously to erythrocytes, as demonstrated by passive hemagglutination with antisera to bacilli and to the purified substance. The antibodies reacting in passive hemagglutination and precipitation tests were the same and the specificity was directed toward polyglycerophosphate. Purified antigens from the two Bacillus species exhibited a single band of identity in immunodiffusion tests. Antibodies to this glycerol teichoic acid occur regularly in several animal species, including man. They were not found in albino mice. In humans and guinea pigs, these antibodies were observed in both IgM and IgG Sephadex fractions but in rabbits they were usually found only in the IgM fraction.
Alteration in Graft vs Host Reactivity and in Hematopoietic Stem Cells of Spleen Cell Inocula from Donor Mice Pretreated with Pertussis Antigen
Authors: Eikman Edward A. , Bowser Robert T.
Published: December 2022

The graft versus host reactivity (GVHR) and hematopoietic stem cells of spleen cell inocula were assayed 3 days after pretreatment of donor mice with heated or unheated pertussis vaccine. Unheated vaccine reduced GVHR to 17% of control reactivity, measured by the Simonsen spleen weight assay, and increased 2.2-fold the proportion of hematopoietic stem cells, assayed as colony forming units (CFU). Heated vaccine, which did not produce the lymphocytosis induced in donors by unheated vaccine, reduced GVHR to 38% of control reactivity, and increased the proportion of CFU 3.8-fold. Alternative mechanisms for these effects are discussed.
Genetic and Cellular Factors in the Immune Response
Authors: Sloan Bruce P. , Gill Thomas J.
Published: December 2022

Immunization with an aggregated antigen decreased the antibody response in the highly responding ACI strain and enhanced the antibody response in the poorly responding F344 strain; the effects occurred with a variety of aggregating agents. These findings are consistent with the hypothesis that a major factor in the action of the aggregate is the prolonged retention and slow release of the antigen. There were more differences in the distribution of antibodies among chromatographic subclasses in the F344 strain than in the ACI strain, and these differences did not always correlate with the net charge of the antigen. Most of the differences in the F344 strain were in the antibodies eluting with 0.01 M buffer (IgG2), whereas the differences in the ACI strain were almost entirely in the antibody eluting with 0.20 M buffer (mainly IgG1). Aggregating the antigen caused some differences in the chromatographic subclasses of the antibody compared to those elicited by the antigen alone, and the differences were also strain-dependent.

The highly responding ACI strain mounted a strong delayed hypersensitivity response against both free and aggregated poly(Glu52Lys33Tyr15), whereas the poorly responding F344 strain was sensitized only by the aggregated polypeptide. Immunization with DNP-poly(Glu52Lys33Tyr15) aggregated with MeBSA or with polylysine sensitized the animals to the antigen and the aggregating agent, but not the hapten.
Presence of the Fc-Fragment Allotypic Determinant, A15, on IgG from an Allotype-Suppressed Aa2 Homozygous Rabbit Lacking the Aa2 Determinants of the Fd Fragment
Authors: Tosi S. Landucci , Mage R. G. , Gilman-Sachs A. , Dray S. , Knight K. L.
Published: December 2022

Approximately 10% to 30% of normal rabbit IgG molecules lack the allotypic specificities, Aa1, Aa2 or Aa3, of the Aa heavy chain locus (1, 2). Suppression of the Aa locus determinants has been observed in Aa locus homozygous rabbits following exposure to anti-allotype antisera in utero and neonatally (3, 4). Approximately 95% of the IgG obtained from an Aa2Aa2 homozygous suppressed rabbit lacked the Aa2 allotypic specificity (5). Recent data (5) on these Aa-negative molecules have shown that: a) Aa-negative IgG molecules carry a “new” set of allotypic determinants, A31, which are present on the Fab portion of the molecule; b) by peptide mapping, the Fab portion of Aa-negative molecules differs from the Fab portion of Aa-positive molecules; and c) by peptide mapping, the Fc portion of Aa-negative molecules is indistinguishable from the Fc portion of Aa-positive molecules.
Immunosuppression in Mice Injected with Heterologous Anti-Immunoglobulin Antisera
Authors: Manning Dean D. , Jutila John W.
Published: December 2022

Total suppression of IgM production, as reflected in serum immunoglobulin level and direct plaque formation, and severe reduction of all other serum immunoglobulins have been effected in mice by neonatal injection of rabbit anti-µ antibodies. Anti-IgG antibodies similarly administered have been shown to reduce serum IgG levels.
Blocking of Autologous and Homologous Leukocyte Responses by Human Alloimmune Plasmas: A Possible in Vitro Correlate of Enhancement
Authors: Buckley Rebecca H. , Schiff Richard I. , Amos D. Bernard
Published: December 2022

Four alloimmune plasmas were found to block the responses of autologous and many homologous lymphocytes to a variety of stimulants, including: phytohemagglutinin P (PHA-P), pokeweed mitogen (PWM), candida antigen, and mitomycin-C-treated allogeneic cells. Three of the plasmas contained lymphocytotoxic activity against the majority of leukocytes in cell panels selected to contain all known HL-A antigens. No cytotoxicity was present in the plasmas against lymphocytes of the plasma donors. The plasmas contained no detectable antibodies to PHA-P and PWM, and candida agglutinin titers were no higher in the blocking plasmas than in non-blocking control plasmas. The blocking of autologous responses by a given plasma showed no correlation with the HL-A types of allogeneic stimulator cells, and no apparent relationship to HL-A type was found in the ability of such a plasma to block homologous cell responses. The blocking was fully effective after heating the plasma to 56°C for 30 min. The effect on homologous responding cells was shown to be reversible, in that such cells could respond to PHA-P and candida antigen if they were washed after a 24-hr incubation period in blocking plasma and then resuspended in media containing autologous plasma. Both the blocking and cytotoxic activities were greatest in pure IgG fractions of the three plasmas that contained both activities. The data are most compatible with a direct inhibitory effect of the blocking plasmas on the responding cells rather than an antigen-masking effect.
Continued Production of a Single Light Chain Population of Rabbit Antibody during Repeated Immunization: Amino Acid Composition and Sequence
Authors: Seon B.-K. , Roholt O. A. , Pressman D.
Published: December 2022

In previous reports, anti-p-azobenzoate antibody produced by a single rabbit for a period of about a year was shown by specific fractionation to consist of principally two molecular species for the first half year after which production of one of the species apparently ceased while production of the other species persisted for at least another half year. Comparative studies were undertaken of the amino acid compositions and the amino-terminal amino acid sequences at the first 11 positions of the light chains from an antibody fraction (BE-1) containing both antibodies and from a fraction (BE-3) containing only one of the antibodies. The amino acid compositions of BE-1 and BE-3 light chains are almost the same except for probable small differences in histidine, aspartic acid and tyrosine. Light chain BE-3, previously shown to give a single band on disc-electrophoresis, also gave a single amino-terminal amino acid sequence, Ala-Val-Leu-Thr-Glx-Thr-Pro-Ser-Ser-Ser-Ser, while light chain BE-1 which showed two bands on disc electrophoresis, showed the amino-terminal amino acid sequence, . At position 5, glutamine (or glutamic acid) and threonine were found in almost equal quantities. The residue at position 6 was not identified unambiguously. Since one of two bands of BE-1 light chain correspond to BE-3 light chain, the other band (slow-moving band) has the amino-terminal amino acid sequence Ala-Val-Leu-Thr-Thr-Thr-( )-Ser-Ser-Ser-Ser. Thus rabbit No. 2663 produced two closely related antibody light chains and one of them disappeared in the latter half of the year. The present sequences are compared with other reported sequences of anti-carbohydrate and anti-azobenzoate antibody light chains. The present amino acid sequence studies were carried out by a modified manual Edman degradation.
Antibody-Mediated Suppression of the Immune Response in vitro
Authors: Feldmann Marc , Diener Erwin
Published: December 2022

The capacity of purified Fab′ and F(ab′)2 pepsin antibody fragments to inhibit the immune response to polymerized flagellin in vitro was investigated. Marked differences between the activities of these fragments were found. Only high doses of univalent Fab′ were suppressive, whereas much smaller amounts of divalent F(ab)′2 were active. Preincubation experiments were performed to determine whether these fragments were capable of potentiating the induction of tolerance in vitro. Only F(ab)′2 fragments facilitated the induction of antibody-mediated tolerance; Fab′ fragments, regardless of dose, were inactive. These findings highlighted several important aspects of the mechanism of antibody-induced suppression. First, the difference in activity of Fab′ and F(ab′)2, despite the same antigen-binding capacity and affinity, demonstrated the existence of at least two different mechanisms of antibody-induced immune suppression in vitro. Second, the importance of divalency for the induction of tolerance strongly supports the antigen cross-linking hypothesis of antibody-mediated tolerance as recently proposed by Diener and Feldmann. Third, the Fc part of the IgG molecule was not involved in the mechanism of antibody-mediated tolerance in vitro.
Neutrophilic Leukocytes in Immunologic Reactions: Evidence for the Selective Release of Lysosomal Constituents
Authors: Hawkins David
Published: December 2022

The release of lysosomal bioactive factors is essential to the role of the neutrophilic leukocyte as a mediator of tissue injury in immunologic inflammatory lesions. The lysosomal release process was studied by exposing rabbit neutrophils to phagocytosable immune complexes, non-phagocytosable immune complexes and antineutrophil antibody and measuring the kinetics of release of a lysosomal marker enzyme (β glucuronidase). All three types of immunologic stimuli produced a rapid, selective release of the lysosomal marker. This release preceded that of a cytoplasmic marker enzyme (lactic dehydrogenase) and was not associated with a general increase in cell membrane permeability as judged by trypan blue dye exclusion. In contrast when neutrophils were exposed to a cytotoxic chemical, N-ethylmaleimide, the opposite pattern was observed. Lactic dehydrogenase was rapidly released while release of β glucuronidase was greatly delayed. These studies indicate that there is a non-cytotoxic, non-cytolytic mechanism for the immunologically induced release of neutrophil lysosomal bioactive factors. This process may be similar to the non-cytolytic release mechanisms described for other cells such as platelets and mast cells.
Transfer of Experimental Allergic Orchitis with Peritoneal Exudate Cells
Authors: Kantor Gary L. , Dixon Frank J.
Published: December 2022

Experimental allergic orchitis (EAO) was transferred to strain 13 guinea pigs by the intratesticular injection of sensitized peritoneal exudate cells (PEC) obtained from syngeneic donors immunized with a testicular antigen in complete Freund's adjuvant (CFA). The lesions were antigen and organ specific, developed within 24 hr of transfer, consisted of macrophages and polymorphonuclear leukocytes (PMN) within the seminiferous tubules, rete testes and interstitial tissues and resulted subsequently in aspermatogenesis. When contrasted to similar lymph node transfer studies, sensitized PEC transfers induced lesions earlier and more frequently, required fewer total cells and lymphocytes per transfer, resulted in a more intense infiltrative process, particularly in the rete testis and produced lesions in irradiated recipients. It appeared that as few as 2 × 105 transferred lymphocytes were needed to induce an immunologic lesion in the presence of phagocytic effector cells.
Immunoglobulin Receptors on a Mouse Mast Cell Tumor
Authors: Cline Martin J. , Warner Noel L.
Published: December 2022

Surface receptors for IgG immunoglobulins have been demonstrated on a mouse mast cell tumor growing in tissue culture. Inhibition of rosette formation between mastocytoma cells and IgG-coated SRBC by purified mouse immunoglobulins indicated receptor specificity for all known IgG globulin classes but not for IgM, IgA or light chain immunoglobulins. Pepsin-digested IgG antibody-coated SRBC did not bind to the mast cell. Mast cell receptors appear to react with immunoglobulin-antigen complexes more strongly than with immunoglobulin alone.

Receptor activity was unaffected by phospholipase D, neuraminidase or azide and was reduced by phospholipase C, iodoacetamide and fluorescein isothiocyanate. Actinomycin D had no effect for periods up to 9 hr. Treatment of mast cells with trypsin enhanced receptor activity but did not alter specificity.

Our initial conclusion is that the mastocytoma IgG-receptor is a stable phospholipid or phospholipoprotein which is protease-resistant.
The Antibody Response of Rabbits to Soluble Mouse H-2 Antigens
Authors: McKenzie Ian F. C. , Painter M. Elisabeth
Published: December 2022

Soluble H-2 transplantation antigens were prepared from H-2d L1210 leukemia cells by sonication and an (NH4)2SO4 precipitation of this soluble material was used to immunize rabbits. The antisera produced reacted broadly in a cytotoxic test with the spleen and tumor cells of many different inbred mouse strains, but, after absorption, reacted specifically with several H-2d alloantigens. “K region” antigens were more immunogenic than “D region” antigens, especially the “private” specificity H-2.31. The rabbit anti-mouse sera contained no detectable non-H-2 antibodies nor antibodies reactive with tumor specific antigens and was unable to form precipitin lines in agar gel. The immunosuppressive activity of this serum, tested by skin grafting, was negligible.
Studies on Immunoglobulins of Mink: Definition of IgG, IgA and IgM
Authors: Coe J. E. , Hadlow W. J.
Published: December 2022

Three major Ig classes, IgG, IgA and IgM were characterized in mink serum. Mink IgM was a β globulin and was changed from 19S to about 6 to 7S after mild reduction and alkylation. IgA was found in serum, urine and saliva, although the greatest concentration of IgA was found in intestinal contents. The IgA's in various fluids appeared antigenically identical, migrated as β proteins and sedimented as 12 to 13S molecules. Only serum IgA, however, was mainly 7S after mild reduction and alkylation.
The Secondary Response to High and Low Dose Priming in Mice
Authors: Grantham William G.
Published: December 2022

Although the level of the antibody response to an antigen is generally considered to be proportional to the antigen dose over a wide dosage range, this is not necessarily true for the secondary response. Sterzl (1) found that neonatal rabbits and piglets primed with a suboptimal dose of sheep red blood cells (SRBC) gave a higher direct and indirect plaque-forming cell (PFC) response to a second antigen injection than did animals that were immunized with a dose optimal for a maximum primary response. Recently, Hanna and Peters (2) reported similar results in adult mice immunized with SRBC. In the primary response, spleens from mice immunized with 2 × 108 SRBC contained approximately twice as many direct PFC and 20 to 30 times as many indirect PFC as mice primed with 2 × 106 SRBC.
Surface Antigens of Immunocompetent Cells. IV. Effect of Anti-b5 Allotype on Immunologic Responses of Homozygous b5 Rabbit Cells in vitro
Authors: Mond J. J. , Luzzati A. L. , Thorbecke G. J.
Published: December 2022

The investigation of surface receptors on cells of lymphoid tissue has been rigorously pursued in recent years. It is now relatively well established that the receptors which are responsible for the specificity of the bone marrow-derived or B-cell in immunologic reactions consist of immunoglobulin (Ig) molecules. This conclusion was mainly derived from two experimental approaches: 1) the study of the ability of these cells to bind antigen (1–3), and 2) the study of their ability to bind anti-Ig (4–6) and to be killed by anti-Ig and complement (C) (7, 8). These approaches have also been applied toward elucidating the identity of the thymus-derived or T-cell receptor, but have provided conflicting results (4, 6, 9).

It has been shown in this laboratory that immune spleen cells from which the majority of B-cells have been killed by anti-Ig and C are severely impaired with respect to their ability to give secondary responses in vitro (10).
DNA Synthetic Response of Thymocytes to a Variety of Antigens
Authors: Krüger Jürgen , Gershon Richard K.
Published: December 2022

The ability of 10 different antigens to stimulate uptake of 125I UDR, as a measure of DNA synthesis in T cells,1 was investigated. There was a good correlation between the stimulatory capacity of the antigens and their antigenicity. Five of the antigens studied have been reported to be “thymus-independent.”. Three of them (brucella, POL and ferritin) were among the best T cell stimulators while the other two (pneumococcal polysaccharide and PVP) were the poorest. This discrepancy suggests that the basis of their reported “thymus-independency” is different. An explanation for the difference is offered.
The Distribution of Alloantigenic Specificities of Native H-2 Products
Authors: Cullen Susan E. , Schwartz Benjamin D. , Nathenson Stanley G.
Published: December 2022

Studies on radiolabeled, detergent-solubilized H-2 alloantigens of two different genotypes with indirect immunoprecipitation show that at least two different antigen molecules can be isolated from cells of each genotype. These H-2 alloantigen molecules may bear the antigenic sites for one, two or more specificities. The findings are consistent with recent views that the H-2 genetic region includes two genes, each of which directs the synthesis of a different H-2 alloantigen molecule.
Studies on the Opsonic Activity of Human Secretory IgA Using an in Vitro Phagocytosis System
Authors: Wilson I. Dodd
Published: December 2022

The potential of human secretory IgA to act as an opsonin for Staphylococcus aureus was tested in an in vitro phagocytosis assay system. Secretory IgA, isolated separately from each of nine mothers and containing agglutinating antibodies against the Staphylococcus used in test, did not have opsonic effect in concentrations between 20 and 100 mg/100 ml. The addition of lysozyme and serum to the test mixture did not affect the results. Whole colostrum potentiated growth of the organisms in the in vitro assay system, suggesting that IgA is not dependent upon other substances present in secretions for opsonic effect. No inhibition or potentiation by secretory IgA of an established opsonic reaction could be demonstrated. These data support the concept that secretory IgA has little, if any, capacity to act as an opsonin for polymorphonuclear leukocytes.
Kinetics and Specificity of Transfer of Immunoglobulin G and Serum Albumin across Rabbit Yolk Sac in Utero
Authors: Sonoda Shunro , Schlamowitz Max
Published: December 2022

The kinetics and specificity of yolk sac transfer of IgG and of serum albumin in vivo, from mother to fetus in the rabbit, were studied by uterine intraluminal administration of 125I-IgG and 125I-serum albumin (125I-RSA).

Methodologic and biologic factors were investigated to assess their effects on the quality of experimental transfer data. The observed range in the data could not be ascribed to problems related to temperature or time of experimental manipulation nor to a failure to extract completely labeled protein from fetal tissue. Metabolic degradation of the labeled proteins was also ruled out as a cause for the observed variation in the data. Factors markedly influencing transfer data and requiring correction include: a) the failure to achieve a uniform distribution of administered protein among the fetuses in a horn, when material was injected through the fallopian tube and cervical canal of the uterine horn, and b) differences in the amounts of endogenous protein in the uterine horns of different rabbits and in the horns of a given rabbit.

The influence of the first two was minimized by modifying the surgical techniques so that we could study transfer of the separate fetuses. The problems due to the second factor were corrected by use of a “dilution factor.”

The transfer reaction of IgG and of serum albumin was shown to follow zero order kinetics. Competition experiments indicated that independent recognition sites on the fetal yolk sac membrane are involved in the transport of each protein. When the observed transfer data were corrected for dilution by endogenous uterine IgG and serum albumin, a preference for the transfer of 125I-RSA over 125I-IgG was seen in most cases.

There is no evidence for a mandatory requirement for a carbohydrate moiety for transfer of protein in this model.

A generalized hypothesis is presented for the materno-fetal (or neonate) passive transfer of IgG, based on ontogenetic and morphologic similarities of the membranes involved in the transfer.
Stimulation of B-Lymphocytes by Endotoxin
Authors: Gery I. , Krüger J. , Spiesel S. Z.
Published: December 2022

Thymus-independent B lymphocytes were shown to play the main role in the transformation response of mouse spleen cultures to endotoxin (LPS). Spleen cells from mice thymectomized at 6 to 8 weeks, lethally irradiated, and reconstituted with syngeneic bone marrow (TxXBM) reacted normally to LPS, while the responses to phytohemagglutinin and ConA were markedly depressed. The reaction to LPS, however, was temporarily depressed in the TxXBM mice and was fully recovered only 4 weeks after treatment. This latent period may be necessary for processing of the reacting cells. Karyotypic analysis of spleen cultures from mouse chimeras with T6T6 thymus grafts showed that only 1.2% of the LPS-stimulated cells were of thymus origin, as compared to 29.7% reacting to PHA and 12.2% to sheep red blood cells.
Ontogenetic Development and Synthesis of Hemolytic C8 by Piglet Tissues
Authors: Geiger Hartmut , Day Noorbibi , Good Robert A.
Published: December 2022

Synthesis of hemolytic C8 was investigated in short-term tissue cultures with different tissues from adult and fetal pigs. Kidney, spleen, liver and lung tissues and very likely the intestine were found to be capable of synthesizing C8. Ontogenetically, the synthesis started in liver, lung and midgut at a very early stage of gestation. The presence of antimetabolites in the tissue culture media and lower incubation temperatures decreased the rate of synthesis markedly.
Hepatitis-Associated Australia Antigen. Protein, Peptides and Amino Acid Composition of Purified Antigen with Its Use in Determining Sensitivity of the Hemagglutination Test
Authors: Vyas G. N. , Williams E. W. , Klaus G. G. B. , Bond H. E.
Published: December 2022

A 0.1% solution of protein from the purified 20-nm particles of hepatitis-associated Australia antigen (HAA) has an optical density of 3.726 at 280 nm. The protein appears to consist of two subunits; it contains a high proportion of proline, leucine and serine and a low content of tyrosine. Depending upon the anti-HAA reagent used, the hemagglutination inhibition assay can detect HAA protein in amounts as low as 1 to 60 ng.
Changes Induced by Lactic Dehydrogenase Virus in Thymus and Thymus-Dependent Areas of Lymphatic Tissue
Authors: Snodgrass M. J. , Lowrey D. S. , Hanna M. G.
Published: December 2022

To examine the effects of lactic dehydrogenase virus (LDV) on stromal and parenchymal cells of the thymus and thymus-dependent regions of lymphatic tissue, virus was injected either i.p. or intrathymically into male BALB/c mice that had been either thymectomized and irradiated, adrenalectomized or not treated. Serum lactate dehydrogenase activity was determined, and the spleen, mesenteric lymph node and thymus were weighed and prepared for histologic and ultrastructural examination.

The principal events of the infection process took place during the first 48 hr after infection and were: 1) virus particles, seen in ultrastructure, consistently associated with the plasma membrane of phagocytic cells of the reticulum, with some virusmembrane configurations suggesting replication by budding from these cells; 2) eventual degeneration of these phagocytic reticular cells; 3) cytotoxic degeneration of thymus-derived lymphocytes and their subsequent phagocytosis by macrophages. The cortex of the thymus was markedly depleted of lymphocytes by 4 days after intrathymic virus injection, as opposed to minimal depletion after i.p. injection. Virus particles were directly associated with the stromal cells but not with the thymusderived lymphocytes, indicating that the latter cytopathic effect was mediated by some intermediate soluble factor. This response also occurred in adrenalectomized animals that were inoculated with LDV, indicating that the intermediate factor is not an adrenal cortical steroid.

Serum lactate dehydrogenase activity increased sharply after i.p. or intrathymic injection of the virus into otherwise untreated animals; it then declined slightly and remained constant throughout the remainder of the experiment. Initially the spleen weights increased, whereas thymus weights decreased, although in thymectomized-irradiated animals the increase in spleen weight was 60% greater, and the serum lactate dehydrogenase activity increased to a peak in half of the time of the controls.
Genetic Control of Immune Response in Mice to a Glutamic Acid, Lysine, Phenylalanine Copolymer
Authors: Merryman Carmen F. , Maurer Paul H. , Bailey Donald W.
Published: December 2022

The ability of inbred mice to make a specific immune response to an optimal dose (10 µg) of the random copolymer GLØ is under the control of a distinct autosomal dominant gene. This gene is associated with the major histocompatibility locus of the species. Mice with the H-2 alleles d and q are responders, whereas those with alleles a, b, k, p and s are non-responders. This association with the H-2 locus was determined by use of recombinant-inbred strains and confirmed by the use of inbred and congenic strains. The application of RI strains to help identify Ir gene associations to H-2 is discussed.
Genetic Control by the H-2 Gene Complex of the Alloantibody Response to an H-2 Antigen
Authors: Stimpfling J. H. , Durham T.
Published: December 2022

The genetic control of the immune response against an H-2 antigenic specificity was investigated. With the aid of five congenic mouse strains, B10.A, C57BL/10, B10.A(2R), B10.A(3R) and B10.A(5R), it was demonstrated that a genetic factor, possibly Ir-1, associated with the H-2(K) subdivision of the H-2b chromosome, exerts a regulatory effect on a recipient's capacity to respond to H-2.2, an antigenic specificity determined by the H-2(D) subdivision of the H-2 gene complex.
The Effect of Antiplasma Cell Sera on the Primary Immune Response
Authors: Harris Nick S. , Jagarlamoody Sreerama M. , McKhann Charles F. , Najarian John S.
Published: December 2022

An antiserum, antiplasma cell sera (APS), prepared against mouse myeloma cells was specifically cytotoxic to myeloma cells; the cytotoxicity could be absorbed out only with myeloma cells. When tested in a hemolytic plaque assay, APS greatly reduced the number of direct (IgM) plaque-forming cells (PFC) when administered 1, 2 or 3 days after immunization with sheep red blood cells (SRBC). The APS was also effective in vitro in reducing the number of direct PFC in spleen cell suspensions from mice that had been immunized 4 days earlier with SRBC. As determined in a skin allografting model, APS had no obvious effects on cellular immunity.
An Adaptation of the Mixed Leukocyte Culture Test for Use in Evaluating Lymphocyte and Macrophage Function
Authors: Twomey J. J. , Sharkey O.
Published: December 2022

The mixed allogeneic leukocyte culture reaction was adapted into a test in which lymphocyte and macrophage function in this system can be assessed individually. In this test, purified lymphocytes from the patient under study were stimulated to incorporate 3H thymidine by irradiated leukocytes from three allogeneic, control donors. Test donors' macrophage function was evaluated by stimulating purified lymphocytes from the three control donors with irradiated leukocytes from the patient under study. The macrophage content in cultures of unpurified leukocytes was also measured. The test proved to be sensitive and reproducible. This report shows that this test can identify specifically lymphocyte unresponsiveness, quantitative and functional macrophage abnormalities, and combined lymphocyte and macrophage defects.
Hemolytic Efficiencies of Genetic Variants of Human C3
Authors: Colten Harvey R. , Alper Chester A.
Published: December 2022

The hemolytic efficiency of the genetic variants of the third component of complement (C3), native C3 in serum and highly purified C3 were estimated. The specific hemolytic activities of each of the partially purified variants were similar to that of C3 in whole serum but only about 20% that of highly purified C3. In the highly purified preparation, approximately 300 C3 molecules were bound for each hemolytic site.
Persistence and Characterization of Idiotypes in Pedigreed Rabbits Producing Antibodies of Restricted Heterogeneity to Pneumococcal Polysaccharides
Authors: Winfield John B. , Pincus Jack H. , Mage Rose G.
Published: December 2022

Anti-idiotype antisera were prepared which reacted with anti-SVIII antibodies of restricted heterogeneity in the sera of two rabbits, each from a different pedigree. No sharing of the idiotypes was detectable in antisera from close relatives of each donor. We looked for shared idiotypic determinants by precipitation tests and by an inhibition of binding assay which we estimate could have detected 1% of antibody with the same idiotype. Antibody to SVIII with idiotypic determinants present during the first course of immunization of an individual rabbit reappeared during the second immunization series 12 months later. During the course of restimulation, the antiserum exhibited increased complexity. By the 4th week, antibody with the original idiotype had decreased in concentration while the total anti-SVIII reached maximum concentration.
The Effects of an Immunoenhancing Bacterial Product on the Adenyl Cyclase Activity of Mouse Spleen Cells
Authors: Winchurch Richard , Actor Paul
Published: December 2022

A somatic antigen fractionated from lysates of Vibrio cholerae was assessed for its ability to enhance responses to sheep and horse red blood cells (sRBC and hRBC). This material proved to possess adjuvant properties when injected with either of these antigens. In addition, this adjuvant substance was found to stimulate both tritiated thymidine uptake and adenyl cyclase activity in normal mouse spleen cell cultures.
Effects of Pyran Copolymer on Oncogenic Virus Infections in Immunosuppressed Hosts
Authors: Hirsch Martin S. , Black Paul H. , Wood Mary L. , Monaco Anthony P.
Published: December 2022

The effects of pyran copolymer on polyoma virus oncogenesis and Rauscher virus leukemogenesis were evaluated in immunosuppressed mice. Mice immunosuppressed by adult thymectomy and antilymphocyte serum were protected by pyran copolymer against tumor development following subsequent polyoma virus inoculation. Similarly immunosuppressed mice were also protected against early erythroblastosis and late lymphomas induced by Rauscher virus. The mechanisms of protection appeared to be secondary to macrophage activation and enhanced phagocytosis induced by pyran copolymer.
Further Purification of Perchlorate-Soluble Antigens from Human Colonic Carcinomata
Authors: Turner Michael D. , Olivares Telba A. , Harwell Lee , Kleinman Martin S.
Published: December 2022

Carcinoembryonic antigen (CEA) from colonic carcinoma extracts was labeled with radioiodine. The 125ICEA appeared homogenous in a number of systems but showed three components on density gradient ultracentrifugation in CsCl. The component of intermediate density appeared to represent purified CEA and reacted vigorously with absorbed rabbit and goat anti-CEA sera. The most dense component reacted with antiserum to blood group A substance as well as with the two anti-CEA sera. Its reactivity with all three antisera was abolished by the addition of blood group A substance. The purified CEA also reacted weakly with anti-blood group A serum. This reaction was abolished by the addition of blood group substance which, however, had no effect on the reaction of the purified CEA with the anti-CEA sera. It was concluded that some preparations of tumor antigen contain a mixture of blood group A substance and the tumor antigen. The rabbit and goat anti-CEA sera appeared to contain a mixture of anti-CEA and anti-A antibodies. CEA molecules may contain both “CEA-determinants” and A-determinants. A sensitive radioimmunoassay for CEA was developed by using 125I-labeled purified CEA.
In Vivo Activity of in Vitro Immunized Lymphocytes
Authors: Rouse Barry T. , Wagner Hermann , Harris Alan W.
Published: December 2022

Mouse thymocytes immunized in vitro against allogeneic spleen cells were assayed for their ability to cause rejection of tumor allografts in vivo. CBA (H-2k) cortisone-resistant thymocytes were immunized by culture for 6 days with mitomycin-treated BALB/c (H-2d) or C57BL (H-2b) spleen cells. The activity of cytotoxic lymphocytes (CL) generated was assayed in vivo by measuring their capacity to prevent tumor growth in lethally irradiated mice and in vitro by measuring their capacity to cause 51Cr release from labeled tumor cells.

Thymocytes immunized in culture had lytic activity against labeled tumor cells of various types (mastocytoma, carcinoma, myeloma and lymphomas) in vitro and were highly effective at rejecting allografts of the same tumors in vivo. Tumor cell lines rejected by the lowest ratio of CL to tumor cells were also the most susceptible to in vitro lytic action.

The protective effect in vivo was immunologically specific in that all H-2d tumors tested were inhibited by H-2d-immunized cells at a CL to tumor cell ratio of 1:1, but an H-2d and H-2k tumor were not inhibited even at a ratio of 10:1. With H-2b-immunized cells, weak reactivity was detected; at a ratio of CL to tumor cells of 10:1 some H-2d tumors were inhibited.

The results support the concept that tumor allograft rejection is a function of T lymphocytes and that no synergism with B lymphocytes is necessary. Furthermore, they indicate that in vitro activated CL can function in vivo as specific effector cells for cell-mediated immunity.
Peptides from the Site of Rabbit Antibody: Anti-Azophenyltrimethylammonium Antibody
Authors: Takeda Yasushi , Friedenson Bernard , Roholt Oliver A. , Pressman David
Published: December 2022

A tyrosine peptide which appears to be derived from the combining site of an anti-azophenyltrimethylammonium antibody was isolated and the amino acid sequence was determined. Binding studies showed that the antibody was inactivated by iodination and that the attack was in the site, since the presence of the hapten, phenyltrimethylammonium ion, during iodination prevented the loss of sites.

The peptide from the site was isolated after a paired labeling procedure as follows. One portion of the antibody was iodinated with 125I-labeled hypoiodite. A second portion was iodinated with 131I-labeled hypoiodite in the presence of hapten to protect the site from iodination. The protected and unprotected preparations were mixed, digested with pepsin and the iodinated peptides separated. The ratio of 125I:131I relative to that of the unfractionated digest was determined for the peptides; a high ratio indicated that the peptide was derived from the antibody site. A high-ratio peptide, equivalent to at least 37% of the total antibody sites present, had the sequence Ile-DIT-Asp. The peptide was shown to be derived from the heavy chain of the antibody and was probably from the variable portion.