bluuebunny/crossref_metadata_2025 huggingface dataset:
A Study of the Association of Human Secretory Component with IgA and IgM Proteins
Authors: Weicker Jena , Underdown Brian J.
Published: December 2022

Human secretory component (SC) was isolated from colostral whey, and the binding of 125I-SC to purified IgA and IgM monoclonal proteins was studied using two methods to separate free from immunoglobulin-bound 125I-SC: a) gel filtration on Sephadex G-200, and b) precipitation of 125I-SC-Ig complexes with anti-Ig antibody. Both IgA dimeric proteins and IgM pentamers bound 125I-SC with approximately one SC-binding site per mole of polymer and similar affinity. Assuming a reversible equilibrium, an apparent association constant ≃ 108 M-1 was calculated to govern the binding of 125I-SC to immunoglobulin polymers. The assignment of a single association constant may be an oversimplication, particularly for the case of IgA polymers, since evidence was obtained that disulfide bonds were formed in the 125I-SC-IgA complex. Despite the complexity of the reaction, binding of 125I-SC to both IgA and IgM polymers could be analyzed by standard methods of saturation analysis, and both were shown to have a similar affinity for 125I-SC. No differences were noted in the affinity of 125I-SC binding to the IgA1 and IgA2 subclasses. Binding of monomeric IgA and IgM proteins could not be measured and was at least 100-fold lower than that found for IgA and IgM polymers. Complexes of 125I-SC with IgA dimers were presumed to involve covalent bond formation, since these complexes did not dissociate in guanidine-HCl. One IgA2 trimer did not form a covalent bond since it was completely dissociated in guanidine. In contrast, 125I-SC-IgM complexes were dissociated in denaturing solvent, indicating that such complexes were held together primarily by non-covalent bonds. Experiments with (Fc)5 µ isolated by high temperature tryptic digestion of IgM showed that binding of 125I-SC was to the Fc region of IgM proteins. It was suggested that the binding of SC with similar affinity to both IgA and IgM polymers may be important in the biologic function of both these immunoglobulin classes.
The Frequency of Phosphorylcholine-Specific B Cells in Conventional and Germfree Balb/c Mice
Authors: Sigal Nolan H. , Gearhart Patricia J. , Klinman Norman R.
Published: December 2022

The frequency of B cells specific for phosphorylcholine (PC) in spleens of nonimmune conventionally-reared and germfree adult Balb/c mice was determined by an in vitro splenic focus technique. Analysis of spleen cells from 11 conventionally reared donors revealed an average of 19.1 PC-specific clonal precursor cells/106 donor B cells when calculated utilizing a cloning efficiency previously determined for the in vitro splenic focus technique of 4%. This number is one-tenth the frequency determined for 2,4-dinitrophenyl- and 2,4,6-trinitrophenyl-specific B cells; however, both idiotypic and isoelectric analysis reveal that the majority of B cells specific for PC yield monoclonal antibodies which share the TEPC 15 idiotype and contain a homogeneous IgM peak with a pK of 4.60. Thus this single clone appears on the average to be represented by more than 103 B cells in conventionally reared nonimmune adults. A similar frequency for precursor cells of this clonotype was found in an analysis of seven germfree adult mice so that the high frequency of B cells of this clonotype appears to be independent of previous antigenic stimulation. Among the 11 conventionally reared donors analyzed, frequencies varied from 1.6 to 52/106 B cells—an extreme degree of variation when compared to the almost invariant frequencies observed for 2,4-dinitrophenyl, 2,4,6-trinitrophenyl, and fluoroscein. This variation is interpreted as a reflection of the life cycle of a single clone with a possible role for antigen implied by smaller variations observed in germfree mice.
Antibody Affinity
Authors: Gopalakrishnan P. V. , Karush Fred
Published: December 2022

The method of fluorescence quenching by bound ligand for the measurement of antibody affinity has been extended to anti-lactose antibodies. The basis for this extension is the use of a lactosyl ligand covalently linked to the 2,4-dinitrophenyl (DNP) group. This group serves as a sensor of the bound state by causing fluorescence quenching similar to that observed with anti-DNP antibody. The synthesis and characterization of this new ligand, N-(Nα-acetyl, Nε -DNP-L-lysyl)-p-aminophenyl-β-lactoside, are described. The affinity of its interaction with three rabbit IgG and one equine IgM antilactose antibody preparations has been measured. The Qmax values range from 24 to 47% and the calculated association constants from 2.5 × 105 M-1 to 7.2 × 105 M-1. For one IgG preparation, measurement of the association constant by equilibrium dialysis with the tritiated ligand gave a value of 6.75 × 105 M-1 compared to the value of 7.2 × 105M-1 obtained by fluorescence quenching. It is evident that with monoclonal anti-lactose antibody the accurate determination of Qmax, which is possible in this system, provides another structurally dependent clonal characterization of the variable region of the antibody. This optical probe along with others can be used for the identification of individual clones of antibody-producing cells through time and inheritance.
Cellular Cooperation During in Vivo Anti-Hapten Antibody Responses
Authors: Janeway Charles A.
Published: December 2022

The effect of x-irradiation on T and B cells involved in an in vivo adoptive anti-hapten antibody response has been studied. In vitro x-irradiation inactivates both B cells and T cells. B cells are somewhat more sensitive than T cells to such treatment, the dose required to reduce B cell activity by 50% being about 50 R, whereas 200 R will reduce helper cell activity by 50%. By contrast, if recipient mice are primed with carrier, they develop helper activity that is totally resistant to 500 R x-irradiation within 3 days of immunization. This activity can not be transferred from irradiated, carrier-primed mice by either spleen cells or serum, although anti-carrier antibody is shown to give a modest helper effect late after boosting. However, this radioresistant helper effect can be reproduced in a cell transfer system by giving carrier-primed spleen cells to normal recipients that are then x-irradiated (500 R) and given a large dose of carrier protein. Neither cells nor carrier alone will give rise to such radioresistant helper effects. Similar helper activity is occasionally observed when normal mice are irradiated 24 hr after being given helper cells, in the absence of carrier, and boosted shortly thereafter. The differences between these radioresistant helper effects, and the radiosensitivity of helper cells transferred after x-irradiation, may be related to x-ray induced changes in cell migration patterns.
Interference of Anti-Allotype Antisera with Antigen-Antibody Binding
Authors: Acuto Oreste , Manzo Ciro , Pane Adriana , Fontana Silvia , Zappacosta Serafino , Tosi Roberto
Published: December 2022

Rabbit anti-b4 antisera are capable of inhibiting the antigen-binding ability of antibodies carrying b4 allotype. The data support the hypothesis of the localization of at least one b group allotypic determinant in the V region.
Compound d+y+ Particles of Australia Antigen
Authors: Mazzur Scott , Burgert Steven , Le Bouvier George
Published: December 2022

Australia antigen (HBs Ag) particles vary in their antigenic structure. They are generally found to carry either determinant d or determinant y, but not both. This report describes seven sera which contain Australia antigen carrying both d and y on the same particle.
Chemotaxis of Basophils by Lymphocyte-Dependent and Lymphocyte-Independent Mechanisms
Authors: Ward Peter A. , Dvorak Harold F. , Cohen Stanley , Yoshida Takeshi , Data Richard , Selvaggio Salvatore S.
Published: December 2022

Guinea pig basophils obtained from blood or bone marrow have been studied for their chemotactic responsiveness. Chemotactic factors for basophils include a substance (lymphokine) present in culture fluids from antigen-stimulated lymphocytes, a material generated in zymosan-activated guinea pig serum, a C5 cleavage factor, and a bacterial factor. When compared with homologous neutrophils and monocytes, basophils respond most rapidly to a chemotactic stimulus. The lymphokine basophil chemotactic factor is physicochemically similar to the previously described monocyte chemotactic factor but appears to be distinct from it as well as MIF and neutrophil chemotactic factor present in the same fluids. Part of the evidence for this is the ability to detect basophil chemotactic factor in the absence of other lymphokine activities under appropriate experimental conditions. More evidence, specifically relating to the monocyte factor, is that monocytes can adsorb basophil chemotactic activity but not vice versa. This latter observation may have implications for the mechanism whereby the accumulation of basophils is controlled and limited in vivo. In addition, it was noted that specific antigen could also suppress basophil chemotaxis. Although the mechanism of this phenomenon is unclear, it could serve as a second means by which basophil accumulation may be controlled in the intact animal. Taken together, these observations provide further definition of the chemotactic behavior of basophils in general, and underscore some of the ways in which lymphocytes can influence basophils through lymphokine-dependent mechanisms.
Characterization of Mouse Cells Releasing or Responding to Mitogenic Factor Induced by Phytomitogens in Vitro
Authors: Jacobsson Hans , Blomgren Henric
Published: December 2022

Mouse lymphocyte populations exposed to mitogen were tested for their capacity to release factors that stimulate other lymphocytes to synthesize DNA or enhance their response to mitogens in vitro. The target lymphocyte for this mitogenic factor(s) (MF) was also characterized.

The results showed that lymph node and spleen cells release more MF than thymocytes upon exposure to Con A, PHA or PWM in vitro. The T cells were found to be largely responsible for MF production, since cell suspensions depleted of phagocytic cells did not exhibit any decreased ability to produce MF and spleen cells from congenitally athymic (nude) mice produced no detectable MF activity. The MF stimulated thymocytes, lymph node cells, and spleen cells to synthesize DNA. Spleen cells from nude mice were also stimulated. The MF released by lymphocytes in response to Con A was found to induce DNA synthesis of lymphocytes in itself and did not require the presence of mitogen.

It is concluded that phytomitogen lectins stimulate T cells to synthesize DNA and to release soluble factor(s) which are mitogenic for both T and B cells. The latter cells may thus be unresponsive to the phytomitogen, but still undergo blast transformation.
Genetic Polymorphism of Properdin Factor B in the Rhesus: Evidence for Single Subunit Structure in Primates
Authors: Ziegler John B. , Watson Lillian , Alper Chester A.
Published: December 2022

Properdin Factor B shows genetic polymorphism in the human and the polymorphic patterns in agarose gel electrophoresis suggest a tetrameric structure. Factor B polymorphism in the rhesus monkey has been demonstrated in the present study to be genetically determined and under the control of a single autosomal locus, rhesus Bf.

Six codominant alleles, BfF, BfG 1, BfG 2, BfS, BfS 1, BfS 2 have been recognized and the first five have been shown to have gene frequencies of 0.307, 0.160, 0.016, 0.377, and 0.139. The electrophoretic appearance of the polymorphic patterns does not suggest a tetrameric structure in the rhesus. Structural studies show purified human factor B to exist as a single subunit of molecular weight 94,000 daltons so that a tetrameric structure appears highly unlikely.
Thymic Involution: Effect on T Cell Differentiation
Authors: Hirokawa Katsuiku , Makinodan T.
Published: December 2022

The thymus of long-lived BC3F1 mice involutes progressively throughout life, beginning at 6 weeks of age. This is manifested by the loss of cortical lymphoid mass and by the degenerative changes in epithelial cells. The purpose of this study was to determine to what extent age-related degenerative changes of the thymus affect its capacity to influence the maturation of thymic-derived (T) cells. Accordingly, thymic lobes of mice ranging in age from 1 day to 33 months were implanted under the kidney capsule of T cell-deprived syngeneic young adult TXB mice, and the emergence of T cells was assessed kinetically by various morphologic and functional indices which may be reflective of different T cell subpopulations. They are: a) histology of the thymus graft, b) lymphocyte repopulation of the T cell-dependent areas of lymph nodes, c) total number of splenic lymphocytes carrying θ antigens (θ+), d) T cell-dependent humoral immune response and e) proliferative response of splenic cells to plant lectins, phytohe-magglutinin (PHA) and succinyl-concanavalin A (s-Con A), and allogeneic lymphocytes. The results revealed that the T cell-transforming influence of thymic tissues generally decreases with increasing age. The difference in the patterns of recovery of the various indices of thymus-grafted TXB mice suggests that the extent to which T cells can mature is dependent upon the degree of involution the thymic tissue has undergone with age. In particular, thymic tissues lose the capacity to influence the following functions with advancing age: 1) lymphocyte repopulation of the T cell-dependent areas of lymph nodes; 2) mitogenic reactivity of splenic cells to T cell-specific mitogens (PHA and s-Con A); 3) number of splenic θ+ lymphocytes and splenic T cell helper function; and 4) mitogenic reactivity of splenic T cells to allogeneic lymphocytes.
Effect of Nonspecific Stimulation on the Defense Mechanisms of Inbred Mice
Authors: Medina Socrates , Vas Stephen I. , Robson Hugh G.
Published: December 2022

Acquired resistance to facultative intracellular parasites was investigated in C57BL/6J inbred mice susceptible to Salmonella typhimurium but inherently resistant to Listeria monocytogenes and in A/J inbred mice which showed the reverse relationship.

A/J but not C57BL/6J mice were protected against S. typhimurium challenge by S. typhimurium phenol vaccine, S. typhimurium purified RNA, or L. monocytogenes purified RNA. C57BL/6J but not A/J could be protected against L. monocytogenes challenge by L. monocytogenes phenol vaccine, S. typhimurium RNA, or L. monocytogenes RNA. Yeast RNA, calf thymus RNA, BCG and poly (I:C) protected A/J against Salmonella and C57BL/6J against Listeria challenge.

Enumeration of viable organisms in the RNA and BCG-stimulated animals after challenge indicated that macrophage activation had probably taken place. The RNA-elicited protection was long lasting and waned after about 60 days. Enhanced resistance to S. typhimurium and L. monocytogenes can be stimulated by nonliving substances bearing no known antigenic relationship to the pathogen. Enhanced resistance is elicited only in the host with a background of inherent resistance to the specific pathogen.
Development of B Lymphocytes
Authors: Sidman Charles L. , Unanue Emil R.
Published: December 2022

The ontogeny of Ig(+), θ(+) and “null” (Ig-, θ-) lymphocytes in the mouse spleen was followed. An event was found at about 10 days after birth when the percentage of Ig(+) lymphocytes in the spleen begins to rise sharply, and the percentage of “null” lymphocytes falls. The Ig(+) lymphocytes before and after this event were then evaluated by four criteria pertaining to B cells. Three of these, the presence of C3 receptors, “capping” behavior, and the amount and/or disposition of surface Ig distinguished the Ig(+) lymphocytes before the event described from those present afterward. The fourth criterion, the presence of Fc receptors, did not distinguish between the early and mature forms of Ig(+) lymphocytes.
Functional Studies of Peyer's Patches: Evidence for Their Participation in Intestinal Immune Responses
Authors: Müller-Schoop Jürg W. , Good Robert A.
Published: December 2022

The pattern of responsiveness of lymphocytes from Peyer's patches, spleen, mesenteric, and inguinal nodes of guinea pigs was compared after oral and parenteral immunization. After oral immunization with BCG, Peyer's patch lymphocytes showed the best in vitro proliferative responses to PPD (tuberculin). Responsiveness to Clostridia, normal constituents of the intestinal microflora, also was significant in Peyer's patches, and low or absent in the other lymphoid tissues examined. In animals immunized parenterally with heat-killed Mycobacterium tuberculosis, lymphocytes from the spleen reacted best to PPD and those from Peyer's patches least. Selective elimination of B or T cells from PPD-reactive Peter's patch lymphocyte populations showed that the T cell-rich fraction remained responsive to PPD, the B cell-rich fraction became unresponsive, but both fractions reacted to Clostridia. We conclude from this experiment that a functional T cell population is present in these organs.

Peyer's patches in the guinea pig are immunocompetent lymphoid organs which seem to be primarily engaged in immune responses to antigens presented from the intestinal lumen.
Effect of Anti-β2-Microglobulin on Antigen and Allogeneic Lymphocyte-Induced Proliferation of Human Lymphocytes
Authors: McCalmon Robert T. , Kubo Ralph T. , Grey Howard M.
Published: December 2022

The effect of anti-β2-microglobulin (β2m) on the mixed lymphocyte reaction (MLR), and an antigen-induced proliferative response was studied. Anti-β2m IgG and Fab′ fragments completely inhibited the MLR. Preincubation of stimulator or responder cells with anti-β2m suggested that the major effect of anti-β2m may be on the responder cell population. A clear-cut effect on responder cells was demonstrated by showing that anti-β2m completely inhibited a MLR in which the stimulator population was a β2m negative lymphoblastoid cell line. Anti-β2m also inhibited PPD-induced proliferation of sensitized lymphocytes. The kinetics of this inhibition indicated that anti-β2m added within the first 18 hr of stimulation was effective in inhibiting the proliferative response. These data are discussed in light of the hypothesis that β2m may be a subunit of an antigen receptor on T cells.
Preventive Effect of Hapten-Reactive Thymus-Derived Helper Lymphocytes on the Tolerance Induction in Hapten-Specific Precursors of Antibody-Forming Cells
Authors: Hamaoka Toshiyuki , Inada Tetsushi , Yamashita Uki , Kitagawa Masayasu
Published: December 2022

The role(s) of helper T lymphocytes in preventing or altering tolerance induction in DNP-specific B lymphocytes was studied. As DNP-reactive helper T cells were reactive against the DNP-portion of the DNP-D-GL molecule, we could probe definitively the physiological role of helper T cells in preventing tolerance induction in B lymphocytes by DNP-D-GL. The results demonstrated that the induction of DNP-specific B cell tolerance by DNP-D-GL can be completely prevented by the presence of DNP-reactive helper T cells, and provide evidence that one critical role of helper T cell participation in humoral responses to antigens is to circumvent the development of a tolerogenic signal that, in the absence of such T cell function, might otherwise ensue after binding of the anti-genic determinants by specific B lymphocytes.
IgE Antibody-Specific Abrogation of an Established Immune Response in Mice by Modified Antigens
Authors: Bach Michael K. , Brashler John R.
Published: December 2022

DBA/1 mice were immunized with low doses of oval-bumin and alumina gel to elicit an IgE response which could be augmented upon reimmunization. Treatment of IgE-producing mice with high doses of chemically modified ovalbumin preparations markedly reduced or eliminated the ability of the mice to respond to subsequent reimmunizations with low doses of ovalbumin. It was found that the primary structural requirement for efficacy of the modified ovalbumins was reduced antigenicity to prevent the anaphylactic demise of the animals. Cumulative doses of 0.5 mg or more given in several increments, i.v., afforded optimal protection. The protection was antigen-specific and persisted for 2 to 3 months regardless of intervening exposures of the animals to low doses of antigen. It could be extended by further courses of treatment. There was no correlation between the levels of circulating hemagglutinating or total antibody and the induction of unresponsiveness. Nor was the passive administration of antibody able to approximate the effects of the modified antigens. It is suggested that the induction of the IgE class specific unresponsiveness in responding animals may take place via effects on T cells.
Receptors for Immunoglobulin and Complement on Human Alveolar Macrophages
Authors: Reynolds Herbert Y. , Atkinson John P. , Newball Harold H. , Frank Michael M.
Published: December 2022

Specific cell membrane surface receptors for immunoglobulin and complement components were identified by rosette fromation on in vitro cultured alveolar macrophages obtained from 24 humans and from rabbits. Respiratory macrophages have easily identified receptors for IgG and the C3b fragment of the third component of complement. A macrophage receptor for the C3d fragment was detected only when purified human complement components were used to form erythrocyte-antibody-C3 immune complexes but was not detected when whole human serum was used as the source of complement. No IgM cell receptor was identified. Thus, with respect to membrane receptors, alveolar macrophages resemble peripheral blood monocytes. These studies in addition emphasize the importance of using a variety of immune reagents, expecially the use of human reagents, to determine correctly these cell receptors.
Prolongation of Allograft Survival in Mice by Administration of Anti-Thy 1 Serum
Authors: Gelfand Michael C. , Paul William E.
Published: December 2022

A single 0.5-ml dose of anti-Thy 1.2 serum raised in AKR (H-2k, Thy 1.1) mice by immunization with C3H (H-2k, Thy 1.2) thymocytes prglonged mean graft survival (MGS) of C57BL/6 (H-2b, Thy 1.2) skin on BALB/c (H-2d, Thy 1.2) mice from 9.2 to 13.0 days (p < 0.01). Absorption of anti-Thy 1.2 serum with C3H but not AKR brain removed this activity. MGS was prolonged by anti-Thy 1.2 serum given as long as 2 weeks before grafting. Four separately raised anti-Thy 1.2 preparations prolonged MGS, one to a dilution of 1:128. Anti-Thy 1.1 serum prolonged MGS on AKR mice but not on BALB/c mice. Thymectomy 3 weeks or more before graft placement or treatment of BALB/c mice with anti-lymphocyte serum (ALS) diminished the effectiveness of anti-Thy 1.2 serum. Furthermore, infusion of 25 × 106 spleen cells from BALB/c mice given anti-Thy 1.2 serum prolonged MGS on syngeneic recipients to 15.6 days whereas spleen cells from mice given no treatment or ALS had no effect. Thymocytes from anti-Thy 1.2 serum-treated donors were slightly less effective and lymph node cells were ineffective in prolonging MGS on transfer. Transfer of serum from anti-Thy 1.2 serum-treated mice also had no effect. The transfer of graft prolongation with lymphoid cells from anti-Thy 1 serum treated mice suggests anti-Thy 1 serum may prolong MGS by activating suppressor T cells.
Physicochemical Characterization of Chemotactic Lymphokines Produced by Human T and B Lymphocytes
Authors: Altman Leonard C. , Chassy Bruce , Mackler Bruce F.
Published: December 2022

Ostensibly pure populations of T and B lymphocytes were separated from peripheral human blood by a sequential rosetting method. These cells, and mononuclear leukocyte (MNL) cell preparations (60 to 85% T cells), were cultured in vitro and the supernatants of these cultures were assayed for chemotactic lymphokine (CTX) activity. B lymphocytes were activated to produce CTX by binding erythrocytes sensitized with antibody and complement (EAC) to their C3 receptors and phytohemagglutinin (PHA) stimulation of isolated T cells was used to produce T cell-derived CTX. Sephadex G-100 gel filtration chromatography of supernatants obtained from T and B cell cultures showed that both cell types produce a CTX of approximate molecular weight 12,500 daltons. This result is identical to that previously obtained in characterizing the CTX produced by buffy coat leukocytes. Isoelectric focusing of lymphocyte supernatants and chemotactically active Sephadex G-100 fractions showed that the CTX produced by B cells and PHA-stimulated MNL contained two peaks of lymphokine activity, a major peak at pH 10.1 and a minor peak at pH 5.6. Additionally, these studies show that the chemotactic complement peptide C5a has an isoelectric point of pH 8.7 and is antigenically distinct from T and B cell-derived CTX. These data indicate that B and T lymphocyte-derived CTX, although relatively homogenous by gel filtration, contain two isoelectrically distinct species. In addition, these findings suggest that T and B cells although possessing different membrane receptors and requiring different mechanisms of activation to initiate lymphokine synthesis, produce lymphokines with identical physicochemical properties.
Naturally Occurring Low Molecular Weight IgM in Patients with Rheumatoid Arthritis, Systemic Lupus Erythematosus and Macroglobulinemia
Authors: Harisdangkul Valee , McDougal J. Stephen , Knapp Michael , Christian Charles L.
Published: December 2022

A method for purification of low molecular weight IgM (LMW IgM) which utilized salt precipitation, gel filtration, and insoluble immunoadsorbents is described. The yield from pathologic sera containing LMW IgM in the range of 15 to 36 mg% was approximately 20%. Antisera prepared against LMW IgM, IgM, and reduced-alkylated IgM recognized common determinants. Purified LMW IgM from seropositive rheumatoid subjects did not have rheumatoid factor activity in agglutination tests but demonstrated binding to 125I-labeled aggregated IgG. LMW IgM from a patient with SLE and vasculitis had anti-nuclear factor activity and anti-native DNA activity. When complexed with reduced-alkylated aggregated IgG, LMW IgM and IgM with anti-IgG activity were essentially equivalent on a weight basis in complement fixation. The source and function of LMW in biologic processes remain unknown.
Detection and Quantitation in Plasma and Synovial Fluid of a Fragment of Human C4 with α Mobility Generated during the Activation of the Complement System
Authors: Perrin Luc H. , Shiraishi S. , Stroud Robert M. , Lambert Paul H.
Published: December 2022

A newly identified fragment of human C4 was detected, using a particular antiserum, in human serum after activation by heat-aggregated immunoglobulins, but not after activation of the complement alternative pathway. This fragment was shown to have a sedimentation velocity of approximately 2.5S, to be heat stable, and to exhibit α mobility in immunoelectrophoresis. This C4α mobility fragment was not generated in human C4 deficient serum but was generated in human C2 deficient serum after incubation with heat-aggregated immunoglobulin. After precipitation of native C4 and its higher molecular weight fragments from serum by polyethylene glycol, it was possible to quantitate the lower molecular weight C4 α mobility fragment by radial immunodiffusion. In kinetic experiments, it was shown that the C4α mobility fragment was generated after some delay when compared to the disappearance of C4 hemolytic activity. Quantitation of the C4α mobility fragment may be of further use in human diseases for the evaluation of the catabolism of C4: joint fluids of patients suffering from rheumatoid arthritis contained high levels of the C4α mobility fragment, and low concentrations were found in patients with degenerative joint disease.
Immunologic Hyporesponsiveness in Man to Group C Meningococcal Polysaccharide
Authors: Artenstein Malcolm S. , Brandt Brenda L.
Published: December 2022

By using a hemagglutination-inhibition assay, certain lots of purified group A meningococcal polysaccharide vaccines were found to contain trace amounts of group C polysaccharide (0.032 to 0.128%). Young adult human volunteers inoculated with these vaccines showed a very poor group C serum anti-polysaccharide antibody response as determined by a radioactive antigen-binding assay. Two weeks later volunteers were given a 50-µg subcutaneous injection of purified group C polysaccharide. Serum specimens obtained 1 month later showed a significant suppression of group C antibody response in comparison with subjects who had initially received group A vaccines which were free of group C antigen. The observed hyporesponsiveness may represent an example of immunologic tolerance in man.
The Control Exerted by Thymic Hormone (THF) on Cellular cAMP Levels and Immune Reactivity of Spleen Cells in the MLC Assay
Authors: Kook Abraham I. , Trainin Nathan
Published: December 2022

We have shown previously that THF induces immunocompetence in lymphoid cells through the control of cellular cAMP levels and DNA synthesis. By the use of a one-way MLC assay it was now shown that the exposure of spleen cells to THF is accompanied by an increase, in the proliferative response of the responding cells. These effects are mediated by regulation of cellular levels of cAMP during the course of the MLC response. Thus, substances which modify cellular levels of cAMP such as DBcAMP, theophylline, and imidazole change the reactivity of cells exposed to THF in the MLC assay. It was also found here that cellular cAMP levels are critical in controlling the reactivity of normal spleen cells in the absence of THF. It is suggested that THF increases the number of competent cells in the spleen cell population capable of responding to antigenic stimulation in the MLC assay.
Antigen-Initiated B Lymphocyte Differentiation
Authors: Schlegel Robert A. , Shortman Ken
Published: December 2022

Distinct sub-populations of B lymphocytes may be fractionated by passage through glass-bead columns, due to a temperature-independent adherence of certain cells. The relative adherence properties of AFC-progenitors from spleens of primed or unprimed CBA mice were compared with an adoptive immune assay employing as antigen the hapten NIP on the carrier polymerized flagellin, under conditions where T lymphocytes did not limit the response.

The AFC-progenitors for an anti-NIP IgM response from unprimed mice, whether conventional, germ-free, or athymic were more adherent than the majority of cells in the spleen. The AFC-progenitors for an IgG response from hapten-primed mice were less adherent than the majority of spleen cells. The results indicate that there are sub-classes of AFC-progenitors, differing in surface properties and probably corresponding to differences between the “antigen-inexperienced” and the “memory” B cell populations.
Isolation of Pure IgG Subclasses from Mouse Alloantiserum and Their Activity in Enhancement and Hyperacute Rejection of Skin Allografts
Authors: Jansen Jan L. J. , Koene Robert A. P. , v Kamp Gerard J. , Tamboer Wim P. M. , Wijdeveld Paul G. A. B.
Published: December 2022

B6AF1 anti-B10.D2 ascites fluid was fractionated by molecular sieving. From the 7S fraction, IgG subclasses were isolated by affinity chromatography with specific antisera against mouse immunoglobulin subclasses coupled to CNBr-activated Sepharose 4B. Thus, 7S IgG1, 7S IgG2, 7S IgG2a, and 7S IgG2b were obtained which were completely pure by criteria of immunodiffusion and immunoelectrophoresis. In the in vitro tests the 7S IgG1 completely lacked cytotoxic activity. All the other subclasses were able to induce lysis of B10.D2 cells in the presence of rabbit complement.

All subclasses were tested for their capacity to induce enhancement and hyperacute destruction of B10.D2 skin grafts in B6AF1 recipient mice, and for the dose-dependency of these effects. 7S IgG1, although showing clearcut enhancing activity, was completely inactive in inducing hyperacute destruction of the grafts, even if high doses were administered. By contrast, 7S IgG2 (IgG2a as well as IgG2b) was not only able to induce enhancement but also hyperacute destruction of the grafts. The results obtained with 7S IgG2 demonstrate that opposite biologic activities can be present within a single subclass.
Structural Studies on Induced Antibodies with Defined Idiotypic Specificities
Authors: Capra J. Donald , Tung Amar S. , Nisonoff Alfred
Published: December 2022

N-terminal amino acid sequence analyses have been performed on three preparations of light chains of A/J mice. Light chains derived from the IgG of unimmunized animals were compared to light chains of anti-p-azophenylarsonate (anti-Ar) antibodies possessing a cross-reacting idiotype (CRI); the latter were derived from the ascites fluid of a single A/J mouse, or from the pooled ascites fluids of 18 A/J mice. The heavy chains of these same two antibody preparations had previously been shown to comprise a single, homogeneous sequence to position 40.

With few exceptions, the first 26 positions of light chains derived from unimmunized animals were extremely heterogeneous; the heterogeneity is comparable to that observed in a composite of sequence data on light chains of BALB/c myeloma proteins. Although the light chains obtained from anti-Ar antibodies possessing the CRI (whether from the pool of 18 A/J mice or from a single mouse) were more restricted in their sequence, at several positions as many as four alternative amino acids were detected.

These studies indicate that an antibody population with defined idiotypic specificity, and very possibly identical heavy chain sequences, may contain at least four distinct light chains. The feasibility of structural studies on antibodies induced in individual mice is further demonstrated.
Inhibition of in Vitro Lymphokine Synthesis by Glucocorticosteroids
Authors: Wahl S. M. , Altman L. C. , Rosenstreich D. L.
Published: December 2022

At pharmacological levels, glucocorticosteroids inhibited two antigen-induced lymphocyte functions, in vitro proliferation and lymphokine synthesis. Lymphocyte production of both macrophage chemotactic factor (CTX) and macrophage inhibition factor (MIF) were decreased in the presence of hydrocortisone. The corticosteroid also blocked the action of MIF on macrophages but did not interfere with the action of CTX on macrophages. Thus, steroids can suppress the immune response at two different stages: by blocking lymphocyte activation and mediator synthesis and also by interfering with the interaction of certain effector molecules with their target cells. Furthermore, these findings suggested that MIF and CTX have distinct mechanisms of action on the effector macrophages, one being sensitive and one resistant to steroids.
Intravascular Effects of IgE Antibody upon Basophils, Neutrophils, Splatelets and Blood Coagulation in the Rabbit
Authors: Halonen Marilyn , Pinckard R. Neal
Published: December 2022

Rabbits synthesizing only IgE antibody to BSA were challenged intravenously with 50 mg BSA and the associated blood cell and coagulation alterations were examined. Basophils decreased by 90% within 1 min of challenge. This was followed by a 70% decrease in neutrophils and a 50% decrease in platelets by 15 min. These changes were found to be highly significant when compared to control unimmunized rabbits similarly challenged. By 60 min, the neutrophils and platelets in the experimental rabbits had returned to 50 and 80% of their prechallenge levels respectively, but no basophils were demonstrable by toluidine blue staining. Lymphocyte counts in the experimental rabbits did not differ from controls at 0, 1, 15, or 60 min after challenge. Blood coagulation alterations also occurred after BSA challenge in the rabbits synthesizing IgE. A significant shortening of the whole blood clotting time in plastic tubes at 25°C occurred in blood samples taken at 30 sec after antigen challenge. Similarly, in vitro addition of BSA to blood samples taken prior to challenge significantly shortened the whole blood clotting time. In addition, significant prolongation of whole blood clotting times was observed in blood obtained 60 min after challenge, but only among the rabbits positive for systemic anaphylaxis. This in vivo study corroborated the known in vitro effects of IgE upon the rabbit blood basophil and platelet. In addition, the results indicate that the intravascular interaction of antigen with specific IgE antibody induced neutrophil and blood coagulation alterations.
A Simple Radio-Immunoassay for the Measurement of Human and Rat IgE Levels by Ammonium Sulfate Precipitation
Authors: Carson Dennis , Metzger Henry , Bazin Herve
Published: December 2022

A simple assay for quantitating IgE levels has been developed based on the observation that antibody-bound IgE is insoluble in 33% saturated (NH4)2SO4 whereas unbound IgE is not. The method has been applied to human and rat IgE. Some preliminary results on IgE levels in parasite-injected rats are presented.
Cell Surface Immunoglobulin. XIV. Synthesis, Surface Expression, and Secretion of Immunoglobulin by Peyer's Patch Cells in the Mouse
Authors: Vitetta Ellen S. , McWilliams Michael , Phillips-Quagliata Julia M. , Lamm Michael E. , Uhr Jonathan W.
Published: December 2022

Peyer's patches (PP)5 have characteristics distinguishing them from both central and peripheral lymphoid tissues. In contrast to central lymphoid tissue, PP are antigen-dependent for their development (1, 2); they contain T as well as B cells (3, 4), and the mitotic rate of their cells is too low for them to be a major source of peripheral B cells during ontogeny (5). Nevertheless, PP also differ from typical peripheral lymphoid tissues in several respects: PP are adjacent to specialized portions of intestinal epithelium (6); they contain few plasma cells (7, 8); they do not make antibody in vivo unless antigen is introduced directly into their parenchyma (9, 10); and they are comparatively rich, in common with other gut-associated lymphoid tissues (4, 11–13), in lymphoid cells bearing surface IgA.

Immunoglobulins produced by PP cells are of interest because of these unusual characteristics and because of the proposed role of the PP as a major source of precursors of IgA-secreting plasma cells (11, 14).
Humoral Immunostimulation
Authors: Shearer William T. , Parker Charles W.
Published: December 2022

The intracellular incorporation of radioactive nucleoside into mouse L cells was stimulated by rabbit anti-L cell IgG antibody and by its divalent pepsin F(ab′)2 fragment but not by univalent pepsin Fab′ or papain Fab fragments. Divalent antibody also caused the cells to grow in an aggregated pattern and undergo capping, but univalent antibody did not. Prevention of aggregation of antibody-treated cells by growth in agar abrogated immunostimulation. Univalent antibody fragments partially blocked stimulation of cells by divalent antibody and binding of 125I-labeled divalent antibody to cells. These results suggest that immunostimulation of cell growth is dependent upon divalent antibody and that aggregation of cells is an associated event.
Enduring Antibody Responses in “Normal” Rabbits to Maternal Immunoglobulin Allotypes
Authors: Adler Frank L. , Noelle Randolph J.
Published: December 2022

A majority of rabbits born to mothers heterozygous with respect to the b locus allotypic markers “spontaneously” develop antibodies against the noninherited maternal allotypes. Such antibodies generally appear in the 4- to 5-month-old rabbit, after what may be a transient period of tolerance, and persist at constant levels for at least 5 to 6 months. The finding of a high incidence of measurable antimaternal allotype responses in “normal” rabbits, added to similar findings by others in mice, pigs, and man, underscores the generality of this rather unexpected phenomenon and invites further inquiries into biological significance.
Influence of Divalent Cations upon Complement-Mediated Enzyme Release from Human Polymorphonuclear Leukocytes
Authors: Goldstein Ira M. , Hoffstein Sylvia T. , Weissmann Gerald
Published: December 2022

The complement component, C5a provokes the selective release of granule-associated enzymes from intact, viable cytochalasin B-treated human polymorphonuclear leukocytes (PMN) in the absence of phagocytosis or cellular adherence to surfaces. Consequently, in this experimental system the influence of divalent cations on these two processes can be disregarded and their effects on enzyme secretion can be studied directly. Cytochalasin B-treated PMN exposed to C5a in calcium and magnesium-free media consistently secreted significant amounts of the granule-associated enzymes, β-glucuronidase and lysozyme. This basal secretory response was not diminished if cells were preincubated with 5.0 mM EDTA, nor was it influenced if 1.0 mM or 2.0 mM EDTA were present in the reaction mixtures. The addition of calcium (up to 1.5 to 2.0 mM) produced a concentration-dependent enhancement of β-glucuronidase release, whereas increasing amounts of calcium (above 2.0 mM) inhibited secretion of this enzyme. Lysozyme release was similarly enhanced by the addition of calcium, but inhibition with high concentrations was not observed. Calcium per se, in the absence of C5a, provoked only the release of lysozyme from these cells. The effects of calcium upon enzyme release were not associated with alterations in the state of assembly of cytoplasmic microtubules. These findings provide another example of the role of calcium in “stimulus-secretion coupling” and provide evidence that exocytosis of various granules in human PMN is regulated by independent mechanisms involving calcium.
Receptors for C3 and IgG on Macrophage, Neutrophil and Eosinophil Colony Cells Grown in Vitro
Authors: Rabellino Enrique M. , Metcalf Donald
Published: December 2022

Macrophage, neutrophil, and eosinophil colony cells from bone marrow culture in semisolid agar medium were studied for membrane C3 and IgG receptors. The capacity of these cells to bind either erythrocytes-19S antibody-complement (EAC) or erythrocyte-7S antibody (EA7S) complexes was measured using the rosette method. Whereas macrophage and neutrophil colony cells showed receptors for both C3 and IgG, eosinophil colony cells appear to bear only IgG receptors. Studies correlating colony age and the presence of receptors showed that 60 to 70% of the cells from 3-day-old macrophage colonies were reactive for EAC and EA7S contrasting with 80 to 90% of the cells from 6- to 12-day-old colonies. Neutrophils behaved somewhat differently: EAC and EA7S reactive cells were seen in colonies after 4 or 5 days in culture and comprised only 50 to 60% of the colony population. Eosinophilic colonies showed 50 to 60% EA7S reactive cells after 6 to 7 days in culture, but no EAC reactive cells were found among these colonies at any time. The characteristics and properties of the receptors detected on colony cells were similar to those on macrophages and neutrophils from normal peritoneal fluid or bone marrow. Most macrophage colony cells were actively phagocytic whereas neutrophils and eosinophilic colony cells failed to show phagocytosis under the same conditions.
Antigen-Antibody Complexes Produced in Chickens Tolerant to Bovine Serum Albumin
Authors: Eardley Diane D. , Tempelis Constantine H.
Published: December 2022

A radioimmunoassay for circulating soluble antigen-antibody complexes is described. The assay detects complexes either in antigen or antibody excess. Soluble complexes were found in the sera of chickens tolerant to bovine serum albumin (BSA). The complexes appeared in the serum as soon as 7 days following neonatal induction of tolerance. The amount of complexes reached a peak between 1 to 2 weeks of age and disappeared by 6 weeks when responsiveness returned. The complexes were found in the bottom third of a 10 to 40% sucrose density gradient and by analytical ultracentrifugation indicated a size of 22.8S.

If tolerant chickens were challenged with BSA at 2, 4, or 6 weeks, the disappearance of complexes was not accelerated, and a proportion of the previously tolerant chickens exhibited a heightened antibody response.
Sera and the in Vitro Induction of Immune Responses
Authors: Shiigi Stanley M. , Fujikawa Leslie S. , Mishell Robert I.
Published: December 2022

The effects of newborn calf (NBC) sera on the in vitro generation of immune responses were studied. NBC sera inhibited the generation of primary humoral responses to SRBC. The inhibitory activity was dose-dependent and diluted out rapidly. Adding inhibitory doses of NBC serum to test cultures at zero or 24 hr resulted in approximately the same amount of inhibition. Pulsing experiments showed that the test cultures were most sensitive to the inhibitory effects of NBC serum during the 24- to 48-hr period. In contrast to its inhibitory effects on primary humoral responses, NBC sera supported the in vitro generation of secondary humoral responses and significantly increased the responses of some of these over control levels.

We found that sera obtained from newborn calves before nursing were not inhibitory for primary responses but sera obtained from the same calves 7 days later did inhibit these responses. We postulate that the inhibitory effects of NBC sera may represent a physiologic control mechanism which becomes active when immunologic maturity develops and that it possibly involves the inactivation of cell mediators.
Alloantibody-Induced Cytotoxicity of Macrophages
Authors: Ptak Wlodzimierz , Hanczakowska Maria
Published: December 2022

Alloantibodies substantially alter the in vitro nonspecific cytotoxic effect of macrophages on bystanding allogeneic and xenogeneic target cells. Although the specific IgM alloantibody increased significantly the ability of parental macrophages to damage target cells, IgG alloantibody had an opposite effect and suppressed the macrophage cytotoxicity. Macrophages of F1 hybrid mice were less affected by this treatment unless alloantibodies against both parental strains were added together. Parental macrophages exposed in vitro to the concomitant action of both IgM and IgG alloantibodies exhibited a diminished cytotoxicity toward target cells. It is proposed that IgM and IgG alloantibodies induce different conformational changes of the macrophage surface.
Subpopulations of Human Thymus Cells Differing in Their Capacity to form Stable E-Rosettes and in Their Immunologic Reactivity
Authors: Galili Uri , Schlesinger Michael
Published: December 2022

The majority of human thymus cells from young donors form stable E-rosettes with sheep red blood cells (SRBC) that do not disintegrate after prolonged incubation at 37°C. With advancing age the proportion of thymus cells forming such rosettes decreases gradually. The thymus of a patient receiving prednisone treatment was found to contain only a few cells that formed stable E-rosettes.

The minor population of thymus cells that fails to form stable E-rosettes (non-rosetting or NR cells) was isolated and tested for its cell surface markers and immunologic reactivity in vitro. Most of the NR-cells were capable of forming regular E-rosettes with SRBC at room temperature. Like the majority of human thymus cells they were sensitive to the cytotoxic effect of normal rabbit serum. Lymphocytes with B cell markers constituted less than 0.2% of the original thymus cell suspensions, but about 1 to 3% of the NR-population.

Thymus cells from donors over the age of 36 and from a prednisone-treated child responded in vitro to stimulation with either phytohemagglutinin (PHA) or concanavalin A (Con A). Unfractionated thymus cells from children up to the age of 14 failed to react to either PHA or Con A, but their NR-population responded vigorously to both lectins. In contrast to unfractionated thymus cell suspensions from children, the NR fraction showed a significant reactivity in mixed lymphocyte cultures with mitomycin-C treated allogeneic lymphocytes.

It is concluded that like the thymus of other species, the human thymus contains a minor population of cortisone-resistant cells endowed with many of the immunologic properties characteristic for peripheral T lymphocytes.