bluuebunny/crossref_metadata_2025 huggingface dataset:
Allorestriction and xenorestriction phenomena in the induction of delayed-type hypersensitivity (DTH) responses in the guinea pig against azobenzenearsonate- (ABA) modified peritoneal exudate cells.
Authors: Sugimoto M , Chiba J , Egashira Y
Published: December 2022

The role of MHC products in the induction of DTH responses Ia antigens play a crucial role in these cells in the recognition of ABA determinants by lymphocytes. Second, JY-1 guinea pigs were employed as responder animals. A clear allorestriction phenomenon was observed again when weakly modified ABA-PEC were used. On the other hand, the allorestriction phenomenon was apparently abrogated when heavily modified ABA-PEC were employed: allogeneic as well as syngeneic ABA-PEC induced strong and comparable DTH responses. Even in this condition, xenogeneic (mouse) ABA-PEC were barely immunogeneic, On the basis of these observations, the cross-reactivity of Ia antigens in the conjunctive recognition of haptenic determinants by T cells is discussed.
Pathophysiologic analysis of peripheral blood lymphocytes from patients with primary immunodeficiency. I. Ig synthesis by peripheral blood lymphocytes stimulated with either pokeweed mitogen or Epstein-Barr virus in vitro.
Authors: Mitsuya H , Osaki K , Tomino S , Katsuki T , Kishimoto S
Published: December 2022

In the present study 5 patients with common variable hypogammaglobulinemia (CVH) and 4 patients with selective IgA deficiency (IgA-D) were analyzed for the cellular defects responsible for impaired Ig synthesis with use of peripheral blood lymphocytes stimulated with either PWM or EBV in vitro. By the use of co-culture with PWM, all the patients examined had intrinsic B cell defects restricted to the synthesis of Ig class corresponding to the low or absent Ig class(es) in the sera. Two types of excessive suppressor T activity were found, which were abrogated by irradiation. One was isotype-nonspecific and the other was IgA-specific. Moreover, failure of IgA-specific helper T activity was demonstrated. The use of EBV as an agent that polyclonally activates B cells independently of T cells and monocytes should allow a clearer delineation of the level of the B cell defects. When co-cultured with EBV, B cells from 3 patients with CVH produced normal to subnormal quantities of IgM although they could produce no IgM upon co-culturing with normal T cells and PWM. B cells from 2 patients with CVH could produce IgM normally by stimulation with either PWM or EBV; however, there was no restoration to produce IgG or IgA in these patients. In addition, B cells from 2 patients with IgA-D produced not only IgG and IgM but also IgA almost normally at 4 days after in vitro stimulation with EBV.
Immunologic studies of chronic relapsing EAE in guinea pigs: similarities to multiple sclerosis.
Authors: Mehta P D , Lassmann H , Wisniewski H M
Published: December 2022

IgG/albumin ratios and presence of oligoclonal bands were studied in neutral pH brain extracts and sera of guinea pigs with chronic relapsing experimental allergic encephalomyelitis (R-EAE). The ratios were found to be significantly increased in central nervous system extracts of animals sacrificed during the first relapse and late stages of the disease relative to those of controls and of animals with acute EAE. These results are consistent with an intracerebral synthesis of IgG found in patients with multiple sclerosis (MS). When the immunofixation (after isoelectric-focusing) patterns of brain extracts and CSF from R-EAE animals were compared with those of controls, R-EAE animals showed distinct oligoclonal IgG bands in the pH region of 7.0 to 9.3, in contrast to diffuse patterns seen in controls. The finding of oligoclonal IgG in brain extracts of these animals further supports the usefulness of R-EAE as an experimental model for MS.
Regulation of in vitro primary anti-DNP antibody production by functional subsets of T lymphocytes in man.
Authors: Morimoto C , Reinherz E L , Schlossman S F
Published: December 2022

A primary in vitro antibody response to DNP-KLH by peripheral blood lymphocytes in culture was developed. Optimal anti-DNP antibody production, as measured by solid phase radioimmunoassay occurred at a cell density of 1 X 10(6), and an antigen concentration of 5 to 10 micrograms per culture when PBL was incubated with the antigen for 5 days and then cultured in the absence of DNP-KLH for 4 additional days. The antibody produced was shown to be primarily of the IgM isotype and was specific for DNP. In the absence of T lymphocytes, no antibody was generated. The regulatory effects of T4+ and T5+/T8+ subsets on antigen-specific antibody production were determined and it was found that T4+ subset alone provided help for anti-DNP antibody production. In contrast, the T8+ subset did not provide help and more importantly, could suppress anti-DNP antibody production in the presence of T4+ inducer T cells. This in vitro antibody forming system should be of considerable use in the analysis of the cellular requirements for antibody production and genetic control of the immune response in man.
Immunogenicity and characterization of supernatant DNA released by murine spleen cells.
Authors: Pancer L B , Milazzo M F , Morris V L , Singhal S K , Bell D A
Published: December 2022

The cellfree supernatant from cultured splenocytes contains an immunogenic molecule capable of inducing anti-DNA antibodies in vitro and in vivo in young preautoimmune NZB/W and some normal mouse strains. The response to this supernatant material does not reflect polyclonal activation, and the molecule responsible for its immunogenic activity appears to be a relatively small (approximately 150 base pairs) DNA molecule, predominantly (75 to 80%) double stranded with tail-end single-strand regions. It does not appear to contain any unusual secondary structure. This immunogenic DNA molecule can be recovered from cultures of some normal as well as young B/W mice. It contains no observable protein moiety, and after phenol-chloroform extraction, the immunogenic DNA migrates on polyacrylamide and agarose gels as a relatively narrow homogeneous band. The anti-DNA antibody response detected in the serum of mice after injection of this DNA molecule appears to have its greatest affinity for the homologous DNA and for undenatured calf thymus DNA. Unique base sequences do not appear to be responsible for its immunogenic effects. Rather, this may depend on its relatively small size, since calf thymus DNA sheared to a similar size and also found incapable of digestion with restriction endonucleases had a similar immunogenic effect.
Cellular synergy in the manifestation of accessory cell activity for in vitro antibody response.
Authors: Inaba K , Nakano K , Muramatsu S
Published: December 2022

The accessory cell (A-cell) activity of murine splenic adherent cells (SAC) and peritoneal exudate cells (PEC) for in vitro anti-SRBC and anti-DNP-KLH antibody responses of spleen lymphoid cells was abolished by the depletion of Ia+ cells from SAC and PEC by treatment with anti-Ia alloantiserum plus complement. The surviving Ia- macrophages in SAC and PEC were incompetent to achieve the A-cell activity. We prepared a crude and a further purified nonmacrophage cell fraction (termed CF and NMF). Nearly 70% of CF cells and more than 90% of NMF cells were Ia+ cells. The mature macrophage content of CF was less than 2%, and that of NMF was virtually negligible; suspected dendritic cells were contained at 5 to 10% in CF and 20 to 30% in NMF. The majority of cells in CF and NMF were lymphoid cells. This offered no problem in these experiments, since CF and NMF were added to lymphoid cells, in order to investigate the A-cell activity, usually in a ratio of 1 to 100. Neither CF nor NMF was adequate by itself to manifest the A-cell activity. However, the combination of either CF or NMF with Ia- macrophages resulted in the development of high A-cell activity. This did not occur with use of CF depleted of Ia+ cells or of NMF treated with anti-Ia without complement. These results indicated that the synergy between Ia- macrophages and Ia+ cells, most probably Ia+ nonmacrophage cells, was effective in developing the A-cell activity. It was also found that the interaction between the lymphoid cell and the Ia+ participant in the A-cell activity was genetically restricted, but Ia- macrophages functioned across the H-2 barrier.
Structural characterization of human melanoma-associated antigen p97 with monoclonal antibodies.
Authors: Brown J P , Nishiyama K , Hellström I , Hellström K E
Published: December 2022

Monoclonal antibodies were used to study p97, a human melanoma-associated antigen (MAA). Four hybridomas, designated 4.1, 96.5, 118.1, and 8.2, were obtained by fusing mouse myeloma cells with spleen cells from mice immunized with human melanoma cells. Antibodies 4.1 and 8.2 were IgG1; antibodies 96.5 and 118.1 were IgG2a. Sequential immunoprecipitation (IP) and sodium-dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that all 4 antibodies recognized the same 97 kilodalton (kD) protein. Binding studies with 125I-labeled antibody showed that antibodies 4.1 and 96.5 bound the same epitope, p97a. Antibodies 118.1 and 8.2 defined epitopes p97b and p97c, respectively. Six monoclonal antibodies (M17, L1, L10, R10, I12, and K5) specific for gp95, a kD melanoma cell surface glycoprotein were also tested. Sequential IP showed that these antibodies bound p97; p97 and gp95 are thus identical. Binding studies showed that antibody m17 bound epitope p971, and antibodies L1, L10, and R19 bound epitope p97c. Antibodies I12 and K5 defined 2 other epitopes, p97d and p97e, respectively. SDS-PAGE under nonreducing conditions indicated that p97 is monomeric, probably with intrachain disulfide bonds. Cell-surface labeling of sialic acid residues and neuraminidase digestion showed that p97 is a sialoglycoprotein. Digestion of p97 with papain or trypsin produced a stable 40 kD fragment, which expressed epitopes p971, p97b, and p97c, but not p97d or p97e.
Stimulation of oxidative metabolism in murine polymorphonuclear leukocytes by unopsonized fungal cells: evidence for a mannose-specific mechanism.
Authors: Danley D L , Hilger A E
Published: December 2022

Murine polymorphonuclear leukocytes (PMN) are stimulated to produce hydrogen peroxide (H2O2) by some unopsonized fungi, including Coccidioides immitis spherules, Candida albicans blastospores, and Saccharomyces sp. blastospores (zymosan). In this communication we have examined the basis for this stimulation by studying the effects of mannan and its primary constituent, D-mannose, on PMN function in vitro, since this polysaccharide is a significant component in many fungal cell walls. Our results show that mannan stimulated H2O2 production and iodination by PMN and enhanced H2O2 production by cells stimulated with zymosan. Moreover, the amount of H2O2 released by PMN was directly related to the concentration of mannan, and mannan obtained commercially or prepared in our laboratory worked equally well. Mannose, on the other hand, inhibited H2O2 release from PMN stimulated with zymosan or mannan and reduced oxygen consumption, carbon dioxide production, superoxide release, and iodination by these cells as well. With respect to specificity of inhibition, 18 different monosaccharides were examined by using 2 different assays for H2O2 release; and only 2-deoxy-D-glucose, a potent glycolytic inhibitor, reduced H2O2 release from zymosan-stimulated PMN. Furthermore, H2O2 release from cells stimulated with phorbol myristate acetate or opsonized Sephadex was not inhibited by mannose whereas H2O2 release was reduced when PMN were stimulated with C. albicans blastospores or C. immitis spherules in the presence of this sugar. From these data we propose that initiation of PMN oxidative metabolic burst in response to some unopsonized fungi occurs through a mannose-specific mechanism.
Lymphocyte transformation and the number of antigen-responsive cells in humans.
Authors: Sohnle P G , Collins-Lech C
Published: December 2022

The lymphocyte transformation response of normal human subjects to the antigen streptokinase/streptodornase (SK/SD) was analyzed by quantitating antigen-responsive cells (ARC), estimating the first generation lymphocyte response (FGLR), and performing standard lymphocyte transformation assays at different time points. Unlike the results obtained with the nonspecific mitogen phytohemagglutinin (PHA), the response to SK/SD appeared to depend almost entirely on the participation of progeny lymphocytes derived from a small number (mean of 21.3 +/- 5.8 per 100,000 mononuclear cells) of cells initially sensitive to the antigen. Similar peak lymphocyte transformation responses were obtained for the 2 stimuli, although those to SK/SD occurred later. Lymphocyte transformation data obtained after a shorter than optimal culture period (6 days) appeared to be a better estimate of ARC than the data obtained at the peak of the response of ARC than the data obtained at the peak of the responses did. However, ARC were best estimated by determining the FGLR or the number of days required for the lymphocyte transformation response to reach its peak.
Increased Fc receptor activity in monocytes from patients with rheumatoid arthritis: a study of monocyte binding and catabolism of soluble aggregates of IgG in vitro.
Authors: Katayama S , Chia D , Nasu H , Knutson D W
Published: December 2022

We studied the binding and catabolic function of adherent monocytes from patients with rheumatoid arthritis (RA) and normal subjects using stable, heat aggregates of IgG125I (A-IgG) as a model for soluble immune complexes. Scatchard plots of 4 degrees C binding data showed that RA monocytes had increased binding avidity and higher maximal binding capacity for A-IgG compared with monocytes from normal subjects. These data suggest that RA monocytes have increased numbers of Fc receptors for IgG, although a concomitant increase in the avidity of individual Fc receptors could not be excluded. At 37 degrees C, RA monocytes; kinetic analysis suggested that increases in catabolized A-IgG were due to increased binding of A-IgG with no change in the fractional rates of catabolism. Latex titers of RA patients correlated with the number of Fc receptors detected on RA monocytes. Mononuclear phagocytes from RA patients are often exposed to endogenous immune complexes that may be present in the blood of such patients; immune complexes may stimulate monocytes and possibly other mononuclear phagocytes to increase their capacity to bind and catabolize soluble immune complexes.
Genetic factors controlling the susceptibility to experimental autoimmune thyroiditis in inbred rat strains.
Authors: Lillehoj H S , Beisel K , Rose N R
Published: December 2022

CDF (high responder) rats were crossed with SHR (low responder) rats to study genetic factors influencing susceptibility to experimental autoimmune thyroiditis (EAT). F1 hybrid rats showed intermediate susceptibility to thyroiditis. Gene segregation patterns of F2 and backcross rats demonstrated that the immune response to rat thyroglobulin (RaTg) is under polygenic control. A main factor controlling immune response to RaTg is associated with the X chromosome. Females of CDF and SHR rats produced higher levels of antibody, suggesting that antibody production to RaTg is a sex-influenced trait. Different genetic control mechanisms are evident in autoantibody production and thyroid lesions. No evidence of linkage of an immune response gene to RaTg to the major histocompatibility complex of the rat (RT1) was observed.
Growth of 17D yellow fever virus in a macrophage-like cell line, U937: role of Fc and viral receptors in antibody-mediated infection.
Authors: Schlesinger J J , Brandriss M W
Published: December 2022

Growth characteristics of 17D yellow fever virus (17D-YF) and conditions for infection were studied in U937, a macrophage-like, Fc receptor-bearing continuous human cell line. Antibody to 17D-YF was obtained by immunization of normal subjects with 17D-YF vaccine. Cells were infected in the presence or absence of immune whole sera or immunoglobulin fractions. Infection of U937 was temperature dependent; the yield of virus was variable but at low temperature viral titers were consistently higher when infection was established in the presence of antibody. Results of infectious center assays indicated that the increased yield of virus was largely or entirely due to an increase in the number of cells producing virus early in the course of infection. Enhancement of viral growth was mediated by IgG but not IgM fractions of immune sera. Trypsinization of U937 resulted in a 90 to 95% reduction of infection in the absence of antibody but in the presence of antibody viral titers were higher in trypsinized than in nontrypsinized cells. Antibody to 17D-YF, contained in the whole IgG fraction of sera, bound to U937 to mediate infection without first being complexed to virus. Preincubation of U937 with IgG1 but not IgG2 myeloma proteins abrogated antibody-mediated infection. This result is compatible with the known affinities of U937 Fc receptors for specific subclasses of IgG and provides evidence for the role of the Fc receptor in antibody-mediated enhancement of viral growth. Persistent infection characterized by a lack of detectable cytopathogenic effect was established in long-term cultures of U937. This pattern of infection might be related to the unique effectiveness of the 17D-YF vaccine.
Macrophage production of fibronectin, a chemoattractant for fibroblasts.
Authors: Tsukamoto Y , Helsel W E , Wahl S M
Published: December 2022

Activation of macrophages results in the production of numerous enzymes and effector molecules. One of these monokines released by macrophages can cause directed migration of connective tissue fibroblasts in vitro. Production of this macrophage-derived chemotactic factor for fibroblasts requires activation of the macrophages either in vivo or in vitro and de novo protein synthesis. The chemotactic activity in the macrophage supernatants could be removed by a fibronectin-specific affinity column and was inhibited in the presence of antibodies to fibronectin. Furthermore, chemotactic activity in the depleted macrophage supernatants could be restored by the addition of exogenous fibronectin. Fibronectin was identified in activated macrophage supernatants by an enzyme-linked immunoassay for fibronectin. From these findings it was concluded that activated macrophages release a chemoattractant for fibroblasts and that the primary chemoattractant molecule is fibronectin. The production of fibronectin by activated macrophages may thus serve as an inflammatory mediator that in addition to its other functions can recruit fibroblasts to an area of damaged tissue, where they can proliferate and form the scar tissue necessary for tissue repair. Furthermore, in chronic inflammation, the prolonged activation of macrophages may be related to the extensive fibroblast infiltration and fibrosis that can accompany these lesions.
Endogenous retroviral envelope antigen in plasma cells.
Authors: Halpern M S , Ewert D L , Flores L J , Lin K Y , England J M
Published: December 2022

Previous observations that endogenous retroviral envelope antigen is expressed in splenic Ig(+) B cells of SRBC-sensitized 15I5 X 71 chickens suggested that expression of this antigen might be a general property of plasma cells in 15I5 and related strains. To investigate this possibility, immunofluorescence assays with antiviral sera were carried out using plasma cells present in Harder glands. Endogenous retroviral envelope antigen was detectable in plasma cells of line 15I5 X 71, 15I5 X 72, and 15I5 chickens but not in plasma cells of line 72, 15B, or 63 chickens. Higher levels of retroviral envelope antigen were detectable in 15I5 X 71 plasma cells as compared to 15I5 X 71 bursal cells. The presence of retroviral envelope antigen in 15I5 plasma cells, coupled with the absence of detectable levels of retroviral p27-related antigen in these cells, suggested that expression of the former is governed by the ev 6 genetic locus.
The role of adherent HLA-DR+ mononuclear cells in autologous and allogeneic MLR.
Authors: Huber C , Fink U , Leibold W , Schmalzl F , Peterson P A , Klareskog L , Braunsteiner H
Published: December 2022

The aim of this study was to define the significance of HLA-DR+ cells for the in vitro induction of human autoreactive T cell proliferation. Proliferation was induced by autologous or allogeneic non-T cells or EB virus-transformed LCL. Pretreatment of stimulator cells with low concentrations of RAHLA-DR and C and with high amounts of RAHLA-DR alone abolished the capacity to stimulate autologous or allogeneic T cells. This suggested that antigens associated with HLA-DR+ stimulator cells or HLA-DR itself are involved in the induction of this proliferation. At the responder cell level the cytotoxic elimination of HLA-DR+ cells as well as depletion of adherent cells by passage through Sephadex G-10 columns reduced both autoreactive and alloreactive responsiveness. Responses of serologically and physically depleted populations were both restored by reconstitution with mitomycin-C treated autologous adherent cells. These cells were resistant to lysis with T cell specific antibodies and preferentially cooperated with T cells of autologous deprivation. Co-cultivation experiments further revealed that such accessory cells were capable of amplifying T cell proliferation in the presence of suboptimal numbers of autologous LCL. We concluded, therefore, that an HLA-DR+-adherent cell, presumably of monocytic origin, is required for optimal proliferative responsiveness of both autoreactive and alloreactive T cells.
Immunochemistry of groups A, B, and C meningococcal polysaccharide-tetanus toxoid conjugates.
Authors: Jennings H J , Lugowski C
Published: December 2022

The successful coupling of the meningococcal groups A, B, and C polysaccharides to tetanus toxoid to yield water soluble conjugates is described. Reactive aldehyde groups were strategically introduced into the terminal residues of the polysaccharides by the controlled periodate oxidation of the native groups B and C polysaccharides and of the group A polysaccharide previously modified by the reduction of its terminal reducing N-acetyl-mannosamine residue. This produced essentially monovalent polysaccharide molecules, which were subsequently covalently linked to tetanus toxoid by means of reductive amination. Although the groups A and C polysaccharides proved to be poor immunogens in rabbits and mice, their tetanus toxoid conjugates produced high levels of polysaccharide-specific antibodies in both animals. By contrast, even in the form of its tetanus toxoid conjugate, the group B polysaccharide failed to elicit homologous polysaccharide-specific antibodies in either animal; a major proportion of the antibodies actually produced had a specificity for the linkage area of the conjugate. This evidence is compatible with the hypothesis of the poor immunogenicity of the group B polysaccharide being structure related. Hyperimmunization of mice with the groups A and C polysaccharide-tetanus toxoid conjugates produced antisera with good bactericidal activity against their respective homologous organisms, and indicated the potential of these conjugates as potential human vaccines.
Most B cells that have switched surface immunoglobulin isotypes generate clones of cells that do not secrete IgM.
Authors: Gearhart P J , Cebra J J
Published: December 2022

The order of genes coding for the constant regions of immunoglobulin heavy chains is 5'-C mu, C delta, C gamma 3, C gamma 1, C gamma 2b, C gamma 2a, C epsilon, C alpha-3'. To try to determine whether switching of the expression of heavy chain genes is irreversible and occurs in a direction from 5' to 3' within a cell line, we have isolated B cells bearing particular membrane isotypes, stimulated them to generate clones, and assayed daughter cells for secreted antibody. B cell with and without surface IgM and IgD, and B cells with and without surface IgG, were isolated by fluorescence-activated cell sorting. Fractionated cells were individually stimulated with phosphorylcholine in a splenic fragment assay, and the antibody secreted by clonal progeny was assayed for several heavy chain isotypes by radioimmunoassay. The results show that 1) B cells bearing IgM and IgD can produce individual clones of cells secreting 3 isotypes of anti-phosphorylcholine antibody--IgM, IgG, and IgA; 2) most B cells expressing surface IgG generate clones that secrete IgG but not IgM; and 3) most B cells from gut-associated lymphoid tissue that do not express surface IgM or IgD generate clones of cells that secrete IgA but not IgM. Therefore, IgM- and IgD-bearing cells can give rise to progeny that secrete IgM, IgG, and IgA. Once B cells switch to a different surface isotype, they appear restricted in isotype potential, since they frequently generate clones of cells that do not secrete IgM. These findings suggest that isotype switching is generally irreversible, and isotypes are expressed in a 5'- to -3' direction that is consistent with the germline gene order.
Studies on immunopathogenesis in epidemic hemorrhagic fever: sequential observations on activation of the first complement component in sera from patients with epidemic hemorrhagic fever.
Authors: Yan D , Gu X , Wang D , Yang S
Published: December 2022

Sequential measurements of activation of the 1st component of complement (C1) in the sera of 29 patients hospitalized with epidemic hemorrhagic fever (EHF) were performed according to a method recently developed. These patients were treated with supportive, routine therapy, but not immunosuppressive agents. This paper describes the kinetic observations on the activation of C1 in the 29 cases. The data confirm that an apparently increased extent of activation occurred in their sera. It was found that the more severely ill the patients were, the more apparent the activation. Additionally, beginning with the 15th day of disease, the extent of C1 activation diminished in most of the moderate and severe types of patients, but not in those with moribund illness and fatal ones. On the basis of the study, it may be reasonably concluded that C1 activation was correlated well with the severity and clinical course of EHF, indicating that the classical C pathway was activated in these patients. We feel that our findings are important to an understanding and elucidation of the immunopathogenetic mechanisms of EHF.
The attack phase of human complement: differentiation between membrane binding and complex formation by the detection of neoantigen expression in situ. A morphometric immunoferritin study.
Authors: Balkarowa-Ständer J , Rother U , Rauterberg E W
Published: December 2022

C5-9 neoantigen was found by morphometric immunoferritin staining on the membrane of guinea pig erythrocytes lyzed by reactive lysis C5b6 or acid-activated C5 + C6 and C7, C8, and C9. Neoantigen first appeared at the C7 step, increasing with C8, and decreasing after the addition of C9. The incubation of E with C5-C8 or C5-C9 at 0 degrees C resulted in EC5-8 without neoantigen. Reincubation of these cells after washing at 37 degrees C for 5 min led to neoantigen expression. An energy-requiring step is postulated after binding of C5-C8 and needed for C9 binding and lysis.
Unique T cell Ia antigen expressed on a hybrid cell line producing antigen-specific augmenting T cell factor.
Authors: Hiramatsu K , Miyatani S , Kim M , Yamada S , Okumura K , Tada T
Published: December 2022

Hybrid cell lines were established by fusion between keyhole limpet hemocyanin(KLH) binding T cells of A/J mice and an AKR T cell tumor line, BW5147. Hybrids were selected for the presence of Ia antigen and KLH-specific augmenting activity of their extracts in the secondary antibody response. The detailed phenotypic and functional analysis of 1 of these clones, FL10, is reported here. The hybrid was positive for both Thy1.1 and Thy1.2 antigens and possessed the Lyt-1+,2-,3- phenotype. Both VH and Ia determinants were detected on their cell surface. The IA locus was mapped in the I-A subregion, but the Ia specificities were serologically distinct from those of B cell Ia antigen. This was demonstrated by the fact that anti-Ia antiserum preabsorbed with B cells could react with the hybrid cells, whereas none of the monoclonal anti-Ia specific for private and public determinations of Iak could. The extract from the cell line specifically augmented the in vitro secondary antibody response against dinitrophenylated KLH, and this activity was removed by absorption with antigen and conventional anti-Ia antisera. The results indicate that the cell line, FL10, carries Ia antigen unique to the T cell, which is associated with the antigen-specific augmenting molecule.
The effects of ultraviolet irradiation on the generation of anti-tumor cytotoxic effector cell responses in vitro.
Authors: Lynch D H , Gurish M F , Daynes R A
Published: December 2022

The effects of irradiation of lymphoid cells in vitro with ultraviolet (UV) light were studied using radiation sources that are widely used both in the study of photocarcinogeneic mechanisms and by commercial tanning parlors (FS40 sunlamps). Low doses of direct exposure of cells was found to abrogate the differentiation events that lead to the generation of functional cytotoxic T lymphocytes (CTL). UV irradiation of cells in vitro induces alterations that, although not immediately obvious, lead to cell death during the next several days in tissue culture. Energy titration experiments demonstrated that both primed T cells and accessory cells were sensitive to the effects of direct UV-exposure; T cells were found to be much more sensitive than accessory cells. A 90% decrease in CTL generation could be affected with as little as 10 sec of UV exposure. These effects appear to be caused by wavelengths in the UVB range (280 to 320 nm), since exposure of cells to much higher doses of energy in the UVA range (320 to 400 nm) were not inhibitory. The spectrum of wavelengths in the UV region is large (200 to 400 nm). The data presented here demonstrate that it is overly simplistic to assume that all wavelengths in the UV spectrum mediate their effects via identical mechanisms. Furthermore, the wavelengths at which a given amount of energy is emitted appears to be much more important than the absolute amount of energy (in joules) which biologic systems are subjected.
Human IgE antibody synthesis in vitro: stimulation of IgE responses by pokeweed mitogen and selective inhibition of such responses by human suppressive factor of allergy (SFA).
Authors: Zuraw B L , Nonaka M , O'Hair C , Katz D H
Published: December 2022

The studies presented herein provide details of an in vitro system in which IgE biosynthesis by human peripheral mononuclear cells can be stimulated by exposure to pokeweed mitogen (PWM). Mononuclear cells of normal donors or patients with hyperimmunoglobulinemia E were cultured in 2 stages, the first stage consisting of exposure of cells to stimulating mitogen; after this first stage incubation, cells were washed and replated in cultures devoid of any additional mitogen for a further 6 days. Under such conditions, human mononuclear cells were shown to synthesize significant quantities of IgE. Both T cells and surface IgE-bearing cells were shown to be necessary in order to obtain an IgE response in this system. Supernatants from 2-way human mixed lymphocyte cultures were shown to contain an IgE class-specific suppressive factor capable of selectively inhibiting in vitro IgE (and not IgG) biosynthesis. This model can be used to compare the regulation of IgE synthesis in normal and atopic donors, and further experiments examining the production and action of this class-specific suppressive factor may have prognostic and therapeutic implications.
Genetic control of antisperm autoantibody response in vasectomized guinea pigs.
Authors: Tung K S , Teuscher C , Goldberg E H , Wild G
Published: December 2022

About 50% of vasectomized and none of sham-operated outbred Hartley GP develop autoantibodies to TSDA, surface antigens on sperm, spermatids, and residual bodies detectable by a sensitive quantitative cellular radioimmunobinding assay. A similar study of anti-TSDA antibody responses on different strains of GP demonstrates that Strain 13 GP are high responders and PLLhr guinea pigs are nonresponders, whereas the responses of Strain 2 and PLLlr GP are intermediate. Furthermore, 100% of vasectomized F1 GP between Strain 13 females and PLLhr males produce anti-TSDA antibody. Of the vasectomized back-cross GP between the F1 and PLLhr, 46% produce anti-TSDA antibody, 54% do not. These data provide strong evidence that autoimmune antibody response to sperm surface antigens in vasectomized GP is controlled by a single dominant autosomal or X-linked gene. Antisera from vasectomized Strain 13, F1, and back-cross GP are found to react with 3 major polypeptides (MW 63, 37, and 22 kd) on sperm surface that can be radioiodinated and extracted by NP-40.
Activated factor B (Bb) of the alternative pathway of complement activation cleaves and activates plasminogen.
Authors: Sundsmo J S , Wood L M
Published: December 2022

Activated Factor B (Bb), the central serine esterase of the alternative pathway of complement activation, exhibits restricted substrate specificity in the complement system for C3 and C5. The results presented here indicate that Bb can cleave and activate plasminogen in an experimental system containing purified plasminogen and Bb; complement cellular intermediate bearing the Bb-enzyme; or cobra venom factor-stabilized Bb-enzyme (CVF,Bb). Cleavage of plasminogen by Factor Bb generated 2 disulfide-linked polypeptides with apparent m.w. of 64,000 and 25,000 to 32,000 (SDS-PAGE). Complement cellular intermediates containing the C3b, Bb-enzyme cleave 40 to 80% of 4.5 micrograms of 125I-labeled plasminogen during 30 min of incubation at 37 degrees C; native Factor B was inactive; and anti-Factor Blg inhibited by 100% the plasminogen cleavage mediated by complement cellular intermediates bearing the Bb-enzyme. Fibrinolytic activity was detected in plasminogen activator (PA) assays when purified plasminogen and 125I-labeled fibrin tubes were incubated with Bb, CVF, Bb, or complement cellular intermediates bearing the C3b,Bb-enzyme: 10 micrograms Bb released 40 to 65% of the 125I-fibrin released by 5 micrograms urokinase in 4 hr at 37 degrees C. Plasminogen activator activity of the C3b,Bb-enzyme was found to be regulated in serum. At dilutions of NHS 1:50, the PA-activity of 1.6 micrograms Bb was 100% inhibited, and at a 1:250 dilution, 50% inhibition was observed. This report describes a novel activity for the Bb-enzyme, which constitutes the C3/C5-convertase of the alternative pathway of complement activation.
Lymphocyte mediation of lipopolysaccharide-stimulated macrophage metabolism.
Authors: Ryan J L , Yohe W B
Published: December 2022

LPS activation of murine macrophage metabolism and arginase production may be mediated by products of B lymphocytes. Splenic nonadherent cells, containing both B and T lymphocytes, splenic T cells, and thymocytes all stimulated macrophage glucose metabolism in co-culture. Supernatants derived from preculturing each of these cells in the absence of serum or other exogenous stimulant were also active in enhancing macrophage glucose utilization. When lipopolysaccharides were used to stimulate the lymphocyte populations, only the B lymphocyte containing NASC and purified B cells exhibited increased stimulatory activity. Thus, it appears that LPS does not directly activate T cells to produce macrophage-activating factors. The converse does not appear to be true, however, because LPS-stimulated macrophages enabled thymocytes to exhibit an enhanced ability to stimulate further macrophage glucose utilization. The active supernatant from NASC was heat resistant and remains to be chemically defined. These experiments clearly demonstrate that LPS-induced macrophage activation may be mediated by the products of lymphocytes, and that products derived from nonactivated lymphocytes are capable of stimulating macrophage metabolism and arginase production.
Characterization of an anti-H-2 monoclonal antibody and its use in large-scale antigen purification.
Authors: Stallcup K C , Springer T A , Mescher M F
Published: December 2022

A rat anti-mouse monoclonal antibody (MAb), M1/42, has been found to react with H-2 antigens from cells of the a, b, d, j, k, s, and u haplotypes (all haplotypes tested). This antibody, when bound to cells and reacted with FITC-conjugated anti-rat Ig, could be used to quantitate H-2 expression on several cell types. The antibody was also useful in comparing the H-2 products precipitated from a variety of haplotypes. M1/42-coupled Sepharose-4B beads were used to purify H-2d antigens by affinity chromatography. Pure H-2 molecules eluted from the column in 0.5% DOC, 0.65 M NaCl, 20 mM Tris, pH 8.0, yielding 110 to 180 micrograms H-2d/10(10) P815 tumor cells. This antibody, when used in series with H-2Kk-specific MAb 11-4.1, allowed purification of Dk and Dd from RDM-4 and YAC cells, respectively. H-2d purified by column chromatography on M1/42 was found to be serologically and biologically active, as determined by MAb rebinding, inhibition of cell lysis by alloantisera plus complement and ability to stimulate alloreactive CTL. This antibody and the described protocols should be useful in the preparation of relatively large quantities of a number of H-2 antigens.
Metastatic tumor cell variants with increased resistance to infection by Semliki Forest virus.
Authors: Barz D , Bosslet K , Schirrmacher V
Published: December 2022

Semliki Forest virus (SFV) is an interesting virus for cell interaction studies because it binds directly to the cells' major histocompatibility antigens. We used this reagent to study the expression and functional properties of H-2 molecules on murine tumor lines that are closely related but differ greatly in metastatic capacity. Tumor cell variants with high metastatic capacity showed an increased resistance to virus infection, an effect that was selective for SFV. Although the high metastatic tumor lines did not express less H-2 antigens than the low metastatic ones, they bound much less of the SFV viral glycoproteins.
Responses of B cells with or without C3 receptors to polyclonal B cell activators.
Authors: Kakiuchi T , Nariuchi H
Published: December 2022

Responses of B cells with or without receptors for C3 (CR) to polyclonal B cell activators (PBA) were studied. Mouse spleen cells were incubated with sheep red blood cells (SRBC) coated with antibody and complement to form rosettes, and they were separated by Ficoll-Hypaque density sedimentation into populations depleted of and enriched with lymphocytes bearing CR (CRL). These 2 populations were cultured with lipopolysaccharide (LPS), purified protein derivative of tuberculin (PPD), or dextran sulfate (DxS) and assayed for anti-TNP PFC. The CRL-depleted population responded well to LPS, poorly to PPD, and it showed practically no response to DxS, whereas the CRL-enriched population seemed to respond poorly to LPS but well to both PPD and DxS. The low responsiveness of the cRL-depleted population to PPD and DxS could not be explained by a shift of time-kinetics, by the dose-response profile of the responding cells, or by the depletion of adherent cells. Suppressor T cells did not take part in the reduced responses, since the treatment of the population with anti-Thy 1.2 plus complement could not restore the responses. These results indicate that B cells with CR [CR(+) B cells] respond well both to PPD and DxS, whereas the cells without CR [CR(-) B cells] respond poorly to PPD and DxS. It was difficult to evaluate the low responsiveness of CR(+) B cells to LPS because of the high background PFC of the cRL-enriched population.
Allergenically active components of cat allergen extracts.
Authors: Anderson M C , Baer H
Published: December 2022

The allergens involved in cat allergy have been studied in pelt extracts, saliva, serum, and urine. Using crossed immunoelectrophoresis (CIE) to examine the antigenic content, and crossed radioimmunoelectrophoresis (CRIE) and RAST to examine the allergenic content, it was found that allergen 1 of Dr. Ohman is the most important allergenic component, whereas albumin and several unidentified proteins play a minor role. Allergen 1 was not detectable in serum and urine. The allergenic and nonallergenic proteins of pelt extract and saliva were identical by CIE, suggesting that pelt extract proteins are mainly of salivary origin.
Unusual phenotypes of human inducer T cells as measured by OKT4 and related monoclonal antibodies.
Authors: Bach M A , Phan-Dinh-Tuy F , Bach J F , Wallach D , Biddison W E , Sharrow S O , Goldstein G , Kung P C
Published: December 2022

OKT4, a murine monoclonal antibody, was previously shown to react with inducer/helper T cells in man. We now report the absence of this reactivity in 2 subjects of African ancestry and the production of 4 new monoclonal antibodies (OKT4A-D) that detect distinct antigens on human inducer/helper T cells.
The fixation of C3b to pneumococcal cell wall polymers as a result of activation of the alternative complement pathway.
Authors: Hummell D S , Berninger R W , Tomasz A , Winkelstein J A
Published: December 2022

The present study was performed in order to determine the identity of the pneumococcal cell wall polymer(s) to which C3b becomes fixed after activation of the alternative pathway. Purified pneumococcal autolysin was used to solubilize pneumococcal cell walls to which C3b had been fixed via activation of the alternative pathway. The resulting soluble cell wall polymers were then examined for the presence of C3b. Chromatographic separation of cell wall digests containing either radiolabeled teichoic acid or radiolabeled C3b demonstrated that although the elution profiles of the 2 radiolabels were similar, they were not identical. In addition, when teichoic acid-containing polymers were removed from solution by immunoabsorption with TEPC-15 myeloma, only 43 to 65% of the C3b was removed. These results demonstrate that C3b activated via the alternative pathway fixes both to teichoic acid-containing pneumococcal cell wall polymers and to other cell wall constituents and/or serum proteins bound to the cell wall.
Regulatory T cells in pregnancy. IV. Genetic characteristics and mode of action of early MLR suppressive T cell subpopulations.
Authors: Chaouat G , Voisin G A
Published: December 2022

The genetic characteristics, surface markers, and mechanisms of action of early-acting, mitomycin-resistant, MLR suppressor T cells from multiparous mouse spleens were studied in MLR of various strain combinations. The cell activity appears to be genetically restricted at the suppressor/reactive cell level, and the major restriction occurs via the IC subregion. However, C3H.OL recombinant pointed toward a possible minor requirement for S-G subregion syngenicity. The suppressor T cell was characterized as an Ia+, Ly1-, Ly2+ subset. The use of Karush double chambers as well as of supernatants of regulated MLR provided evidence that the suppressor cells act via soluble product(s).
Spontaneous autoimmune thyroiditis in chickens. II. Evidence for autoresponsive thymocytes.
Authors: Livezey M D , Sundick R S , Rose N R
Published: December 2022

A major factor involved in the spontaneously occurring autoimmune thyroiditis of OS chickens is the presence of thyroid reactive cells in the thymus of young OS chicks. This was determined by transferring OS and normal strain thymocytes to T cell-depleted histocompatible normal chickens and examining the recipients for thyroiditis. Although both thymocyte populations reconstituted T cell function, only the OS thymocytes induced thyroiditis. Additional evidence for hyperactivity of the young OS thymus was provided by studying the in situ uptake of a radiolabeled thymidine analogue.
A variant alpha-chain in an Ia molecule from the I-A subregion of the mouse major histocompatibility complex: a possible intragenic recombination.
Authors: Rose S M , Cullen S E
Published: December 2022

The recombinant MHC haplotype t1, present in mouse strain A.TL and B10.TL, encodes an I-A molecule with a variant alpha-chain that was recognized when tryptic peptide digests from t1 and the nominal parental haplotype (a1 or k) were compared by double-label reverse phase HPLC. The t1 I-A alpha-chain possesses a tryptic peptide absent from a1 and k I-A alpha-chains, and lacks several peptides present in a1 and k. The peptide that differs in t1 is not a mannose-bearing peptide, and thus its altered mobility is probably not due to carbohydrate modification. The alteration in t1 could result from mutation or intragenic recombination, but the unique t1 peptide migrates in a position identical to a peptide found in haplotype s, a partner in the (s X a1) cross that generated the t1 recombinant. If this apparently shared peptide indicates an intragenic recombination, this places the I-A alpha-gene centromeric to the other I-A subregion gene, and together with the data of Jones, suggests a gene order of I-A alpha, I-A beta, I-E beta (A alpha, A beta, Ae).
Interaction between Schistosoma mansoni and the complement system: role of IgG Fc peptides in the activation of the classical pathway by schistosomula.
Authors: Ouaissi M A , Auriault C , Santoro F , Capron A
Published: December 2022

It was previously reported that Schistosoma mansoni schistosomula in the presence of IgG activate complement through the classical pathway (CCP). In the present work we have demonstrated that schistosomula incubated with IgG peptides resulting from IgG hydrolysis by schistosomula proteases are able to initiate complement activation through CCP, since a marked consumption of C1, C4, and C2 was observed. Our results eliminate an eventual direct action of the substances released by schistosomula on the activation of CCP. The first step of CCP activation required the preliminary binding of IgG peptides on the schistosomula surface. This interaction induced an increase in the C1q uptake by schistosomula. Since the involvement of the Fc portion of IgG molecules has been clearly evidenced, in this case the peripheral globular subunits of C1q recognize the CH2 region on IgG peptides or intact IgG molecules. By this mechanism, the local consumption of complement around schistosomula could contribute to its survival in the host.
T cells with Fc receptors for IgA: induction of T alpha cells in vivo and in vitro by purified IgA.
Authors: Hoover R G , Dieckgraefe B K , Lynch R G
Published: December 2022

Previous studies demonstrated that BALB/c mice with the IgA-secreting plasmacytomas MOPC-315 (alpha lambda 2), MOPC-167 (alpha kappa), McPC-603 (alpha kappa) and TEPC-15 (alpha kappa) developed large numbers of T cells with surface membrane receptors for IgA (T alpha cells). The lack of T alpha cell expansion in mice with variant plasmacytomas that were nonsecretors or low secretors of IgA implied that elevated serum IgA contributed to the increase in T alpha cells. The present studies show that normal BALB/c mice that were given daily injections of 30 mg of IgA (M315 protein) develop a marked increase in the number of T alpha cells. These studies also show that the T alpha cells induced by injection of IgA are Lyt1-2+ T cells. In addition, the data presented demonstrate that nylon wool nonadherent T cells, treated with purified polymeric IgA (M315 protein) in vitro, develop a marked increase in the number of T alpha cells. The in vitro induction of T alpha cells by IgA requires DNA and protein synthesis. These findings indicate that the T alpha cell expansion observed in mice with IgA myeloma is related to the high serum level of IgA and not to the myeloma tumor per se. In addition, these observations have a more general relevance to the issue of B cell regulation because they demonstrate that secreted immunoglobulin can directly induce expansion of immunoregulatory T cells.