bluuebunny/crossref_metadata_2025 huggingface dataset:
Hapten-specific T cell response to azobenzenearsonate-N-acetyl-L-tyrosine in the Lewis rat. III. Effects of peptide-spacer structure on eliciting ABA-specific helper activity with TNP-haptened ABA-peptide-Ficoll.
Authors: Komatsu T , Lawn C Y , Amsden A , Leskowitz S
Published: December 2022

A series of antigens was synthesized in which peptide spacers were inserted between the ABA group and TNP-Ficoll. When these antigens were used to assess helper activity in Lewis rats immunized with ABA-tyr, it was found that an increase in the ABA-epitope density and an increase in the peptide spacer size both increased the efficacy of the antigen in eliciting ABA-specific help, manifest by an enhancement of anti-TNP PFC. Substitution of D- for L-amino acids progressively decreased the ability of these conjugates to elicit help until D-tyr-D-ala-D-ala, which when used for coupling ABA to the TNP-Ficoll resulted in a nonimmunogenic molecule. When these same Ficoll conjugates were used to study ABA-specific in vitro proliferative responses, it was found that introduction of even a single D-amino acid into the spacer greatly reduced reactivity. By contrast the ABA-peptides, free of the Ficoll backbone, were all equivalent on a molar basis in their ability to elicit ABA-specific in vitro proliferation, regardless of their content of D-amino acids. These results suggest some form of digestion is required in the processing of these antigens before they can elicit helper activity. This processing can occur only if one or more L-amino acid residues are present. If the ABA-peptides are free of the Ficoll backbone, they are all capable of stimulating T cell proliferation without apparent further processing.
Activation of antigen-enriched B cells. II. Role of linked recognition in B cell proliferation to thymus-dependent antigens.
Authors: Snow E C , Noelle R J , Uhr J W , Vitetta E S
Published: December 2022

The responsiveness to T-dependent (TD) and T-independent (TI) TNP-antigens of murine splenic B cells previously enriched for antigen-binding cells (ABC) was examined. TNP-TI antigens induced B cell proliferation. TNP-TD antigens did not induce a proliferative response regardless of the physical form or nature of the TNP-TD antigen (e.g., soluble vs particulate, low or high haptenation of carrier, TNP on various insoluble matrices, etc.). TNP-TD antigens were effective in enhancing the response of the TNP-ABC to all concentrations of lipopolysaccharide (LPS) tested, indicating that binding of antigen to surface immunoglobulin alters the LPS responsiveness of the cell. Irradiated, keyhole limpet hemocyanin- (KLH) primed T cells induced a threefold to fourfold greater B cell proliferative response with TNP-KLH than with fluoresceinated KLH (FLU-KLH) or FLU-KLH together with TNP-human serum albumin (TNP-HSA). Therefore, linked recognition appears essential for optimal T cell-mediated B cell proliferation, whereas the induction of B cell proliferation via nonlinked, carrier-activated T cells is a minor component of the response.
Selective activation of antigen-specific human B cells in recently immunized individuals by nonspecific factors in the absence of antigen.
Authors: Peters M , Fauci A S
Published: December 2022

The in vitro effect of nonspecific factors (derived from mixed lymphocyte culture [MLC] supernatants) on human B cell responses was studied in individuals recently immunized in vivo to keyhole limpet hemocyanin, tetanus toxoid, and/or diphtheria toxin. In T cell-depleted fractions of peripheral blood mononuclear cells, nonspecific factors alone, without antigen, selectively induced a specific antibody response to the antigen to which the individual had been recently immunized, at dilutions that did not generate a significant polyclonal response in the remainder of the B cell repertoire. The source of these factors, with respect to MLC donors, did not affect the antibody response. Supernatants of MLC from nonimmunized individuals induced a specific antibody response as effectively as supernatants of MLC from immunized individuals, when added to B cells plus monocytes from recently immunized individuals. Studies in which the same individuals were followed over time showed that these factor-sensitive B cells are seen in the peripheral blood of recently immunized individuals for only a finite period of time. Thus, in vivo immunization with a specific antigen results in the transient appearance in the peripheral blood of B cells that are specific for the antigen in question. These B cells are probably preactivated in that nonspecific factors selectively induce in vitro their further differentiation into antibody-secreting cells, in the absence of added antigen or mitogen. These studies may add further insight into our understanding of the sequential steps involved in the activation and differentiation of human B lymphocytes and provide a model for the combined in vivo and in vitro study of human B cell physiology.
Antigen-presenting capabilities of human monocytes correlates with their expression of HLA-DS, an Ia determinant distinct from HLA-DR.
Authors: Gonwa T A , Picker L J , Raff H V , Goyert S M , Silver J , Stobo J D
Published: December 2022

Utilizing a monoclonal antibody (Mac-120) specific for 40 to 60% of peripheral blood adherent mononuclear cells (M phi), we were able to separate M phi into two populations based on their reactivity with the antibody. Both populations, Mac-120+ and Mac-120- cells, were then compared for a) their ability to present antigen to T cells, b) their display of HLA-DR determinants, c) their ability to stimulate in an autologous and allogeneic mixed lymphocyte reaction, and d) their display of an Ia molecule, HLA-DS, which is distinct from HLA-DR and which is homologous with murine I-A. Our findings indicate that a) only Mac-120+ cells can present antigen, b) Mac-120+ and Mac-120- cell populations are equivalent in terms of the number of HLA-DR+ cells and in the mean density of HLA-DR determinants per cell, c) although Mac-120+ and Mac-120- cells are equivalent in their ability to serve as stimulators in an allogeneic mixed lymphocyte reaction, Mac-120+ cells are better stimulators in an autologous mixed lymphocyte reaction, and d) only Mac-120+ cells display HLA-DS. These studies demonstrate that peripheral adherent mononuclear cells exhibit heterogeneity with regard to their display of Ia antigens. Furthermore, they provide functional data to support the existence of a human Ia determinant, HLA-DS, which is distinct from HLA-DR and which is important in antigen presentation and stimulation in the autologous mixed lymphocyte reaction.
Cellular interactions in marrow-grafted patients. I. Impairment of cell-mediated lympholysis associated with graft-vs-host disease and the effect of interleukin 2.
Authors: Mori T , Tsoi M S , Gillis S , Santos E , Thomas E D , Storb R
Published: December 2022

Forty patients with hematologic malignancy or aplastic anemia were given allogeneic marrow after conditioning with high-dose cyclophosphamide alone or in combination with total body irradiation. Between 28 and 3857 days after transplantation, their peripheral blood mononuclear leukocytes were tested for reactivity in indirect cell-mediated lympholysis against normal leukocytes from unrelated individuals, and the results were compared to those with cells from their healthy marrow donors. An impairment of cell-mediated lympholysis was found with cells from most patients with acute and chronic graft-vs-host disease (GVHD) whereas cells from most short-term and long-term patients without GVHD had cell-mediated lympholysis reactivity comparable to that of cells from the marrow donors. When interleukin 2 was added to the mixed leukocyte cultures during the sensitization phase, the impaired cell-mediated immunity of cells from most short-term patients with acute GVHD, but not that of cells from most patients with chronic GHVD, could be restored to normal levels. These results suggest the impairment of cell-mediated immunity seen in cells of short-term patients with acute GVHD is attributable to helper cell defects or to ineffective communication between antigen-presenting cells and helper T cells. The impairment in cell-mediated immunity seen in patients with chronic GVHD, however, may reside on the effector cells (or their precursors) or may be due to the presence of suppressor cell activity.
Serologic aspects of IgG4 antibodies. I. Prolonged immunization results in an IgG4-restricted response.
Authors: Aalberse R C , van der Gaag R , van Leeuwen J
Published: December 2022

Labeled antigen-binding tests were used to determine quantitatively the contribution of IgG4 antibodies to the total IgG antibody response in humans. In agreement with literature, we found no IgG4-restricted antibody responses with tetanus toxoid or streptococcal carbohydrate. In the serum of individuals immunized for several years with phospholipase (PLA) from honey bee venom, grass pollen allergen, or house dust mite allergen, we often found that more than 50% of the total antigen-binding capacity was due to IgG4 antibodies. In the case of beekeepers, it could clearly be shown that during prolonged immunization a shift in the IgG4:IgG1 antibody ratio occurs that finally results in an IgG4-dominated antibody response. Evidence is provided that antigen-binding assays may even underestimate the contribution of IgG4 antibodies, because in contrast to IgG1 antibodies, IgG4 antibodies act as monovalent antibodies in being unable to cross-link immunosorbent-bound antigen and radiolabeled antigen.
Antigens of Ambrosia elatior (short ragweed) pollen. III. Crossed radioimmunoelectrophoresis of ragweed-allergic patients' sera with special attention to quantification of IgE responses.
Authors: Løwenstein H , Marsh D G
Published: December 2022

Short ragweed allergenic extract has been studied by means of crossed radioimmunoelectrophoresis (CRIE) with the use of sera from 37 allergic patients and the relevant control sera. In this study 22 of 52 antigens, detectable in crossed immunoelectrophoresis (CIE) against polyspecific rabbit anti-ragweed IgG, were able to bind specific human IgE to their corresponding immunoprecipitates. This binding was semiquantified by comparison with the binding of a standard serum pool. Nine antigens were identified as important allergens, including the previously isolated components, AgE, AgK, and Ra6. Certain allergens (e.g., AgE, AgK, and Ag 31) bound IgE in almost all patients' sera, whereas others showed a bimodal distribution for sera of responder and nonresponder patients. The total CRIE score was found to correlate significantly both with ragweed-specific serum IgE antibody determined by RAST (rs = 0.88; p less than 0.001) and with total IgE level (rs = 0.55; p less than 0.01). Patient's CRIE scores to AgE also correlated significantly with their specific IgE antibody to AgE measured by RIA (r = 0.47; p less than 0.01) and with skin-test sensitivity to AgE (r = 0.44; p less than 0.05). It was concluded that CRIE is well suited for identification of important ragweed allergens without the previous need for laborious isolation procedures.
Generation of alloreactive cytotoxic T lymphocytes: evidence for a differentiation factor distinct from IL 2.
Authors: Finke J H , Scott J , Gillis S , Hilfiker M L
Published: December 2022

In this report we provide evidence that the generation of cytotoxic T lymphocytes (CTL) involves a differentiation signal in addition to a proliferative signal, and that the differentiation signal is mediated by a factor distinct from interleukin 2 (IL 2). Generation of CTL from nylon wool-nonadherent splenocytes stimulated by heat-treated thymocytes did not correlate with the addition of IL 2-containing supernatants to this factor-dependent bioassay. IL 2 from two different sources and prepared by different methods (purified IL 2 from the LBRM33 cell line and partially purified IL 2 from the EL4 cell line) was not sufficient to generate a cytotoxic response in the bioassay. The inability of IL 2 to generate a cytotoxic response was not due to the use of heat-treated stimulator cells because similar results were obtained with UV-treated cells. As we previously reported, supernatant fluids containing a differentiation/activation factor did generate CTL from nylon wool-nonadherent splenocytes. Supernatant fluid from co-cultures of activated T cells and M phi as well as from cultures of activated M phi contained this CTL helper factor; however, only the former supernatant contained IL 2 activity. These results suggest a factor distinct from IL 2 is providing a differentiation/activation signal that is necessary for CTL development. Although IL 2 was not effective at generating CTL from nylon wool-nonadherent splenocytes, it was capable in some, but not all, experiments of generating a cytotoxic response from unseparated splenocytes. The ability of IL 2 to induce a cytotoxic response from unseparated splenocytes appeared to correlate with the presence of a weak background response seen when unseparated cells were used as responder cells.
Limiting dilution analysis of the frequency of human T cells and large granular lymphocytes proliferating in response to interleukin 2. II. Regulatory role of interferon on proliferative and cytotoxic precursors.
Authors: Vose B M , Riccardi C , Bonnard G D , Herberman R B
Published: December 2022

Limiting dilution analysis has shown that large granular lymphocytes (LGL) and small T cells undergo proliferation in response to lectin-free IL 2. The latter is critically dependent on prior stimulation with lectin. Depending on conditions, alpha or beta interferons (IFN) were found to have either of two opposing effects on the frequency of proliferating cells. Pretreatment of responder cells with IFN resulted in dose-dependent augmentation of proliferative progenitors such that at 500 IU/ml, a fivefold increase of progenitor frequency was apparent. Under such conditions, approximately 5% of LGL could be expanded, and at least a proportion of the cultured cells killed the NK-sensitive K562 cell line. Similar results were apparent with T cell responders: whatever the initial frequency of proliferation (dependent on the PHA dose), augmentation was obtained with IFN pretreatment, although no killing of K562 was induced. In contrast, when IFN was present throughout the 7-day assay period, proliferative frequencies were reduced. This inhibitory effect was mediated through the presence of irradiated T cells in the feeder populations. With purified monocytes as feeder cells, little reduction was seen, whereas addition of T cells resulted in a 19-fold inhibition. Separation of OKT4+ and OKT8+ subsets demonstrated that both were essential for optimum induction of suppression. These data indicate that IFN has a powerful immunodulatory role on both NK and T cell proliferation, and that it may function both by induction of IL 2 receptors and activation of suppressive T cells. The interaction between different soluble mediators provides a complex immunoregulatory circuit in vitro.
Staphylococcus aureus protein A induces the production of interferon-alpha in human lymphocytes and interferon-alpha/beta in mouse spleen cells.
Authors: Smith E M , Johnson H M , Blalock J E
Published: December 2022

Human and mouse lymphocytes produced IFN-alpha and IFN-alpha/beta, respectively, when incubated with highly purified Staphylococcus aureus protein A (SPA). Maximum IFN induction occurred when lymphocytes were incubated with SPA at 250 to 500 micrograms/ml for 24 hr. The SPA induced-IFN were identified by neutralization of antiviral activity with specific antisera. A commercial preparation of SPA was found to induce both HuIFN-alpha and HuIFN-gamma, with the latter comprising over 90% of the antiviral activity. The SPA preparation that induced IFN-gamma was shown by SDS-polyacrylamide gel electrophoresis and an enzyme-linked immunosorbent assay to be contaminated with 5% S. aureus enterotoxin A (SEA), a potent T lymphocyte mitogen and IFN-gamma inducer. Preincubation of the commercial SPA with a specific antiserum to SEA blocked the induction of IFN-gamma and had no effect on IFN-alpha production. SEA is a very potent IFN-gamma inducer, effective at concentrations of 1 ng/ml or less. Thus, very minor contamination of SPA by SEA or other IFN-gamma inducers could be responsible for previous reports that SPA induces IFN-gamma in lymphocytes or that SPA behaves as a T cell mitogen.
A newly defined murine alloantigen controlled by the S region of the H-2 complex: molecular association with the fourth component of complement.
Authors: Spinella D G , Passmore H C
Published: December 2022

A previous study has shown that prolonged immunization of strain A.TBR16 mice (H-2 haplotype at16) with lymphoid tissue derived from A.TBR13 (at13) donors results in an antiserum that defines a new H-2-associated allotype with a strain distribution antithetical to H-2.7. The present report describes a sensitive ELISA that demonstrated that this specificity represents an allotypic determinant(s) on the fourth component of complement (C4). Studies with Sephadex G-200 fractionated plasma and serum proteins suggest that, as is the case for H-2.7, the new specificity appears to reside on the C4d fragment after C activation. We therefore propose the tentative nomenclature C4d.2 for this specificity and suggest revision of the nomenclature for H-2.7 to C4d.1. Also described in this report is the formal mapping of the genetic control of the C4d.2 allotype to the H-2S region.
Immune enhancement in mice by ara-A.
Authors: Hinrichs J , Kitz D , Kobayashi G , Little J R
Published: December 2022

Ara-A in single, apparently nontoxic doses stimulates murine immune responses to several well-defined antigens. Both humoral and cell-mediated responses are enhanced by ara-A, but not all mouse strains respond equally well. In certain mouse strains, ara-A also promotes increased (or decreased) survival during experimental infection with Cryptococcus neoformans, suggesting that clinically significant effects on host resistance may result from the use of ara-A as an antiviral therapeutic agent. The biochemical mechanisms of action of the aranucleosides suggest that the immunostimulation produced by ara-A depends on suppressor cell toxicity.
Hormonal activation of adenylate cyclase in macrophage membranes is regulated by guanine nucleotides.
Authors: Verghese M W , Snyderman R
Published: December 2022

Many macrophage functions such as chemotaxis, phagocytosis, enzyme secretion, and cytotoxicity are influenced by intracellular cyclic nucleotide levels, but the regulatory mechanisms involved are poorly defined. We have developed methods that allowed us to study the activation of AC in isolated guinea pig (g.p.) macrophage membranes. AC in these membrane preparations could be stimulated approximately twofold by guanine nucleotides. We could not obtain any hormonal activation of membrane-bound AC in the absence of guanine nucleotides. In the presence of GTP, however, the hormones isoproterenol and PGE1 elicited an additional threefold rise in AC activity, which subsided after approximately 15 min. As little as 10(-8) M concentrations of these two hormones induced significant elevations of AC activity. Replacement of GTP by its nonhydrolyzable analogue Gpp(NH)p resulted in a persistent hormone-independent activation of AC, and addition of hormones enhanced this level of activation. Thus, GTP-ase activity is present in macrophage membrane preparations and serves to regulate AC activation. Hormonal stimulation of AC was receptor mediated, because the effect of the beta-adrenergic agonist isoproterenol, but not PGE1, was inhibited by the beta-adrenergic blocker propranolol. In addition, the potency series of PG corresponded to that observed for stimulation of cAMP production in intact g.p. macrophages, i.e., PGE1 = PGE2 greater than PGA1 greater than PGF2 alpha. AC activation by PG in the membrane preparation was inhibited by an alpha-adrenergic agonist, thus demonstrating one means for down regulating cAMP production in g.p. macrophages. Our studies also showed that certain hormones (e.g., beta-adrenergic agonists, PG) can exert their effect on cAMP production by stimulation of membrane-bound AC, whereas other agents such as lectins or arachidonic acid require additional intracellular components to elevate cAMP levels in macrophages. The mechanism of activation of AC by hormones in g.p. macrophage membranes appears to fit the model of a ternary complex, the components of which include the hormone receptor, AC, and guanine nucleotide regulatory protein, which transmits the signal from the receptor to AC.
A new lymphocyte cell surface antigen, Ly-22.2, controlled by a locus on chromosome 4 and a second unlinked locus.
Authors: Meruelo D , Offer M , Flieger N
Published: December 2022

A serum raised by immunizing (B10.A(4R) X B10.HTT)F1 mice against A.AL lymphocytes detects a new antigenic determinant designated Ly-22.2. Ly-22.2 expression is under the control of two independently segregating genes, one of which maps to chromosome 4 adjacent to a locus affecting XenCSA expression. Ly-22.2 is present in varying amounts in all lymphoid organs, but appears to be expressed primarily on T lymphocytes. Ly-22.2 is not detectable in brain, kidney, lung, liver, or erythrocytes. Strain distribution studies show Ly-22.2 is present in all strains examined except B10-derived congenic strains. It is of interest that C57BL/6 and C57BL/10 mice differ in the expression of this antigen.
Influence of dietary restriction and aging on natural killer cell activity in mice.
Authors: Weindruch R , Devens B H , Raff H V , Walford R L
Published: December 2022

Natural killer cells (NK) are believed to defend against tumor growth. Because rodents subjected to dietary restriction without malnutrition live longer and develop spontaneous tumors less often or later in life than unrestricted controls, we measured NK activity in restricted and in unrestricted mice. An age-related decline in NK responses to YAC-1 tumor target cells was detected in both groups. NK responses for control mice were highest in 2- to 3-mo-old mice, sharply reduced in middle-age mice (14 to 15 mo), and slightly reduced further in old mice (30 to 33 mo). At all ages the response of restricted mice was less than that of controls. However, after injection with Poly I:C (which increases NK activity), old restricted mice showed NK cytolysis not different from young mice on either diet, and substantially higher responses than old unrestricted mice. In addition, restricted mice showed increased in vitro generation of cytotoxic T lymphocytes (CTL) to YAC-1 and P815 compared with age-matched controls. Restricted mice may better resist cancer via an NK system very responsive to induction signals coupled with a CTL system more effective than that of unrestricted controls.
Pteridines as a new marker to detect human T cells activated by allogeneic or modified self major histocompatibility complex (MHC) determinants.
Authors: Huber C , Fuchs D , Hausen A , Margreiter R , Reibnegger G , Spielberger M , Wachter H
Published: December 2022

Evidence is presented that activation of T cells by allogeneic or modified autologous cells leads to the specific production of two distinct molecular compounds of the PT class. One of these two PT has previously been identified as neopterin. The structure of the other is still under investigation. This conclusion was based on the following findings: a) In vitro both PT were produced by T cells proliferating in response to HLA incompatible allogeneic or to autologous EB-virus-transformed or TNP-haptenized cells. b) In vitro stimulation of T cells with mitogenic lectins or protein antigens, of macrophages with zymosan-complement, of B cells with PWM or EB-virus, and of NK effector cells with tumor targets failed to induce comparable release of these PT. c) In vivo increased amounts of both PT were excreted via the urine during allograft rejection and viral infections. It thus appears that the production of these PT is primarily a feature of activated T cells involved in the control of self integrity.
T cell regulation of B cell activation: antigen-specific and antigen-nonspecific suppressor pathways are mediated by distinct T cell subpopulations.
Authors: Asano Y , Hodes R J
Published: December 2022

The present studies were carried out to characterize the cellular events involved in the induction and function of carrier-specific Ts cells, which selectively regulate the generation of IgG responses by Lyb-5- B cells. It was demonstrated that this regulation is in fact mediated by two distinct suppressor pathways. In one pathway, carrier-primed Lyt-1 + 2 - Ts cells are specifically activated by in vitro reexposure to the priming antigen. After this specific activation, these Lyt-1 + 2 - Ts cells are able to suppress IgG responses in an antigen-nonspecific manner. This suppression requires the participation of unprimed Lyt-1 - 2 + T cells, and is effective in both the early and the late phases of antibody responses. A second suppressor pathway requires the antigen-specific activation of primed Lyt-1 - 2 + Ts cells. Suppression of antibody responses by activated Lyt-1 - 2 + Ts cells is highly carrier specific, in contrast to the nonspecific effector function of Lyt-1 + 2 - Ts cells, appears to act without requirement for additional T cell populations; and is effective only early in the course of the antibody response. Thus, it appears that two Ts cell populations may function through distinct mechanisms to regulate the generation of IgG Lyb-5- B cell responses.
The origin of cytotoxic T lymphocyte precursors (CTL-P): MHC-restricted and alloreactive CTL-P in the spleen during regeneration after a sublethal dose of cyclophosphamide (Cy).
Authors: Sihvola M , Hurme M
Published: December 2022

The regenerating spleen 8 days after an injection of a sublethal dose (300 mg/kg) of cyclophosphamide (Cy) had a defective capacity to give rise to cytotoxic T lymphocytes (CTL) in response against allogeneic cells, whereas the cytotoxicity against 2,4,6-trinitrophenyl-(TNP) modified syngeneic cells was at the normal level. Alloresponse was first obtained 2 wk after the Cy treatment. The limiting dilution analysis showed this at the clonal level: the frequency of anti-TNP-specific CTL precursors (CTL-P) in the spleen treated with Cy 8 days previously was the same as the frequency in the normal spleen. The defective alloresponse was due to a decreased number of allospecific CTL-P that was later increasing. The regenerative capacity was not abolished by adult thymectomy or treatment of the mice with a bone marrow-seeking isotope, 89Sr, suggesting that these CTL-P are derived from Cy-resistant splenic precursors rather than from the thymus or bone marrow. These precursors have probably been under thymic education: the dominance of H-2k-restricted CTL over H-2d-restricted CTL in the response of (H-2k X H-2d)F1 mice to TNP-self is known to be influenced by the H-2 genotype of the thymus, and this dominance was also demonstrated with anti-TNP CTL derived from these F1 mice pretreated with Cy. The CTL-P in the regenerating spleen (day 8) were not hydrocortisone sensitive, and nylon wool-purified T cells from this stage had a lactate dehydrogenase (LDH) isoenzyme pattern of the mature T cell type (rather than of the thymocyte type). Thus, in these aspects the T cells of the regenerating spleen resembled normal splenic cells. These data suggest that the Cy-resistant spleen population contains cells that can give rise to CTL-P that have a defective specificity repertoire at the beginning of the regeneration, but later mature to a normally alloreactive population.
Correlation between differentiation, expression of monocyte-specific antigens, and cytotoxic functions in human promyelocytic cell lines treated with leukocyte-conditioned medium.
Authors: Dayton E T , Perussia B , Trinchieri G
Published: December 2022

Human promyelocytic cells lines treated with conditioned medium from PHA-stimulated leukocytes acquire several phenotypic and functional markers of differentiated monocytes. In this paper, we demonstrate that promyelocytic cells treated with conditioned medium express, among other markers, monocyte-specific and HLA-DR antigens absent from the parental cells and become potent effectors of antibody-dependent cell-mediated cytotoxicity against erythrocytes and tumor cells. In cultures of promyelocytic cell lines maintained in the presence of conditioned medium, an equilibrium between proliferation and differentiation is established, and two cell populations can be separated on the basis of expression of differentiation surface markers. One population has a differentiated morphology, expresses nonspecific esterase activity, Fc receptors, C receptors, monocyte-specific and HLA-DR antigens, is able to mediate antibody-dependent cytotoxicity, and has a limited ability to proliferate. A second population retains the phenotype of undifferentiated promyelocytes and continues to proliferate. The differentiated monocyte-like cells originate from a proportion of the proliferating promyelocytes that respond to the differentiation inducers contained in the conditioned medium.
Estimation of polymeric IgA in human serum: an assay based on binding of radiolabeled human secretory component with applications in the study of IgA nephropathy, IgA monoclonal gammopathy, and liver disease.
Authors: Newkirk M M , Klein M H , Katz A , Fisher M M , Underdown B J
Published: December 2022

Binding of 125I-human secretory component (SC) to human polymeric immunoglobulin A (pIgA) was employed to measure quantitatively the pIgA present in human sera. Interference by IgM in some sera was prevented by removal of IgM with glutaraldehyde polymerized anti-IgM antibodies. 125I-SC complexed to pIgA was measured by precipitation with anti-IgA antibodies and the quantity of pIgA in human serum was estimated by comparing the quantity of 125I-SC bound by several dilutions of human serum to that bound by standard quantities of human monoclonal pIgA proteins. The assay was specific for pIgA because heat-aggregated monomeric IgA or hypogammaglobulinemic serum did not bind 125I-SC greater than a precipitate formed with human monoclonal IgG and anti-IgG. Moreover, analysis of a series of IgA myeloma sera indicated no correlation between the IgA content of the serum and the quantity of pIgA measured. The quantity of pIgA found in 30 normal human sera was 0.13 +/- 0.08 mg/ml (1S.D.), which consisted of 11.3 +/- 5.3% (1 SD) of the total IgA. Patients with IgA monoclonal gammopathy were most often found to have predominantly monomeric IgA. Patients with IgA nephropathy also showed an elevation of pIgA, but this appeared to be a consequence of an overt IgA elevation. IgA nephropathy patients with elevated serum IgA in fact showed a significant elevation of monomeric IgA. Selective elevation of pIgA was observed in patients with primary biliary cirrhosis and alcoholic liver disease. A comparison of this assay with other assays to measure pIgA is discussed.
Immune islet killing mechanisms associated with insulin-dependent diabetes: in vitro expression of cellular and antibody-mediated islet cell cytotoxicity in humans.
Authors: Charles M A , Suzuki M , Waldeck N , Dodson L E , Slater L , Ong K , Kershnar A , Buckingham B , Golden M
Published: December 2022

Because immune mechanisms are associated with insulin-dependent diabetes, multiple organ specific and xenogeneic cytotoxicity assays were developed. Human cellular and antibody effector systems were incubated with 51Cr-labeled dispersed normal rat islet target cells. In eight of 11 diabetic patients, nonenriched mononuclear cells incubated with islet target cells were more cytotoxic than cells from age- and sex-matched controls (p less than 0.01). When non-T cell-enriched mononuclear cells were used at diabetes onset, seven of 11 patients' cells showed excessive islet cytotoxicity (p less than 0.05). In four patients showing elevated cytotoxicity at diabetes onset, cytotoxicity decreased to control levels during diabetes remission. Islet specificity was suggested in that mononuclear cells derived from diabetic subjects did not mediate cytotoxicity against rat spleen or macrophage target cells. Three cytotoxic antibody mechanisms were also evaluated. C-dependent antibody-mediated cytotoxicity with the use of patient serum-coated islet target cells was elevated above control levels in four of 14 patients. Antibody-dependent cellular cytotoxicity exceeded control values in only two of 16 patients, although four assay systems were evaluated. C-augmented antibody-dependent cellular cytotoxicity was elevated in three of 14 patients. No differences were observed for antibody-mediated mechanisms in four patients evaluated at both diabetes onset and remission. Cytotoxic antibody was present in only about one-half of the patients showing increased cellular cytotoxicity, whereas most patients expressing increased cytotoxic antibody had cellular cytotoxicity. Islet cell cytotoxicity assays with the use of effector systems from patients with recent onset insulin-dependent diabetes suggest that direct cellular cytotoxicity is more active than antibody-mediated cytotoxic mechanisms, and that cellular cytotoxicity can correlate with disease activity.
Detection of autoimmune cells proliferating to myelin basic protein and selection of T cell lines that mediate experimental autoimmune encephalomyelitis (EAE) in mice.
Authors: Ben-Nun A , Lando Z
Published: December 2022

A procedure is described for the detection of an in vitro proliferative response to the autologous mouse myelin basic protein in mice injected with mouse spinal cord homogenate (MSCH) or with myelin basic proteins (BP) of mouse (MBP) or rat (RBP) origin. The administration of MSCH, but not of MBP or RBP, in a suitable adjuvant could produce a reproducible clinical disease. Nevertheless, a proliferative response to the autologous MBP could not be detected after either inoculation. Only the removal of the adherent cell fraction from the immunized cell population and its replacement with fresh naive accessory cells could reveal a proliferative response to the autologous MBP and to the heterologous RBP. A heteroclitic cross-reactivity between MBP and RBP is demonstrated. The possibility of detecting an in vitro proliferative response to BP allowed the selection and propagation in vitro of cells specific to BP. T cell lines were established that specifically proliferated in response to BP and mediated EAE in normal mice. Intravenous inoculation of as few as 10(6) line cells was capable of producing clinical signs of EAE in normal recipients within 5 to 6 days. Thus, although an inoculation of MSCH was required for active induction of the disease, T cells specifically reactive against BP are sufficient for mediation of EAE in mice.
Interferon-induced enhancement of macrophage Fc receptor expression: beta-interferon treatment of C3H/HeJ macrophages results in increased numbers and density of Fc receptors.
Authors: Vogel S N , Finbloom D S , English K E , Rosenstreich D L , Langreth S G
Published: December 2022

Previous studies demonstrated that the augmentation of Fc receptor-(FcR) mediated phagocytosis and binding of opsonized sheep erythrocytes is cytokine mediated. Specifically, beta-interferon- (beta-IFN) and gamma-IFN-rich preparations were shown to increase binding and phagocytosis of opsonized erythrocytes by C3H/HeJ macrophages. In this report we extend these studies by examination of the mechanisms by which highly purified beta-IFN increases FcR capacity in C3H/HeJ macrophages. Our findings indicate that beta-IFN augments FcR function for both IgG2a and IgG2b receptor subclasses. Moreover, this increased ability to bind and phagocytose opsonized erythrocytes is associated with a concomitant increase in both the number and surface membrane density of FcR.
Potentiation of the cytolytic activity of peripheral blood monocytes by lymphokines and interferon.
Authors: Fischer D G , Golightly M G , Koren H S
Published: December 2022

Human peripheral blood monocytes obtained by EDTA-reversible adherence to plastic surfaces precoated with autologous serum can rapidly lyse a variety of tumor cells. That the effector cells in this system are indeed monocytes has been demonstrated (1). Using a short-term (3 to 4 hr) 51Cr-release assay and the single cell conjugate cytotoxic assay, we studied the effects of lymphokine-rich supernatants containing gamma-interferon and partially purified fibroblast interferon on the monocyte cytolytic activity. Overnight incubation of the monocytes in fetal bovine serum-containing medium resulted in a relatively small decrease in cytotoxic activity compared to the one obtained with monocytes incubated in autologous serum. The addition of lymphokines or interferon under both incubation conditions resulted in augmented activity as measured in the 51Cr-release assay. However, the proportions of binding and cytotoxic monocytes, determined in the single cell conjugate assay, did not increase. These results suggest that augmented activity is not due to recruitment of inactive cells. Kinetics studies of tumor cell lysis indicate the increase in killing efficiency is probably due to both an increase in the rate of killing and in the recycling ability of the cytotoxic cells. Using the conjugate/agarose technique, we also demonstrated that excess tumor cells could impair the lytic machinery of freshly isolated monocytes, whereas monocytes treated with lymphokines or interferon partially lost their sensitivity to this inhibitory effect. The ability of tumor cells to impair the lytic machinery of monocytes could be one of the mechanisms by which tumors escape immunosurveillance.
Characterization of human B cell growth factor (BCGF) from cloned T cells or mitogen-stimulated T cells.
Authors: Yoshizaki K , Nakagawa T , Fukunaga K , Kaieda T , Maruyama S , Kishimoto S , Yamamura Y , Kishimoto T
Published: December 2022

An assay system for the measurement of human BCGF was established by employing anti-id-stimulated B-CLL cells, anti-mu-stimulated normal B cells, or a transformed B cell line. PHA-Sup from PHA-stimulated T cells induced proliferation of anti-lg-stimulated B cells or augmented proliferation of a B cell line, and the activity was dose-dependent. BCGF, IL 2, and BCDF activities in PHA-Sup were eluted in a 17K fraction by gel filtration in 0.5 M NaCl. BCDF activity (pl 5.8) was isolated by chromatofocusing, but BCGF and IL 2 activities (pl 6.5 and 6.9) were not separated. Absorption with an IL 2-dependent T cell line or anti-id-stimulated B-CLL cells showed BCGF and IL 2 were distinct molecules. An IL 2-dependent helper T cell clone, d4, secreted BCGF with the m.w. of 50K. The 50K-BCGF fraction had no IL 2 activity and was eluted in a 19K fraction by gel filtration in 4 M urea and 1 M NaCl. PMA stimulation of normal T cells induced 50K-BCGF, the m.w. of which was reduced to 19K in 4 M urea and 1 M NaCl. 50K-BCGF from d4 cells and 17K-BCGF from PHA-Sup showed a synergistic effect on the proliferation of anti-lg-stimulated B cells. The results show the existence of two distinct kinds of BCGF and the presence of the synergism between them.
Anaphylatoxin-mediated regulation of the immune response. II. C5a-mediated enhancement of human humoral and T cell-mediated immune responses.
Authors: Morgan E L , Thoman M L , Weigle W O , Hugli T E
Published: December 2022

C5a derived from the fifth component of human complement was found to potentiate both specific and non-specific human in vitro humoral immune responses. In addition, C5a is capable of potentiating antigen- and alloantigen-induced T cell proliferative responses. In contrast to these results, C5a was unable to modulate mitogen-induced B and T cell proliferative responses. The carboxyterminal arginine of C5a is not essential in order for C5a to enhance immune responses. C5ades Arg was found to augment the immune response to the level of C5a-mediated enhancement. The serum carboxypeptidase inhibitor, 2-mercaptomethyl-5-quanodinopentanoic acid, which prevents cleavage of the terminal arginine, allowed us to assay the effects of C5a on in vitro immune responses in the presence of serum. Helper T cells are involved in C5a-mediated immuno-potentiation. Substitution of T cells by soluble T cell-replacing factors, (Fc)TRF, rendered lymphocyte cultures refractory to the enhancing properties of C5a.
Presence of an intrastrain cross-reactive idiotype on A/J antibodies of the IgE class specific for the p-azophenylarsonate group.
Authors: Hirano T , Kojima S , Hirayama N , Nelles M J , Inada T , Nisonoff A , Ovary Z
Published: December 2022

An intrastrain cross-reactive idiotype, CRIA, is associated with a large proportion of the anti-p-azophenylarsonate (anti-Ar) antibodies of A/J mice. The present experiments, in which the methods of direct and reverse passive cutaneous anaphylaxis (RPCA) were used, indicate that IgE anti-Ar antibodies are produced in A/J mice upon stimulation with a protein-Ar conjugate and that a large proportion of these antibodies express CRIA. The use of monoclonal anti-CRIA for RPCA eliminated the strong nonspecific reactions previously observed. The results provide a basis for studying factors that regulate the switch to IgE biosynthesis during an immune response.
Studies of the Epstein-Barr virus receptor found on Raji cells. I. Extraction of receptor and preparation of anti-receptor antibody.
Authors: Simmons J G , Hutt-Fletcher L M , Fowler E , Feighny R J
Published: December 2022

Raji, a human lymphoblastoid cell line, expresses a membrane receptor (EBVR) specific for Epstein Barr virus (EBV). A component that binds EBV was extracted from this cell line by treatment of the cells for 3 hr on ice with Tris buffer containing 10% glycerol. The treatment reduced the capacity of the cells to bind virus, and after concentration the receptor extract (RE) inhibited both EBV binding and superinfection of fresh Raji cells. Similarly prepared extracts of EBVR- cells lacked such activity. An antibody was made to the extract (anti-RE), which after absorption with EBVR- cells, bound to the same percentages of EBVR+ lymphoblastoid cell lines, EBVR+ human/mouse somatic cell hybrids, and fresh peripheral B cells as the virus did. In reciprocal assays, preincubation of EBVR+ cells with anti-RE inhibited virus binding. Doubly stained patches were observed on membranes of EBVR+ cells that had been incubated simultaneously with virus and anti-RE and stained respectively with rhodaminated and fluoresceinated reagents. The major polypeptide immunoprecipitated by anti-RE from radiolabeled Raji cells had an approximate calculated m.w. of 150,000.
Schistosoma mansoni adult microsomal antigens, a serologic reagent. II. Specificity of antibody responses to the S. mansoni microsomal antigen (MAMA).
Authors: Tsang V C , Hancock K , Kelly M A , Wilson B C , Maddison S E
Published: December 2022

The purified Schistosoma mansoni adult microsomal antigen, MAMA, was used in the quantitative single-tube kinetic dependent enzyme-linked immunosorbent assay (k-ELISA) to measure antibody levels of various human patient sera. The 511 serum specimens tested were from patients with both homologous and heterologous infections. Sera from U.S., Egyptian, Brazilian, and Puerto Rican patients infected with S. mansoni reacted strongly with MAMA. Chinese patients infected with S. japonicum, and Nigerians or Egyptians infected with S. haematobium produced much lower responses to this antigen than those infected with S. mansoni. Sera from patients with echinococcosis, filariasis, paragonimiasis, clonorchiasis, trichinosis, amebiasis, and hepatitis and from healthy uninfected control individuals generally contained no detectable antibodies against this antigen. The S. mansoni adult microsomal antigen, MAMA, therefore, appears to be a highly potent and specific reagent for the serodiagnosis of S. mansoni infections.
Anti-Lyt-1 monoclonal antibody inhibits induction of anti-self-TNP but not of alloreactive cytotoxic T lymphocytes.
Authors: Guimezanes A , Schmitt-Verhulst A M
Published: December 2022

The role of Lyt-1+ T cells in the induction of anti-self + TNP and anti-allogeneic CTL was investigated by the use of monoclonal antibodies directed against these molecules. When present during the 5 days of in vitro induction of CTL, anti-Lyt-1.1 mAb partially inhibited the induction of anti-self + TNP CTL but did not affect the induction of alloreactive CTL. The inhibitory effect was dose-dependent, Lyt-1 allotype-specific, and directed at the responding cells. The inhibition could be abolished by the addition of interleukin 2-containing supernatants. These results suggest that under some experimental conditions, Lyt-1-specific mAb can be shown to affect a T cell function, probably a helper function. Possible differences in the induction mechanisms of anti-self + TNP and alloreactive CTL are discussed.
Cytolytic T lymphocyte clones that proliferate autonomously to specific alloantigenic stimulation. II. Relationship of the Lyt-2 molecular complex to cytolytic activity, proliferation, and lymphokine secretion.
Authors: Glasebrook A L , Kelso A , MacDonald H R
Published: December 2022

Previous analyses of the inhibitory effects of anti-Lyt-2 monoclonal antibodies (mAb) on cytolytic activity suggested that Lyt-2/3 antigens expressed on the surface of murine cytolytic T lymphocytes (CTL) are involved in antigen recognition. In the present study, we investigated the effects of anti-Lyt-2 mAb (in the absence of complement) on the functional activities of H-2K/D-specific Lyt-2+ CTL clones that proliferate to antigenic stimulation in the absence of helper T cells or added interleukin 2 (IL 2) and secrete lymphokines. For those clones that were inhibited in cytolysis by anti-Lyt-2 mAb, a parallel inhibition of antigen-dependent proliferation and lymphokine secretion (interferon, macrophage-activating factor) was observed. Inhibition of proliferation or lymphokine secretion could be overcome by the addition of IL 2 or lectin, respectively. Collectively, these results would strongly suggest that anti-Lyt-2 mAb were inhibiting CTL antigen recognition. Not all CTL clones, however, were inhibited in cytolysis by anti-Lyt-2 mAb, in which case proliferation and lymphokine secretion were similarly unaffected. This heterogeneity of Lyt-2+ CTL clones in their susceptibility to inhibition of cytolytic activity, proliferation, and lymphokine secretion by anti-Lyt-2 mAb is discussed in the context of a model proposing that Lyt-2/3 molecules function to stabilize the interaction between CTL receptors and the corresponding target/stimulating cell antigens. Such a stabilization may be required by CTL possessing few and/or low affinity receptors.
Analysis of murine T lymphocyte markers during the early phases of GvH-associated suppression of cytotoxic T lymphocyte responses.
Authors: Hurtenbach U , Shearer G M
Published: December 2022

Graft-vs-host- (GvH) associated immune-suppression was generated by injecting nonirradiated (B6 X C3H)F1 hybrid mice with 40 to 60 X 10(6) B6 or C3H parental spleen cells. Susceptibility to suppression as well as the potential of parental donor lymphocytes to generate suppressor activity was investigated in allogeneic cytotoxic T cell responses by mixing spleen cells from the inoculated F1 recipients with syngeneic responder cells from untreated mice. The susceptibility to suppression in F1 recipients, as well as the potential of parental lymphocytes to induce suppression in F1 mice, varied with the age of the mice. Cellular analysis revealed that GvH-suppression was mediated by 500 R radiosensitive parental T cells during the period tested (from day 5 until day 8 after injection of B6 parental spleen cells). In contrast, selected F1 lymphocytes from recipients injected with parental spleen cells failed to exhibit suppressor cell activity. The sensitivity of parental GvH suppressor cells to anti-Lyt reagents was time dependent. Thus, suppressor cells were sensitive to monoclonal anti-Lyt-1 or anti-Lyt-2 and C (usually until day 7 after injection of parental spleen cells); subsequently, suppression became insensitive to anti-Lyt-1, but remained sensitive to anti-Lyt-2 and C treatment.