bluuebunny/crossref_metadata_2025 huggingface dataset:
Release of histamine from human leukocytes stimulated with the tumor-promoting phorbol diesters. II. Interaction with other stimuli.
Authors: Schleimer R P , Gillespie E , Daiuta R , Lichtenstein L M
Published: December 2022

The tumor promoter and irritant, 12-O-tetradecanoyl-phorbol-13-acetate (TPA), previously shown to be a potent histamine releaser, has been used to further probe the mechanism of histamine release from human basophils. TPA and the calcium ionophore, A23187, produced a synergistic response in which subeffective concentrations of each stimulus (which alone produced less than 3% release) together produced over 70% histamine release. TPA also synergized with the IgE cross-linking stimulus anti-IgE. Desensitization of cells by incubation with anti-IgE in the absence of calcium rendered the cells unresponsive to anti-IgE and super-responsive to TPA. This marked increase in the TPA response was the result of an increase in the rate of TPA-induced histamine release, and occurred in the absence of extracellular calcium. The ability of various concentrations of anti-IgE to "sensitize" cells to TPA paralleled their ability to produce histamine release in untreated cells rather than their ability to desensitize the cells. These results suggest that in the absence of calcium, anti-IgE induces desensitization of some activation of other elements of the histamine-release process. The anti-IgE dose-response pattern of this activation event further suggests that it is an integral part of the anti-IgE-induced release process itself.
Chido and Rogers antigenic determinant on the fourth component of human complement.
Authors: Chu V F , Marsh W L , Gigli I
Published: December 2022

The blood group substances Chido (Cha) and Rogers (Rga) represent two electrophoretic variants of human C4. Based on the observation that anti-Cha and anti-Rga antisera agglutinated human red blood cells prepared in sucrose-activated autologous serum (LIS cells) at 37 degrees C, it has been assumed that the Cha and Rga antigenic determinants reside in the C4d fragment of C4. Here, we present evidence indicating that C4d is not present on those cells. In order to identify structurally the C4 fragments deposited, LIS cells were prepared at 37 degrees C and 4 degrees C in autologous serum to which 125I-C4 was added. Membranes of LIS cells were solubilized and analyzed by SDS-PAGE in 5 to 15% gradient gels followed by autoradiography. C4d was not deposited on LIS cells prepared at 37 degrees C, whereas C4c (beta, gamma, alpha 3 alpha 4) was. Cells prepared at 4 degrees C carried C4d (alpha 2) and C4c. Anti-Cha and anti-Rga antisera agglutinated both cell types, although C4d was not present on the cells prepared at 37 degrees C. Purified C4, C4c, C4d, and alpha-, beta- and gamma-chains of C4, as well as alpha 3 and alpha 4, were used to neutralize these antisera. C4 and the alpha-chain C4d and alpha 4 fragment of C4c, but neither the alpha 3 fragment nor the beta- or gamma-chains, were capable of neutralizing anti-Cha and anti-Rga antisera. These results strongly suggest that C4d and alpha 4 share an antigenic determinant, both of which are recognized by anti-Cha and anti-Rga antisera.
Biochemical and immunologic heterogeneity of Ia glycoproteins isolated from a chronic lymphocytic leukemia.
Authors: Letarte M , Falk J
Published: December 2022

Ia glycoproteins have been isolated from human chronic lymphocytic leukemic cells (CLL) by Lens culinaris chromatography and by filtration on ACA-34 Ultrogel. Ia antigenic activity, measured by inhibition of the cellular radioimmunoassay, was separated by gel filtration into 2 fractions, peak I and peak II. Monoclonal antibodies, produced against peak II glycoproteins, appear to recognize different antigenic determinants of Ia molecules. Monoclonal antibody 18a4 reacted with Ia molecules of peaks I and II, whereas monoclonal antibodies 18c2 and 18d5 reacted almost exclusively with peak II molecules both in the cellular radioimmunoassay and by immunoprecipitation. In addition to antigenic differences, minor variations in the apparent m.w. of the Ia polypeptide chains were observed between peaks I and II. These results indicate the existence of antigenically distinct subsets of Ia molecules that are separable by gel filtration.
Clonal analysis of restriction antigens for human TNP-specific cytotoxic T lymphocytes.
Authors: Neefe J R , Sullivan J E , Hartzman R J
Published: December 2022

Previous studies of populations of human TNP-specific cytotoxic T lymphocytes have shown a preference for HLA-matched, TNP-modified targets. However, substantial killing of modified targets mismatched for serologically defined HLA antigens is also observed. The present report concerns oligoclonal and monoclonal populations of TNP-specific cytotoxic lymphocytes from a single donor whose lymphocytes were primed in vitro to TNP-modified autologous lymphocytes and frozen in aliquots. Conditions for the expansion of precursors of cytotoxic cells to sufficient numbers for assay are described. Expansion required IL 2-containing supernatants and stimulator or feeder lymphocytes in great excess; PHA, present in the IL 2-containing supernatants, was not required. TNP modification of the stimulators in secondary cultures was not required but appeared to enhance activity. Oligoclonal populations showed marked preference for TNP-modified targets sharing any of 3 of the 4 HLA-A and -B antigens of the donor (Aw24, B8, Bw44) when compared with targets sharing only the 4th antigen (A1) or no antigens. Monoclonal TNP-specific populations showed marked preference for certain TNP-modified targets when tested against a panel, but the pattern often did not correspond with the serologically defined HLA-A and -B antigens. The data suggest that most or all TNP-specific cytotoxic clones are restricted in their killing patterns by determinants associated closely with but not necessarily identical to the serologically defined HLA-A and -B antigens.
Immunoglobulin expression by human B lymphocytes clonally transformed by Epstein Barr virus.
Authors: Brown N A , Miller G
Published: December 2022

Epstein Barr virus (EBV) immortalizes human B lymphocytes, resulting in long-term lymphoblastoid cell lines. To date, such cell lines have typically been derived by suspension culture of mixed mononuclear cells, infected in vivo or in vitro with EBV. We have derived 208 lymphoblastoid lines by clonal transformation of peripheral blood lymphocytes from 3 neonates, 3 normal adults, and 1 patient with systemic lupus erythematosus (SLE), utilizing colony formation in agarose and then separate propagation of the colonies in liquid medium. A total of 180 lines have been analyzed for expression of cytoplasmic and/or secreted immunoglobulin isotypes. Such analysis reveals that 167 (92%) of the lines are monoclonal with respect to isotype expression. All clonally derived lines from neonates produce only IgM. Most clonal transformants from normal adults also produce IgM (81%), but clones producing IgG (16%) or IgA (3%) were also found. Clonal transformation of lymphocytes from the SLE patient revealed a shift in isotype expression, with increased numbers of IgG-secreting clones (48%) and IgA-secreting clones (13%). The data support the use of EBV-mediated clonal transformation as a probe for the analysis of states of isotype differentiation among circulating B cells, and as a method for deriving immortalized, monoclonal human B cell lines.
Defective regulation of B lymphocyte colony formation in patients with systemic lupus erythematosus.
Authors: Kumagai S , Sredni B , House S , Steinberg A D , Green I
Published: December 2022

We have succeeded in generating B cell colonies from freshly separated human peripheral blood by the double agar layer technique and expanding them in liquid culture for several weeks. Populations of cells enriched in B cells were placed into the upper layer with or without bacterial LPS. Although B cell colonies were observed without LPS, the addition of LPS to the cells in the upper agar layer increased the number of colonies. Also necessary for optimal B cell colony formation was the presence in the lower agar layer of either culture supernatant of PHA-stimulated mononuclear cells or purified T cells and PHA. Without such helper factors in the lower layer, only a few B cell colonies were observed in the upper layer. By using these techniques, peripheral blood lymphocytes from normal individuals and patients with active systemic lupus erythematosus (SLE) were studied for the generation of B cell colonies. Significantly more B cell colonies were observed in patients with SLE, using either kind of "helper" factor. In addition, T cells from normal individuals and patients were tested for their ability to help B cell colony formation. The SLE T cells were found to be defective in this function relative to normal T cells. Thus, although patients with SLE were hyperactive in forming B cell colonies, thier T cells were defective in supporting the formation of B cell colonies from normal individuals.
Immunochemical studies on thymosin: radioimmunoassay of thymosin alpha 1.
Authors: McClure J E , Lameris N , Wara D W , Goldstein A L
Published: December 2022

A sensitive and specific radioimmunoassay (RIA) has been developed to measure thymosin alpha 1 in human blood and tissue extracts. Thymosin alpha 1, originally isolated from bovine thymosin fraction 5, has an identical primary structure in bovine and human species. Antiserum to synthetic thymosin alpha 1 was prepared in rabbits. A synthetic analogue, N-Ac-(Tyr1)thymosin alpha 1 was radioiodinated by utilizing Na125I and soluble lactoperoxidase. The RIA is capable of detecting as small an amount as 40 pg thymosin alpha 1 in serum/plasma and does not significantly cross-react with other putative thymic hormones or other purified blood proteins that have been assayed. Proteolytic degradation of thymosin alpha 1 does not occur during the collection, storage, or assay of serum or plasma. The concentration of thymosin alpha 1 in the blood is highest in utero, decreases sharply after birth, and appears to remain fairly constant during the first 15 yr of life. Maternal blood levels of thymosin alpha 1 appear to be above normal during pregnancy.
Delayed hypersensitivity granuloma formation and modulation around Schistosoma mansoni eggs in vitro. II. Regulatory T cell subsets.
Authors: Doughty B L , Phillips S M
Published: December 2022

Schistosoma mansoni granuloma modulation was first described in an in vivo murine model of schistosomiasis and was characterized as a diminished cellular responsiveness to newly formed eggs in chronic infections. This phenomenon of modulation was studied using an in vitro model of granuloma formation. Negative selection procedures using anti-Lyt-1.2, 2.2, and Qa-1 sera and complement on spleen cell populations from normal or S. mansoni-infected C57BL/6 or (C57BL/6 X A/J)F1 mice were examined for their effect on in vitro granuloma formation or modulation. These data suggest that there is a Lyt-1+2-, Qa-1+ T cell present in chronically infected spleen cells that is capable of inducing feedback suppression. In addition, selective removal of a population of Lyt-1-2+ T cells from chronically infected spleen cells augmented the ability of that population to form granulomas in vitro. The selective removal of Lyt-1+2- cells ablated the ability of chronically infected spleen cells to form in vitro granulomas. Granuloma formation is primarily dependent on a population of Lyt-1+, Qa-1+ cells, and modulation is dependent on 2 populations, 1 acting directly (Lyt-1-2+) and the other through a (Lyt-1+2-, Qa-1+) feedback mechanism of suppression.
The selective role of membrane IgG in the antigen-induced inhibition of human in vitro antibody synthesis.
Authors: Stevens R H , Benveniste E , Pineda E
Published: December 2022

IgG-anti-tetanus toxoid-producing B cell precursors from recently immunized individuals can be stimulated by pokeweed mitogen and T cells to produce IgG-Tet antibodies in vitro. Treatment of these cells with tetanus toxoid selectively inhibits the synthesis of IgG-Tet. Because the IgG-Tet precursors display heterogeneity in their surface isotypes (mu, delta, gamma), the aim of this study was to determine the surface isotype responsible for transmitting this antigen-induced inhibitory signal to the B cell. B cells were fractionated by rosetting techniques on the basis of surface IgM or IgD. The 4 resulting B cell subsets (SIgM+, sIgM-, sIgD+, sIgD-) were found to be equally susceptible to antigen-induced inhibition. Experiments were then performed with anti-isotype antibodies to investigate further the roles of sIgM, sIgD, and sIgG in antigen inhibition. Treatment of peripheral blood lymphocytes with 10 micrograms/ml of anti-gamma antibody inhibited IgG-Tet antibody production, whereas the addition of up to 100 micrograms of anti-mu or anti-delta antibodies did not. Tetanus toxoid and anti-gamma antibody inhibition of IgG-Tet synthesis in vitro followed similar temporal kinetics, with the IgG-Tet precursors being sensitive to inhibition by either tetanus toxoid or anti-gamma antibodies during the first 3 days of culture and not becoming totally resistant to these agents until day 5 of culture. Capping off of sIgG receptors, with the subsequent addition of tetanus toxoid, abolished the antigen-induced inhibition of IgG-Tet seen previously, whereas capping off of sIgD or sIgM did not affect antigen inhibition of IgG-Tet antibodies. These results implicate sIgG molecules as those responsible for rendering IgG-Tet precursors susceptible to antigen-induced inhibition.
Arming of mononuclear phagocytes by eosinophil peroxidase bound to Staphylococcus aureus.
Authors: Ramsey P G , Martin T , Chi E , Klebanoff S J
Published: December 2022

When eosinophil peroxidase (EPO) was incubated with Staphylococcus aureus (staph) approximately 40% remained firmly bound to the bacteria as indicated by guaiacol assay. The staph-EPO complex was killed readily by concentrations of H2O2 and a halide that did not alter the viability of control (non-EPO-coated) organisms or organisms pretreated with another basic protein, protamine. This toxic effect was abolished by omitting either H2O2 or halide and was inhibited by azide, cyanide, or aminotriazole. Peroxidase activity was evident on the surface of the organism after ingestion by rabbit blood monocytes. The staph-EPO complex was killed at a greater rate by both rabbit blood monocytes and alveolar macrophages than were control organisms, an effect that did not appear to be secondary to improved phagocytosis. The peroxidase inhibitor azide inhibited the killing of staph-EPO by these mononuclear phagocytes without affecting the killing of control staphylococci; in the presence azide, the killing curve with staph-EPO returned to that seen with control organisms. Further, aerosolized EPO-coated organisms were cleared more rapidly from rat lung than were uncoated organisms or organisms pretreated with protamine. These findings suggest mononuclear phagocytes can utilize EPO bound to the surface of ingested microorganisms to enhance their microbicidal activity.
Genetic control of C6 polymorphism and C6 deficiency in rabbits.
Authors: Goldman M B , Cohen C , Stronski K , Bangalore S , Goldman J N
Published: December 2022

The genetic control of the sixth component of complement (C6) in rabbits has been studied by quantitation of C6 functional and antigenic levels and identification of polymorphism by isoelectric focusing (IEF) in gels. Patterns of inheritance of C6 variants in families carrying a silent gene for C6 were examined, and it was found that 3 common plasma phenotypic variants, C6 A, C6 B, and C6 QO were under the genetic control of allelic genes, C6*A, C6*B, and C6*QO. In IEF patterns, C6 A could be identified by its isoelectric point that was slightly more acidic than that of C6 B. C6 QO was undetectable because it lacked functional and antigenic activity. The C6*A/C6*B genotype displayed a mixed IEF pattern with bands characteristic of both C6 A and C6 B. Functional and antigenic levels of C6 that were found in heterozygous C6*A/C6*QO and C6*B/C6*QO rabbits were approximately one-half of the C6 levels found in the corresponding homozygous animals. The phenotypic variation closely resembles that previously observed in humans and rhesus monkeys, as well as preliminary data in rabbits. The patterns of inheritance indicated that the two common C6 structural genes and the deficiency gene were allelic variants at the same genetic locus.
A cross-reactive idiotype, QUPC52 IdX, present on most but not all anti-alpha (1 replaced by 6) dextran-specific IgM and IgA hybridoma antibodies with combining sites of different sizes.
Authors: Sharon J , D'Hoostelaere L , Potter M , Kabat E A , Morrison S L
Published: December 2022

Seven BALB/c IgM, 4 BALB/c IgA, and 1 C57BL/6 IgA anti-alpha (1 replaced by 6) dextran hybridoma antibodies were characterized idiotypically. Five of the 7 IgM and all 4 BALB/c IgA proteins bear a cross-reactive idiotype present on the anti-alpha (1 replaced by 6) dextran BALB/c myeloma protein QUPC52 and on a majority of anti-alpha (1 replaced by 6) dextran antibodies in BALB/c mice. Of these 9 monoclonal antibodies, some have combining sites as large as 6 glucose residues, and some have combining sites as large as 7 glucose residues. Individual idiotypes present on QUPC52 are differentially expressed on the 9 hybridoma proteins that bear the cross-reactive idiotype. One BALB/c IgM hybridoma protein and the C57BL/6 IgA hybridoma protein did not react with anti-QUPC52 idiotypic antibodies; another BALB/c IgM hybridoma antibody showed only marginal reactivity.
Role of I-region gene products in T cell activation. I. Stimulation of T lymphocyte proliferative responses by subcellular membrane preparations containing Ia alloantigens.
Authors: Germain R N , Mayer S V , Mescher M F
Published: December 2022

A model system has been developed for exploring the requirements for activation of T cells by subcellular forms of Ia alloantigen. Lymph node cells from mice recently primed subcutaneously with viable allogeneic cells show strong proliferative responses in vitro to membrane preparations derived from cells bearing the appropriate I-region-encoded glycoproteins. This stimulation shows kinetics characteristic of a secondary response, with a peak at 24 to 48 hr. Primary responses to alloantigen-bearing membranes are weak or absent under these conditions. The predominant cell type involved in the secondary response is the Lyt-1+ T lymphocyte, and the major antigenic stimulus is the I-A subregion-encoded Ia glycoprotein. Syngeneic Ia+ accessory cells do not appear necessary for activation to occur. Detergent solubilized reconstituted membrane vesicles also will stimulate primed T lymphocytes to respond by proliferation. The applications of this approach to the study of T cell recognition of antigen and the role of nonspecific lymphokines in T cell triggering are discussed.
Idiotypic analysis of hybridoma antibodies to branched synthetic polymer (Tyr, Glu)-Ala-Lys: idiotypic relationship with antibodies to linear random polymer (Glu60, Ala30, Tyr10).
Authors: Ju S T , Pincus S H , Stocks C J , Pierres M , Dorf M E
Published: December 2022

The expression of three anti-GAT idiotypes, CGAT, Gte, and GA-1, on 17 C57BL/10 and four C3H.SW hybridoma anti-(T,G)-A--L antibodies was analyzed. These hybridoma anti-(T,G)-A--L antibodies exhibited two patterns of fine antigen binding specificity. The majority of the hybridoma antibodies bound the (T,G)-A--L, GT, and GAT polymers but not the GA polymer, and were designated as GT-reactive hybridoma antibodies. A minor population of hybridoma anti(T,G)-A--L antibodies bound to (T,G)-A--L but not to GT, GAT, or GA, i.e., (T,G)-A--L-specific. A complete correlation between fine antigen binding pattern and the expression of CGAT idiotype was demonstrated. None of the 21 hybridoma anti-(T,G)-A--L antibodies expressed the GA-1 idiotype. All of the GT-reactive and none of the GT-nonreactive hybridoma anti-(%,G)-A--L antibodies expressed the CGAT idiotype. Furthermore, the Gte idiotype was found on the majority of CGAT+-bearing C57BL/10 hybridoma anti-(T,G)-A--L antibodies. These results indicate that C57BL/10 anti-(T,G)-A--L antibody repertoire can be grouped into a minimum of three families; i.e., CGAT+ Gte+, CGAT+ Gte-, and CGAT- Gte- families, with the CGAT+ Gte+ family as the major compartment. This is confirmed by the high percentage idiotype binding of serum anti-(T,G)-A--L antibodies with anti-CGAT idiotypic antisera. Finally, anti-idiotypic antisera made against CGAT+ hybridoma anti-GAT or anti-(T,G)-A--L antibodies crossreact extensively with other CGAT+ hybridoma anti-GAT and anti-(T,G)-A--L antibodies. However, additional experiments demonstrated that CGAT+ hybridoma anti-(T,G)-A--L antibodies also possess private idiotypes.
Anti-DNA autoantibodies in (NZB X NZW)F1 mice are clonally heterogeneous, but the majority share a common idiotype.
Authors: Marion T N , Lawton A R , Kearney J F , Briles D E
Published: December 2022

Monoclonal anti-DNA antibodies from 13 cloned hybridoma cell lines have been used to analyze the clonal heterogeneity of autoimmune anti-DNA antibodies. The hybridomas were obtained by fusing spleen cells from a single, autoimmune (NZB X NZW)F1 mouse. With the use of the criteria of isotype, isoelectric focusing patterns, fine specificity for various nucleic acids, and idiotype, the monoclonal anti-DNA antibodies were found to be heterogeneous in that 12 different clonotypes were identified among the 13 monoclonal antibodies studied. However, eight out of the 13 monoclonal antibodies have similar antigen specificity and share a common idiotype. This suggests that although autoimmune anti-DNA antibodies within a single (NZB X NZW)F1 mouse are the products of a large number of different antibody-producing clones, many of the clones produce antibodies with similar antigen-binding regions. Furthermore, in the majority of (NZB X NZW)F1 mouse sera the appearance of detectable levels of the common idiotype coincides with the appearance of anti-DNA autoantibody.
Genetic control of eosinophilia in mice: gene(s) expressed in bone marrow-derived cells control high responsiveness.
Authors: Vadas M A
Published: December 2022

A heterogeneity in the capacity of strains of mice to mount eosinophilia is described. BALB/c and C3H are eosinophil high responder strains (EO-HR) and CBA and A/J are eosinophil low responder strains (EO-LR), judged by the response of blood eosinophils to Ascaris suum, and the response of blood, bone marrow, and spleen eosinophils to keyhole limpet hemocyanin given 2 days after 150 mg/kg cyclophosphamide. Some of the gene(s) for high responsiveness appear to be dominant because (EO-HR X EO-LR)F1 mice were intermediate to high responders. This gene is expressed in bone marrow-derived cells because radiation chimeras of the type EO-HR leads to F1 were high responders and EO-LR leads to F1 were low responders. This description of a genetic control of eosinophilia in mice may be useful in understanding the role of this cell in parasite immunity and allergy.
Immunoglobulin production in human mixed lymphocyte cultures: implications for co-cultures of cells from patients and healthy donors.
Authors: Rümke H C , Terpstra F G , Huis B , Out T A , Zeijlemaker W P
Published: December 2022

When human peripheral blood lymphocytes (PBL) are cultured in the presence of irradiated allogeneic lymphocytes, the resulting mixed lymphocyte reaction (MLR) leads to the secretion into the supernatant of substantial amounts of IgM and IgG, derived from nonirradiated responder B lymphocytes. Our data indicate that stimulation to Ig production by responder B cells may result from different types of interactions. First, B cells and monocytes among the irradiated stimulator cells activate T cells in the responder population, which in turn trigger responder B cells to produce Ig; second, "responder" B cells activate irradiated "stimulator" T cells, leading to a "helper" signal, back to the responder B cells and leading to Ig production. The latter system is radiosensitive, because allogeneic T cells, irradiated at a dose of 4000 rad or more, failed to induce Ig production by responder B cells. In some combinations of human allogeneic lymphocytes, the co-culture of the cells leads to inhibition of Ig production, both in the presence and in the absence of PWM. Thus, co-culture of allogeneic cells may cause "positive" as well as "negative" allogeneic effects. The implications of these findings for the interpretation of co-cultures that are aimed at establishing defects in lymphocytes from patients with, for example, immunodeficiencies, who fail to produce Ig in the presence of PWM are discussed.
Antigen-specific suppression of type II collagen-induced arthritis by collagen-coupled spleen cells.
Authors: Schoen R T , Greene M I , Trentham D E
Published: December 2022

The induction of arthritis in Wistar-Lewis rats was suppressed by i.v. injections of spleen cells coupled to native type II collagen, beginning 14 days before intradermal injection of native type II collagen in incomplete Freund's adjuvant and continuing at 7-day intervals for the next 3 weeks. Cells coupled to native type I collagen or to denatured type II collagen were ineffectual, suggesting antigen-relevant cells were required for suppression. The phenomenon appeared to be mediated by a humoral effector mechanism; IgG antibody titers but not hemagglutinating antibody titers or delayed-type hypersensitivity to collagen were decreased by the injection of type II collagen-coupled cells. These data demonstrate the potential of antigen-specific immunotherapy for autoimmune disease.
Interaction of specific and innate factors of immunity: IgA enhances the antimicrobial effect of the lactoperoxidase system against Streptococcus mutans.
Authors: Tenovuo J , Moldoveanu Z , Mestecky J , Pruitt K M , Rahemtulla B M
Published: December 2022

The antimicrobial effect of the lactoperoxidase (LPO) system (enzyme with the thiocyanate ion and hydrogen peroxide) on Streptococcus mutans NCTC 10449 (serotype c) was significantly enhanced when the system was combined with secretory IgA. Similar enhancement was observed with LPO-myeloma IgA1 or IgA2 combinations. This enhancement of the antimicrobial efficiency was not dependent on the presence of specific antibodies to S. mutans in the IgA preparation, but seemed to require binding between LPO and immunoglobulin. However, neither human polyclonal nor myeloma IgG or IgM nor rabbit IgG enhanced the antibacterial activity of the LPO system. None of the immunoglobulins, when added alone, produced antimicrobial effects. LPO was shown to bind to colostral secretory IgA, myeloma IgA1, IgA2, and to a lesser degree to monoclonal and polyclonal IgG and monoclonal IgM. This binding had a stabilizing effect on the enzyme activity. Our results suggest that IgA significantly enhances the antibacterial efficiency of one of the innate immune factors--the LPO system.
Specific thymus-derived (T) cell recognition of papova virus-transformed cells.
Authors: Greene M I , Perry L L , Kinney-Thomas E , Benjamin T L
Published: December 2022

As a first step toward identifying papova virus-specific transplantation antigens and characterizing the immune response to these antigens, a series of congenic B10 mouse lines transformed by polyoma virus and SV40 has been obtained. B10.D2 (H-2d) mice immunized with x-irradiated polyoma-transformed B10.D2 cells (PyB10.D2) contained within their spleens a population of T cells that upon restimulation in vitro preferentially lyse 51Cr-labeled PyB10.D2 compared with other allogeneic polyoma-transformed target cells or SV40-transformed B10.D2 cells. Twenty hours after acute infection with polyoma virus, SV10.D2 cells express cell surface structures recognizable by polyoma-specific CTL. This CTL activity is therefore both polyoma specific and H-2 restricted, indicating associative recognition of H-2 molecules along with the virus-specific antigen.
Functional heterogeneity of helper T cells: two distinct helper T cells are required for the production of T cell growth factor.
Authors: Larsson E L
Published: December 2022

A rat monoclonal antibody, obtained after immunization with cells from a cytolytic, TCGF-dependent T cell clone, was found to react with 10 to 20% of murine splenic and lymph node cells and with 65 to 70% of thymocytes. Addition of this antibody to normal spleen cell cultures, or cytotoxic elimination of the cells recognized by the antibody before culture, resulted in the abolishment or profound inhibition of the Con A-induced proliferative responses. The antibody did not react with or inhibit the proliferating, TCGF-reactive T cells, but it inhibited lectin-dependent TCGF production. An analysis of this inhibition, using purified cell populations and the macrophage-derived factor LAF, demonstrated that the cell removed by antibody cytotoxicity was neither the macrophage nor the TCGF-producing T cell, indicating that this antibody eliminates a second helper T cell required for lectin-dependent production of LAF by the accessory cells. These results extend previous observations on the T cell dependence of Con A-induced LAF production and identify a fourth cell type involved in the collaborative processes leading to TCGF-dependent T cell proliferation.
T cell-specific suppressor factor(s) with regulatory influence on interleukin 2 production and function.
Authors: Kramer M , Koszinowski U
Published: December 2022

In this study we report that alloantigen-activated spleen cells produce both amplifying and suppressive factors under the same conditions. Both types of soluble mediators--as detected in different assay systems--were present in the supernatants of in vivo sensitized and in vitro restimulated spleen cell populations and were separable by gel filtration. As shown by others, the amplifying factor (IL 2) was eluted in the size range of 30,000 m.w. The suppressive factor(s) (SF) was eluted in the size range of 10,000 m.w. SF was shown to inhibit the proliferative response of T cells to alloantigen, as well as the generation of regulatory T cells and cytotoxic T cells from their precursors when added at the beginning of the in vitro culture. Furthermore, SF inhibited the release of IL 2 from producer T cells but had no detectable effect on the interaction of IL 2 with receptive T cells. In addition it was shown that SF does not affect the generation of PFC from their precursors after activation by T cell-independent antigens. The results indicate that SF selectively acts on T cells and that it is involved in the regulation of the immune response by modulating early events in T cell activation.
Detection of at least two distinct mouse I-E antigen molecules by the use of a monoclonal antibody.
Authors: Ozato K , Sachs D H
Published: December 2022

In an attempt to determine whether the expression of more than a single Ia antigen is determined by the I-E subregion of the mouse major histocompatibility complex (MHC), sequential immunoprecipitation analyses were performed by using a monoclonal antibody and alloantisera reactive with I-E subregion products. 3H-leucine-labeled glycoprotein preparations obtained from H-2d spleen cells were precleared with the monoclonal antibody 14-4-4S and then examined for residual Ia activity precipitable by an alloantiserum detected by SDS-polyacrylamide gel electrophoresis. Residual Ia activity was observed for all three strains of the H-2d haplotype tested. The residual Ia activity could be detected by sera absorbed with B10.A spleen cells, indicating that products of the I-E subregion rather than of the I-C subregion were responsible for this activity. No separable I-Ek molecules were detected in products of B10.A cells with the use of combinations of two monoclonal antibodies (including 14-4-4S) and several appropriate alloantisera. These findings indicate the presence of at least two similar but distinct I-E antigens encoded by the H-2d haplotype.
Characterization and functional properties of tumor cell lines in accessory cell replacement assays.
Authors: Walker E B , Lanier L L , Warner N L
Published: December 2022

This study reports the initial phenotypic and functional characterization of a series of cloned, murine, myelomonocytic tumors and their parent cell line WEHI-3, and a group of murine B lymphoma tumors. The tumor cell lines of the myelomonocytic lineage demonstrated the ability to reconstitute a macrophage-depleted, primary in vitro anti-SRBC PFC response, but only marginally enhanced an accessory cell-depleted, ova-primed, lymphocyte proliferation in vitro response. The B lymphoma tumors displayed exactly the reverse functional profile, being highly efficient in reconstitution of the proliferation response, but not supporting the SRBC PFC response. Detailed analysis of the cell surface phenotype of the various B lymphoma tumors used in this study show they display cell surface markers characteristic of normal B lymphocytes but are heterogeneous in their various stages of differential arrest. Future work will concentrate on the orchestration of Ia-mediated and soluble factor-mediated (IL 1) modalities of antigen-triggering of lymphocytes by these B lymphoma and myelomonocytic tumor cell lines.
The use of conglutinin in a quantitative assay for the presence of cell-bound C3bi and evidence that a single molecule of C3bi is capable of binding conglutinin.
Authors: Brown E J , Gaither T A , Hammer C H , Hosea S W , Frank M M
Published: December 2022

We have developed a quantitative assay for cell surface C3bi using 125I-labeled conglutinin. Conglutinin was purified to homogeneity from bovine serum and radiolabeled with 125I Bolton Hunter reagent. Conditions of time, temperature, ionic strength, and cell concentration that optimized the binding of conglutinin to erythrocytes bearing C3bi were then determined. The interaction between conglutinin and C3bi under these conditions was highly specific, since EAC4b3b, EAC4b3d, EAC4b3b-beta IH, and EAC4b treated with serum did not bind radioconglutinin significance better with EA or EAC4b. Using this assay, was examined the kinetics of inactivation of both human and guinea pig C3b bound to erythrocytes and showed that, for both, maximum conglutinin binding occurred after EAC4b3b had been incubated with a source of beta 1H and C3INA for 10 to 20 min at 37 degrees C.l We showed a linear relationship between the number of molecules of C3bi per erythrocyte and the amount of conglutinin bound for both guinea pig and human C3bi. The affinity of conglutinin for cell-bound C3bi was shown to be independent of C3bi density on the erythrocyte surface, and the Kd for conglutinin binding to erythrocytes bearing human C3bi was determined to be 1.3 X 10(-8) M. The number of conglutinin binding sites per erythrocyte as calculated from Scatchard plots was equal to the number of C3bi molecules on the cell surface as determined by direct assay using 125I-labeled C3. Moreover, for both human and guinea pig C3bi, the plot of log (cell surface C3bi) vs log (conglutinin bound) had a slope of 1. These findings imply that a single molecule of C3bi is capable of binding a molecule of conglutinin under the conditions of our assay.
Feedback regulation of immune responses by immune complexes; possible involvement of a suppressive lymphokine by FcR gamma-bearing B cell.
Authors: Miyama-Inaba M , Suzuki T , Paku Y H , Masuda T
Published: December 2022

Inoculations of antigen-antibody complexes (immune complexes) with the intact Fc portion generates suppressor cells in vivo by binding to FcR gamma on B cells via Fc portions. The cell type responsible for the suppression appears to be B cells bearing FcR gamma. Neither T cells nor macrophages participate in both the inductive and effective phases of this type of regulation. The suppression caused by splenic B cells, previously stimulated with immune complexes in vivo, is mediated by humoral factor(s) released from them. The suppressive factor(s) have H-2 gene product(s) coded by the right-hand side of the H-2 gene complex, but not for FcR gamma themselves or immunoglobulins. It has shared component(s) with suppressive B cell factor (SBF) released from FcR gamma + B cells stimulated with immune complexes in vitro, and it resembles SBF in its mode of action. These findings indicate that immune complexes, the final products of antibody responses, control the immune responses by stimulating surface FcR gamma on B cells. It is of interest that this type of regulation functions in vivo.
Abnormal anti-Epstein Barr virus antibodies in carriers of the X-linked lymphoproliferative syndrome and in females at risk.
Authors: Sakamoto K , Seeley J K , Lindsten T , Sexton J , Yetz J , Ballow M , Purtilo D T
Published: December 2022

The asymptomatic hemizygous female carriers of the X-linked lymphoproliferative syndrome (XLP) have abnormal antibody responses to EBV. This suggests partial expression of the defect that leads to EBV-provoked life-threatening diseases in their affected sons. EBV specific antibodies were measured in 65 serum samples of 12 obligate carrier females and seven of their daughters (females at risk) during periods ranging from 1 to 5 yr. Abnormal qualitative antiviral capsid antigen (VCA) IgG titers were nearly fourfold higher than normal controls, two carriers had persistent IgM anti-VCA antibody, two-thirds had persistent IgA anti-VCA antibody, and half of the women had titers to early antigen (EA). Five of seven females exhibited a similar persistent pattern. In contrast, none of the unaffected family members nor 23 normal controls expressed IgA or IgM titers to VCA even with high exposure to the virus, and anti-EA was detected in only one control. Therefore, these findings may prove useful for detecting carriers of the syndrome. Abnormal anti-EBV titers similar to the carrier pattern have been reported in patients and other immunosuppressed individuals, and are indicative of active viral infection.
Transfer of allergic encephalomyelitis with spleen cells from donors sensitized with myelin basic protein in incomplete Freund's adjuvant.
Authors: Namikawa T , Richert J R , Driscoll B F , Kies M W , Alvord E C
Published: December 2022

Myelin basic protein (BP) emulsified in incomplete Freund's adjuvant (BP/IFA) is relatively nonencephalitogenic in Lewis rats. Furthermore, repeated injections of BP/IFA prevent subsequent induction of experimental allergic encephalomyelitis (EAE) by BP emulsified in complete Freund's adjuvant (BP/CFA). In spite of this, spleen cells from rats injected repeatedly with BP/IFA transfer EAE after they are cultured with BP almost as effectively as BP/CFA spleen cells. However, unlike the latter, BP/IFA spleen cells do not proliferate in response to BP in culture. Furthermore, BP/IFA spleen cells are unable to transfer EAE after culture with concanavalin A (Con A), in contrast to BP/CFA spleen cells. Both populations of spleen cells undergo a strong proliferative response to Con A in culture. For BP/IFA cells, at least, a proliferative response to BP in vitro is not a prerequisite for enhanced transfer of EAE in Lewis rats.
OT-CLL: a human T cell chronic lymphocytic leukemia that produces IL 2 in high titer.
Authors: Friedman S M , Thompson G , Halper J P , Knowles D M
Published: December 2022

This report identifies and describes a human T cell chronic lymphocytic leukemia, OT-CLL, which can be triggered by selected mitogens (either PHA or Con A) to produce IL 2 in high titer. Optimal IL 2 production requires culturing OT-CLL cells at 2 to 5 X 10(6)/ml for 24 hr in the presence of 1 to 2% PHA-M. Under these conditions, the titer of IL 2 generated is greater than 20-fold that obtained from conventional sources, e.g., from mitogen-activated tonsillar lymphocytes. Two lines of experimental evidence suggest that the tumor cell product(s) is IL 2. First, in functional assays, suprenatants derived from cultures of PHA-activated OT-CLL cells trigger the proliferation and long-term growth of IL 2-dependent human TCL cells. Second, a partial biochemical purification of the active moiety(ies) derived from OT-CLL demonstrates marked similarity to conventional human IL 2. Thus, the biologically active material(s) precipitates in 50 to 70% saturated (NH4)2 SO4 solutions; elutes from DEAE-Sepharose in the presence of 0.04-0.08 M NaCl; and has an apparent m.w. of approximately 14,000, as determined by Sephadex G-100 gel filtration. In addition, analysis of OT-CLL cells by indirect immunofluorescence, utilizing a panel of monoclonal antibodies, confirms not only that these tumor cells are of T cell lineage but that they display surface antigens that define the normal human peripheral T cell subset subserving helper or inducer function: OKT3+, OKT4+ , OKT8-.
Identification of human CML target. HLA-B locus (B12) antigen variants defined by CTL generated between B locus-identical (B12) responder-stimulator pairs.
Authors: Kato S , Iványi P , Lacko E , Breur B , Du Bois R , Eijsvoogel V P
Published: December 2022

The genetics of human CML targets were studied by seven CTL generated in combinations iun which the responder/stimulator difference was limited to one (or two) HLA-A, -B, or -C antigens. Unstimulated peripheral blood lymphocytes were used as targets. CTL sensitized against antigens A11, Aw31, or B17 lysed all cells bearing the respective target from a large panel of cells from unrelated individuals. Hence, at least one CTL clone was directed against the HLA antigen molecule. However, all CTL also exerted cross-kill to cells not sharing the stimulating HLA antigen. For two CTL, the target of the cross-kill was not clarified. Five CTL were found where the cross-kill was directed against antigen HLA-B12 (Bw44 and Bw45). All these cTL were generated in R/S pairs identical for B locus antigens (Bw44/Bw35 heterozygotes). The individual CTL lysed different parts of the panel of B12-positive target cells. The interpretation is that these CTL detect subtypes of HLA antigens, but alternative possibilities are also considered. Four B12 subtypes are described, tentatively designated as B12-related CML targets. Identification of HLA-B-related CML targets represent CML "typing" of HLA-antigen differences that were not detected serologically. The subtypes can now be tested for their possible functional significance.
Anti-idiotype as antibody against the formyl peptide chemotaxis receptor of the neutrophil.
Authors: Marasco W A , Becker E L
Published: December 2022

Anti-idiotypic antibodies have been produced in mice, guinea pigs, and goat against rabbit antibodies to the chemoattractant f Met-Leu-Phe. The anti-idiotypic antibodies were demonstrated by their capacity in the presence of preimmune rabbit serum to block the binding of 125I anti-f Met-Leu-Phe to f Met-Leu-Phe-human transferrin absorbed to microtiter plates. Goat anti-idiotypic antibodies were further demonstrated by their ability, when absorbed to Staphylococcus aureus, to selectively bind 125I anti-f Met-Leu-Phe. In addition, some of the goat anti-idiotypic IgG was able to bind nearly all rabbit and rat anti-f Met-Leu-Phe antibodies examined. This was shown by 1) the ability of antibodies produced in rabbits immunized with either f Met-Leu-Phe conjugated to goat IgG or f Met-Leu-Phe conjugated to keyhole limpet hemocyanin (KLH) when adsorbed to S. aureus to bind the 125I F(ab')2 goat anti-idiotype, and 2) the ability of various rat anti-f Met-Leu-Phe antibodies to block the binding of 125I rabbit anti-f Met-Leu-Phe to goat anti-idiotype absorbed to S. aureus. Finally, goat anti-idiotypic antibodies can also cross-react with the receptor on the rabbit neutrophil for the formyl peptides as evidenced by 1) the direct binding of goat anti-idiotypic IgG to the PMN, 2) the ability of F(ab')2 fragments to goat anti-idiotype to partially inhibit the binding of f Met-Leu-(3H)Phe to rabbit PMN, and 3) loss of anti-idiotype and anti-PMN receptor activity after passage over an anti-f Met-Leu-Phe column. These data support Jerne's hypothesis of "internal image" and suggest a feasible experimental approach for producing anti-cell receptor antibody without purifying the receptor.
Modulation of the immune response to lipopolysaccharide.
Authors: Hiernaux J R , Baker P J , Delisi C , Rudbach J A
Published: December 2022

Mice given a single optimally immunogenic dose (20 micrograms) of bacterial LPS make an antibody response that is characterized by cyclic (oscillatory) patterns. This kinetic pattern is dose as well as mouse strain dependent. LPS can induce a secondary response in BALB/c mice that is also cyclic, although it differs in amplitude as well as periodicity from a primary response. To rule out the involvement of B cell mitogenic activity of LPS in the expression of such cyclic patterns, the responses produced by both C3H/HeN and C3H/HeJ mice to a single injection of LPS were compared; the latter do not give B cell proliferative responses to LPS and also have been reported to give low LPS-specific responses in vivo. Both strains gave an oscillatory kinetic pattern. By contrast, BALB/c nu/nu mice given an optimal immunogenic dose of LPS (2 micrograms) do not present a cyclic immune response. This indicates that mature T cells are required for the generation of an oscillatory anti-LPS response.
Activation of Fc receptor-bearing lymphocytes by immune complexes. I. Stimulation of lymphokine production by nonadherent human peripheral blood lymphocytes.
Authors: Neville M E , Lischner H W
Published: December 2022

Activation of human peripheral blood lymphocytes by incubation with particulate immune complexes or aggregated human gamma-globulin was studied by measuring the release of leukocyte migration inhibitory factor (LIF) activity. LIF-active supernatants were consistently produced when nonadherent lymphocytes containing less than 1% surface immunoglobulin-bearing cells and less than 0.2% nonspecific esterase-positive monocytes were incubated in the presence of RBC sensitized with rabbit or human antibodies or with pooled heat-aggregated human gamma-globulin. This immune complex-induced lymphokine production (ICLP) was dependent on the presence of cells bearing receptors for the Fc portion of IgG (Fc gamma). ICLP could not be demonstrated with lymphocyte preparations enriched for B cells even though the latter showed vigorous LIF production in the presence of complement-sensitized erythrocytes. ICLP was dependent on the concentration of lymphocytes and of stimulant as well as on the duration of coincubation, and it required active metabolic processes and RNA and protein synthesis but not DNA synthesis. Ca++ but not Mg++ was obligatory. ICLP by non-B Fc gamma receptor-bearing lymphocytes may play a role in antibody-dependent protective inflammation and immunologic injury phenomena, which is similar to that of lymphokine release by antigen-activated T cells in delayed hypersensitivity responses.
Immunotherapy of a chemically-induced sarcoma in rats: characterization of the effector T cell subset and nature of suppression.
Authors: Fernandez-Cruz E , Gilman S C , Feldman J D
Published: December 2022

Chemically-induced sarcomas (BC5), established subcutaneously and growing progressively in BN rats, were completely eliminated by i.v. infusion of syngeneic effector cells. The effector cells were generated from BN spleen cells immune to BC5 in a mixed lymphocyte-tumor cell culture (MLTC). Generation of a high yield of effector cells that were efficacious in eliminating BC5 in vivo necessitated: depletion of macrophages from immune spleen populations, before preparation of MLTC; addition back to MLTC of a small number of macrophages from normal spleens to attain a level of 0.5% of the spleen cells in culture; and addition of T cell growth factor on day 5 of MLTC. With these conditions a subset of T cells was expanded. They were blast cells with surface markers W3/25 and Ia, which exhibited no cytotoxic activity in vitro, and probably functioned as a helper or amplifier element in the tumor-bearing host. Effector cells generated in MLTC with BC5, treated with mitomycin C, were specific for BC5 in vivo and did not affect growth of a viral-induced BN tumor.