bluuebunny/crossref_metadata_2025 huggingface dataset:
Nonresponsiveness to the male antigen H-Y in H-2 I-A-mutant B6.C-H-2bm12 is not caused by defective antigen presentation.
Authors: de Waal L P , de Hoop J , Stukart M J , Gleichmann H , Melvold R W , Melief C J
Published: December 2022

The B6.C-H-2bm12 (bm12), H-2 I-Ab mutant, originated in the C57BL/6 (B6, H-2b) strain, is a nonresponder to the male antigen H-Y in both T cell proliferation and cytotoxic T lymphocyte (CTL) responses. Defective H-Y presentation by bm12-adherent cells, as a cause of the CTL nonresponsiveness, can be excluded, because 1) CTL from primed responder/nonresponder F1 female mice (B6/bm12 F1) were activated by H-Y antigen on antigen-presenting cells (apc) from either parent; 2) T cells from primed nonresponder bm12 females did not generate CTL to H-Y presented by responder B6 apc, but conversely, CTL from B6 females could be activated by antigen on bm12 apc; and 3) adherent cell-depletion experiments indicated that male adherent cells are necessary for generation of optimal H-Y-specific CTL responses, male adherent bm12 cells being equally as efficient as male adherent B6 cells in presentation to F1 T cells. The need for T helper cell activation by H-Y-presenting adherent cells during secondary in vitro restimulation can be circumvented by interleukin 2 (IL 2), because IL 2 completely restored the H-Y-specific CTL response of B6/bm12 F1 cells in cultures depleted of adherent cells. However, addition of IL 2 during in vitro restimulation of bm12 cells did not result in an H-Y-specific CTL response, indicating that H-Y-specific, I-A-restricted T helper cells are probably needed in vivo during priming for the generation of sufficient numbers of Db-restricted CTL memory cells. The data are compatible with the concept that H-Y antigen is presented in the context of the I-A alpha, beta molecule on the surface of adherent cells to T helper cells and in the context of the Db molecule on the surface of nonadherent as well as adherent cells to CTL precursors. The most likely explanation for the difference in H-Y response between B6 and bm12 include positive selection of the responder phenotype by the I-Ab alpha, beta molecule of the B6 strain or generation of suppressor T cells in the bm12 strain.
Fc dependence of macrophage accumulation and subsequent injury in experimental glomerulonephritis.
Authors: Holdsworth S R
Published: December 2022

An IgG-initiated, macrophage-mediated model of experimental glomerulonephritis was induced in rabbits. Experiments were designed to determine the importance of the Fc portion of this disease-initiating IgG antibody in inducing macrophage accumulation and subsequent proteinuria and histologic injury. The model was a passive model of the autologous phase of anti-glomerular basement membrane antibody-induced glomerulonephritis. Leukocyte depletion with nitrogen mustard prevented the development of proteinuria, macrophage accumulation, and histologic injury, but decomplementation with cobra venom had no effect, confirming leukocyte dependence but complement independence of injury in this model. The effect of equimolar kidney binding quantities of the intact disease-initiating IgG and an F(ab')2 fraction of this same antibody were compared. Intact IgG deposition was associated with heavy proteinuria (630 mg/24 hr, mean +/- 106 SD). A diffuse endocapillary proliferative glomerulonephritis with prominent macrophage accumulation (54 +/- 21 macrophages/glomerulus) developed. Deposition of the F(ab')2 fraction was associated with only minimal proteinuria (28 +/- 7 mg/24 hr). Histologic appearances showed no significant glomerulonephritis, and macrophage accumulation (4.1 +/- 0.6 macrophages/glomerulus) was substantially prevented. Thus, macrophage accumulation and subsequent injury in this model are dependent on the Fc portion of the disease-initiating IgG molecule. These data suggest immune adherence is an important mechanism for macrophage accumulation in antibody-initiated glomerulonephritis. Furthermore, the prevention of the Fc-dependent macrophage accumulation and simultaneous abrogation of injury suggest the lesion is mediated by macrophages.
Biosynthesis of C4 by mouse peritoneal macrophages. II. Comparison of C4 synthesis by resident and elicited cell populations.
Authors: Newell S L , Atkinson J P
Published: December 2022

Five elicited macrophage populations synthesized one-third to one-tenth as much hemolytically active C4 when compared with resident peritoneal macrophages. This decrease in functional C4 activity was not caused by inhibitors or protease activity in the elicited macrophage supernatants. Analysis of C4 antigen indicated a similar reduction in the elicited cells compared with the resident macrophages. A defect in precursor processing or secretion was deemed unlikely because the intracellular C4 precursor was appropriately reduced in the elicited cells. We postulate that mouse resident peritoneal macrophages are a pluripotent cell population with broad capabilities in regard to the initiation of the inflammatory response. In contrast, elicited or activated macrophages may be more specialized cells, with one manifestation of this being the "down" regulation of C4 synthesis.
Characterization of the lysis of fresh human solid tumors by autologous lymphocytes activated in vitro with phytohemagglutinin.
Authors: Mazumder A , Grimm E A , Rosenberg S A
Published: December 2022

Human peripheral blood lymphocytes (PBL), obtained from patients with a variety of malignancies, when incubated in vitro with phytohemagglutinin (PHA), lysed fresh autologous tumors during a short-term 51Cr-release assay. These PHA-activated killer (PAK) cells produced maximum lysis of tumor cells by 4 to 8 hr of co-cultivation. PHA incubation induced the generation of cells lytic for autologous and allogeneic tumors but not autologous or allogeneic PBL. Cold target inhibition studies demonstrated that autologous and allogeneic tumors of various histologic types all shared the determinants recognized by PAK cells. Some adherent cells were necessary for generation of these PAK, but higher numbers were suppressive. Augmentation of tumor cell lysis was seen when adherent cells were partially removed before PHA activation. The PAK effector cell was OKT3+, OKT8+. The precursor cell of the PAK was separated from natural killer (NK) cells on Percoll gradients and was generated from thoracic duct lymphocytes, a population devoid of NK cells. Furthermore, the phenotype of the majority of the precursor cells was OKT3+, OKM1-. Activation by PHA for 2 days was found to be an efficient and convenient method for generating lymphoid cells lytic for fresh autologous human tumor. The biologic and possible therapeutic role of these cells is currently being investigated.
Natural killer (NK) cells as a responder to interleukin 2 (IL 2). I. Proliferative response and establishment of cloned cells.
Authors: Suzuki R , Handa K , Itoh K , Kumagai K
Published: December 2022

The proliferative response of murine natural killer (NK) cells to a T cell growth factor, interleukin 2 (IL 2) was investigated by using the NK-enriched, low-density fractions (F.1) and NK-depleted, T cell-enriched, and high-density fractions (F.3) of normal spleen cells after Percoll-discontinuous density gradient centrifugation. When F.1 and F.3 cells were cultivated with IL 2-containing medium without addition of any other factors, like lectins, feeder layer, or macrophage products, F.1 but not F.3 cells showed an IL 2-dependent proliferative response. Growing populations showed a potent NK activity and consisted largely of cells with a morphology of large granular lymphocyte (LGL) and a surface marker profile characteristic for murine NK cells. By limiting dilution in the liquid culture and colony formation in the soft agar medium, F.1 but not F.3 cells showed a high frequency of clonal replication in IL 2. Almost all growing colonies in either medium showed a typical morphology of LGL. Further, F.1 cells treated with anti-asialo GM1 antibody and C, from which NK cells and cytolytic activity were almost completely abrogated and for which T cells were enriched, conversely formed a few IL 2-dependent colonies. From the colonies in soft agar, 11 clones, all of which showed a typical morphology (LGL), a typical surface profile with the exception of lacking IgG-Fc receptors for NK cells (Thy-1 +/-, Ly-5+, ASGM1+, Lyt-1-, Lyt-2-, Fc gamma R-), and a cytotoxic activity of activated nature, were established and maintained for a period of longer than 1 yr in the medium containing IL 2 alone. These results indicate that a major population in the naive spleens of normal mice that respond directly to IL 2 and clonally replicate without other stimulating factors may be NK cells.
Relapsing murine experimental allergic encephalomyelitis induced by myelin basic protein.
Authors: Fritz R B , Chou C H , McFarlin D E
Published: December 2022

Experimental allergic encephalomyelitis (EAE), an experimental autoimmune disease of the central nervous system (CNS), is readily induced in many mammalian species by immunization with CNS tissue or myelin basic protein (MBP) purified from the CNS. EAE has been frequently used as a model for multiple sclerosis (MS). However, EAE generally presents as an acute monophasic disease in the adult animal after immunization with MBP. After recovery, the animal is resistant to rechallenge with encephalitogen (1). Two exceptions to these observations have been reported. McFarlin et al. (2) reported that a variable number of Lewis rats showed signs of a single, mild relapse about a week after recovery from MBP-induced acute EAE. Panitch and Ciccone (3) have reported induction of recurrent EAE in rats immunized with human MBP. Chronic, relapsing EAE has been induced in the mouse; however, an apparent requirement for CNS tissue had been noted (4, 5). Recently, during the course of a series of experiments on the induction of EAE in SJL/J, PL/J, and (SJL/J X PL/J)F1 (SPL F1) mice, it was observed that the F1 mice frequently had paralytic relapses after recovery from MBP-induced symptoms. Experiments were initiated to examine this phenomenon, and the findings are presented below.
Idiotype-specific T lymphocytes. I. Regulation of antibody production by idiotype-specific H-2-restricted T lymphocytes.
Authors: Yamamoto H , Bitoh S , Torii M , Fujimoto S
Published: December 2022

Cytotoxic T lymphocytes (CTL) specific for MOPC-104E myeloma cells of BALB/c origin could be induced in BALB/c, (BALB/c X BALB.B)F1, and (BALB/c X BALB.K)F1 mice. (BALB/c X BALB.B)F1 CTL activity specific for MOPC-104E was effectively inhibited by anti-H-2d but not by anti-H-2b alloantiserum. However, the activity was hardly blocked by specific anti-idiotypic antibodies to MOPC-104E. For further analysis of the recognition of idiotype on target cells by CTL, the effect of those lymphocytes on anti-dextran B1355S antibody-producing B lymphocytes, which have a cross-reactive idiotype to MOPC-104E, was investigated. Lymphocytes from the CTL population did inhibit antibody production by dextran-immune spleen cells, but those from the CTL population specific for irrelevant myeloma cells (MOPC-167) did not. The (BALB/c X BALB.K)F1 CTL population suppressed the antibody production of BALB/c but not of BALB.K. This indicates that F1 cells can preferentially see H-2 antigens of immunizing myeloma cells on target B lymphocytes. The inhibition of antibody production was antigen specific and was only restricted to the PFC that were inhibitable by anti-idiotypic antibodies. The surface phenotypes of the cells that inhibited the antibody production were Thy-1+, Lyt-1-, Lyt-2+, and I-J-. These results strongly suggest that CTL specific for MOPC-104E recognize self H-2 antigens simultaneously with idiotypic determinants on B lymphocytes. Possible immunoregulatory roles of idiotype-specific CTL on antibody production systems are also suggested.
Polyclonal activation of murine B lymphocytes by immune complexes.
Authors: Morgan E L , Weigle W O
Published: December 2022

Murine splenic B lymphocytes are stimulated by homologous immune complexes to proliferate and secrete polyclonal antibody. The use of antibody from whole serum or monoclonal antibodies to form complexes resulted in the stimulation of mouse B lymphocytes. The ratio of antibody to antigen appears to be critical for the generation of the polyclonal antibody response. Because antigen and antibody are added independently at culture initiation, the exact nature of the complex is unknown, but optimal polyclonal antibody formation occurs in slight antigen excess. Immune complex-induced polyclonal antibody production requires the presence of both macrophages and T cells, whereas B cell proliferation requires only macrophages. The role of the macrophage appears to be to cleave a low m.w. (17,000) fragment from the complex, which is responsible for lymphocyte activation.
Murine T cell suppression demonstrable in the absence of cytotoxicity and the effect of Cyclosporin A on this system.
Authors: Hutchings P R , Cooke A , Gunn H C
Published: December 2022

In vitro culture of murine spleen cells in FCS without prior immunization or allogeneic stimulation leads to the development of spontaneous cytotoxicity. This cytotoxicity is not H-2 restricted and can affect any subsequent in vitro assays using syngeneic cells, especially if those assays include prolonged culture in FCS. Studies on murine spleen cells cultured in NMS, however, led to the detection of a suppressor system that did not display cytotoxic effects. Furthermore, it was found that this suppression, in contrast to the cytotoxicity and suppression generated during culture in FCS, was not sensitive to CYA. The suppressor cell may be an effector or an inducer of suppression and is sensitive to treatment with anti-Thy-1.2 and complement. It is suggested that some in vitro suppression is really due to cytotoxicity that may be directed toward FCS determinants adsorbed onto syngeneic targets.
Is the delayed-type hypersensitivity observed after a low dose of antigen mediated by helper T cells?
Authors: Milon G , Marchal G , Seman M , Truffa-Bachi P , Zilberfarb V
Published: December 2022

In mice receiving, i.v., a dose of antigen optimal for antibody response, no delayed-type hypersensitivity (DTH) reaction is detectable. In contrast, in mice receiving a dose of antigen too small to induce B cell activation, a DTH reaction is elicitable shortly and transiently after immunization. Using a sensitive titration assay of DTH-mediating T lymphocytes, this reciprocal relationship between antibody production and DTH responses was reinvestigated. The absence of peripheral DTH reactivity in mice primed i.v. with a high dose of antigen (10(9) heterologous red blood cells) does not result either from the absence of activation and clonal expansion of DTH-mediating cells or from induction of suppressive mechanisms but results from a decreased circulation of DTH-mediating cells. The present studies show that DTH-mediating cells disappear from blood to enter the spleen only when specific B lymphocytes are present and activated by a high dose of antigen. These results are compatible with the hypothesis that T cells activated by antigen can function either as helper cells for B lymphocytes or as DTH-mediating cells, depending on the environment they reach during their migration. In order to demonstrate that the same cell may support the two functions, monoclonal T lymphocytes were assayed for their helper function and for their ability to transfer a DTH reaction.
Growth at limiting dilution of human T cell colonies from T cell-depleted peripheral blood leukocytes.
Authors: Moreau J F , Miller R G
Published: December 2022

Using a new culture system, we found that up to one-fourth of nylon-wool nonadherent human peripheral blood lymphocytes (NAd) could give rise to a colony containing T cells. Even after NAd cells were depleted of T cells (rosette-forming and/or OKT3+ cells and/or OKT11+ cells), up to one-ninth of cells could still give rise to a colony containing T cells. Colonies were grown in microwell liquid cultures in a vol of 0.02 ml/culture seeded with from 2 to 40 cells. Each cell concentration was set up with 60 or more replicates, and the results were analyzed using limiting dilution theory. Growth had an absolute requirement both for lymphokine(s) present in the supernatant of PHA-stimulated peripheral blood leukocytes and for PHA. A fit to limiting dilution theory could be obtained only when heavily irradiated (3000 rad) peripheral blood leukocytes or NAd cells were also included, 300 cells/well being optimum. The system supported the development of OKT3+ and OKT11+ cells from OKT3-, OKT11-, OKM1-, and OKla1- precursors. All colonies contained some OKT3+ and OKT11+ cells, usually more than half the colony cells carrying the markers. Some colonies also contained OKTM1+ cells and OKla1+ cells at a frequency such that some colony cells must carry more than one marker.
Analysis of Fc (IgG) receptors on human peripheral blood leukocytes by dual fluorescence flow microfluorometry. II. Quantitation of receptors on cells that express the OKM1, OKT3, OKT4, and OKT8 antigens.
Authors: Titus J A , Sharrow S O , Segal D M
Published: December 2022

Human peripheral blood mononuclear cells were analyzed for the expression of Fc(IgG) receptors (FcR) by a quantitative dual fluorescence flow microfluorometric (FMF) technique. Mononuclear cells from all donors tested were distributed into three distinct subsets on the basis of FcR expression: cells expressing high, intermediate, or background levels of FcR. To characterize these subsets, cells were doubly labeled for FcR and with OKM1, OKT3, OKT4, or OKT8 hybridoma antibodies by using nonoverlapping red and green fluorophores. Each cell was then analyzed for the presence of both labels with a dual laser flow cytometer. Cells that did not express FcR were heterogeneous for OKT3, OKT4, and OKT8, did not express OKM1, and were primarily T cells on the basis of rosette formation with sheep erythrocytes (E+ cells). Most of the cells that expressed intermediate levels of FcR (an average of 1.3 x 10(4) FcR/cell) also expressed high levels of OKM1 but did not bind OKT3, OKT4, or OKT8 hybridoma antibodies. These cells were characterized as monocytes because they were removed by passage over Sephadex G-10 columns and because they were larger than the other cells as judged by light scatter. The majority of cells with the highest FcR density (an average of 4.5 x 10(4) FcR/cell) formed rosettes with E and were, by definition, T gamma cells. These cells expressed negligible levels of OKT4 and intermediate levels of OKM1, levels that clearly distinquished them from monocytes. The T gamma cells were heterogeneous in the expression of both OKT3 and OKT8, with approximately 20% being positive for OKT3 and OKT8. The level of OKT3 expression for the OKT3 and OKT8. The level of OKT3 expression for the OKT3+ T gamma cells was about the same as for as for OKT3+ T nongamma cells. In contrast, the OKT8 T nongamma cells expressed lower levels of OKT8 than did the OKT8+ T nongamma cells. By compaing our results with those obtained by other invesigators, we tentatively assigned functions to the subsets of cells that we describe here.
Lymphokine production by human T cell hybridomas.
Authors: Le J , Vilcek J , Sadlik J R , Cheung M K , Balazs I , Sarngadharan M G , Prensky W
Published: December 2022

Human T cell hybridomas were generated by hybridization of SH9 cells, the 6-thioguanine-resistant variant of human T lymphoma Hut102-B2, with concanavalin A-stimulated human peripheral blood lymphocytes. The hybrid nature of the established cell lines was documented by the difference in D14S1 restriction fragment length polymorphism between SH9 and the hybridoma cell DNA, and by the expression of OKT11 antigen on hybrid cells. T cell growth factor, macrophage growth factor (MGF), and interferon (IFN) activities have been demonstrated in the supernatants of different hybrid cultures, but not in SH9 cell cultures. Substantial quantities of MGF were secreted by several hybrids including the L23 line. MGF activity was dose-dependent, heat-labile, and synergistic with indomethacin. High titers of IFN activity were found in the cultures of hybridoma L415 and its subclones. Neutralization with specific antisera showed the IFN synthesized by L415 clones was immune interferon (IFN-gamma). Like the parental SH9 line, all of the hybridomas producing these lymphokines exhibited a cell surface phenotype typical for helper T cells. The hybridoma system therefore shows potential for the study of various lymphokines produced by human helper T lymphocytes.
Cyclic AMP as a mediator of prostaglandin E-induced suppression of human natural killer cell activity.
Authors: Goto T , Herberman R B , Maluish A , Strong D M
Published: December 2022

The role of cyclic AMP in mediating the prostaglandin (PG) E2-induced suppression of natural killer (NK) cell function was studied. Highly purified preparations of human large granular lymphocytes (LGL; which have been shown to be closely associated with human NK activity) and T lymphocytes were obtained by using Percoll gradients. Basal cyclic AMP was similar in both cell populations. With LGL, PGE2 (but not PGF2 alpha) suppressed NK activity (75%) and enhanced cellular cycle AMP (600%). In contrast, in small, high density T lymphocytes, PGE2 caused very small increases in cyclic AMP (20%). Phosphodiesterase inhibitors (PDEI) such as isobutyl methylxanthine and theophylline also increased cyclic AMP and suppressed NK activity in LGL. The effect of PGE2 and PDEI, in combination on cellular cyclic AMP and NK activity, was greater than the effect of each agent alone. Finally, exogenous derivatives of cyclic AMP also suppressed NK activity. These results indicate that in LGL, increased cellular cyclic AMP mediates the action of PGE2 on the suppression of NK activity.
Schistosoma mansoni adult microsomal antigens, a serologic reagent. I. Systematic fractionation, quantitation, and characterization of antigenic components.
Authors: Tsang V C , Tsang K R , Hancock K , Kelly M A , Wilson B C , Maddison S E
Published: December 2022

A systematic and quantitative search was conducted to identify and isolate a serologically pertinent antigen with high specific activity and low cross-reactivity from adult worms of Schistosoma mansoni. Adult worms of S. mansoni quickly thawed from liquid N2 temperatures were disrupted by controlled homogenization in isotonic buffered sucrose. Differential centrifugation of the homogenate yielded three particulate and one soluble fractions: the 480 x G pellet (nuclear), the 7650 x G pellet (mitochondrial), the 360,000 x G pellet (microsomal), and the 360,000 x G supernatant (cytosol). Quantitative analysis indicated a major concentration of specific antigenic activities in the microsomal fraction. Further purifications of the urea-solubilized, n-butanol-treated microsomal particles by gel filtration and ionic-exchange chromatography resulted in a microsomal antigen (MAMA) possessing high specific activity and low cross-reactivity. The final purification post-ionic exchange chromatograph showed a 30-fold increase of specific antigen activity over that of the cytosol fraction. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunotransfer blot by the "Western blot" technique (EITB) indicated specific antigenic activities in association with several different m.w. bands (heterogeneous m.w. by extrapolation = 4.3 to 11.9 x 10(6), 2.0 x 10(5), and 2.0 x 10(4) daltons) of the MAMA fraction. When compared with other reported serologic antigens, MAMA showed substantially higher specific activity and lower cross-reactivity.
Immunoregulation in experimental filariasis. II. Responses to parasite and nonparasite antigens in jirds with Brugia pahangi.
Authors: Lammie P J , Katz S P
Published: December 2022

Chronic B. pahangi infection (greater than or equal to 5 mo) in the jird, Meriones unguiculatus, leads to the induction of adherent nonspecific suppressor cells that are capable of modulating the in vitro mitogen responsiveness of spleen cells. In the present studies, a correlation between suppression of mitogen responsiveness and lack of reactivity to B. pahangi antigens was observed in vitro with splenic lymphocytes from chronically infected animals. However, the ability of jirds with a chronic B. pahangi infection to develop in vivo humoral responsiveness to SRBC and DTH to DNFB was comparable to that of uninfected controls. Analysis of the relationship between the development of antigen-specific and nonspecific immunoregulatory activity over the course of the infection was undertaken, too. Altered in vitro responsiveness of spleen cells from infected jirds to mitogens and B. pahangi antigens was associated with the onset of microfilaremia (8 wk post-infection). A transient lack of reactivity to SRBC was observed after the development of a patent infection in jirds. However, nonspecific suppressor cells capable of modifying the in vitro mitogen responsiveness of normal lymphocytes were not observed in the spleens of B. pahangi-infected animals exhibiting a lack of reactivity to SRBC. The relationship of antigen-specific suppressor cells to immunoregulation in experimental filariasis is discussed.
Studies of a high molecular weight human melanoma-associated antigen.
Authors: Hellström I , Garrigues H J , Cabasco L , Mosely G H , Brown J P , Hellström K E
Published: December 2022

Hybridomas were generated by fusing SP2/0 mouse myeloma cells with spleen cells from mice that had been immunized with cultured human melanoma cells. One of the hybridomas secreted a monoclonal IgG1 antibody, 48.7, which binds to a cell surface antigen of cells from human melanomas and compound nevi. The presence of the target antigen in vivo was demonstrated immunohistologically by staining frozen sections of primary and metastatic melanoma by the peroxidase anti-peroxidase technique. Weak staining of some blood vessel cells was also seen, but other normal cells, including skin melanocytes, were unstained, as were cells from other tumor types. Antibody 48.7 immunoprecipitated polypeptides with apparent m.w. on sodium dodecyl sulfate-polyacrylamide gel electrophoresis of 250,000 and greater than 400,000.
Cross-reactivity of suppressor T cells specific for lactate dehydrogenase B and IgG2a myeloma protein.
Authors: Nagy Z A , Baxevanis C N , Klein J
Published: December 2022

The major histocompatibility complex-controlled immune responsiveness of mice to two unrelated antigens, lactate dehydrogenase B (LDHB) and IgG2a myeloma protein is remarkably similar (1, 2). The similarity is explained by our recent work, demonstrating that both antigens, when presented in the context of cell surface Ek (Ek alpha Ek beta) molecules, generate strong suppression that causes nonresponsiveness to all mouse strains expressing this molecule. The suppression is mediated by antigen-specific, Ek-restricted Lyt-1+2+ suppressor T (Ts) cells, which act by inhibiting the proliferation of A(A alpha A beta)-restricted, Lyt-1+2- (possibly T helper [Th]) cells specific for the same antigen (3-5). However, the question of why the Ek molecule, in combination with two structurally unrelated proteins, preferentially induces Ts cells has remained unanswered. We demonstrate here that the anti-LDHB and anti-IgG2a Ts cells are fully cross-reactive, which indicates that the LDHB + Ek and IgG2a + Ek determinants recognized by these cells are very similar or identical.
Expression of Ia antigen on epidermal keratinocytes after the grafting of normal skin to nude mice.
Authors: Daynes R A , Emam M , Krueger G G , Roberts L K
Published: December 2022

During the course of experimentation designed to evaluate the migration of host Langerhans cells (LC) into normal skin grafted onto nude mice, we observed that the epidermal cells of these grafts were induced to express la determinants solely of graft origin. The data presented herein indicate that the expression of la by normal epidermal cells correlates with the infiltration of host LC into the graft. This la expression is restricted to the keratinocytes within the epidermis of the grafted skin, is first observed 7 to 9 days post-grafting, and persists within the grafted skin for greater than 12 wk. The induction of la expression by keratinocytes appears to be the result of the environment that is provided by the nude mouse host and is independent of both passenger lymphocytes within the skin graft and allogeneic differences between graft and host. We strongly believe that these studies provide the basis for the development of an experimental animal model system for investigating the potential role that la expression by epidermal cells may play in enhancing the immune response to antigens encountered in the skin.
Monoclonal anti-Lyt-2 antibodies block the activation of cytolytic T lymphocyte precursors.
Authors: Glasebrook A L , MacDonald H R
Published: December 2022

In the present study, we reinvestigated the effects of anti-Lyt-2 monoclonal antibodies (mAb) on the generation of H-2K/D-specific Lyt-2+ cytolytic T lymphocytes (CTL). A sensitive limiting dilution microculture system supplemented with a source of interleukin 2 (IL 2) was employed to provide a minimal estimation of the frequency of CTL precursors (CTL-P) in normal spleen or mixed leukocyte culture (MLC) populations. The addition of anti-Lyt-2 (but not anti-Thy-1) mAb at culture initiation caused an average 78% reduction in the number of CTL-P within spleen cell populations that were activated by allogeneic stimulation. Most CTL that were generated from Lyt-2+ CTL-P in the presence of anti-Lyt-2 mAb were not inhibited in cytolysis by such mAb, whereas the majority of CTL generated in the absence of anti-Lyt-2 were inhibitable. In contrast to normal spleen, the frequency of (operationally defined) CTL-P that had been activated for 5 days in primary MLC was unaffected by the presence of anti-Lyt-2 mAb. Collectively, these results demonstrate that anti-Lyt-2 mAb are capable of inhibiting the antigen-dependent activation of normal splenic precursors of Lyt-2+ CTL. The fact that IL 2 could not overcome the inhibition suggests that anti-Lyt-2 mAb may have affected the expression of IL 2 receptors as a direct consequence of inhibiting CTL-P antigen recognition.
Suppression of lymphocyte proliferative responses: demonstration of two stages occurring in the in vitro generation of suppressor macrophages.
Authors: Stout R D , Fisher M
Published: December 2022

The suppressor macrophages generated by in vitro culture of spleen cells are shown to be derived from nonadherent splenic precursors. The induction of suppressor macrophage generation requires the presence of both plastic-adherent and plastic-nonadherent spleen cells during the first 24 to 48 hr of culture. After this induction period, the primed nonadherent cells can continue to generate suppressor macrophages through several serial transfers. The primed nonadherent spleen cell population is predominantly comprised of cFcR+ cells (40 to 50%) and Thy-1+ cells (40%), many of which appear to be undergoing blastogenesis. The generation of suppressor macrophages appears to proceed in two stages. The first stage is radiosensitive and has a minimum duration of 1 to 2 days. Although this stage initially coincides with the inductive period, it is also required for the continuous propagation of the suppressor macrophages upon serial transfer of the nonadherent spleen cells. The second stage is radioresistant and has a maximum duration of 2 to 3 days. The in vitro events thus bear similarities to events occurring during in vivo differentiation and activation of effector macrophages. It is suggested that the in vitro generation of suppressor macrophages may be one component of syngeneic mixed lymphocyte responses when spleen cells are used as the responding population, and that the suppressors represent part of a regulatory system controlling lymphocyte and macrophage differentiation and activation.
Treatment of autoimmune MRL/Ipr mice with monoclonal antibody to Thy-1.2: a single injection has sustained effects on lymphoproliferation and renal disease.
Authors: Seaman W E , Wofsy D , Greenspan J S , Ledbetter J A
Published: December 2022

MRL/Mp-lpr/lpr (MRL/lpr) mice spontaneously develop an autoimmune disease characterized by anti-DNA antibodies, immune-complex glomerulonephritis, and massive proliferation of a distinct population of T cells. The proliferating T cells have the phenotype Thy-1.2+, T200+, Lyt-1+,2-,3-, but Thy-1.2 and Lyt-1 are expressed in abnormally low density. These cells appear to function as helper cells, and neonatal thymectomy prevents both lymphoproliferation and autoimmunity, which suggests that autoimmunity in MRL/lpr mice is secondary to T cell proliferation. We therefore attempted to reduce lymphoproliferation by treating MRL/lpr mice with a single injection of rat monoclonal antibody (MAb) to Thy-1.2 (30-H12, IgG2b). Mice were treated at 8 wk, before the onset of overt disease. We found that MRL/lpr mice were resistant to depletion of circulating T cells (CTC) by anti-Thy-1.2; 0.6 mg of antibody totally depleted CTC from normal mice, but had little or no effect on CTC in MRL/lpr mice. However, treatment with 6 mg of MAb against Thy-1.2 reduced CTC in MRL/lpr mice by over 70%. Moreover, this single treatment markedly reduced the proliferation of CTC over the ensuing 3 mo, despite clearance of the anti-Thy-1.2 from the circulation within 3 wk. Treated mice maintained better renal function than untreated controls, as assessed by levels of blood urea nitrogen (BUN), although anti-DNA antibodies were not significantly reduced. The effect of anti-Thy-1.2 was specific; treatment with rat MAb to the common leukocyte antigen T200 produced only a transient effect on circulating lymphocytes and did not reduce renal disease. The prolonged effects of a single injection of anti-Thy-1.2 suggest that the MAb produces a sustained alteration in immune regulation. The improvement in renal disease is in accord with evidence that autoimmune disease in MRL/lpr mice is T cell dependent. Monoclonal anti-lymphocyte antibodies may be useful in the treatment of autoimmunity.
Effects of aging on cell cooperation and lymphocyte responsiveness to cytokines.
Authors: Rosenberg J S , Gilman S C , Feldman J D
Published: December 2022

Functional activities and cell cooperation of macrophages (Mphi), T cells, and B cells of young and old Lewis rats were compared. Splenic M phi from young and old rats provided accessory help for T cell mitogenesis and B cell mitogenesis, provided accessory help for generation of PFC, and produced IL 1 equally well as measured in costimulator assays. Splenic T cells of aged Lewis rats, however, were poorly responsive in mitogen assays and did not respond to supplemental IL 2 and antigen with blast transformation and with increased help for B cells to produce PFC. "Old" B cells did not respond in vitro to mitogens with help from M phi and T cells, nor did they respond to B cell helper factor with increased PFC. The data indicate that hyporesponsiveness of the immune system, especially of B cells, in aged rats is due in part to defective reactivity to interleukins and cytokine(s) and to defective cell-cell cooperation.
Lymphokine regulation of activated (G1) lymphocytes. I. Prostaglandin E2-induced inhibition of interleukin 2 production.
Authors: Walker C , Kristensen F , Bettens F , deWeck A L
Published: December 2022

PGE2-induced inhibition of the proliferatory response of PHA-stimulated human PBL and Con A-stimulated murine thymocytes was analyzed by flow cytometry. It was found that the activation process (G0-G1a transition) was not influenced by PGE2 over a wide range of concentrations (10(-10) to 10(-6) M), nor was the formation of IL 2 receptors inhibited. Similarly, the viability of human lymphocytes was practically unaltered. In contrast, the IL 2-dependent cell cycle event (G1a-G1b transition), which is required for proliferation, was inhibited in a dose-dependent fashion. The addition of IL 2-containing supernatants to such cultures prevented the PGE2-mediated block in the G1a phase and reconstituted a normal lymphocyte proliferation. Furthermore, lower IL 2 titers were measured in supernatants from PHA-stimulated human PBL treated with PGE2. These findings strongly suggest that PGE2 primarily exerts its inhibitory effect on lymphocyte proliferation through an inhibition of IL 2 production.
Cross-reactive idiotypic determinants on antibodies and antigen-specific helper T cell continuous lines.
Authors: Lifshitz R , Apte R N , Mozes E
Published: December 2022

Anti-idiotypic antibodies produced in C57BL/6 mice (H-2b, Igh-1b) against (T,G)-A--L-specific antibodies of C3H.SW mice (H-2b, Igh-1j) were used to probe (T,G)-A--L-specific helper T cell lines and clones for the expression of idiotypic determinants on the cell surface of the monoclonal functional T cells. By using the fluorescence-activated cell sorter (FACS II), anti-idiotypic sera of individual mice that specifically bind C3H.SW anti-(T,G)-A--L antibodies were shown to stain significantly cells of the E-9M(+) continuous T helper line originated from C3H.SW (T,G)-A--L "educated" T cells. The same antisera did not react with a helper T cell line of C3H.SW origin specific to human gamma-globulin. They also did not stain a (T,G)-A--L-specific helper T cell line derived from CWB (H-2b, Igh-1b) mice, which differ from C3H.SW mice only in their heavy chain allotypes. Thus, the expression of the idiotypic determinants on the T cell lines appears to be antigen-specific and linked to the heavy chain allotypic marker as shown for the specific antibodies. Different clones derived from the E-9 M(+) line were tested their reactivity with the individual anti-idiotypic sera. All clones but one (1.11) were stained significantly. The clones were tested for their biologic activity and all of them except clone 1.11 were found to exert helper activity specific to (T,G)-A--L. Thus, individual anti-idiotypic sera against C3H.SW anti-(T,G)-A--L antibodies recognize cross-reactive idiotypic structures on the surface of antigen-specific monoclonal helper T cells.
Biosynthesis of glycosaminoglycans by epithelial and lymphocytic components of murine thymus.
Authors: Britz J S , Hart G W
Published: December 2022

Interactions between the epithelial and lymphocytic components of the thymus are required for T cell maturation, yet the molecular bases for these interactions remain elusive. In the development and function of other endodermally derived organs, glycosaminoglycan-containing proteins are known to play a critical role. In contrast, virtually nothing is known about the macromolecules that are major constituents of thymic interstitial spaces. For these reasons, we undertook metabolic labeling studies in vitro with D-(6-3H)glucosamine and 35SO4(-2) to begin to characterize systematically the relative amounts and types of glycosaminoglycans made by enriched subpopulations of cells within the thymus. Hydrocortisone, which depletes the thymus of 90% of its lymphocytes, was used both to enrich for epithelium-derived glycoconjugates and to determine if significant alterations in glycoconjugate metabolism accompany drug-induced premature thymic involution. Results indicate: 1) glycosaminoglycans account for a substantial proportion of the total glycoconjugates synthesized by both thymocytes and epithelium; 2) Glycosaminoglycans show a tissue-specific distribution. Hyaluronic acid is the major glycosaminoglycan synthesized by thymic epithelium, whereas it accounts for less than 15% of the total glycosaminoglycans made by thymocytes; 3) Similar proportions of sulfated glycosaminoglycans are made by thymic epithelium and thymocytes. Chondroitin sulfates predominate (75 to 90%) over heparan sulfates (10 to 25%). Chondroitin sulfates from both nonstimulated thymocytes and epithelium are nearly exclusively sulfated at the 4-position of their N-acetylgalactosamine residues; 4) The major high m.w. glycoconjugate of thymocytes, however, is nonsulfated and is resistant to pronase, hyaluronidase, chondroitinase ABC, nitrous acid, keratanase, and neuraminidase; 5) Although hydrocortisone treatment causes a dramatic inhibitory effect on the incorporation of radioactivity into smaller oligosaccharide side-chains by "cortisone-resistant" thymocytes, the drug exerts negligible effects on the metabolism of glycoconjugates by epithelium. These data, which quantify and categorize the complex arrays of glycoconjugates synthesized by the major cell types of the thymus, establish the necessary foundations for further investigations into the functional roles of these glycoconjugates in thymic epithelium-induced maturation of T lymphocytes.
SFR3-DR5, a monoclonal antibody with HLA-DR5 specificity.
Authors: Radka S F , Machamer C , Cresswell P , Kostyu D D , Ward F E , Amos D B
Published: December 2022

We have developed a monoclonal antibody with HLA-DR5 serologic specificity. The antibody, SFR3-DR5, binds specifically to DR5-positive lymphoblastoid cell lines, and immunoprecipitates alpha- and beta-chains characteristic of DR antigens from them. Cytotoxic activity of the antibody segregates with the DR5-bearing haplotype in a family. The antibody reacted with the cells of 16 of 17 DR5 individuals and was negative on all DR5-negative cells tested. SFR3-DR5 reacted weakly with PWM-activated cells of the single DR5 individual whose B lymphocytes were unreactive with the monoclonal antibody by cytotoxicity. Possible interpretations of these results are discussed.
Salmonellae activate complement differentially via the alternative pathway depending on the structure of their lipopolysaccharide O-antigen.
Authors: Liang-Takasaki C J , Grossman N , Leive L
Published: December 2022

Differences in the O-antigen polysaccharide structure of lipopolysaccharide were previously shown to affect the rate of phagocytosis of Salmonellae strains by the murine macrophage-like cell line J774. Phagocytosis required a serum factor(s) that is labile to heat (56 degrees C for 30 min) and to zymosan treatment, which indirectly suggested the participation of C. We now show, using guinea pig serum, that these bacteria activate C3 at different rates, and this activation is proportional to the later rate of phagocytosis. Activation is predominantly via the alternative pathway, because C4 is not consumed and the reaction proceeds equally well in the serum of C4-deficient guinea pigs. Because the extent of activation of C3 and the subsequent rate of phagocytosis are inversely proportional to virulence, we propose that virulence of a strain may be influenced by the ability of the polysaccharide structure of its lipopolysaccharide to activate the alternative pathway of C, destining it for subsequent phagocytosis.
Functional properties of membrane-associated complement receptor CR1.
Authors: Iida K , Nussenzweig V
Published: December 2022

It was previously shown that membrane receptors for C3b (CR1) purified from human erythrocytes were powerful inhibitors of the complement cascade and that they encompass the regulatory functions of the serum proteins beta 1H (H) and C4-binding protein (C4bp). In the present report we study the functional properties of membrane-associated CR1. When tonsil lymphocytes, which contain between 30 and 60% of CR1-bearing B cells, are incubated with the red cell complement intermediate EAC14oxy2lim or EAC14oxy23lim, they inhibit both C42 and C423 in a dose-dependent manner. These effects are mediated by membrane-associated molecules. Indeed, mild trypsinization of the lymphocytes abolishes their activity, and formaldehyde-fixed cells are as effective as viable cells. The inhibitory effects are in part mediated by CR1. The lymphocyte activities are reversed about 60% if monoclonal antibodies to CR1 or fluid phase C3b are present in the incubation medium. Moreover, upon addition of C3b-inactivator (l), lymphocytes release C3c fragments from EAC14oxy23b. The release of C3c was also abolished by antibodies to CR1. These results support the idea that CR1, as well as other molecules from the lymphocyte membrane, can function as inhibitor(s) of complement activation in their vicinity.
The biologic activity of mast cell granules. IV. The effect of complement depletion on rat cutaneous late phase reactions.
Authors: Lemanske R F , Joiner K , Kaliner M
Published: December 2022

Cutaneous late phase reactions (LPR) in rats can be induced by the intradermal injection of anti-IgE antibody or isolated rat peritoneal mast cell granules. Rat LPR are characterized by neutrophil-rich infiltrates at 2 to 8 hr followed by mononuclear cell-rich infiltrates thereafter. Rat Arthus reactions are histologically similar and are complement (C) dependent. To determine the importance of C in the pathogenesis of rat LPR compared with its role in Arthus reactions, rats were treated with cobra venom factor (CVF) (250 U/kg i.v.), and the effects of this treatment on total hemolytic complement (CH50), C3 titers, LPR, and Arthus reactions were assessed. CVF treatment produced profound decreases in both CH50 (from 197 +/- 20 to less than 1.0 U/ml) and C3 (from 44,240 +/- 2840 to less than 5 U/ml) titers after 6 hr, which persisted through at least 30 hr. The inflammatory intensity of heterologous reverse passive Arthus reactions was significantly decreased in CVF-treated animals. In contrast, the intensity of LPR was unaffected by CVF treatment. Therefore, although LPR and Arthus reactions share certain histologic characteristics, these similarities are not due to a mutual requirement for the presence of C.
IgE-mediated release of histamine from human cutaneous mast cells.
Authors: Tharp M D , Suvunrungsi R T , Sullivan T J
Published: December 2022

We investigated the ability of antigen-IgE interactions to stimulate histamine release from human infant cutaneous mast cells. Skin obtained at circumcision contained numerous perivascular mast cells, as assessed by light and electron microscopy. The histamine content of this tissue averaged 17.7 ng (+/- 1.5 SEM)/mg wet weight. Challenge of 200-microns thick sections of unsensitized skin with varying concentrations of monoclonal murine antibodies to human IgE caused no net release of histamine. After skin sections were incubated in the presence of 5 micrograms/ml of human myeloma IgE (S) for 120 min at 37 degrees C, monoclonal anti-IgE challenge resulted in 40.1% (+/- 6.0 SEM) histamine release. Similar passive sensitization with 1/20 dilutions of serum from humans expressing IgE to purified Juniperus sabinoides (JS) antigen rendered the tissue responsive to specific antigen challenge. Dose-related histamine release occurred over 30 min with optimal release of 12.6% (+/- 2.4 SEM) after stimulation with 100 ng/ml of JS antigen. This reaction required sensitization with serum containing IgE to JS and was antigen-specific. Optimal reactions to antigen occurred at 3 mM added Ca++, 34 degrees C to 37 degrees C, pH 7.2. Antigen-induced release was markedly influenced by the added Ca++ concentration; no release occurred in the absence of Ca++, 54% of the optimal response was observed at 2 mM Ca++, and 28% of the optimal response occurred at 4 mM Ca++. The addition of Mg++ did not influence antigen-induced release. The results of this study provide functional evidence that 1) human infant cutaneous mast cells express Fc-epsilon receptors; 2) these receptors are largely unoccupied in vivo; and 3) stimulation of passively sensitized infant mast cells with anti-IgE or specific antigen leads to immediate histamine release. This new system should permit detailed in vitro studies of immediate hypersensitivity reactions in human skin.