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Biomedical subjects

R S Geha

Publications and source records attributed to R S Geha.

At least 163 records · Page 9Linked to original sources
PubMed1989

An immunodeficiency characterized by defective signal transduction in T lymphocytes.

We studied a nine-year-old boy with severe, recurrent infections. The patient was exposed in utero to azathioprine and prednisone. He had autoimmune hemolytic anemia, bronchiectasis, and Hodgkin's disease. The patient's circulating lymphocytes were normal in number and phenotype, but stimulation of the T-cell receptor by antigens, mitogens, and monoclonal antibodies failed to induce interleukin-2-receptor expression, interleukin-2 synthesis, or lymphocyte proliferation. The early biochemical events necessary to initiate lymphocyte activation--accumulation of the second messenger diacylglycerol, activation of the enzyme protein kinase C, and elevation of the free intracellular calcium concentration--failed to occur in this patient's lymphocytes. The defect in the lymphocyte could be corrected in vitro by two agents that bypass the receptor-mediated signal mechanism (the diacylglycerol analogue phorbol and the calcium ionophore ionomycin). Further studies localized the defect in signal transduction to the interaction between cell-surface receptors and the guanine nucleotide-binding protein. We conclude that this patient's immunodeficiency was caused by a defective coupling of surface receptors to signal-transducing proteins in his T lymphocytes, resulting in failure of lymphocyte activation.

Calcium↗
PubMed1989

Intracellular cleavage of newly synthesized low affinity Fc epsilon receptor (Fc epsilon R2) provides a second pathway for the generation of the 28-kDa soluble Fc epsilon R2 fragment.

It has been reported that the 45-kDa low affinity Fc epsilon R (Fc epsilon R2) on B cells is cleaved spontaneously from the cell surface to release a 28-kDa soluble fragment (sFc epsilon R2). This study demonstrates an additional mechanism by which B cells generate this fragment. Data from 35S methionine pulse-chase experiments with the Fc epsilon R2 bearing human B lymphoblastoid cell line, RPMI 8866, and immunoprecipitations of cell lysates and culture supernatants with an Fc epsilon R2 specific mAb, mAb 25, demonstrates the existence of a cell-associated 28-kDa Fc epsilon R2 fragment. This fragment was shown by partial amino(NH2)-terminal sequence analysis to be identical to the previously described 28-kDa sFc epsilon R2. The resistance to cell treatment with trypsin indicated that it was located intracellularly. Its appearance early in the biosynthesis of the Fc epsilon R2 (within a 10-min pulse), before the Fc epsilon R2 reached the cell surface, suggested that some of this fragment was generated intracellularly. Neutralization of acidic organelles with NH4Cl inhibited the formation of this intracellular fragment, strongly suggesting that it was a produce of intracellular cleavage of the Fc epsilon R2. Finally, this 28-kDa intracellular fragment was shown to be released into the culture supernatant, suggesting an intracellular mechanism by which the cells generate sFc epsilon R2.

Antigens, Differentiation, B-Lymphocyte↗
PubMed1989

Endogenous interleukin 6 plays an obligatory role in interleukin 4-dependent human IgE synthesis.

The lymphokine interleukin (IL) 4 plays a crucial role in the regulation of IgE synthesis. In the present study, the cellular and cytokine requirements for the IL4-dependent induction of IgE synthesis in humans were analyzed. Recombinant IL4 could induce IgE synthesis by peripheral blood mononuclear cells and autologous T/B cell mixtures, but not by highly purified B cells. IgE induction by IL4 was strongly decreased in monocyte-depleted peripheral blood mononuclear cells. These results show that the induction of IgE synthesis by recombinant IL4 is T cell dependent and optimal in the presence of monocytes. IL5 and IL6, but not IL2, IL1 and tumor necrosis factor-alpha, strongly up-regulated the IL4-dependent synthesis of IgE, with modest effects on cell proliferation. An anti-IL6 polyclonal antibody strongly inhibited IL4-driven IgE production. Endogenous IL6 plays, therefore, an obligatory role in the IL4-dependent induction of IgE. However, a combination of IL4, IL5 and IL6 (with or without IL1) at optimal concentrations could not induce IgE synthesis by purified normal B cells, indicating that cytokine-mediated signals, although essential, are not sufficient for the IL4-dependent induction of IgE synthesis.

B-Lymphocytes↗
PubMed1989

Toxic shock syndrome toxin 1 binds to major histocompatibility complex class II molecules.

Toxic shock syndrome toxin 1 (TSST-1) is a 22-kDa exotoxin produced by strains of Staphylococcus aureus and implicated in the pathogenesis of toxic shock syndrome. In common with other staphylococcal exotoxins, TSST-1 has diverse immunological effects. These include the induction of interleukin 2 receptor expression, interleukin 2 synthesis, proliferation of human T lymphocytes, and stimulation of interleukin 1 synthesis by human monocytes. In the present study, we demonstrate that TSST-1 binds with saturation kinetics and with a dissociation constant of 17-43 nM to a single class of binding sites on human mononuclear cells. There was a strong correlation between the number of TSST-1 binding sites and the expression of major histocompatibility complex class II molecules, and interferon-gamma induced the expression of class II molecules as well as TSST-1 binding sites on human skin-derived fibroblasts. Monoclonal antibodies to HLA-DR, but not to HLA-DP or HLA-DQ, strongly inhibited TSST-1 binding. Affinity chromatography of 125I-labeled cell membranes over TSST-1-agarose resulted in the recovery of two bands of 35 kDa and 31 kDa that comigrated, respectively, with the alpha and beta chains of HLA-DR and that could be immunoprecipitated with anti-HLA-DR monoclonal antibodies. Binding of TSST-1 was demonstrated to HLA-DR and HLA-DQ L-cell transfectants. These results indicate that major histocompatibility complex class II molecules represent the major binding site for TSST-1 on human cells.

Animals↗
PubMed1989

Constitutive and stimulus-induced phosphorylation of CD11/CD18 leukocyte adhesion molecules.

The leukocyte CD11/CD18 adhesion molecules (beta 2 integrins) are a family of three heterodimeric glycoproteins each with a distinct alpha subunit (CD11a, b, or c) and a common beta subunit (CD18). CD11/CD18 mediate crucial leukocyte adhesion functions such as chemotaxis, phagocytosis, adhesion to endothelium, aggregation, and cell-mediated cytotoxicity. The enhanced cell adhesion observed upon activation of leukocytes is associated with increased surface membrane expression of CD11/CD18, as well as a qualitative upregulation of CD11/CD18 functions. To elucidate the nature of the qualitative modifications that occur, we examined the phosphorylation status of these molecules in resting human leukocytes and upon activation with PMA or with the chemotactic peptide F-met-leu-phe (FMLP). In unstimulated cells, all three CD11 subunits were found to be constitutively phosphorylated. In contrast, phosphorylation of the common CD18 subunit was minimal. PMA induced rapid and sustained phosphorylation of CD18 that occurred at high stoichiometry, but had only minimal effects on phosphorylation of the associated CD11 subunits. FMLP also induced rapid phosphorylation of CD18, but the effect was of short duration. FMLP-induced phosphorylation of CD18 was not related to its Ca++-mobilizing effect, as CD18 phosphorylation was not observed upon treatment of leukocytes with the Ca++ ionophore, ionomycin. Phosphoamino acid analysis of CD11/CD18 in PMA- or FMLP-treated monocytes revealed a predominance of phosphoserine residues in all CD11/CD18 subunits. A small component of phosphothreonine was present in CD11c and CD18 and a minor component of phosphotyrosine was also detected in CD18 upon leukocyte activation may regulate the adhesion functions mediated by the CD11/CD18 family of molecules.

Amino Acid Sequence↗
PubMed1989

Interleukin 4 dependent induction of IgE synthesis and CD23 expression by the supernatants of a human helper T cell clone.

The supernatants of the human alloreactive helper T cell clone A1, which secretes interleukins (IL) 4 and 5, could induce IgE synthesis by normal peripheral blood mononuclear cell and expression of Fc epsilon R2/CD23 on normal human monocytes. Both effects were IL-4 dependent, since they could be induced by recombinant IL-4 alone and were inhibited by an anti-IL-4 antibody. The activation of such clones may contribute to the high IgE production and the high percentage of circulating CD23+ monocytes observed in allergic disorders.

Antigens, Differentiation, B-Lymphocyte↗
PubMed1989

Regulation of human IgE synthesis.

Allergic diseases result from the interaction with IgE bound to cell surface receptors. Therefore, rational therapeutic approaches to allergic diseases would be aimed at decreasing IgE and/or at blocking the binding of IgE to effector cells such as mast cells and monocytes. Our investigation of the mechanism of IgE synthesis in man shows that IgE synthesis by peripheral blood mononuclear cells (PBMC) absolutely requires the presence of IL-4 and requires endogenous IL-6, because antibody to IL-6 inhibits IgE production completely. IgE synthesis requires T/B cell contact and involves interactions between B cell surface MHC Class II molecules and T cell surface receptors, as antibodies to both of these cell surface molecules inhibit IgE synthesis. Furthermore, alloreactive T cell clones which are unable to engage the B cell MHC Class II molecules fail to induce IgE synthesis in spite of their ability to secrete IL-4. Studies on the immunoglobulin sites that are involved in IgE binding to high affinity receptors on mast cells and basophils have used recombinant fragments of IgE to block mast cell binding. These studies suggest that a stretch of 76 amino acids which straddles the C epsilon 2 and C epsilon 3 domains is essential for this binding. Parallel studies on IgE binding to low affinity receptors on monocytes and B cells suggest that sequences within C epsilon 3 are involved in this binding. Peptides or analogues that inhibit IgE binding to its cellular receptors may be useful in the treatment of allergic diseases.

B-Lymphocytes↗
PubMed1989

The IgE system.

The data presented herein indicate that IgE synthesis results from a complex interaction between T cells, B cells, and monocytes, under the control of T cell-derived and monocyte-derived lymphokines (IL-4, IL-5, and IL-6). In addition to their direct effects on IgE synthesis, these lymphokines have other functions, many of which are related to IgE. IL-4 is not only a crucial signal for the induction of IgE synthesis, but also a growth factor for murine mast cells together with IL-3, a product of the same T cell clones. Furthermore, IL-4 is a potent inducer of Fc epsilon R2b expression on both B cells and monocytes. On the other hand, IL-4-secreting T cells also produce IL-5, which promotes B cell maturation and induces differentiation of eosinophil precursors. Eosinophils, in turn, express CD23 when activated; thus, the activation of the IgE system leads to increased IgE synthesis and to enhanced expression of Fc epsilon R1 (on mast cells) and Fc epsilon R2 (on B cells, monocytes, and eosinophils). This in turn results in increased receptor-ligand interactions, with release of different chemical mediators involved in the pathogenesis of allergy. Such considerations suggest that the regulation of IgE synthesis should be viewed in the wider context of the IgE system, a multi-component network in which different cell types are functionally integrated mainly through lymphokine-mediated signals.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Hyper immunoglobulin E syndrome.

The Hyper Immunoglobulin E(HIE) Syndrome is a rare disorder of unknown etiology associated with extremely high serum IgE levels, serious recurrent infections and chronic dermatitis. The current paper will review the clinical and laboratory manifestations of the HIE Syndrome and discuss them in relationship to underline immunologic abnormalities found in this syndrome.

Adolescent↗
PubMed1988

Functional heterogeneity among human inducer T cell clones.

Analysis of mouse CD4+ inducer T cells at the clonal level has established that a dichotomy among CD4+ T cell clones exists with regard to types of lymphokines secreted. Mouse T cell clones designated Th1 have been shown to secrete IL-2 and IFN-gamma, whereas T cell clones designated Th2 have been shown to produce IL-4 but not IL-2 or IFN-gamma. To determine if such a dichotomy in the helper inducer T cell subset occurred in man, we examined a panel of human CD4+ helper/inducer T cell clones for patterns of lymphokine secretion and for functional activity. We identified human T cell clones which secrete IL-4 but not IL-2 or IFN-gamma, and which appeared to correspond to murine Th2 clones. In marked contrast to murine IL-2 secreting Th1 clones which do not produce IL-4 or IFN-gamma, we observed that some human T cell clones secrete IL-2, and IFN-gamma as well as IL-4. Southern blot analysis indicated that these multi-lymphokine-secreting clones represented the progeny of a single T cell. IL-4 secretion did not always correlated with enhanced ability to induce Ig synthesis. Although one T cell clone which secreted IL-2, IL-4, and IFN-gamma could efficiently induce Ig synthesis, another expressed potent cytolytic and growth inhibitory activity for B cells, and was ineffective or inhibitory in inducing Ig synthesis. These results indicate that although the equivalent of murine Th2 type cells appears to be present in man, the simple division of T cells into a Th1 and Th2 dichotomy may not hold true for human T cells.

Adult↗
PubMed1988

Phosphorylation of T cell membrane proteins by activators of protein kinase C.

Activation of the enzyme protein kinase C (PKC) plays an important role in T cell activation. We investigated the phosphorylation of CD2, CD3, CD4, CD5, CD7, CD8, CD28 (Tp44), CD43 (sialophorin, gp115), and LFA-1 after incubation of human PBMC with the (PKC) activator PMA. These proteins were chosen for their role in transmembrane signal transduction (CD2, CD3, CD5, CD28, CD43), cell-cell interaction and adhesion (CD2, CD4, CD8, and LFA-1), or involvement in immunodeficiency states (CD43, CD7). CD5, CD7, CD43, and the alpha-chain of LFA-1 were found to be constitutively phosphorylated. PMA induced rapid hyperphosphorylation of CD5, CD7, and CD43, but not of the LFA-1 alpha-chain, and induced the phosphorylation of CD3, CD4, CD8 and of the LFA-1 beta-chain. PMA did not cause the phosphorylation of CD2 and CD28. PMA-induced phosphorylation was partially inhibited by the PKC inhibitor 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine dihydrochloride. Finally, the T cell activator Con A, which binds to the CD3/TCR complex was shown to induce a profile of protein phosphorylation similar to that observed with PMA. We conclude that PKC-mediated phosphorylation of T cell Ag may represent an important regulatory mechanism that governs the process of T cell activation.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1988

Human recombinant interleukin 4 induces Fc epsilon R2/CD23 on normal human monocytes.

rIL-4 (B cell stimulatory factor 1) induces the expression of Fc epsilon R2/CD23 on normal human monocytes (Mo). Fc epsilon R2/CD23 induction was detectable both by flow cytometry using anti-CD23 mAbs as well as soluble IgE, and by the immunoprecipitation with CD23-specific mAb or IgE of a 45-kD band from 125I-lactoperoxidase-labeled Mo. Fc epsilon R2/CD23 was fully expressed after a 24-h incubation with rIL-4, and was still detectable after 72 h from the addition of IL-4. This effect was specific, because none of the other rILs tested (IL-1, IL-2, IL-3, IL-5, B cell stimulatory factor 2, granulocyte-macrophage colony stimulating factor, and IFN-gamma) could induce FC epsilon R2/CD23, either alone or in various combinations. No synergism was observed between IL-4 and other ILs. IFN-gamma was not able to inhibit the IL-4-induced expression of Fc epsilon R2/CD23 on Mo, neither when added to the culture together with IL-4, nor when added 36 h earlier.

Gene Expression Regulation↗
PubMed1988

Toxic shock syndrome toxin-1 induces inositol phospholipid turnover, protein kinase C translocation, and calcium mobilization in human T cells.

Toxic shock syndrome toxin-1 (TSST-1) is a 22-kDa exotoxin produced by most Staphylococcus aureus strains responsible for toxic shock syndrome. TSST-1 is a mitogen for human T cells. The mechanism of T cell activation by TSST-1 was investigated. TSST-1 induced IL-2R expression, IL-2 synthesis, and proliferation in T cells in a monocyte-dependent fashion. Neither IL-1 nor IL-2, alone or in combination, substituted for monocytes in supporting TSST-1-induced mitogenesis. We investigated the mechanism by which TSST-1 induces initogenesis. TSST-1 failed to induce ADP-ribosylation of T cell membrane proteins. However, the toxin induced transient translocation of protein kinase C from cytosol to plasma membranes and also induced the mobilization of cellular Ca2+ stores in both PBMC and the Jurkat human tumor T cell line, suggesting that TSST-1 triggered inositol phospholipid turnover. This was directly demonstrated to be the case in both cellular preparations studied. TSST-1 induced the increased synthesis of the inositol phospholipid phosphatidyl inositol, phosphatidyl inositol-4 phosphate, and phosphoinositol inositol-4,5-bisphosphate, and induced the breakdown of inositol phospholipid as evidence by the accumulation of phosphatidic acid and inositol phosphates. We conclude that the action of TSST-1 involves the induction of inositol phospholipid turnover, protein kinase C activation, and mobilization of cellular Ca2+ stores. This effect is similar to that of mitogenic lectins and of anti-CD3 antibodies.

Adenosine Diphosphate Ribose↗
PubMed1988

Increased in vitro bone resorption by monocytes in the hyper-immunoglobulin E syndrome.

Children with the hyper-immunoglobulinemia E syndrome are prone to bone fractures. We determined bone density in six patients with hyper-IgE syndrome using photon absorptiometry. All six patients had significantly reduced bone density compared with age- and sex-matched controls. The capacity of peripheral blood monocytes to degrade 45Ca-labeled bone in vitro was studied. The mean percent 45Ca release in four hyper-IgE patients (40.8 +/- 8.6) was significantly higher (p less than 0.01) than in ten age-matched healthy control subjects (7.1 +/- 2.6), seven age-matched patients with recurrent infections (7.9 +/- 1.6), or nine patients with severe atopic dermatitis and elevated serum IgE levels (5.8 +/- 1.3). Monocytes from four of four patients studied spontaneously released abnormally high levels of prostaglandin E2. Bone degradation by these monocytes was significantly reduced in the presence of 10(-6) M indomethacin in vitro. Administration of aspirin in vivo to two hyper-IgE patients reduced boned degradation by their monocytes to normal levels. These results suggest that monocytes from patients with hyper-IgE syndrome are activated to resorb bone via products of the prostaglandin synthase (cyclooxygenase) pathway. The activation of cells in the monocyte/macrophage family to resorb bone may contribute to the osteopenia observed in hyper-IgE syndrome.

Aspirin↗
PubMed1988

Regulatory effects of human IgE-binding factors in the IgE synthesis by human and rat lymphocytes.

We have previously established human T cell hybridomas which produce IgE-binding factors. Incubation of one of the T cell hybridomas, 166A2, with human IgE dimer in the presence of 1 microgram/ml bradykinin resulted in the formation of IgE-binding factors having affinity for lentil lectin. The factors selectively enhanced both IgE-forming cell responses of rat mesenteric lymph node (MLN) cells and spontaneous IgE synthesis by human peripheral blood B cells of atopic patients, without affecting the IgG response. The same factors that enhanced IgE synthesis of B cells from atopic patients also enhanced IgE synthesis induced under bystander conditions by activated alloreactive T cells. Fractionation of the affinity-purified IgE-binding factors by gel filtration revealed three molecular mass species, i.e., 60 kDa, 30 kDa and 15 kDa. The 60-kDa and 15-kDa IgE-binding factors selectively enhanced both the spontaneous IgE synthesis by B cells of atopic patients and IgE response of rat MLN cells. In contrast, the 30-kDa IgE-binding factors had only marginal enhancing effects on the IgE synthesis by both human B cells and rat MLN cells. When the 166A2 hybridoma cells were incubated with IgE dimer in the presence of glycosylation-inhibiting factor (GIF), essentially all IgE-binding factors formed by the cells had affinity for peanut agglutinin (PNA) but for neither lentil lectin nor concanavalin A. All of the 60-kDa, 30-kDa and 15-kDa species, having affinity for PNA, selectively suppressed the potentiating factor-enhanced IgE response of rat MLN cells. The factors also suppressed the IgE synthesis of human B cells from atopic patients when the synthesis was enhanced by IgE-potentiating factor. The results indicate that human IgE-binding factors regulate IgE synthesis by both human and rat lymphocytes.

Animals↗
PubMed1988

Recognition by a human alloreactive T-cell clone of an HLA-DP-associated epitope on monocytes and fibroblasts but not on B cells.

We examined the ability of a panel of allospecific (N = 9) and of TT-specific (N = 15) human inducer T-cell clones to respond to antigen presented by B cells or by monocytes. With one exception all T-cell clones responded equally well to antigen presented by monocytes, by lightly irradiated (1000 rads) peripheral blood resting B cells, or by heavily irradiated (7500 rads) Epstein-Barr virus (EBV)-transformed B cells. One alloreactive human T-cell clone, Clone A1, which recognized an HLA-DP-associated antigen proliferated in response to allogeneic monocytes and gamma-interferon-treated fibroblasts but not in response to allogeneic B cells even in the presence of autologous monocytes. Nonspecific conjugate formation between B cells and Clone A1 was normal. Yet in contrast to allogeneic monocytes, allogeneic B cells failed to induce a rise in the intracellular calcium ion concentration and failed to cause interleukin 2 (IL2) receptor expression in Clone A1. Neither interleukin 1 (IL1) nor phorbol myristate acetate (PMA) reversed the inability of Clone A1 to proliferate to allogeneic B cells. The failure of allogeneic B cells to stimulate A1 was not due to their lack of expression of the HLA-DP gene product recognized by Clone A1 or to excessive sialation of this product. These results suggest that Clone A1 recognizes an epitope associated with HLA-DP which is expressed on monocytes and on gamma-interferon-treated fibroblasts but which is either absent or altered on B cells.

Adult↗