Search PubMed⌕ Search

Biomedical subjects

T Chatila

Publications and source records attributed to T Chatila.

At least 37 records · Page 2Linked to original sources
PubMed1993

Microbial superantigens induce NF-kappa B in the human monocytic cell line THP-1.

Staphylococcal superantigens bind to MHC class II molecules and induce transcriptional activation of IL-1 beta and TNF-alpha genes in human monocytic cells. The understanding of the mechanisms by which superantigens activate cytokine gene expression is incomplete. In this study, we demonstrate that toxic shock syndrome toxin-1 (TSST-1) and staphylococcal enterotoxins A and B induce the activation of NF-kappa B, a transcriptional enhancer that binds to sequences found in both the IL-1 beta and TNF-alpha promoters. Electrophoretic mobility-shift assays showed a rapid induction of nuclear proteins that bound to the consensus kappa B motif. Furthermore, TSST-1 potently stimulated chloramphenicol acetyltransferase (CAT) expression by THP-1 cells transfected with a consensus NF-kappa B-promoter CAT construct, indicative of induction of NF-kappa B enhancer function. Induction of both NF-kappa B DNA-binding proteins and NF-kappa B enhancer function was down-regulated by inhibitors of protein kinase C and protein tyrosine kinase, indicating a role for these protein kinases in the induction of NF-kappa B by MHC class II ligands. Using neutralizing antibodies, we demonstrated that after the stimulation of cells with TSST-1, TNF-alpha, but not IL-1 beta, acted to up-regulate binding of NF-kappa B to DNA and the activation of the NF-kappa B-promoter CAT construct. These results indicate that induction of NF-kappa B by superantigens is up-regulated in part by an autocrine loop involving TNF-alpha.

Antigens, Bacterial↗
PubMed1993

Defective expression of the CD40 ligand in X chromosome-linked immunoglobulin deficiency with normal or elevated IgM.

B lymphocytes from patients with X chromosome-linked immunoglobulin deficiency with normal or elevated serum IgM are unable to switch from the synthesis of IgM/IgD to that of other immunoglobulin isotypes. Isotype switch recombination was evaluated in three affected males by examining interleukin 4-driven IgE synthesis. T-cell-dependent IgE synthesis was completely absent in the B lymphocytes of the patients. In contrast, CD40 mAb plus interleukin 4 induced the patients' B cells to synthesize IgE and to undergo deletional switch recombination. Because interaction between CD40 and its ligand on activated T cells is critical for T-cell-driven isotype switching, we examined CD40 ligand expression. In contrast to normal T cells, lymphocytes from the patients expressed no detectable CD40 ligand on their surface after stimulation with phorbol 12-myristate 13-acetate and ionomycin, although the mRNA of the ligand was expressed normally. These results suggest that defective expression of the CD40 ligand underlies the failure of isotype switching in this disease.

Adolescent↗
PubMed1993

The T cell receptor associated CD3-epsilon protein is phosphorylated upon T cell activation in the two tyrosine residues of a conserved signal transduction motif.

Signal transduction through the T cell receptor for antigen, the TcR/CD3 complex, involves phosphorylation of tyrosine residues in the CD3-zeta chain. Since both CD3-epsilon and the zeta chain contain a tyrosine-based signaling motif, we examine phosphorylation of CD3-epsilon in human T cells. Engagement of the TcR/CD3 complex induced tyrosine phosphorylation of CD3-epsilon in vivo. Induction of CD3-epsilon phosphorylation followed similar kinetics to that of the zeta chain phosphorylation. In contrast to zeta, CD3-epsilon phosphorylation was strictly dependent upon cell surface expression of this member of the TcR/CD3 complex. Chemical and proteolytic cleavage combined with peptide-specific Western blotting established that CD3-epsilon phosphorylation occurred in the two tyrosine residues located in the signal transduction motif in the C-terminal portion of the molecule. Taken together, these data indicated that phosphorylation of CD3-epsilon by tyrosine protein kinases may serve to couple the TcR/CD3 complex to other effector molecules in the signaling cascade.

CD3 Complex↗
PubMed1993

Activator protein-1 (AP-1) is stimulated by microbial superantigens in human monocytic cells.

Microbial superantigens bind to major histocompatibility complex (MHC) class II molecules and activate gene transcription in monocytes. In search of transcription factors that potentially mediate the effects of superantigens at the nuclear level, we examined the capacity of staphylococcal superantigens to stimulate the activity of the transcriptional promoter factor activator protein-1 (AP-1). Electrophoretic mobility shift assays showed an increase in nuclear proteins that bound to the consensus AP-1 motif within 5 min following the stimulation of the monocytic cell line THP-1 with toxic shock syndrome toxin-1 (TSST-1) or staphylococcal endotoxin A. We show that mRNA levels for the subunits that compose AP-1, the protooncogenes c-fos and c-jun, are upregulated by stimulation of THP-1 cells with TSST-1. The activated AP-1 complexes were functional, as evidenced by the capacity of TSST-1 to stimulate the expression of an AP-1-driven reporter gene construct transfected into THP-1 cells. These results establish that the engagement of MHC class II molecules by superantigens increases the activity of functional AP-1 complexes and that this may proceed in part by transcriptional activation of c-fos and c-jun protooncogenes.

Antigens, Bacterial↗
PubMed1993

Staphylococcal superantigens as inducers of signal transduction in MHC class II-positive cells.

Staphylococcal superantigens (SEs and TSST-1) interact with and potentially activate two of the main subsets of the immune system: T lymphocytes and MHC class II-positive cells. Since the interaction of SEs and TSST-1 with MHC class II molecules is the first step in triggering immune cells activation, a detailed understanding of the nature of this interaction is essential for understanding its effect on the immune system and for designing therapeutic strategies for SEs and TSST-1-mediated injury. A series of events is induced in MHC class II-positive cells (B cells, activated T cells, monocytes, and synoviocytes) upon engagement with superantigens. Some of these events require monomeric forms of superantigens, whereas others are critically dependent on cross-linking of toxin-bound MHC class II molecules by a biochemical agent (biotin-avidin) or a natural physiological one such as the TCR. The ability of superantigens to induce polyclonal activation of MHC class II-positive cells may confer to the superantigen its capacity to trigger autoimmune diseases.

Animals↗
PubMed1993

Severe combined immunodeficiency with selective T-cell cytokine genes.

A 4-y-old female with severe combined immunodeficiency disease had normal numbers of T cells in her circulation and normal T-cell subsets. However, her T cells proliferated poorly to mitogens and did not proliferate to antigens or to anti-CD3 MAb. IL-2 receptor expression was normal, but IL-2 synthesis was undetectable. The addition of recombinant IL-2 to a mitogen-stimulated culture resulted in normalization of the proliferative response. Northern blot analysis of total RNA derived from the patient's T cells revealed a weak or absent expression of mRNA coding for IL-2, IL-3, IL-4, and IL-5. In contrast, there were normal amounts of mRNA coding for granulocyte-macrophage colony-stimulating factor. Tumor necrosis factor and IL-6 production were also normal. Nuclear run-on transcriptional assays revealed markedly decreased levels of newly initiated nuclear transcripts coding for IL-2, IL-3, IL-4, and IL-5 and normal levels of granulocyte-macrophage colony-stimulating factor transcripts in the patient relative to control lymphocytes. Gel retardation assays suggest that the NFAT-1 nuclear transcription complex is abnormal in this patient. These results indicate that the patient suffers from a defect that affects the transcription of multiple T-cell lymphokines and suggest that abnormalities affecting the production of T-cell lymphokines may underlie some of the primary immunodeficiency diseases.

Bone Marrow Transplantation↗
PubMed1992

Role of protein kinase activation in the induction of B cell adhesion by MHC class II ligands.

Engagement of MHC class II (Ia) molecules on B cells induces tyrosine phosphorylation, phosphoinositide turnover, elevation of intracellular calcium concentrations, and a rise in cAMP levels. However, a role for these biochemical signals in mediating functional responses induced by Ia ligands remains largely undefined. In this study, we utilized the induction of B cell adhesion by Ia ligands to demonstrate a role for signals transduced via Ia molecules in the generation of a functional response. Ia ligands that induced B cell aggregation induced tyrosine phosphorylation, whereas Ia ligands that did not induce B cell aggregation failed to induce any detectable tyrosine phosphorylation. Ia-induced B cell aggregation and tyrosine phosphorylation were inhibited by genistein and by herbimycin A, inhibitors of tyrosine kinases (PTK). Sphingosine and calphostin C, inhibitors of protein kinase C (PKC), also inhibited Ia-induced adhesion whereas HA1004, an inhibitor of cyclic nucleotide-dependent kinases, did not. Ia ligands induced both LFA-1-dependent and LFA-1-independent B cell adhesion. These two pathways of cell adhesion differed in their requirement for activation signals. PKC activation was sufficient for LFA-1-dependent adhesion, whereas LFA-1-independent adhesion required independent phosphorylation events mediated by PKC and by PTK. These results provide functional relevance for biochemical signals transduced via Ia molecules by demonstrating that Ia-induced B cell adhesion is mediated by the activation of PKC and by one or more PTK.

B-Lymphocytes↗
PubMed1992

Signals delivered via MHC class II molecules synergize with signals delivered via TCR/CD3 to cause proliferation and cytokine gene expression in T cells.

We examined the role of MHC class II molecules in transducing signals to activated human T cells. Cross-linking of MHC class II molecules synergized with submitogenic amounts of anti-CD3 mAb in causing proliferation and secretion of the cytokines IL-2, IL-3, IFN-gamma, and TNF-alpha by MHC class II-alloreactive T cell lines. Signaling via MHC class II molecules in T cells resulted in activation of tyrosine kinases, in generation of inositol phosphates, and in Ca2+ mobilization that was abrogated by the tyrosine kinase inhibitor herbimycin A. Thus, like signaling via TCR/CD3, signaling via MHC class II molecules involved tyrosine kinase-dependent activation of phospholipase C, resulting in phosphoinositol turnover and Ca2+ flux. However the signaling pathways coupled to MHC class II molecules and to TCR/CD3 differed, because engagement of the transmembrane phosphatase CD45 inhibited Ca2+ fluxes triggered via TCR/CD3 but not Ca2+ fluxes triggered via MHC class II molecules.

Antigens, CD↗
PubMed1992

Engagement of MHC class I molecules induces cell adhesion via both LFA-1-dependent and LFA-1-independent pathways.

We here demonstrate that ligand binding to MHC class I molecules induces homotypic cell adhesion of lymphocytes and monocytes. mAb to beta 2-microglobulin caused sustained, largely LFA-1-independent adhesion whereas mAb to the MHC class I alpha H chain caused transient LFA-1-dependent adhesion. Both the protein kinase C inhibitor sphingosine and the tyrosine kinase inhibitor genistein abrogated MHC class I-mediated cellular adhesion. These results indicate that MHC class I molecules transduce signals that induce cell adhesion and suggest that interaction between MHC class I-restricted T cells and APC may result in reciprocal enhanced adhesiveness of these cells.

Animals↗
PubMed1992

Association of neuronal pp60c-src with growth cone glycoproteins of rat brain.

Tyrosine phosphorylation and protein tyrosine kinase (PTK) activity in the growth cone membrane-associated glycoprotein (GCGP) fraction of 1-day-old rat brain were examined. Using immunoblotting and immunoprecipitation techniques, pp60c-src was identified as one of the major PTKs associated with GCGPs. Furthermore, only GCGP-associated src that was also tyrosine phosphorylated was active. Immunoprecipitation experiments using various src antibodies revealed that pp60c-src contributed partially to the PTK activity detected in GCGPs, and that it is associated with several proteins of Mr 140 K, 120 K, 85 K and 50 K. This association of src protein with GCGPs was specific, and another src family member p59fyn, which is also abundant in the brain, did not exhibit such an association. In addition to pp60c-src, the GCGP fraction contained several major phosphotyrosine-containing proteins of Mr 140 K, and a 97/90 K doublet that corresponded to the beta subunits of IGF-I/insulin receptors. These studies show that pp60c-src associated with GCGPs is an active PTK that could be involved in neuronal growth and development, transmembrane signalling, and in recognition and/or adhesive events.

Animals↗
PubMed1992

Superantigens.

Over the past three years superantigens have come to the forefront of immunological research. Studies in a number of laboratories have indicated that superantigens play a central role in shaping the T-cell repertoire in the development of tolerance, autoimmunity and immunodeficiency. This is in addition to their classic role in the pathogenesis of a number of infectious diseases.

Animals↗
PubMed1991

Expression of a neuronal Ca2+/calmodulin-dependent protein kinase, CaM kinase-Gr, in rat thymus.

The regional and tissue-specific expression of the Ca2+/calmodulin-dependent protein kinase, CaM kinase-Gr, were examined. The Mr 65,000 alpha-polypeptide of CaM kinase-Gr is expressed ubiquitously in different anatomical regions of rat brain, whereas an additional Mr 67,000 beta-polypeptide is observed solely in the cerebellum. The alpha-polypeptide appears in the neonatal rat forebrain and cerebellum, whereas the beta-polypeptide appears by the second postnatal week and may reflect cerebellar granule cell differentiation. Most peripheral tissues do not express either CaM kinase-Gr polypeptide. However, rat thymus and thymocytes derived therefrom express CaM kinase-Gr at levels comparable to those of the central nervous system. The identity of the enzyme in rat thymus was corroborated by immunoblot assays, Northern blots, and direct enzyme purification. Rat spleen and testis also produce CaM kinase-Gr, but at lower levels than either thymus or brain. These observations demonstrate selective regional and developmental expression of CaM kinase-Gr polypeptide in brain, and suggest that it may participate in Ca2+ signalling in cells derived both from the immune system as well as the central nervous system.

Animals↗
PubMed1991

Transcriptional activation of IL-1 beta and tumor necrosis factor-alpha genes by MHC class II ligands.

Ligands that bind MHC class II (Ia) molecules, including staphylococcal exotoxins (SE) and mAb induce IL-1 and TNF secretion in human monocytes and monocytic cell lines. In this study, we have analyzed the mechanisms by which SE induce IL-1 beta and TNF-alpha production. Treatment of human peripheral blood monocytes with staphylococcal exotoxin B and toxic shock syndrome toxin-1 resulted in a biphasic increase of IL-1 beta mRNA that lasted more than 12 h and in a more transient rise in TNF-alpha mRNA. A F(ab) preparation of the anti-HLA DR mAb L243 also caused a significant increase in monokine mRNA levels. Stimulation of a monocytic cell line, THP-1, with staphylococcal exotoxin B and toxic shock syndrome toxin-1 induced a rapid rise in IL-1 beta and TNF-alpha mRNA levels. This response peaked at 1 to 3 h poststimulation and remained detectable at 12 h. Nuclear run-on transcription assays demonstrated that SE cause transcriptional activation of the IL-1 beta and TNF-alpha genes. This transcriptional activation did not require de novo protein synthesis as it was not inhibited by the protein synthesis inhibitor cycloheximide. These results define an important function for Ia molecules as regulators of cytokine gene expression and add to the understanding of the changes in cellular function induced by SE.

Bacterial Toxins↗
PubMed1991

Engagement of MHC-class II molecules by staphylococcal exotoxins delivers a comitogenic signal to human B cells.

The staphylococcal exotoxin toxic shock syndrome toxin-1 (TSST-1) has been demonstrated to bind to monomorphic determinants on MHC-class II molecules. In this study, we have used TSST-1 to probe the role of MHC-class II molecules in the activation and differentiation of resting human B cells. Highly purified B cells were stimulated with TSST-1, alone or in combination with PMA or with anti-human IgM antibodies (anti-mu) and the resulting B cell proliferation and Ig production were monitored. On its own, TSST-1 failed to induce B cell proliferation or Ig production. However, TSST-1 synergized with PMA and with anti-mu in inducing B cell proliferation in the absence of any added T cells or T cell factors. TSST-1 did not induce or potentiate early activation events associated with anti-mu treatment such as phosphoinositide hydrolysis and Ca2+ mobilization. Also, TSST-1 did not potentiate the capacity of anti-mu to induce the transcription of early activation genes such as c-myc. Finally, in contrast to its capacity to promote mitogen-trigered B cell proliferation, TSST-1 failed to induce the differentiation of B lymphocytes into Ig-secreting cells in the absence of added T cells. These results indicate that TSST-1 delivers a comitogenic signal to B cells via MHC-class II molecules that is distinct from signals delivered via surface mu and further strengthens the role of MHC-class II molecules as signal transducing structures.

Antibodies, Anti-Idiotypic↗
PubMed1991

Novel immune deficiencies: defective transcription of lymphokine genes.

A 4-year-old female with severe combined immunodeficiency (SCID) had normal numbers of T cells in circulation and normal T cell subsets. However, her T cells proliferated poorly to mitogens and did not proliferate to antigens or to anti-CD3 mAb. Interleukin-2 (IL-2) receptor expression was normal but IL-2 synthesis was undetectable. The addition of recombinant IL-2 to a mitogen-stimulated culture resulted in normalization of the proliferative response. Northern blot analysis of total RNA derived from the patient's T cells revealed a weak or absent expression of mRNA coding for IL-2, IL-3, IL-4, and IL-5. In contrast, there were normal amounts of mRNA coding for granulocyte-macrophage colony-stimulating factor (GM-CSF). Tumor necrosis factor and IL-6 production was also normal. Nuclear run on transcriptional assays revealed markedly decreased levels of newly initiated nuclear transcripts coding for IL-2, IL-3, IL-4, and IL-5 and normal levels of GM-CSF transcripts in patient relative to control lymphocytes. These results indicate that the patient's T cells suffered from a defect affecting the transcription of multiple T cell lymphokines and suggest that abnormalities affecting the production of T cell lymphokines may underlie some of the primary immunodeficiency diseases.

Bone Marrow Transplantation↗