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Biomedical subjects

S Sriram

Publications and source records attributed to S Sriram.

At least 73 records · Page 4Linked to original sources
PubMed1994

Myelin basic protein peptide specificity and T-cell receptor gene usage of HPRT mutant T-cell clones in patients with multiple sclerosis.

Characterization of T cells responding to autoantigens is central to understanding autoimmune disease. We have used somatic mutation at the hypoxanthine guanine phosphoribosyltransferase (HPRT) gene as an index of T-cell amplification in vivo. With this strategy we previously showed that myelin basic protein-reactive T cells can be isolated only from the HPRT mutant T-cell population cultured from the peripheral blood of multiple sclerosis patients and not from normal individuals. In this study, 165 HPRT mutant and 104 wild-type clones were examined for their reactivity to myelin basic protein and overlapping peptides of myelin basic protein. Five HPRT mutant clones that recognized myelin basic protein and myelin basic protein peptides along with three clones that responded to myelin basic protein peptide alone were isolated. All but one of the eight clones recognized peptides derived from the carboxy terminus of myelin basic protein (p84-168). Sequence analysis showed heterogeneous expression of T-cell receptor V alpha and V beta genes and CDR3s. These studies showed that in vivo amplified autoimmune T cells from patients with long-standing disease use diverse T-cell receptor elements in the recognition of C-terminal myelin basic protein peptides.

Adult↗
PubMed1994

Homologies between T cell receptor junctional sequences unique to multiple sclerosis and T cells mediating experimental allergic encephalomyelitis.

The selection of T cell clones with mutations in the hypoxanthine guanine phosphoribosyltransferase (hprt) gene has been used to isolate T cells reactive to myelin basic protein (MBP) in patients with multiple sclerosis (MS). These T cell clones are activated in vivo, and are not found in healthy individuals. The third complementarity determining regions (CDR3) of the T cell receptor (TCR) alpha and beta chains are the putative contact sites for peptide fragments of MBP bound in the groove of the HLA molecule. The TCR V gene usage and CDR3s of these MBP-reactive hprt-T cell clones are homologous to TCRs from other T cells relevant to MS, including T cells causing experimental allergic encephalomyelitis (EAE) and T cells found in brain lesions and in the cerebrospinal fluid (CSF) of MS patients. In vivo activated MBP-reactive T cells in MS patients may be critical in the pathogenesis of MS.

Amino Acid Sequence↗
PubMed1994

Longitudinal study of frequency of HPRT mutant T cells in patients with multiple sclerosis.

Analysis of somatic mutation in circulating peripheral blood T cells can be used as an index of in vivo T-cell amplification. The ability to assess the frequency of T cells that survive in vitro in the presence of 6-thioguanine is an index of mutation at the HPRT gene locus. In the absence of exposure to mutational agents, elevations of the HPRT mutant frequency (mF) of T cells reflects errors in DNA replication and repair that have become fixed during the process of cell division. We estimated the mF of T cells in MS patients and controls over a period of 36 months and found that the mF was consistently elevated in MS patients of all clinical subgroups. In the chronic progressive group of MS, the mF increased over a 3-year period and appeared to correlate with the clinical worsening of the disease.

Adult↗
PubMed1993

Long-term treatment of chronic relapsing experimental allergic encephalomyelitis by transforming growth factor-beta 2.

It had been demonstrated previously that the administration of transforming growth factor-beta 1 (TGF-beta 1) reduced the clinical severity of experimental allergic encephalomyelitis (EAE). Treatment with the related immunosuppressive molecule, TGF-beta 2, resulted in similar inhibition of T cell activation and proliferation in vitro. Long-term treatment was effective in reducing clinical severity of EAE and the number of relapses in mice receiving either myelin basic protein- or peptide-91-103-specific T cell lines. When examined histologically, mice that had received TGF-beta 2 demonstrated significantly less inflammation and demyelination in the central nervous system. Examination of other organs demonstrated no pathology or deleterious side effects from long-term TGF-beta 2 therapy. These findings have relevance for the use of TGF-beta 2 as a therapeutic agent for the human demyelinating disease, multiple sclerosis.

Amino Acid Sequence↗
PubMed1993

Experimental allergic encephalomyelitis: neutralizing antibody to TGF beta 1 enhances the clinical severity of the disease.

Experimental allergic encephalomyelitis (EAE) is a well established model for the human autoimmune disease multiple sclerosis. Recently, we and others have shown that the administration of TGF beta is therapeutically effective in reducing incidence and severity of EAE. Here we show that the addition of anti-TGF beta 1 to myelin basic protein (MBP)-activated lymph node cells enhance the T cell proliferative response by 28% in vitro and in vivo and that injections of anti-TGF beta 1 antibody worsen EAE both in incidence and severity. Further, an inverse relationship was observed in the amount of IL-2 and TGF beta detected in MBP stimulated culture supernatants. We show that IL-2 decreases from 248 U/ml at 48 h to non-detectable at 96 h, while TGF beta increases from 0.5 ng/ml to 1.2 ng/ml, respectively. These observations further indicate a role for endogenous TGF beta 1 in the immunoregulation of EAE.

Animals↗
PubMed1993

Lumbosacral radiculoplexopathy as a manifestation of Epstein-Barr virus infection.

We report the clinical features of five patients with lumbosacral radiculoplexopathy (LSRP) and one patient with a femoral neuropathy, all of whom had serologic evidence of a recent Epstein-Barr virus (EBV) infection. After a thorough investigation, no other etiology was apparent. Pain was a prominent feature in all cases, and the prognosis was generally good, with recovery in weeks to months. We conclude that LSRP may occur as a postinfectious process following recent EBV infection.

Adult↗
PubMed1992

Transforming growth factor-beta 1 differentially regulates proliferation and MHC class-II antigen expression in forebrain and brainstem astrocyte primary cultures.

To facilitate investigation of cytokine regulation of reactive astrogliosis, primary astrocyte cultures from neonatal murine forebrain and brainstem were established. Forebrain and brainstem astrocytes proliferated at a similar rate under basal culture conditions, and both were growth-inhibited by treatment with recombinant murine interferon-gamma. The growth of cultured brainstem astrocytes was significantly enhanced by exposure to recombinant human transforming growth factor-beta 1. In contrast, proliferation of forebrain astrocytes was not significantly affected by transforming growth factor-beta 1. The disparate responses of brainstem and forebrain astrocytes to transforming growth factor-beta 1 treatment were not limited to effects on cell growth, since transforming growth factor-beta 1 could block interferon-gamma-induced MHC class-II antigen expression on cultured brainstem astrocytes but not on forebrain cells. Results could not be attributed to use of an heterologous cytokine/cellular target system, since similar variability in transforming growth factor-beta 1 modulation of major histocompatibility complex antigen expression could be demonstrated using two human astrocytoma cell lines. This report is the first to document mitogenic response to transforming growth factor-beta 1 for neuroepithelial cells. The role of transforming growth factor-beta 1 in regulating aspects of reactive astrogliosis, particularly in the context of inflammatory demyelination, requires further investigation. Furthermore, these studies may provide insight into regional variability in the sequelae of inflammation within the central nervous system.

Animals↗
PubMed1992

Analysis of TCR V beta gene usage and encephalitogenicity of myelin basic protein peptide p91-103 reactive T cell clones in SJL mice: lack of evidence for V gene hypothesis.

We have analyzed the epitope specificity and encephalitogenicity of peptides that span the C terminus of MBP, p84-103. Our studies show that multiple antigenic epitopes with disease-inducing capacity exist in SJL mice. Three peptides that span this region were examined and found to be immunogenic. However, the mode of immunization (active or passive) determined the incidence and severity of EAE. In our experiments adoptive transfer of p91-103-reactive T cell lines was most consistent in the development of disease. Interestingly, the response to peptides p89-101, p91-103, and p84-102 was absent following immunization with MBP. This suggests that although p91-103 and p89-101 were encephalitogenic they were not the major immunogenic epitopes following immunization with MBP. Analysis of a panel of eight p91-103-reactive T cell clones showed significant heterogeneity in the fine specificity, the TCR V beta gene usage, and in their ability of transfer EAE. These studies suggest that in SJL mice the epitopes involved in the pathogenesis of disease are multiple and there is no clear correlation between encephalitogenicity and TCR V beta gene usage. These observations argue against the presence of a dominant TCR V beta gene in the pathogenesis of EAE in SJL mice.

Amino Acid Sequence↗
PubMed1991

Antigen-specific therapy of experimental allergic encephalomyelitis by soluble class II major histocompatibility complex-peptide complexes.

Experimental allergic encephalomyelitis is a T-cell-mediated, major histocompatibility complex (MHC) class II gene-linked autoimmune demyelinating disease of the central nervous system. To develop therapies that will specifically inactivate only the autoantigen-reactive T cells, mice were treated with soluble MHC class II molecules that had been complexed with encephalitogenic peptides. Intravenous injections of 300 micrograms of complexes consisting of encephalitogenic peptide 91-103 of myelin basic protein plus I-As protein on day 0, 4, and 7 were effective in preventing experimental allergic encephalomyelitis. Similarly, administration of 45 micrograms of I-As protein complexed to peptide 139-151 from proteolipoprotein on day 1, 4, and 7 prevented mortality and significantly reduced paralysis induced by immunization with the encephalitogenic proteolipoprotein peptide. Histological examination of sections of animal brains revealed that treatment with I-As protein plus myelin basic protein 91-103 peptide prevents the development of inflammatory lesions characteristic of experimental allergic encephalomyelitis. Thus, treatment with MHC-self-peptide complexes could serve as a highly specific therapeutic modality in treating autoimmune disease when the putative autoantigen and the MHC restricting elements are known.

Animals↗
PubMed1991

Successful treatment of experimental allergic encephalomyelitis with transforming growth factor-beta 1.

Transforming growth factor-beta 1 (TGF-beta 1) is a multifunctional cytokine with immunosuppressive effects on T cells in vitro. Experimental allergic encephalomyelitis is an archetypal T cell-mediated autoimmune demyelinating disease of the central nervous system that often serves as a model for multiple sclerosis. In vivo administration of TGF-beta 1 into SJL mice was successful in reducing the incidence of clinical disease and the histologic severity of inflammation and demyelination in the brain and spinal cord. Immunohistochemical studies performed on control animals showed that TGF-beta-1, -2, and -3 were present in inflammatory perivascular lesions in the brain. The use of a naturally occurring cytokine with immunoregulatory functions in the treatment of an autoimmune disease is novel. However, potential long term complications of such therapy must be addressed before its use in human autoimmune disease such as multiple sclerosis.

Animals↗
PubMed1991

Haplotype-specific inhibition of homing of radiolabeled lymphocytes in experimental allergic encephalomyelitis following treatment with anti-IA antibodies.

In vivo treatment with anti-IA antibodies has been shown to induce a haplotype-specific inhibition of EAE when the disease was following passive transfer of MBP-sensitized T cells. In order to determine the mechanism by which anti-IA antibody prevents passively transferred EAE, the homing of radiolabeled cells to the brain following anti-IA therapy was studied. Administration of anti-IA antibodies at the earliest onset of clinical signs of EAE prevented the homing of radio-labeled cells to the brain. In F1 (Balb/c x SJL/J) mice that developed EAE and received anti-IAs antibody there was a decreased homing of radiolabeled cells when compared to animals that received anti-IAd antibody. In addition, there was preferential expression of IAs antigen, over IAd antigen on capillary endothelium of the brain. The differential expression of IA antigens and the homing of radiolabeled cells in F1 (SJL x Balb/c) mice could in part explain the haplotype-specific suppression of disease following treatment with anti-IA antibodies.

Animals↗
PubMed1991

Treatment of chronic relapsing experimental allergic encephalomyelitis with the intravenous administration of splenocytes coupled to encephalitogenic peptide 91-103 of myelin basic protein.

Chronic relapsing experimental allergic encephalomyelitis (CR-EAE) is an autoimmune demyelinating disease of the central nervous system and serves as an experimental model of human multiple sclerosis. Amino acid residues p91-103 of myelin basic protein are encephalitogenic in SJL mice and transfer of T cell lines that recognize this epitope results in CR-EAE. We show here that coculture of T cells in the presence of p91-103 that has been chemically cross-linked to the antigen presenting cells renders the T cell lines tolerant to the antigen. Injection of p91-103 coupled splenocytes into animals that had received encephalitogenic p91-103 reactive T cells significantly reduced the incidence and severity of EAE. Furthermore, treatment of mice with a single injection of antigen coupled splenocytes after they had recovered from their initial paralytic attack prevented the development of subsequent clinical relapses in all animals. These studies indicate that this effect is long lasting and can be successfully accomplished in an established autoimmune disease. Hence this form of immunotherapy may be considered as a therapeutic modality in the treatment of autoimmune diseases when the autoantigens are known.

Animals↗
PubMed1991

In vivo administration of TNF-alpha prevents EMC-M virus induced viral encephalitis.

EMC-M virus causes a monophasic paralytic syndrome characterized by encephalitic lesions in the brain and patchy demyelinating lesions in the spinal cord and nerve roots of BALB/c mice. Since the replication of EMC virus in vitro is inhibited by tumor necrosis factor (TNF)-alpha we have studied the effect of in vivo administration of this cytokine on the acute disease. Our studies show that periodic administration of TNF-alpha to animals infected with EMC-M reduces viral titers in the brain, and decreases the degree of clinical paralysis and the severity of the inflammatory lesions in the brain.

Animals↗
PubMed1991

Indirect role of T cells in development of polioencephalitis and encephalomyelitis induced by encephalomyocarditis virus.

Infection of female BALB/c mice with encephalomyocarditis virus results in the development of a paralytic syndrome in 7 to 10 days postinoculation. Previous studies had suggested the involvement of an immune component in the development of central nervous system pathology. We have examined the effects of T-cell depletion on the development of polioencephalitis (neuronal necrosis and inflammation of the brain and brain stem) and the relative contribution of the CD4+ and CD8+ subsets following the establishment of viremia. We show that monoclonal antibody depletion of T cells is effective in the reduction of polioencephalitis when given prior to viral inoculation. However, administration of the antibodies 12 h or more after viral inoculation failed to alter the development of polioencephalitis or encephalomyelitis. We conclude that T cells are involved in the development of central nervous system disease during the initial stages of infection but are not responsible for the later progression of disease.

Animals↗
PubMed1990

T cells responsive to myelin basic protein in patients with multiple sclerosis.

Gene mutation in vivo in human T lymphocytes appears to occur preferentially in dividing cells. Individuals with multiple sclerosis (MS) are assumed to have one or more populations of diving T cells that are being stimulated by autoantigens. Mutant T cell clones from MS patients were isolated and tested for reactivity to myelin basic protein, an antigen that is thought to participate in the induction of the disease. The hypoxanthine guanine phosphoribosyltransferase (hprt) clonal assay was used to determine mutant frequency values in MS patients with chronic progressive disease. Eleven of 258 thioguanine-resistant (hprt-) T cell clones from five of the six MS patients who were tested proliferated in response to human myelin basic protein without prior in vitro exposure to this antigen. No wild-type clones from these patients, nor any hprt- or wild-type clones from three healthy individuals responded to myelin basic protein. Thus, T cell clones that react with myelin basic protein can be isolated from the peripheral blood of MS patients.

Adult↗
PubMed1989

Spontaneous development in vitro of a myelin basic protein-specific suppressor T cell line.

T cell lines to myelin basic protein (MBP) developed following in vitro culture cause experimental allergic encephalomyelitis (EAE) upon transfer into naive recipient mice. We have, however, repeatedly observed that MBP-specific T cell lines lose their ability to transfer EAE after 40 days in culture. Analyses of such cell lines failed to show any differences in their proliferative responses to antigen, or in the secretion of interleukin-2 (IL-2) and/or IL-4 when compared to their encephalitogenic counterparts. In contrast, examinations of T cell receptor (TCR) beta-chain gene rearrangement patterns showed sequential changes in the clonal population of cells concomitant with the loss of encephalitogenic function. Furthermore, transfer of a non-encephalitogenic, genotypically altered cell line after long-term in vitro culture into mice challenged with MBP suppressed the development of EAE. These findings suggest that the development of such putative regulatory cells in vivo may be involved in the recovery in EAE.

Animals↗
PubMed1989

Treatment of encephalomyocarditis virus-induced central nervous system demyelination with monoclonal anti-T-cell antibodies.

Infection of BALB/c mice with the M variant of encephalomyocarditis virus resulted in the development of a paralytic syndrome in 7 to 10 days. The paralysis was maximal during the period of viral clearance; most of the animals recovered from the initial deficit and showed no delayed recurrences. Pathologically, the white matter of brain and spinal cord showed well-demarcated areas of perivascular cuffing, demyelination, and, during recovery, remyelination by oligodendrocytes--all suggestive of postinfectious encephalomyelitis. Depletion of either the CD4 or CD8 subset of T cells in vivo with the appropriate monoclonal antibody, GK1.5 or 2.43, respectively, administered one day (24 h) prior to infection was sufficient to limit the development of the paralytic syndrome by 79% (GK1.5) and 82% (2.43).

Animals↗