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Biomedical subjects

R Martin

Publications and source records attributed to R Martin.

At least 253 records · Page 14Linked to original sources
PubMed1997

Differential expression of cyclic nucleotide phosphodiesterase 3 and 4 activities in human T cell clones specific for myelin basic protein.

Little is known concerning the relative distribution and function of the different cyclic nucleotide phosphodiesterases (PDEs) in lymphocytes. Recent reports, however, have indicated that specific PDE4 inhibitors were effective in treatment of experimental allergic encephalomyelitis, an animal model of multiple sclerosis. The therapeutic effect of PDE4 inhibitors is thought to be related to inhibition of autoreactive CD4+ T cells specific for myelin basic protein (MBP) or other myelin proteins. Human autoreactive CD4+ T lymphocyte clones (TCC), specific for the immunodominant MBP epitope (amino acids 83-99), contain PDE3 and PDE4, two PDEs that exhibit a high affinity for cAMP. Amplification of TCC mRNA by reverse transcription-PCR indicated that TCC PDE3 mRNA was of the PDE3B, not PDE3A, subtype. Different TCC contained different proportions of PDE3 and PDE4, and their activities increased during Ag (MBP) stimulation. Specific PDE3 (cilostamide) and PDE4 (rolipram) inhibitors suppressed [3H]thymidine incorporation in TCC. Since it is believed that many autoimmune diseases are at least partially mediated by autoreactive CD4+ T cells, these observations may have important implications not only for the treatment of multiple sclerosis but also for other autoimmune diseases.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1997

Immunodominance of a low-affinity major histocompatibility complex-binding myelin basic protein epitope (residues 111-129) in HLA-DR4 (B1*0401) subjects is associated with a restricted T cell receptor repertoire.

The pathogenesis of multiple sclerosis (MS) is currently ascribed in part to a T cell-mediated process targeting myelin components. The T cell response to one candidate autoantigen, myelin basic protein (MBP), in the context of HLA-DR15Dw2, has been previously studied in detail. However, the characteristics of cellular immunity in the context of other MS-associated HLA-DR haplotypes are scarcely known. MBP-specific T cell lines (TCL) were generated from HLA-DR4 (B1*0401)-positive MS subjects. Out of 275 MBP-specific TCL, 178 (64. 7%) specifically recognized region MBP(111-129), predominantly in the context of DRB1*0401. The major T cell epitope for MBP recognition corresponded to residues MBP(116-123). These TCL expressed disparate profiles of cytokine secretion and cytotoxicity. T cell receptor analysis, on the other hand, revealed a strikingly limited heterogeneity of rearrangements. In contrast to MBP(81-99), which binds with high affinity to HLA-DR15 and is recognized by a diverse T cell repertoire, MBP(111-129) binds weakly to DRB1*0401, suggesting that only high affinity T cell receptors might be able to efficiently engage such unstable MHC/peptide complexes, thus accounting for the T cell receptor restriction we observed. This study provides new insight about MBP recognition and proposes an alternative mechanism for immunodominance of self-antigen T cell epitopes in humans.

Adult↗
PubMed1997

Identification of high potency microbial and self ligands for a human autoreactive class II-restricted T cell clone.

CD4+ class II-restricted T cells specific for self antigens are thought to be involved in the pathogenesis of most human autoimmune diseases and molecular mimicry between foreign and self ligands has been implicated as a possible mechanism for their activation. In this report we introduce combinatorial peptide libraries as a powerful tool to identify cross-reactive ligands for these T cells. The antigen recognition of a CD4+ T cell clone (TCC) specific for myelin basic protein peptide (MBP) (86-96) was dissected by the response to a set of 220 11-mer peptide sublibraries. Based on the results obtained with the libraries for each position of the antigen, artificial peptides were found that induced proliferative responses at much lower concentrations than MBP(86-96). In addition stimulatory ligands derived from protein sequences of self and microbial proteins were identified, some of them even more potent agonists than MBP(86-96). These results indicate that: (a) for at least some autoreactive CD4+ T cells antigen recognition is highly degenerate; (b) the autoantigen used to establish the TCC represents only a suboptimal ligand for the TCC; (c) a completely random and unbiased approach such as combinatorial peptide libraries can decrypt the spectrum of stimulatory ligands for a T cell receptor (TCR).

Amino Acid Sequence↗
PubMed1997

Modifications of peptide ligands enhancing T cell responsiveness imply large numbers of stimulatory ligands for autoreactive T cells.

In this report, we demonstrate for autoreactive T cell clones that single amino acid modifications of the antigenic ligand can result in not only abrogated, decreased, or unmodified, but also increased, T cell responsiveness (superagonist ligands). We further studied the effects of combinations of multiple substitutions with different effects in single peptides. Experiments with peptides carrying multiple amino acid exchanges revealed that the final outcome of TCR ligation by a given ligand is the integration of negative, neutral, and positive effects of each single residue. In addition, the introduction of superagonist substitutions together with nonconservative modifications of primary and secondary TCR contacts resulted in stimulatory ligands. These findings indicate that: 1) the specificity of a single TCR is highly degenerate; 2) ligands exist for autoreactive T cells that have higher agonist activity than the autoantigen itself; 3) the rules to search for cross-reactive epitopes in autoimmunity should take into account that amino acids at certain positions within an antigenic peptide may exert superagonist activity and compensate for the negative effects of residues at other positions that would otherwise not be tolerated.

Amino Acid Sequence↗
PubMed1997

Human autoreactive CD4+ T cell clones use perforin- or Fas/Fas ligand-mediated pathways for target cell lysis.

It is well established that target cell lysis by MHC class I-restricted CD8+ T cells is an important defense mechanism during infections with intracellular pathogens or against tumor targets. On the other hand, little is known about the physiologic role and the mechanisms of cytotoxicity of CD4+ MHC class II-restricted T cells. We have recently demonstrated that human autoreactive CD4+ T cells specific for one candidate autoantigen of multiple sclerosis, myelin basic protein, can mediate cytotoxicity. In the present report, we analyze the cytolytic mechanisms employed by these cells. We show that individual T cell clones, regardless of their cytokine phenotype, can be noncytotoxic or lyse target cells via either perforin- or Fas/Fas ligand-mediated cytotoxicity.

Autoantigens↗
PubMed1997

Analysis of a 35.6 kb region on the right arm of Saccharomyces cerevisiae chromosome XV.

We report the sequence of a 35,600 bp fragment covering the PET123 region on the right arm of chromosome XV from Saccharomyces cerevisiae. This region contains 19 possible open reading frames (ORFs) of which 16 are non-overlapping ORFs. Eight ORFs correspond to the SPP2, SMP3, PDR5, NFI1, PUP1, PET123 and MTR10 loci, described previously. Two ORFs correspond to yeast homologues of genes from other organisms: O3530 is a member of the large ribosomal subunit protein L13 family and O3560 (SME1 gene) is a 94-codon ORF and is a homologue of the mammalian SmE spliceosomal core protein. Three ORFs (O3513, O3521, O3548) present significant similarities to proteins of unknown function and three ORFs (O3510, O3536, O3545) lack homology to sequences within the databases screened.

Amino Acid Sequence↗
PubMed1997

HLA-DR-restricted presentation of purified myelin basic protein is independent of intracellular processing.

Antigen presentation to CD4+ T cells involves intracellular antigen processing and loading of peptides onto newly synthesized major histocompatibility complex (MHC)-class II molecules. Some antigens, such as the lipid-bound, native form of myelin basic protein (LB-MBP) can also be presented by recycling of cell surface MHC class II molecules. The data reported here demonstrate that a preparation of highly purified, delipidated MBP (HP-MBP) follows yet another presentation pathway. Similar to LB-MBP, presentation of HP-MBP to HLA-DR1-restricted T cells was independent of HLA-DM, of newly synthesized proteins, and of invariant chain expression. However, in contrast to LB-MBP, presentation of HP-MBP was also independent of internalization of surface HLA-DR molecules. The different requirements for the presentation of the two molecular forms of MBP were further confirmed by use of the protease inhibitor E64: presentation of LB-MBP but not of HP-MBP was inhibited after treatment of target cells with E64. Furthermore, intact HP-MPB bound to isolated HLA-DR molecules in vitro with an association rate that was considerably faster than that of short peptides. These results show that presentation of HP-MBP is independent of intracellular processing and suggest that it may be presented to T cells by direct binding to surface HLA-DR molecules.

Antigen Presentation↗
PubMed1997

Tumor-associated proteolytic factors uPA and PAI-1 in endometrial carcinoma.

The levels of plasminogen activator urokinase (uPA) and of its inhibitor (PAI-1) were measured by use of ELISA in the cytosol of tissue homogenates obtained from endometrial carcinomas and the marginal, tumor-free endometrium of postmenopausal patients (n = 64). Significantly higher median levels of uPA and PAI-1 were found in malignant endometrium (uPA 1.89 ng/mg, PAI-1 3.04 ng/mg) compared to tumor-free endometrium (uPA 0.84 ng/mg, PAI-1 1.01 ng/mg). Concerning uPA, no significant differences were found in dependence on histomorphological prognostic factors (staging, grading), but the median level of PAI-1 was significantly higher in G2/G3 carcinomas compared to G1 tumors (5.08 ng/mg vs 2.19 ng/mg). Because of the good prognosis of operated patients with endometrial carcinomas, the prognostic value of uPA and PAI-1 can only be decided by a larger number of patients and a long observation time.

Aged↗
PubMed1997

Genetics of multiple sclerosis--how could disease-associated HLA-types contribute to pathogenesis?

Multiple sclerosis is a chronic demyelinating disease of the central nervous system in young adults. It is considered a T cell-mediated autoimmune disease which is probably triggered by exogenous events, e.g. infectious agents, in susceptible individuals. Population, family and twin studies indicate that genetic factors and most likely several genes are associated with disease, but it is clear from the concordance rates of identical twins (25-30%) that genetic background as well as exogenous or somatic events are required to develop disease. Among many candidate genes which have been analyzed during recent years, the strongest association was shown for genes of the HLA-class II complex, in particular HLA-DR15 Dw2 and -DQw6. At present, it is not clear how the expression of a particular HLA-class II gene translates into susceptibility to develop an organ-specific autoimmune disease. Potential explanations how this could occur will be discussed.

Adult↗
PubMed1997

Immunological aspects of experimental allergic encephalomyelitis and multiple sclerosis and their application for new therapeutic strategies.

The etiology of multiple sclerosis (MS), a demyelinating disorder of the central nervous system (CNS), is not yet known. Immunological, clinical and pathological studies suggest, however, that T lymphocytes directed against myelin antigens are involved in the pathogenesis of MS. The examination of an experimental animal model for MS, experimental allergic encephalomyelitis (EAE), demonstrated that myelin basic protein-(MBP) or proteolipidprotein-(PLP) specific T cells mediate the destruction of CNS myelin. In recent years, elegant studies in EAE showed that encephalitogenic T cells recognize short peptides of MBP or PLP in the context of MHC/HLA-class II molecules, express a restricted number of T cell receptor (TCR) molecules and secrete interferon-gamma and tumor necrosis factor-alpha/beta. Understanding the pathogenetic steps of demyelination at the molecular level led to highly specific immunotherapies of EAE targeting each individual molecule. MBP- and PLP-specific T cells with similar properties could also be isolated from MS patients and control individuals. Due to their heterogeneity in terms of specificity, function and TCR usage, it was difficult, however, to draw definite conclusions from these results, so far. The recent approval of interferon-beta, a cytokine that antagonizes a number of the effects of interferon-gamma, for the treatment of MS has raised great interest in examining novel strategies for immunotherapies in MS. The basic concepts as well as the current candidates for such new immunotherapies will be outlined in this brief article.

Animals↗
PubMed1997

[Multiple sclerosis. New therapeutic strategies in the experimental stage].

Extensive research in the field of multiple sclerosis (MS) has lead to a preliminary pathogenetic concept without solving the etiopathogenesis. According to animal experiments and human in vitro studies MS is a T cell-mediated autoimmune disease. Experimental therapeutical strategies are aiming at the inhibition of T cell activation, transmigration through the blood brain barrier and local inflammation and demyelination. Several substances are already being tested in clinical studies with magnetic resonance imaging as a tool to quantify inflammatory lesions and to shorten the study course. This review will give a summary about actual experimental therapies resulting from the current pathogenetic concept.

Animals↗
PubMed1997

Intragastric distribution and gastric emptying assessed by three-dimensional ultrasonography.

BACKGROUND & AIMS: Three-dimensional (3D) ultrasound imaging of the total stomach volume has not yet been achieved. The aim of this study was to investigate whether a magnetic position sensor system for acquisition of 3D ultrasonograms could be used to determine gastric emptying rates and intragastric distribution. METHODS: A system for position and orientation measurement was interfaced to an ultrasound scanner. In vitro accuracy was evaluated by scanning a porcine stomach. Fourteen volunteers, with a median age of 35 years, were scanned fasting and postcibally by two-dimensional (2D) and 3D ultrasound after ingesting a 500-mL soup meal. RESULTS: This 3D system yielded a strong correlation (r = 0.997) between true and estimated volumes in vitro. The limits of agreement were -9.1:70.1 mL in the volume range 1200-1900 mL. The intersubject variability of the total gastric volumes ranged from 12.5% to 46.0%, less than for antral area variability. The average half-emptying time was 22.1 +/- 3.8 minutes. Intragastric distribution of the meal, expressed as proximal distal volume, varied on average from 3.6 +/- 2.1 (5 minutes postpradially) to 2.7 +/- 1.9 (30 minutes postprandially). CONCLUSIONS: This 3D ultrasound system using magnetic scanhead tracking showed excellent in vitro accuracy, calculated gastric emptying rates more precisely than by 2D ultrasound, and enabled estimation of intragastric distribution of a soup meal.

Adult↗
PubMed1997

Incidence of premature birth and neonatal respiratory disease in infants of HIV-positive mothers. The Pediatric Pulmonary and Cardiovascular Complications of Vertically Transmitted Human Immunodeficiency Virus Infection Study Group.

OBJECTIVE: We sought to determine the prematurity rate in infants of HIV-positive mothers and to characterize the incidence and severity of neonatal respiratory disease in this population. STUDY DESIGN: From 1990 to 1994, 600 live-born infants of HIV-infected mothers were enrolled prenatally (73%) or postnatally (27%) from five U.S. centers. Logistic regression was used to determine the association of HIV status in the infant with prematurity (< or = 37 weeks), low birth weight (< or = 2.5 kg), and very low birth weight (< or = 1.5 kg) rates. The incidence of respiratory distress syndrome (RDS), bronchopulmonary dysplasia, meconium aspiration syndrome, and neonatal pneumonia was compared with anticipated rates for gestational age and birth weight. RESULTS: Very high rates of prematurity (19%), low birth weight (18.3%), and very low birth weight (3.3%) were found in the infants of HIV-positive mothers; and HIV infection in the infant was associated with younger gestational age. The overall incidence of RDS was 3% (17/600), which coincided with the anticipated rate, after adjusting for prematurity and birth weight. Only five infants (all < or = 1.5 kg) had bronchopulmonary dysplasia, and none required assisted ventilation beyond 14 days. Three term infants had mild meconium aspiration syndrome, and there were no cases of documented neonatal pneumonia. CONCLUSION: Infants born to HIV-positive mothers exhibited high prematurity and low birth weight rates, and the odds of prematurity were higher in infants who were infected with HIV. Despite the high incidence of prematurity and perinatal risk of this population, incidence and severity of neonatal respiratory disease were not higher than would be expected from available neonatal data in populations not exposed to HIV.

Adult↗
PubMed1997

Autosomal recessive inheritance of affective disorders in families of responders to lithium prophylaxis?

In this paper we report the results of a study of the mode of inheritance in affective disorders responsive to lithium. Earlier we described a series of 71 families in which the genetic transmission was compatible with a single-gene model. We have now carried out an independent replication study on 25 newly recruited families in a different geographical location. The autosomal recessive model from our original study could not be rejected with the new data. In a subsequent analysis of the pooled sample of 96 families, a recessive model with a common predisposing allele (q = 0.16) and sex-specific penetrance (0.35 in males, 0.66 in females) fitted the data best. On the other hand, X-chromosome and polygenic models could be rejected. The finding of a major-gene effect represents a specific hypothesis that can be tested by molecular genetic techniques.

Adult↗
PubMed1997

2',3'-cyclic nucleotide 3'-phosphodiesterase: a novel candidate autoantigen in demyelinating diseases.

Autoaggressive T-cells specific for myelin proteins like proteolipid protein (PLP) and myelin basic protein (MBP) are thought to play a major role in the pathogenesis of demyelinating diseases of the central nervous system (CNS). 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) is the third most abundant myelin protein in the CNS. Due to lack of supply with enough CNPase of sufficient purity its immunologic properties have not been studied yet. We subcloned a human CNPase cDNA and expressed human recombinant CNPase (rh-CNPase) in E. coli. Purification of the protein was achieved by Ni(2+)-chelating chromatography. Furthermore we describe for the first time several rh-CNPase specific T-cell lines from a multiple sclerosis patient and a healthy control.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗
PubMed1997

In vitro modulation of human, autoreactive MBP-specific CD4 + T-cell clones by cyclosporin A.

Cyclosporin A (CsA) is a potent immunosuppressant affecting many components of cellular and humoral immunity. Its main action probably results from inhibition of T-lymphocyte activation and interference with secretion of cytokines like IL-2, IL-4, IFN-gamma and TNF-alpha. Correspondingly, CsA has beneficial effects on the course of several autoimmune diseases thought to be mediated by T-lymphocytes, including a mild effect on multiple sclerosis. We exposed CD4 + cytotoxic T-lymphocytes specific for myelin basic protein, a putative target autoantigen in MS, to CsA in vitro, and determined the drug's effects on proliferation, expression of high affinity IL-2R, secretion of the proinflammatory cytokines IFN-gamma and TNF-alpha as well as on the secretion of the chemokines MIP-1 alpha and MIP-1 beta. In all instances, we observed a partial to complete inhibition. In contrast, the response of activated cells to IL-2 was resistant to CsA. Our observations are in line with results obtained in different experimental systems. The discrepancy between the profound inhibition of T-cells and the modest therapeutic effects on MS is discussed.

CD4-Positive T-Lymphocytes↗