Search PubMed⌕ Search

Biomedical subjects

H Wekerle

Publications and source records attributed to H Wekerle.

At least 145 records · Page 8Linked to original sources
PubMed1989

Resistance to experimental autoimmune encephalomyelitis induced by neonatal tolerization to myelin basic protein: clonal elimination vs. regulation of autoaggressive lymphocytes.

The target autoantigen of experimental autoimmune encephalomyelitis (EAE), myelin basic protein (MBP), appears late in ontogeny. In the rat MBP is expressed first on days 2-3 post partum, at a development stage, when self tolerance to most other autoantigens has already developed. To shed light on the cellular mechanisms that lead to immunological self tolerance to MBP, we treated neonatal rats with high doses of MBP before ontogenetic appearance of this autoantigen. We found that high doses are required to confer MBP-specific tolerance lasting until the adult life. Neonatally tolerized, adult rats are completely resistant to induction of EAE by injection of MBP in complete Freund's adjuvant (CFA). Upon MBP CFA challenge, these animals develop a limited humoral response to MBP, but are completely unreactive to MBP on the T cell level. The function of antigen-presenting cells is unchanged by neonatal tolerization, and there is no evidence for the induction of suppressive mechanisms. Transfers of large numbers of tolerized lymphocytes to normal hosts fails to interfere with EAE inducibility. Moreover, neonatally tolerized lymphocytes do not reduce MBP reactivity of primed lymph node cells or T line cells in vitro. Finally, neonatally tolerized rats are susceptible to EAE transferred by activated primed lymphocytes or by in vitro-activated MBP-specific T line cells. The apparent deletion of MBP-specific T lymphocytes in neonatally tolerized rats is in striking contrast to the physiological self tolerance to MBP, which is characterized by the presence of MBP-specific clones in the normal immune repertoire.

Animals↗
PubMed1989

Clonotypic chromosomal aberrations in long-term lines of myelin-specific rat T lymphocytes.

A panel of 16 long-term rat T lymphocyte lines and clones were screened for cytogenetical abnormalities using chromosomal banding techniques. All T lines were CD4+, recognizing the relevant antigen in the molecular context of major histocompatibility complex (MHC) class II determinants. With one exception (an ovalbumin-specific line), all lines were specific for myelin proteins, and apart of one BS rat-derived T line and its clones, all lines were selected from the Lewis strain of rat. After in vitro culture of more than 1 year, all lines and clones exhibited subtle but definite chromosomal aberrations, which included deletions, enlargement, translocations and formation of isochromosomes. All lines were near diploid, structural chromosomal changes being more frequent than numerical abnormalities. Each T line investigated had an individual pattern of chromosomal changes. In our analysis, 16 of the 22 different chromosomes had changes in at least one line. Chromosome 9 and the X chromosome appeared to have an enhanced susceptibility of alterations. In two cases, chromosomal markers could be traced through different stages of in vitro culture of the T lines.

Animals↗
PubMed1989

Immunotherapy of experimental autoimmune encephalomyelitis (EAE): differential effect of anti-IL-2 receptor antibody therapy on actively induced and T-line mediated EAE of the Lewis rat.

Treatment of Lewis rats with monoclonal anti-interleukin-2 receptor (IL-2 R) antibody ART-18 is highly efficient in protecting the recipients from T-line transferred experimental autoimmune encephalomyelitis (tEAE) in vivo. In contrast, ART-18 did not affect the development of EAE actively induced (aEAE) by immunization with myelin basic protein (MBP) in complete Freund's adjuvant (CFA). ART-18 caused a slight delay in the development of aEAE only in combination with a subtherapeutic dose of cyclosporine A (Cy-A), but failed to influence duration or severity of clinical signs. The discrepancy in therapeutic efficiency of ART-18 in tEAE and aEAE could be due to a different intensity of IL-2 R-expression on in vitro- and in vivo-activated MBP-specific T cells. Our results therefore caution against a general therapeutic application of anti IL-2 R-directed therapy in all manifestations of T-cell-mediated autoimmunity.

Animals↗
PubMed1989

Autoimmune T lymphocytes in myasthenia gravis. Determination of target epitopes using T lines and recombinant products of the mouse nicotinic acetylcholine receptor gene.

Oligoclonal and cloned T lines from peripheral blood or thymuses of patients with myasthenia gravis (MG) were selected for reactivity against nicotinic acetylcholine receptors (AChR) from Torpedo california, or against a recombinant fusion peptide, X4, representing the extracellular portion of the mouse AChR alpha-chain. All cell lines expressed the CD4 membrane phenotype, and their antigen reactivity was blocked by antibodies against monomorphic HLA DR/DP determinants. Using a panel of fusion proteins of different, overlapping mouse AChR alpha-chain sequences, a major T cell epitope was localized between amino acid positions 85 and 142. This determinant was distinct from the humoral main immunogenic region, which has been identified on the sequence 61-76. The response pattern of uncloned T lines from three patients with different HLA haplotypes suggests, however, that in any one MG patient T lymphocytes may recognize more than one autoantigenic epitope on the AChR alpha-chain, and that the T lymphocyte response profiles vary among individual patients.

Adult↗
PubMed1988

Suppression of experimentally induced autoimmune encephalomyelitis by cytolytic T-T cell interactions.

Down-regulatory phenomena have been described in several experimental models of tissue-specific, T-cell-mediated autoimmunity. For example, resistance to active induction of experimental autoimmune encephalomyelitis (EAE) can be induced by pretreating animals with non-pathogenic inocula of autoantigen or effector cells. Moreover, animals that have recovered from one EAE episode are resistant to subsequent induction of EAE. In some models, resistance to EAE has been transferred with immune cells to naive recipients. These experiments, which were based on transfers of unseparated immune cell populations, are difficult to interpret. Immune suppression circuits are known to be complex and involve various distinct cellular subsets. To further complicate the issue, resistance to EAE can be transferred not only by suppressor cells, but also by encephalitogenic effector cells injected in 'subclinical' doses. We describe now the isolation of homogeneous T lymphocyte lines from the spleens of Lewis rats that had recovered from T-cell-mediated EAE (tEAE) caused by the MBP-specific T cell line S1. These spleen-derived T line cells express the CD8 phenotype and specifically respond to determinants on the inducing S1 line, but not to the autoantigen MBP. Furthermore, the anti-S1 cells selectively lyse the encephalitogenic S1 T line in vitro and efficiently neutralize their encephalitogenic capacity in vivo.

Animals↗
PubMed1988

B-T lymphocyte interactions in experimental autoimmune myasthenia gravis: antigen presentation by rat/mouse hybridoma lines secreting monoclonal antibodies against the nicotinic acetylcholine receptor.

Many, but not all rat/mouse B hybridoma cells, producing monoclonal antibodies against determinants on the nicotinic acetylcholine receptor (AChR) of the electric organ of Torpedo californica, were able to immunologically present antigen to AChR-specific, Ia-compatible CD4+ T lymphocyte lines. Most of the hybridomas presented AChR in a privileged manner, i.e. they present AChR even more effectively than macrophages or dendritic cells. However, in the presentation of antigens other than AChR, they were inferior to macrophages. Moreover, some hybridomas were able to present AChR not only in soluble state, but also in membrane vesicles. Privileged presentation of AChR by hybridomas depended on the reactivity of the secreted immunoglobulins with epitopes of the AChR alpha chain, and on the expression of major histocompatibility complex class II antigens on the hybridoma cell surface. There was, however, no quantitative correlation between the actual AChR presentation capacity of one clone and the density of its surface Ia. Neither fine specificity nor isotype of hybridoma immunoglobulin are critical in determining privileged AChR presentation. We postulate that different hybridomas vary in their ability to take up soluble and particulate antigen, to process and to re-express them on the cellular membrane. This capacity may determine their efficiency to present antigen to T cells.

Animals↗
PubMed1988

T cell epitopes in experimental autoimmune myasthenia gravis of the rat: strain-specific epitopes and cross-reaction between two distinct segments of the alpha chain of the nicotinic acetylcholine receptor (Torpedo californica).

T cell epitopes on the nicotinic acetylcholine receptor (A ChR) of Torpedo californica were analyzed using T cell lines isolated from Lewis, BN, and (Lewis X BN)F1 rats. All lines selected for reactivity against either native or denatured AChR or for 6 selected synthetic peptides of the AChR alpha chain expressed the CD4 membrane phenotype and recognized their antigen in the context of major histocompatibility complex class II determinants. They were tested in vitro for reactivity with each of these antigens. The results indicate that parental Lewis and BN rat T lymphocytes recognize distinct molecular epitopes on the AChR protein, whereas (Lewis X BN)F1 hybrids respond against both sets of epitopes. Two peptides (P10 and P11) which represent distinct amino acid sequences on the putatively extracellular part of the AChR alpha chain, and which share only 4 common amino acids, two of them contiguous, showed an unexpected cross-reactivity in the Lewis rat. T cells selected for either peptide co-recognize the other peptide in vitro. In addition, these cells are responsive against full length AChR. P11, in particular, appears to be a major epitope for Lewis rats immunized with AChR.

Amino Acid Sequence↗
PubMed1988

Regulatory circuits in autoimmunity: recruitment of counter-regulatory CD8+ T cells by encephalitogenic CD4+ T line cells.

In this study, pretreatment of Lewis rats with a syngeneic encephalitogenic T cell line (S1) was found to be able to constantly induce resistance to the subsequent induction of transferred experimental autoimmune encephalomyelitis (tEAE). This treatment was capable of protecting recipient animals for at least 2-4 months. Here we show an enhanced suppressor T(anti-S1) cell activity, which can be readily detected in the lymphoid organs of animals which recovered from S1-induced tEAE, or from rats pretreated with attenuated (irradiated, fixative treated or water-lysed) S1 cells. Anti-S1 cells, which uniformly express the CD8 phenotype, were selectively stimulated to grow and expand into lines by confronting primed lymphoid cells with irradiated S1 cells in culture. The proliferative response of anti-S1 cells was independent of myelin basic protein and antigen-presenting cells, and the responses against unrelated encephalitogenic T cell lines were minimal. It was also found that none of the monoclonal antibodies tested (including CD8 and MHC class I antigen-specific antibodies) was able to block S1/anti-S1 interactions. These cells are functionally suppressive to the proliferation of S1 cells in vitro, are specifically cytolytic directed against the EAE-inducing S1 cells and are able to antagonize encephalitogenic capacity of S1 cells in vivo. In vivo elimination of the CD8+ T subset from Lewis rats, using a combined treatment of thymectomy and OX-8 antibody injection before the initial cell transfer, totally blocked the induction of resistance. Our experiments document that induction of functionally active suppressor T cells is responsible for the induced resistance observed in tEAE.

Animals↗
PubMed1988

Thymus in myasthenia gravis. Isolation of T-lymphocyte lines specific for the nicotinic acetylcholine receptor from thymuses of myasthenic patients.

The thymus is believed to play a central role in the pathogenesis of Myasthenia gravis (MG). According to a previous hypothesis, MG is initiated within the thymus by immunogenic presentation of locally produced nicotinic acetylcholine receptor (AChR) to potentially autoimmune T cells. Data of 10 consecutive MG patients demonstrate two critical features of MG thymuses that support the concept of intrathymic activation of autoreactive, AChR-specific lymphocytes. Morphologically, the thymuses showed lympho-follicular hyperplasia in nine cases and benign thymoma in one case. The paramount feature revealed by immunohistological double marker analyses was the intimate association of myoid cells (antigen producing) with interdigitating reticulum cells (potentially antigen presenting cells), both of which were surrounded by T3+ lymphocytes in thymus medulla. All 10 thymuses contained T lymphocytes reactive with AChR. This was in contrast to the peripheral immune compartment (blood) where in only 3 of 10 patients, significant T cell responses to AChR were observed. AChR-specific T cell lines could be established from 8 of 10 thymuses, all members of the helper/inducer subset as indicated by the expression of markers T3 and T4.

Adolescent↗
PubMed1988

Pathogenesis of myasthenia gravis. Acetylcholine receptor-related antigenic determinants in tumor-free thymuses and thymic epithelial tumors.

The authors describe an immunohistologic study of acetylcholine receptor (AChR)-related antigenic determinants in tumor-free thymuses of myasthenia gravis (MG) patients (13 cases) and nonmyasthenic controls (10 cases) and in thymic epithelial tumors of patients with MG (8 cases) and without MG (6 cases). Monoclonal antibodies (MAbs) to the cytoplasmic part and to the extracellular main immunogenic region (MIR) of the alpha subunit of AChRs were used. Their intrathymic binding sites were defined by double-immunostaining, and compared with alpha-bungarotoxin (alpha-Bgt) labeling demonstrated by fluorescence microscopy. Tumor-free thymuses of MG patients and control patients contained cytoplasmic AChR epitopes and alpha-Bgt binding sites on myoid cells and some epithelial cells. Only myoid cells also expressed extracellular MIR epitopes, suggesting that they bear complete AChRs, and are important targets for the autoimmune attack in tumor-free MG thymus. Evidence that AChR-related antigenic determinants of epithelial cells are also significant for MG is provided by our findings in thymic epithelial tumors. All eight tumors with MG but only two out of six tumors without MG showed cytoplasmic AChR epitopes and alpha-Bgt binding sites on neoplastic epithelial cells. Myoid cells and MIR epitopes did not occur in the neoplasms, but in some tumor-free thymic remnants beside thymomas. It is assumed that nonneoplastic and neoplastic thymic epithelial cells contain only incomplete AChRs or AChR-like molecules. The different expression of AChR epitopes in thymic epithelial tumors and tumor-free thymuses might explain some of the heterogeneous region specificities of anti-AChR antibodies in sera of MG patients with and without thymoma.

Adolescent↗
PubMed1987

Expression of apolipoprotein E by mouse brain astrocytes and its modulation by interferon-gamma.

We have found that mouse brain astrocytes in culture synthesize and secrete apolipoprotein E (ApoE) and that its expression is modulated by interferon-gamma, which influences the functional state of astrocytes. Interferon-gamma, which induces expression of Ia determinants on the surface of the astrocyte, reduced secretion of ApoE while enhancing intracellular expression of ApoE. Our data suggest that the synthesis and secretion phases of the ApoE pathway are regulated independently. It is possible that, by the production of ApoE, astrocytes may perform functions that are usually ascribed to macrophages and that may be important in immune reactivity in the brain.

Animals↗
PubMed1987

Synthesis and cell surface display of class II determinants by long-term propagated rat T line cells.

We have investigated the capacity of the encephalitogenic BS rat T cell line bs 83 and its variant clone bs 83.III.C6 to synthesize and express RT1.B-specific class II molecule subsets defined by monoclonal antibodies (mAb) MRC-OX6 and MRC-OX3. Earlier studies had indicated that mAb MRC-OX6 recognizes three distinct molecular species: an immature oligomeric polypeptide chain complex comprised of the polymorphic subunits alpha, beta and the invariant proteins of the gamma group; a biosynthetic intermediate composed of post-translationally modified alpha, beta and gamma chain (denoted p35) and a fully glycosylated alpha, beta two-chain complex derived from the plasma membrane. MRC-OX3 was shown to recognize a serologically distinct alpha, beta two-chain complex that coexists with the MRC-OX6-specific heterodimer at the cell surface. Here we show that premutant bs 83 cells were unable to synthesize class II molecules of either set. In contrast endogeneous synthesis by mutant cells of MRC-OX6-specific molecules was demonstrated. Unlike control spleen cells variant cells failed to synthesize the mature MRC-OX3-reactive class II subset. Instead a three-polypeptide chain complex comprised of the terminally glycosylated subunits alpha, beta and invariant chain p35 was present at the cell surface. This complex appears to represent the preserved biosynthetic intermediate that failed to release invariant chain p35 upon its transit into the plasma membrane. These latter observations support our notion of gamma chain-induced epitope diversification during post-translational maturation of RT1.B-specific class II molecules.

Animals↗
PubMed1987

Ultrastructural immunocytochemistry of glia cells. Double labeling studies using LR White embedding and colloidal gold.

The introduction of acrylate resins (Lowicryl K4M, LR White) into electronmicroscopic immunocytochemistry applied to embedded tissue (post-embedding method) has improved the localization of antigens because of a satisfactory preservation of both ultrastructure and antigenicity of tissues. Here we describe a method that allows double staining of intracellular and membranous determinants in ultrathin sections of nervous tissue and cultures of peripheral nervous system cells. Ultrathin sections of the rat central nervous system fixed on uncoated grids were stained first for MBP selectively on the one face, then the opposite face was stained for GFAP using monoclonal antibodies and indirect immunogold staining method (IGS). Cultured Schwann cells induced to express major histocompatibility complex (MHC) class II antigens were stained for class II antigens by pre-embedding method then followed by post-embedding IGS for the other intracytoplasmic antigens.

Acrylates↗