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H Offner

Publications and source records attributed to H Offner.

At least 127 records · Page 7Linked to original sources
PubMed1989

PHA activation of encephalitogenic T cells: in vitro line selection overcomes splenic suppression.

In this study we compared myelin basic protein (MBP) and phytohemagglutinin (PHA) for their ability to induce proliferation and experimental autoimmune encephalomyelitis (EAE) transfer activity in mixed cell cultures obtained from spleen and lymph nodes versus highly selected MBP-specific T cell lines and clones. Established MBP-specific cells derived initially from immune lymph nodes attained both proliferative and EAE-transfer activities after in vitro activation with either MBP or PHA. In contrast, PHA was unable to induce immune spleen cells to transfer EAE, in spite of its potent mitogenic activity. On the basis of these results, we evaluated the in vitro proliferation and differentiation responses of MBP-specific T cells during the line selection process using cells derived from both immune lymph node and immune spleen. During the initial selection process with MBP, proliferation of MBP-specific T cell precursors from immunized spleen populations was reduced relative to lymph node cells. After antigen-dependent selection the encephalitogenic cells from either organ exhibited identical in vitro response characteristics. Freshly isolated immune spleen cells were potent suppressors of MBP-specific T cell proliferation suggesting that the in vitro differences between the two organs was due to splenic suppression of the encephalitogenic cells.

Animals↗
PubMed1989

T cell determinants of myelin basic protein include a unique encephalitogenic I-E-restricted epitope for Lewis rats.

The major encephalitogenic epitope for Lewis rats is the 72-89 sequence of guinea pig basic protein (GP-BP) or rat basic protein (Rt-BP). T cells responsive to this epitope are I-A restricted and preferentially express the V alpha 2:V beta 8 gene combination in their TCR. In this work, we describe for the first time the delayed appearance of T cells specific for additional discrete determinant of BP, the nonencephalitogenic 55-68 sequence of GP-BP restricted by I-A, and the encephalitogenic 87-99 sequence of Rt-BP restricted by I-E. The TCR V alpha 2:V beta 8 gene combination was expressed by both encephalitogenic GP-BP S72-89 and Rt-BP S87-99 T cell specificities but not by GP-BP 44-68-specific T cells. This is the first demonstration of I-E-restricted encephalitogenic T cells in Lewis rats and supports the conclusion that the I-E class II locus is involved in autoimmune diseases.

Amino Acid Sequence↗
PubMed1989

Both rat and mouse T cell receptors specific for the encephalitogenic determinant of myelin basic protein use similar V alpha and V beta chain genes even though the major histocompatibility complex and encephalitogenic determinants being recognized are different.

Prospects for specific immune intervention in T cell-mediated autoimmune disease via anti-idiotypic regulation depend on the degree of diversity of the responder cell antigen receptor repertoire. A highly heterogenous response against self epitopes offers little chance for such regulation. We report here that the Lewis rat autoimmune disease experimental allergic encephalomyelitis, generally considered to be a model of human multiple sclerosis, is caused by T cells that use a limited set of TCR V genes. We have cloned the rat TCR alpha and beta chain cDNAs from the Lewis rat x mouse T cell hybridoma 510, which retains the rat specificity for the encephalitogenic determinant of myelin basic protein (MBP). Using Northern blot analysis of T cell RNA with the cloned V region probes, we have found a specific, and near perfect, correlation between expression of TCR message hybridizing to the V alpha 510 and VB510 probes and specificity for the encephalitogenic determinant of MBP in both T cell hybridomas and encephalitogenic T cell clones. This restricted V gene usage provides a basis for observed idiotypic regulation of auto-reactive T cells, and possible therapy for autoimmune disease. A curious and unexplained observation is that the Lewis rat V alpha/V beta combination that dominates the encephalitogenic response to the 68-88 peptide of MBP is precisely the same V alpha/V beta combination used by the B10.PL mouse response to the encephalitogenic response to the 1-9 peptide of MBP.

Amino Acid Sequence↗
PubMed1989

Selection of encephalitogenic rat T-lymphocyte clones recognizing an immunodominant epitope on myelin basic protein.

Using the soft agar-cloning technique, we isolated 13 T-cell clones from guinea pig basic protein (GP-BP)-specific T-cell lines derived from Lewis rats. The clonal frequency was approximately 2.5 x 10(-5). Each of these clones had a similar but not identical pattern of response to a battery of synthetic peptides representing overlapping epitopes in the encephalitogenic region for Lewis rats (69-89 sequence). All clones responded to the minimal encephalitogenic sequence (residues 72-84) restricted by I-A but not I-E molecules, and all transferred clinical experimental autoimmune encephalomyelitis (EAE) and delayed-type hypersensitivity (DTH) reaction to naive rats. Phenotypically, the clones were W3/13+ (total T), W3/25+ (T helper), and OX-22+ (DTH associated). This report demonstrates for the first time the applicability of the soft agar-cloning technique for obtaining encephalitogenic T-cell clones. The exclusive recovery of 72-84-specific T-cell clones after only two rounds of stimulation with GP-BP indicates the immunodominance of this epitope and the power of the line selection technique for obtaining encephalitogenic T-cell specificities.

Agar↗
PubMed1989

Human T lymphocyte response to myelin basic protein: selection of T lymphocyte lines from MBP-responsive donors.

The goal of this study was to delineate the importance of blood T lymphocyte responses to several myelin basic protein (MBP) preparations in the ultimate selection of MBP-specific T lymphocyte lines. Proliferation responses to human myelin basic protein (MBP) were assessed in blood samples from 27 multiple sclerosis (MS) patients, 20 patients with other neurologic diseases (OND), and 26 normal subjects, using five MBP preparations with different histories and electrophoretic characteristics to enhance the spectrum of epitopes represented. Substantial variations were observed in the ability of different MBP preparations to induce blood T cell proliferation in a given donor. However, four out of five of the MBPs induced modest but significant proliferation in the MS study population relative to normal individuals, with intermediate responses occurring in OND patients. Positive responses occurred more frequently in MS patients (78%) than in normal donors (31%), and were an important prerequisite for the successful selection of MBP-specific T cell lines.

Adult↗
PubMed1989

Response of human T lymphocyte lines to myelin basic protein: association of dominant epitopes with HLA class II restriction molecules.

In animals, the selection in vitro of T cell lines to myelin basic protein (MBP) can define immunodominant and encephalitogenic epitopes which are preferentially associated with class II major histocompatibility (MHC) molecules. These principles were used to evaluate the specificity and MHC restriction of 14 human MBP-reactive T cell lines selected from normal individuals and patients with multiple sclerosis (MS) and other neurological diseases (OND). The four normal T cell lines recognized single, separate immunodominant MBP epitopes which were restricted by MHC molecules from the DR or in one case the DP class II locus. In contrast, the MS and OND T cell lines recognized multiple MBP epitopes, each in association with a discrete class II MHC molecule from the DR or DQ locus. Overall, HLA-DR molecules were used preferentially to associate with epitopes on human MBP, restricting 26/33 responses. As predicted from animal studies, T cells from genetically disparate individuals responded to different immunodominant epitopes on human MBP in association with distinct MHC class II molecules. HLA-DR2, which is overrepresented in MS patients, possessed an unusual capacity to restrict all eight epitopes identified on MBP in this study. These data provide the first evidence of genetically restricted human T cell recognition of potentially encephalitogenic epitopes of MBP.

Adult↗
PubMed1989

Defective T helper cell epitope responsible for the failure of region 69-84 of the human myelin basic protein to induce experimental allergic encephalomyelitis in the Lewis rat.

Studies from our laboratory have shown that region 69-84 (synthetic peptide S49S) of myelin basic protein (MBP) defines an encephalitogenic sequence for experimental allergic encephalomyelitis (EAE) in Lewis rats. The most potent EAE inducers are the guinea pig MBP (Gp-MBP) and region 69-84, known as synthetic peptide Gp-S49S: (See text: formula). Human (H-MBP) was considerably less potent than Gp-MBP, and region 69-84 (H-S49S) of H-MBP did not induce hind leg paralysis or any histological signs of EAE. Since the development of EAE requires the expression of specific T and B cell epitopes, sequence analysis of H-S49S and Gp-S49S revealed phylogenetic variations in the H-S49S sequence, characterized by positions 77 and 78, and substitution of Ser with Thr at position 80: (See text: formula). Like Gp-S49S, peptide H-S49S induced the formation of antibodies with specificities directed against the C-terminal of the H-S49S, Gp-S49S, and homologous sequences. In contrast to Gp-S49S, neither II-S49S nor shorter peptides induced clonal T cell expansion when either of the peptides was added to encephalitogenic T cell clone D in culture. Clone D, which expresses T helper phenotype, was selected from encephalitogenic peptide-primed Lewis rats. The results of the study show that the failure of H-S49S to induce EAE is related to sequence alterations in the T helper cell epitope but not in the B cell epitope located in the N- and C-terminal portions of the S49S sequence, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1989

Specific ganglioside binding to receptor sites on T lymphocytes that couple to ganglioside-induced decrease of CD4 expression.

The binding of different gangliosides to rat T-helper lymphocytes was characterized under conditions that decrease CD4 expression on different mammalian T-helper lymphocytes. Saturation binding by monosialylated [3H]-GM1 to rat T-lymphocytes was time- and temperature-dependent, had a dissociation constant (KD) of 2.2 +/- 1.4 microM and a binding capacity near 2 fmoles/cell. Competitive inhibition of [3H]-GM1 binding demonstrated a structural-activity related to the number of unconstrained sialic acid moieties on GM1-congeneric gangliosides. A comparison between the results of these binding studies and ganglioside-induced decrease of CD4 expression demonstrated that every aspect of [3H]-GM1 binding concurs with ganglioside modulation of CD4 expression. It is concluded that the specific decrease of CD4 expression induced by pretreatment with gangliosides involves the initial process of ganglioside binding to specific sites on CD4+ T-helper lymphocytes.

Animals↗
PubMed1989

T cell lines selected with synthetic peptides are highly encephalitogenic in SJL/J mice.

T cell lines were selected from basic protein (BP)-immunized SJL/J mice using synthetic peptides encompassing the major SJL/J encephalitogenic determinant. Synthetic peptide-derived T cell lines proliferated in response to BP, the 89-169 peptidase fragment of BP and the synthetic peptides, pM87-99, pM90-99 and pM91-99. These lines transferred a demyelinating and chronic relapsing form of experimental autoimmune encephalomyelitis (EAE) into naive mice, and EAE induced by synthetic peptide-derived lines was more severe than that induced by whole BP-derived lines. This study demonstrates that T cell lines selected with synthetic peptides are encephalitogenic in SJL/J mice and offers an improved means for selecting SJL/J encephalitogenic T cell lines.

Animals↗
PubMed1989

Murine monoclonal anti-myelin basic protein (MBP) antibodies inhibit proliferation and cytotoxicity of MBP-specific human T cell clones.

Myelin basic protein (MBP)-specific T cell clones, isolated from two patients with multiple sclerosis, expressed the CD4+ phenotype and induced MBP-dependent cytolysis of autologous Epstein-Barr virus (EBV)-transformed B cells. The proliferation and cytolytic activity of the T cell clones were inhibited by four of a panel of five murine monoclonal anti-MBP antibodies in a dose-dependent manner. An isotype-matched antibody with an irrelevant specificity did not have such an effect. These MBP-specific monoclonal antibodies did not block phytohemagglutinin-induced T cell proliferation or allospecific cytotoxicity. These results suggest that some antibodies directed at the autoantigen MBP may play a regulatory role in T cell activation, rather than a pathogenic role, for which there is currently little supporting evidence.

Autoantibodies↗
PubMed1989

Lymphocyte vaccination against experimental autoimmune encephalomyelitis: evaluation of vaccination protocols.

Repeated vaccination with encephalitogenic but not other T cell lines could effect marked resistance to 'active' experimental autoimmune encephalomyelitis (EAE) induced by injection of GP-BP in adjuvant. Partial resistance to active EAE was observed in rats recovered from 'passive' line-mediated EAE and in rats vaccinated with T cells attenuated by irradiation or ganglioside treatment. However, no resistance was observed in animals given low doses of activated encephalitogenic T cells. Treatment with hydrostatic pressure alone was found to be ineffective as a means of attenuation, and vaccination with pressure-treated encephalitogenic T cells actually induced mild signs of EAE. However, vaccination with cells that were first pressure treated and then irradiated prevented both clinical and histologic signs of active EAE. In contrast, protection against passive EAE appeared to be clonotypic. Lymphocyte vaccination induced delayed type hypersensitivity (DTH) reactions against autologous T cells, mostly to shared antigens, demonstrating the immunogenicity of multiple antigens on the vaccinating cells.

Animals↗
PubMed1988

Encephalitogenic T cell clones with variant receptor specificity.

We explored antigenic differences between guinea pig (GP)-basic protein (BP), rat (Rt)-BP, and respective peptides from the encephalitogenic region for Lewis rats by comparing the fine specificity of T lymphocyte lines and clones selected from animals primed with these Ag. Encephalitogenic T cell lines specific for GP-BP or Rt-BP predictably recognized the corresponding 72-89 and to a lesser degree the 72-84 (S55S) amino acid sequence. T cell lines selected from rats primed with GP-S55S responded preferentially to GP-S55S compared to other peptides. A T cell line raised to Rt-S55S, however, initially recognized the S55S and S72-89 peptides but were nearly unresponsive to the intact GP-BP or Rt-BP. T cell clones selected from the Rt-S55S line at that point had two distinct patterns of response: clones that recognized both of the BP and the S55S peptides adoptively transferred delayed-type hypersensitivity and experimental autoimmune encephalomyelitis. These clones also recognized residues 69-81 (S67) but not peptide S75-89. In contrast, T cell clones that responded only to synthetic peptides GP-S55S and Rt-S55S but not to the parent BP adoptively transferred delayed-type hypersensitivity but not disease in Lewis rats. The same clones failed to respond to either the S67 or the S75-89 sequences. These results demonstrate that the encephalitogenic Rt-S55S sequence houses a minimum of two T cell epitopes with differing specificities and functions. One epitope is immuno-dominant and resembles the encephalitogenic region of the intact BP molecule. The second non-encephalitogenic epitope is restricted to the S55S sequences and is not shared by the parent BP, the S67, or the S75-89 sequences. Both types of Rt-S55S-specific clones differ in fine specificity from encephalitogenic clones selected from GP-BP immunized rats, thus indicating that uniformity of T cell recognition of the encephalitogenic epitope is not an absolute condition for T cells to be encephalitogenic.

Amino Acid Sequence↗
PubMed1988

Basic protein-specific T-cell lines that induce experimental autoimmune encephalomyelitis in SJL/J mice: comparison with Lewis rat lines.

Myelin basic protein (BP)-specific T-cell lines were selected from SJL/J mice using techniques to select similar lines from Lewis rats. SJL/J BP-specific T-cell lines were composed of T cells with the helper/inducer phenotype (Lyt 1.2+, 2.2- and L3T4+) and proliferated in response to both the 1-37 and the 89-169 fragments of guinea pig BP. BP-specific T-cell lines transferred delayed-type hypersensitivity (DTH) responses to BP that persisted for over 60 days. Most recipient animals (32/41) developed acute experimental autoimmune encephalomyelitis (EAE), and most survivors (19/24) developed chronic relapsing EAE. Spinal cords of animals during both the acute and the chronic phases of illness contained plaques of demyelination and infiltrates of lymphocytes and macrophages. These findings differed from those of Lewis rat BP-specific lines which respond to a different region of BP, transfer DTH responses that last less than 12 days, and induce acute EAE in which demyelination does not occur.

Animals↗
PubMed1988

The autoreactive T cell population in experimental allergic encephalomyelitis: T cell receptor beta-chain rearrangements.

T cell receptor beta-chain gene rearrangements were examined in myelin basic protein (MBP)-reactive T cell lines and hybridomas from Lewis (Lew) rats. Nearly 75% of the cloned hybrids were specific for the major encephalitogenic determinant residues 68-88 of guinea pig (GP) MBP; three fine specificities could be distinguished. Southern blot analysis of receptor beta-chain genes revealed polyclonality and shared rearrangements not seen in non-68-88-specific clones. Generation of short-term, encephalitogenic Lew T cell lines revealed rearrangements shared with the 68-88-specific hybrids, indicating that the hybrids were representative of the whole antigen-specific population and suggesting restricted V beta gene usage within this polyclonal population.

Animals↗
PubMed1987

Gangliosides induce selective modulation of CD4 from helper T lymphocytes.

The cluster designation (CD)4 molecule is one of several nonpolymorphic T lymphocyte surface proteins that have been implicated in T cell-target cell interactions, and is thought to play an important role in regulating T helper cell function. Previously, we found that gangliosides inhibited the function of rat T helper cell lines, and simultaneously inhibited the expression of the rat CD4 molecule identified by the W3/25 antibody. We have now evaluated the generality and mechanism(s) of ganglioside-induced modulation of CD4 expressed by mouse, rat, and human T helper lymphocytes. Ganglioside pretreatment induced rapid and selective disappearance of the CD4 molecule from T helper cells of all three species. The ganglioside effect was temperature- and dose-dependent, reversible within 24 hr of ganglioside removal, azide-insensitive, and was neutralized completely by 10% serum. CD4 modulation appeared to be a general property of gangliosides since the effect could be induced similarly by highly purified individual gangliosides with varying amounts of sialic acid, or by mixed gangliosides. The activity of gangliosides appeared to require both the lipid and sialated oligosaccharide moieties. Gangliosides did not inactivate antibody function, but prevented binding at the cell surface by 12 different monoclonal antibodies specific for a variety of different CD4 epitopes. Preclearance of CD4 by antibody-mediated capping reduced binding of 3H-GM1 to T helper cells. Labeled GM1 bound to several detergent-extracted and transblotted lymphocyte-associated proteins, but apparently did not bind directly to the CD4 molecule under these conditions. These results indicate that gangliosides induce a profound change in the molecular orientation of CD4 within the T helper cell membrane which renders epitopes on the CD4 molecule inaccessible to antibody. This ganglioside effect represents a novel pathway which may contribute to the understanding of the role of CD4 as a regulatory molecule and as a specific receptor for the acquired immune deficiency syndrome virus.

Animals↗