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Biomedical subjects

B Fleischer

Publications and source records attributed to B Fleischer.

At least 109 records · Page 6Linked to original sources
PubMed1994

Differential expression of mRNA encoding interleukin-12 p35 and p40 subunits in situ.

Interleukin-12 (IL-12) is a heterodimeric cytokine that plays an important role in the regulation of the immune response. For biological activity the expression of both subunits of IL-12, p35 and p40, is required. Moreover, in the mouse the p40 chain of IL-12 specifically inhibits the effects of the IL-12 heterodimer. In the present study we have analyzed by in situ hybridization the expression of the p35 and p40 mRNA in the spleens of BALB/c and mutant (SCID, nude, beige) mice, unstimulated and after in vivo stimulation with lipopolysaccharide (LPS) and with staphylococcal enterotoxin B (SEB). In unstimulated spleens of BALB/c mice p35 and p40 mRNA were only detectable in a few strongly stained single cells, p35 mRNA was expressed in addition weakly in the B cell areas. After injection of LPS or SEB, p40 mRNA was strongly induced in the T cell areas all over the spleen, whereas expression of p35 mRNA and its distribution pattern did not change. Surprisingly, most of the mRNA for p35 and p40 was localized in different areas of the spleen and was apparently produced by different cells. In macrophage-depleted spleens the increased expression of p40 mRNA in response to LPS was reduced but still detectable, demonstrating that other cells besides macrophages can up-regulate IL-12 p40 mRNA. Nude mice showed a stronger expression of p35 mRNA, SCID mice lacked the weak p35 staining of the B cell areas but showed a strong basal expression of both p35 and p40 mRNA and a focal response to LPS. The pattern of IL-12 mRNA expression in beige mice was the same as in normal mice. These data demonstrate a spatial dissociation of expression of the two chains of IL-12 and are compatible with a regulatory role of the isolated IL-12 p40 chain in vivo. In addition, they indicate that the demonstration of mRNA for both chains of IL-12 in whole tissues or cell mixtures is not necessarily indicative of functional IL-12.

Animals↗
PubMed1994

Macrophages are dispensable for superantigen-mediated stimulation and anergy induction of peripheral T cells in vivo.

Bacterial superantigens provoke T lymphocyte activation by cross-linking the variable part of the T cell receptor (TCR) beta-chain with MHC class II molecules on antigen-presenting cells. Although the molecular mechanisms of this interaction are well characterized, the in vivo accessory cell requirements for this stimulation of T lymphocytes by bacterial superantigens remain unknown. In the present study we have addressed the role of splenic macrophages in the activation of V beta 8+ peripheral T cells by staphylococcal enterotoxin B (SEB) in BALB/c mice. SEB-triggered clonal expansion and subsequent induction of unresponsiveness of both CD4+ and CD8+ T cells were investigated in naive animals, or in mice injected intravenously with dichloromethylene diphosphonate-containing liposomes. Such a treatment resulted in the complete and long-lasting elimination of the splenic macrophage population. Remarkably, however, this complete depletion of peripheral macrophages had only a rather minor effect on the superantigen-induced T cell response in the spleen, and macrophage-depleted animals exhibited overall the same magnitude and kinetics of SEB-mediated T cell activation and anergy-induction as their nondepleted counterparts. Our data thus exclude an essential role of peripheral macrophages or macrophage-secreted cytokines in the systemic T cell activation caused by bacterial superantigens in vivo.

Animals↗
PubMed1994

Human T helper cells reactive with somatic bacterial antigens belong to the Th1 subset.

The aim of this study was to characterize the cytokine secretion patterns of human T helper cells from healthy donors reactive with somatic antigens from various bacteria, the nematode Anisakis and tetanus toxoid. From the peripheral blood of four healthy donors we have established 70 T cell lines reactive with antigens from Yersinia, Salmonella, Morganella, Klebsiella, Serratia, Escherichia, Chlamydia, Shigella, Streptococcus, tetanus toxoid and Anisakis, respectively. Our results show that all T cells reactive with bacteria produce interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha), but no interleukin (IL)-4 and no or very little IL-2 and IL-10 and, thus, belong to the Th1 subset, while T cells reactive with tetanus toxoid or Anisakis belong to the Th0 subset with production of IFN-gamma, IL-2, IL-4, IL-10 and TNF-alpha. In summary, our data further substantiate the concept of a functional diversity of human T helper cells with respect to their cytokine profiles. Furthermore, they indicate that a Th1 cytokine profile is not restricted to intracellular bacteria.

Animals↗
PubMed1994

Major histocompatibility complex class II binding site for streptococcal pyrogenic (erythrogenic) toxin A.

Streptococcal pyrogenic exotoxin A (SPEA) is an important pathogenicity factor of group A streptococci. It is a member of the family of "superantigens" produced by Staphylococcus aureus and Streptococcus pyogenes and its T lymphocyte stimulating activity is involved into the pathogenesis of certain diseases caused by pyogenic streptococci. In this study we have produced and characterized recombinant SPEA molecules in Escherichia coli. These molecules are indistinguishable from natural SPEA in both T cell stimulatory and HLA class II binding activities. Human class II molecules are more efficient than mouse class II molecules in presenting SPEA to T cells. In binding tests to major histocompatibility complex class II-positive cells SPEA competes with staphylococcal enterotoxin B and A but not with toxic shock syndrome toxin-1.

Animals↗
PubMed1994

Superantigens produced by infectious pathogens: molecular mechanism of action and biological significance.

"Superantigens" have in common an extremely potent stimulatory activity for CD4+, CD8+, and some gamma delta+ T lymphocytes. Superantigens use a unique mechanism: they crosslink variable parts of the T cell receptor with MHC class II molecules on accessory or target cells. The interaction site on the T cell receptor is the variable part of the beta-chain (V beta). There are several reasons why these molecules have aroused such tremendous interest in recent years. First, they have provided key information on tolerance mechanisms, both on the deletion of T cells in the thymus and on the induction of peripheral tolerance by anergy and apoptosis. Second, of all polyclonal T cell stimulators they are the ones that most closely mimic the recognition of specific antigen. Finally, they have been recognized as important factors in the pathogenicity of the producing pathogens, inducing shock and immunosuppression. Moreover, it has been postulated that superantigens could be involved in the pathogenesis of certain human diseases.

Enterotoxins↗
PubMed1994

CD26: a surface protease involved in T-cell activation.

CD26 is a proteolytic enzyme (dipeptidylpeptidase IV) with a wide tissue distribution and a unique specificity. Recent developments indicate that CD26 is a multifunctional molecule that may have important functions in the immune system. Here, Bernhard Fleischer reviews the current knowledge of CD26 and discusses the possible functions of this molecule in T lymphocytes.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1994

Role of bacteria-specific T cells in the immunopathogenesis of reactive arthritis.

Reactive arthritis is a usually self-limited sterile inflammation of joints that follows certain bacterial gastrointestinal or urogenital infections. The immunopathogenesis involves CD4+ T cells, which mediate an antigen-specific TH1 response to bacterial constituents within the joint. Properties of the arthritogenic bacteria and the physicochemical characteristics of the bacterial antigens may contribute to the development of reactive arthritis.

Animals↗
PubMed1994

Separation of mitogenic and pyrogenic activities from so-called erythrogenic toxin type B (Streptococcal proteinase).

It is well-established that three types of erythrogenic toxins (ETA, ETB, ETC) are produced by Streptococcus pyogenes (group A streptococci) strains. Culture filtrate concentrates from Streptococcus pyogenes strains T19P (T19, ETA+, ETB+, ETC-), 27337 (T12, B3264, ETA-, ETB+, ETC+), 27252 (T4, ETA-, ETB+, ETC+) and 27195 (T8, ETA-, ETB+, ETC-) were analyzed by preparative isoelectric focusing. These concentrates and the purified erythrogenic toxin type B (ETB) isolated by ion exchange chromatography had mitogenic and pyrogenic activity. Now, it has been found that the mitogenic activity and the pyrogenic activity of this ETB can be separated by preparative isoelectric focusing in Sephadex gels. This means that ETB is not a superantigen as described in literature. The mitogenic and biological activity is caused by traces of ETA (strain T19P), ETC (strains 27252 and 27337) and/or by unknown mitogen(s) (MX, strain 27195) which preferentially stimulate V beta 8+ T cells. The differentiation between ETA (stimulating V beta 12+ but not V beta 8+ or V beta 2+), ETC (stimulating V beta 2+ but not V beta 8+), and MX (stimulating V beta 8+) was done using established leukemic cell lines.

Animals↗
PubMed1994

A superantigen as virulence factor in an acute bacterial infection.

This study addresses the role of a bacterial superantigen as a potential virulence factor during an acute systemic infection. BALB/c mice were intravenously infected with a recombinant Staphylococcus aureus strain capable of producing plasmid-encoded staphylococcal enterotoxin B (SEB) or with the SEB plasmid-deficient parental strain. Infection with SEB-producing bacteria resulted in an initial expansion and subsequent decrease of circulating V beta 8+ T lymphocytes. This numeric decrease was accompanied by a SEB-specific state of hyporesponsiveness of splenic T cells. In parallel with SEB-triggered unresponsiveness of a large proportion of T lymphocytes, a weakening of the overall T cell responsiveness towards the invading bacteria was found. Furthermore, the production of SEB altered the kinetics of bacterial clearance: Animals infected with the SEB-producing variant showed a significantly elevated bacterial burden and could less efficiently clear the bacteria. However, the overall effect of SEB production on the course of bacterial infection was surprisingly weak, suggesting that the superantigen was only a minor virulence factor for the bacterium.

Acute Disease↗
PubMed1994

T cells involved in psoriasis vulgaris belong to the Th1 subset.

Although the pathogenesis of psoriasis vulgaris is still unknown, several characteristics point to an immunologically mediated process. Epidermal psoriatic lesions are characterized by a hyperproliferation of keratinocytes and an infiltration of T lymphocytes and granulocytes. Because the former may be mediated in part by lymphokines secreted by T cells, we have focused our interest on the in vivo and in vitro cytokine secretion patterns of T lymphocytes from psoriatic lesions. In five patients T lymphocytes were obtained from epidermal specimens. The cells were propagated with lectin and irradiated feeder cells and subsequently cloned by limiting dilution. The resulting T-cell clones were phenotypically and functionally characterized. Our data show that the majority of T-cell clones were CD4+ (74%), whereas only 25% were CD8+ and 1% were CD4-/CD8-. Also, we have further investigated the cytokine secretion pattern of T-cell lines or CD4+ T-cell clones, respectively. All cells tested produced interferon-gamma whereas only a minority secreted interleukin (IL)-4. Moreover, these cells produced high amounts of IL-2 but only little or no IL-10 or tumor necrosis factor-alpha. To correlate these data with the in vivo situation, biopsies from psoriatic lesions of five patients were investigated for the presence of the mRNA of IL-4, IL-10, and interferon-gamma using the polymerase chain reaction. In these biopsies only the mRNA for the Th1 cytokine interferon-gamma but not for the Th2 cytokines IL-4 and IL-10 could be detected. Identical experiments were performed to test the in vivo cytokine production of synovial fluid mononuclear cells of two patients with arthropathia psoriatica. Again, only the mRNA for interferon-gamma but not IL-4 could be detected. This indicates that T cells involved in psoriasis exhibit a Th1-like cytokine secretion profile.

Adult↗
PubMed1994

Differential effects of IL-10 on proliferation and cytokine production of human gamma/delta and alpha/beta T cells.

Gamma/delta TCR bearing T lymphocytes represent a T-cell subset whose functional relevance remains unclear. Nevertheless these T cells may play a role in the early immune response against bacteria. Until now the regulatory mechanisms on this response have not been investigated. The study described here evaluated the immunoregulatory effects of Interleukin-10 on gamma/delta and alpha/beta TCR-positive T-cell clones and freshly isolated peripheral-blood mononuclear cells (PBMC). IL-10 has been shown previously to inhibit lectin and antigen-induced proliferation and cytokine production by alpha/beta T cells. The results outlined below show that rhIL-10 strongly inhibits lectin-induced production of IFN-gamma, TNF-alpha, IL-2, and to a lesser degree proliferation and IL-4 production of both T-cell subsets. As IL-10 did not inhibit proliferation but at the same time strongly suppressed cytokine production in various experiments, the hypothesis that it could function as a growth factor for human T cells as has been described for murine thymocytes was tested. The data demonstrate that, although the gamma/delta T-cell clones tested do not produce IL-10 they can use it as a growth factor in combination with IL-2, IL-4 or alone. Furthermore, IL-10 has the same properties on human alpha/beta T-cell clones and PBMC. In summary, it is shown that IL-10 has pleiotropic effects on gamma/delta and alpha/beta TCR+ T cells by inhibiting lectin-induced cytokine production and by acting as a growth factor for these cells alone or in combination with IL-2 or IL-4.

CD3 Complex↗
PubMed1994

Phenotypical analysis and cytokine release of liver-infiltrating and peripheral blood T lymphocytes from patients with chronic hepatitis of different etiology.

Cytokines released by infiltrating T cells may contribute to the hepatic injury in chronic hepatitis. Therefore, we characterized peripheral blood- and liver-infiltrating T cells from patients with chronic hepatitis of different etiology and determined the T cell phenotypes and the cytokine release. Liver tissue and peripheral blood-derived T cells from patients with autoimmune hepatitis and primary biliary cirrhosis predominantly expressed CD4-molecules and the alpha- and beta-chains of the T cell receptor (TCR). In chronic viral hepatitis B and C, liver- and blood-derived T cells were preferentially CD8+ T cells expressing the alpha beta TCR. Mitogenic stimulation with irradiated Daudi lymphoma cells and phytohemagglutinin led to a strong release of interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) and interleukin-2 (IL-2) by T cells in patients with chronic hepatitis and in healthy controls. T cells from patients with primary biliary cirrhosis and some patients with autoimmune hepatitis showed a significantly higher secretion of interleukin-4 (IL-4) and interleukin-10 (IL-10) than T cells from patients with chronic viral hepatitis or healthy controls. Histologic inflammatory activity did not correlate with the amount of cytokines released after mitogenic activation. In conclusion, liver tissue and peripheral blood T cells of patients with autoimmune hepatitis and primary biliary cirrhosis were dominated by CD4+ TCR alpha beta+ T helper/inducer cells, whereas in chronic viral hepatitis an enrichment of CD8+ TCR alpha beta + cytotoxic/suppressor T cells was observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1994

Superantigens.

"Superantigens" is the term for a group of molecules that have in common an extremely potent stimulatory activity for T lymphocytes of several species. They stimulate CD4+, CD8+ and gamma delta + T cells by a unique mechanism: they cross-link variable parts of the T-cell receptor (TCR) with MHC class II molecules on accessory or target cells. The interaction site on the class II molecule and on the TCR is different from the peptide binding site; on the TCR it is the variable part of the beta chain (V beta). The prototype superantigen is the staphylococcal enterotoxin B (SEB), member of a family of genetically related proteins produced by Staphylococcus aureus and Streptococcus pyogenes. These are soluble exotoxins of approximately 27 kd molecular mass. It is intriguing that this molecular mechanism of T-cell stimulation has been independently produced at least three times in evolution. Other pathogens producing superantigens are retroviruses (the Mouse Mammary Tumor Viruses) and a mycoplasma (Mycoplasma arthritidis). Many additional candidate superantigens have been proposed, but in most cases unequivocal evidence for superantigen activity is still missing. There are several reasons why these molecules have aroused such tremendous interest in recent years. First, they have provided key information on tolerance mechanisms, both on the deletion of T cells in the thymus and on the induction of peripheral tolerance by anergy and apoptosis. Second, of all polyclonal T-cell stimulators they are the ones that most closely mimic the recognition of specific antigen. Finally, they have been recognized as important factors in the pathogenicity of the producing pathogens, inducing shock and immunosuppression. Whilst there is evidence that superantigens could be involved in the pathogenesis of certain human diseases, in most cases this is still very preliminary and indirect.

Animals↗
PubMed1994

Bacteria-specific cytotoxic CD8+ T cells: a missing link in the pathogenesis of the HLA-B27-associated spondylarthropathies.

The term seronegative spondylarthropathies is used for an entity of rheumatic syndromes of peripheral joints and the spine (ankylosing spondylitis, reactive arthritis, Reiter's syndrome, arthritis in psoriasis and in inflammatory bowel disease) which are strongly associated with the MHC class I molecule HLA-B27. However, the mechanisms whereby HLA-B27 confers disease susceptibility have so far remained unknown. There is strong evidence that gut inflammation and infection with gram-negative bacteria play a role in the induction of B27-associated disease. HLA-B27, like other MHC class I molecules, physiologically binds antigenic peptides in its binding groove and presents them to CD8+ T lymphocytes. Consequently, if the disease association with HLA-B27 arises from its role as a T-cell restriction element, synovial fluid CD8+ rather than CD4+ T cells should play a prominent pathogenetic role and should be detectable within the affected joints. In this paper, recent studies on bacteria-specific cytotoxic T cells and on peptide binding to HLA-B27 are reviewed. Particular emphasis is laid on the role of HLA-B27 restricted synovial CD8+ T cells with specificity for bacterial antigens or autoantigens. These cytotoxic T cells could provide a missing link in the pathogenesis of the spondylarthropathies and could now serve as tools to identify the critical antigenic epitopes of bacterial and self peptides which are involved in disease induction.

Animals↗
PubMed1994

Superantigens.

Explore the source record for details and available documents.

Animals↗
PubMed1993

Distribution and kinetics of superantigen-induced cytokine gene expression in mouse spleen.

The polyclonal stimulation of T cells by bacterial superantigens is involved in the pathogenesis of the toxic shock syndrome in certain staphylococcal and streptococcal infections. Here we describe the onset and kinetics of superantigen-induced cytokine production in situ in spleens of normal BALB/c mice monitored at the level of cytokine mRNA expression by in situ hybridization. Messenger RNAs for interleukin 2 (IL-2), interferon gamma, and tumor necrosis factors (TNF) alpha and beta were not expressed at detectable levels in spleens of unstimulated animals but became visible already 30 min after intraperitoneal application of 50 micrograms staphylococcal enterotoxin B. All mRNA levels showed peak expression approximately 3 h after injection and a slow decrease up to 24 h after injection. Expression of the mRNAs was restricted to the T cell-dependent area of the periarteriolar lymphatic sheets of the spleen. Interestingly, TNF-alpha mRNA showed a biphasic response, the early appearing mRNA had the same localization as the other mRNAs, whereas after 3 h TNF-alpha mRNA showed a broader distribution indicating a second cell population producing TNF-alpha. The expression of IL-2 and TNF proteins in the serum increased in parallel to the observed mRNA changes with a slight delay. The presence of macrophages was not required for the expression of the cytokine mRNAs in the spleen as the expression was unchanged in macrophage-depleted mice. Only the second phase of TNF-alpha mRNA expression was abrogated in such animals. The expression of all mRNAs was completely suppressed by prior administration of cyclosporin A. These data show that nonphagocytic cells are the essential superantigen-presenting cells in vivo and indicate that at least part of the pathogenetic TNF-alpha is T cell derived.

Animals↗
PubMed1993

HLA-B27-restricted CD8 T cells derived from synovial fluids of patients with reactive arthritis and ankylosing spondylitis.

Ankylosing spondylitis and seronegative spondylarthropathies such as Reiter's syndrome and reactive arthritis are strongly associated with HLA-B27. However, the mechanisms by which HLA-B27 is involved in disease susceptibility and pathogenesis are unknown. If the disease association is a consequence of HLA-B27's physiological function in antigen presentation, the disease should be mediated by cytotoxic T lymphocytes (CTLs) that recognise bacterial or self peptides presented by HLA-B27. Proof of this arthritogenic peptide model requires isolation of B27-restricted CD8 T cells from arthritic joints of patients with spondylarthropathies. An important question is whether "arthritogenic" bacteria such as yersinia or salmonella can generate HLA-B27-restricted bacteria-specific CTLs. We describe such HLA-B27-restricted CTLs. We tested a panel of 354 alpha beta-TCR CD8 T lymphocyte clones (TLCs) that had been derived from the synovial fluid of 4 patients with reactive arthritis and 2 patients with ankylosing spondylitis. In 1 patient with yersinia-induced arthritis, 2 TLCs were identified that killed specifically yersinia-infected B27 target cells. In another patient with salmonella-induced arthritis, 1 B27-restricted CD8 TLC that recognised both salmonella and yersinia was identified. In 5 of the 6 patients autoreactive CTLs were found, 5 of which showed B27-restricted killing of uninfected cell lines. B27-restricted CTLs with specificity for arthritogenic bacteria or autoantigens provide a missing link in the pathogenesis of the HLA-B27-associated spondylarthropathies.

Adult↗