- Email: [email protected]
Activated T cells in patients with SLE
308
Pathogenesis of autoimmunity
Conclusions: Our results suggest a role for the Tin2 cell balance in RA. In view of the Tl cell predominance intra-articularty our results may be explained by selective Tl cell migration into the joint or peripheral suppression of Tl cell activity.
P.5.11.07
T ceils expressing 76 receptors and Fca receptors in Behcet’s disease
F. Fortune ‘, J. Freysdottk ‘, J. Walker2. ’ Unit of Medicine in Relationto Oral Disease, UMDS, Guy’.. Hospital, London, 2Department of Otal Immunology, UMDS, Guys Hospital, London Introduction: The onset of Behcet’s disease is associated with epithelial lesions affecting the oral and genital mucosa and the skin. T cells with y6 receptor and increased secretory IgA concentrations have been credited with playing an important role in surveillance and protection of epithelial surfaces from microbial infections. Indeed, T cells expressing the Fca receptors may upregulate IgA synthesis. Aim: The purpose to this investigation was to examine both resting and activated x6 T cells in patients and controls for Fen receptor expression. Materlab and Methods: Venous blood was collected from Behcet’s patients and healthy controls. The cells were separated by density gradient centdfugation and then by using Nylon wool columns. Oral biopsy specimens were taken from both patients and controls. Both resting and activated cells (using anti-y6 antibody and rll-2) were double immunofluorescent labelled and analysed by flow cytometry in order to determine the proportion of T cells expressing or@ or yS receptor, the proportion of CD4 or CD8 cells expressing class II and the proportion of yS T cells expressing the Fco receptor. The proportion of y& T cells expressing the Fca receptor in the oral biopsies was determined by double immunoenzyme staining. Reeults: Patients with Behcet’s disease had increased proportion of CD8+ yS T cells compared to healthy controls, whereas no difference was observed for CD8+ ~rfi T cells between the patient and control groups. T cells showed increased expression of class II in the patient group compared to the normal group. When the cells were stimulated an increased proportion of T ceils expressed the yS receptor in the patient group compared to the normal group. Furthermore, a higher proportion of stimulated T cells expressed both yS and Fan receptors in the patient group compared to the normal group. This increased proportion of T cells expressing y6 and Fca receptors was confirmed in the oral biopsies. Conduslon: We have shown that increased proportion of resting and activated cells from patients with Behcet’s disease express the y6 receptor and that raised proportion of yS T cells from Behcet’s patients also express the Fen receptor. The activation linked expression of Feel receptor and the y6 T cell receptor on lymphocytes may provide an insight into potential mechanisms for associating the functional activities of y6 T cells with immune reactions that involve IgA.
R5.11.08
Programmed ceil death and its role in pathogenesis of SLE
D. Dim&ova, A. Michailova, D. Martnova, E. Naumova, G. Stefanova. Division of Clinical and Transplantation Immunobg~ Medical Universi& Sofia, Bulgaria Programmed cell death involves an orderly process of morphological disintegration that has been termed apoptosis. Unlike cell degradiation or necrosis, programmed cell death is an active cell suicide process regulated by signals provided by the environment. In general, cells undergoing apoptosis display profound structural changes including a rapid blebbing of the plasma membrane and nuclear disintegration. The nuclear collapse is associated with extensive damage to chromatin and DNA cleavage into oligonucleosomal lenght DNA fragment. The enrichment of mono-and digonucleosomes in the cytoplasm of the apoptotic cells is due to the fact that DNA degradation occurs several hours before plasma membrane breakdown. Apoptosis plays an important role.in the normal immune system development and homeostasis, tumorogenesis, viral infection and autoimmune disorders. To investigate the potential role of apoptosis in SLE pathogenesis, we used flow cytometric analysis of peripheral blood lymphocytes from 15 patients aged 18 to 60 years with different disease activity. The patients were compared with 15 random heatthy individuals. Freshly isolated peripheral blood lymphocytes were stained with Propidium iodide (PI) and Fluorescein-12dUTP. We observed increased number of apoptotic lymphocytes in SLE patients compared with the control group. Approximately 15% of peripheral blood lymphocytes in SLE patients were apoptotic. Our preliminary data support the hypothesis that the production of antinuclear antibody in SLE might be due to the released of intact nuclear antigens (oligonucleosomes) from apoptotic cells.
24 June 1997 - Poster presentations
1P.5.11 .O9 ( Activated T ceils in patlents with SLE A. Michailova, D. Dimitrova, P. Spasova, D. Madnova, E. Naumova, G. Stefanova. Division of Clinical and Tmnsplantation Immunology Medical Universiu Sofia Bulgaria Increased values of activated lymphocytes is one of the prominent features of SLE. There are, however, conflicting data on values of activated cells in SLE patients with different disease activity. Using 2-color flow cvtometric analvsis (FACSCalibur. Becton Dickinson, USA) we investigated C&+89+, CD&25+,’ CDI+HLA-DR+, CD&HlA-DR+ and CD8+38+ cells in 14 patients with SLE with different disease activity. At the time of study 5 patients.were without immunosuppressive treatment, 9 were receiving corticosteroids alone or in combination with Cyclophosphamid. We found in our SLE patients a rising of activated CD69+ T cells, compared with the control values, which was more significant in active disease. Increased values of CDSHLA-DR+ and CD&HLA-DR+ cells were established in active SLE, compared with controls and inactive SLE. Although CD38 antigen is not directly linked to antigen-driven stimulation, elevated orcoortions of CD8+38+ cells were found in active SLE. The values of activated T.celts bearing CD25+ antigen did not differ significantly from the controls. Our data suaoest that for the clinical follow UDof SLE. the measurement of CD25+ and CD69+ activated cells is of limited values. Therefore, it appears that the enumeration of HLA-DR+ T cells could be a practical marker for monitoring the clinical course of SLE.
1P.5.11.10 ] Selective stimulation of type-l or type-2 responses precedes and matches the autoimmune diseases trlggered by the chemicals streptozotocin and Hg’Z R. Albers, C. de Heer, R. Bleumink, W. Seinen, R. Pieters. RlTOXsection Immunotoxiwlog~ Utmcht Universi& Box 80.176.3606 TD Utrecht. The Netherlands _Introduction: Induction of (auto)immune responses by drugs or environmental chemicals requires costimulatory signals in addition to release of (auto)anttgens. Previous studies have indicated that chemicals can induce this costtmulation, and that the type of response elicited is influenced by genotype and relates to susceptibility to the adverse immune effects of the compound. Here, we assessed the modulatory role of the stimulating chemical itself. Materials and Methods: The prototype autoimmunogenic chemicals streptozotocin (STZ) and HgCls that induce distinct adverse immune effects in the same mouse strain, were w-injected with the reporter antigen TNP-OVA into the footpad of BALB/c mice. Next, the type of immune response elicited to the reporter antigen was assessed by (intracellular) flowcytometry, TNP-specific ELISPOT assays, and immuno histochemistry. ResuL: A single injection of STZ induced an increase of IFN-y producing Th (CD4+) and Tc (CD8+) cells in the draining lymph node (data on IL-4 and IL-10 producing cells are pending). Moreover, the total number of CD4+ cells was decreased, whereas CD8+ cells, macroohaoes and aooototic cells were increased in conjunction with enhanced IgG2,‘andlgGsr, production to TNP-OVA. Injection of HgC12on the other hand, reduced the number of IFN-Y producing cells, inducedaccumulation of B cells and of CD4+ and CD8+ T cells, enhanced IgGr and IgE production to TNP-OVA, and primed for secondary IgG, and IgE production as well as for DTH reactions. Conclusion: Together these results indicate that STZ stimulates type-l responses, whereas HgCls enhances mixed type-1 and 2 responses in BALB/c mice. Importantly, these responses stimulated by a single injection of chemical match the (auto)immune effects elicited to unknown (auto) antigens following multiple injections of these compounds, suggesting that induction of selective costimulation by chemicals is an important factor provoking the onset of autoimmunity in susceptible individuals.
P.5.11 .l1
The use of reporter antigens In the popliteai lymph node assay to assess immunomoduiation by chemicals
Ft. Albers, E. van ‘t Erve, A. Broeden, A. van der Pijl, W. Seinen, R. Pieters. RITOX, lmmunotaxicoogy Section, Utrecht Univetsib PO Box 80.176, 3.506 TD Utrecht, The Netherlands Introduction: Various drugs and other low molecular weight chemicals can induce T cell-dependent B cell activation which may cause allergic or autoimmune-like diseases. Because the nature of the relevant (neo-) antigens or hapten-carder complexes are generally not (in case autoimmunity) or not alwavs (in case of allerov) known. we exolored the ootential use of well-defined reporter antigens to d&mine T cell-dependent B’cell activation by chemicals. TNP-ficoll and TNP-OVA were used as reporter antigens because they are recognized by the same TNP-specific B cells. For specific antibody production. however, distinct co-stimulation is required.
Pathogenesis of autoimmunity
Conclusions: Our results suggest a role for the Tin2 cell balance in RA. In view of the Tl cell predominance intra-articularty our results may be explained by selective Tl cell migration into the joint or peripheral suppression of Tl cell activity.
P.5.11.07
T ceils expressing 76 receptors and Fca receptors in Behcet’s disease
F. Fortune ‘, J. Freysdottk ‘, J. Walker2. ’ Unit of Medicine in Relationto Oral Disease, UMDS, Guy’.. Hospital, London, 2Department of Otal Immunology, UMDS, Guys Hospital, London Introduction: The onset of Behcet’s disease is associated with epithelial lesions affecting the oral and genital mucosa and the skin. T cells with y6 receptor and increased secretory IgA concentrations have been credited with playing an important role in surveillance and protection of epithelial surfaces from microbial infections. Indeed, T cells expressing the Fca receptors may upregulate IgA synthesis. Aim: The purpose to this investigation was to examine both resting and activated x6 T cells in patients and controls for Fen receptor expression. Materlab and Methods: Venous blood was collected from Behcet’s patients and healthy controls. The cells were separated by density gradient centdfugation and then by using Nylon wool columns. Oral biopsy specimens were taken from both patients and controls. Both resting and activated cells (using anti-y6 antibody and rll-2) were double immunofluorescent labelled and analysed by flow cytometry in order to determine the proportion of T cells expressing or@ or yS receptor, the proportion of CD4 or CD8 cells expressing class II and the proportion of yS T cells expressing the Fco receptor. The proportion of y& T cells expressing the Fca receptor in the oral biopsies was determined by double immunoenzyme staining. Reeults: Patients with Behcet’s disease had increased proportion of CD8+ yS T cells compared to healthy controls, whereas no difference was observed for CD8+ ~rfi T cells between the patient and control groups. T cells showed increased expression of class II in the patient group compared to the normal group. When the cells were stimulated an increased proportion of T ceils expressed the yS receptor in the patient group compared to the normal group. Furthermore, a higher proportion of stimulated T cells expressed both yS and Fan receptors in the patient group compared to the normal group. This increased proportion of T cells expressing y6 and Fca receptors was confirmed in the oral biopsies. Conduslon: We have shown that increased proportion of resting and activated cells from patients with Behcet’s disease express the y6 receptor and that raised proportion of yS T cells from Behcet’s patients also express the Fen receptor. The activation linked expression of Feel receptor and the y6 T cell receptor on lymphocytes may provide an insight into potential mechanisms for associating the functional activities of y6 T cells with immune reactions that involve IgA.
R5.11.08
Programmed ceil death and its role in pathogenesis of SLE
D. Dim&ova, A. Michailova, D. Martnova, E. Naumova, G. Stefanova. Division of Clinical and Transplantation Immunobg~ Medical Universi& Sofia, Bulgaria Programmed cell death involves an orderly process of morphological disintegration that has been termed apoptosis. Unlike cell degradiation or necrosis, programmed cell death is an active cell suicide process regulated by signals provided by the environment. In general, cells undergoing apoptosis display profound structural changes including a rapid blebbing of the plasma membrane and nuclear disintegration. The nuclear collapse is associated with extensive damage to chromatin and DNA cleavage into oligonucleosomal lenght DNA fragment. The enrichment of mono-and digonucleosomes in the cytoplasm of the apoptotic cells is due to the fact that DNA degradation occurs several hours before plasma membrane breakdown. Apoptosis plays an important role.in the normal immune system development and homeostasis, tumorogenesis, viral infection and autoimmune disorders. To investigate the potential role of apoptosis in SLE pathogenesis, we used flow cytometric analysis of peripheral blood lymphocytes from 15 patients aged 18 to 60 years with different disease activity. The patients were compared with 15 random heatthy individuals. Freshly isolated peripheral blood lymphocytes were stained with Propidium iodide (PI) and Fluorescein-12dUTP. We observed increased number of apoptotic lymphocytes in SLE patients compared with the control group. Approximately 15% of peripheral blood lymphocytes in SLE patients were apoptotic. Our preliminary data support the hypothesis that the production of antinuclear antibody in SLE might be due to the released of intact nuclear antigens (oligonucleosomes) from apoptotic cells.
24 June 1997 - Poster presentations
1P.5.11 .O9 ( Activated T ceils in patlents with SLE A. Michailova, D. Dimitrova, P. Spasova, D. Madnova, E. Naumova, G. Stefanova. Division of Clinical and Tmnsplantation Immunology Medical Universiu Sofia Bulgaria Increased values of activated lymphocytes is one of the prominent features of SLE. There are, however, conflicting data on values of activated cells in SLE patients with different disease activity. Using 2-color flow cvtometric analvsis (FACSCalibur. Becton Dickinson, USA) we investigated C&+89+, CD&25+,’ CDI+HLA-DR+, CD&HlA-DR+ and CD8+38+ cells in 14 patients with SLE with different disease activity. At the time of study 5 patients.were without immunosuppressive treatment, 9 were receiving corticosteroids alone or in combination with Cyclophosphamid. We found in our SLE patients a rising of activated CD69+ T cells, compared with the control values, which was more significant in active disease. Increased values of CDSHLA-DR+ and CD&HLA-DR+ cells were established in active SLE, compared with controls and inactive SLE. Although CD38 antigen is not directly linked to antigen-driven stimulation, elevated orcoortions of CD8+38+ cells were found in active SLE. The values of activated T.celts bearing CD25+ antigen did not differ significantly from the controls. Our data suaoest that for the clinical follow UDof SLE. the measurement of CD25+ and CD69+ activated cells is of limited values. Therefore, it appears that the enumeration of HLA-DR+ T cells could be a practical marker for monitoring the clinical course of SLE.
1P.5.11.10 ] Selective stimulation of type-l or type-2 responses precedes and matches the autoimmune diseases trlggered by the chemicals streptozotocin and Hg’Z R. Albers, C. de Heer, R. Bleumink, W. Seinen, R. Pieters. RlTOXsection Immunotoxiwlog~ Utmcht Universi& Box 80.176.3606 TD Utrecht. The Netherlands _Introduction: Induction of (auto)immune responses by drugs or environmental chemicals requires costimulatory signals in addition to release of (auto)anttgens. Previous studies have indicated that chemicals can induce this costtmulation, and that the type of response elicited is influenced by genotype and relates to susceptibility to the adverse immune effects of the compound. Here, we assessed the modulatory role of the stimulating chemical itself. Materials and Methods: The prototype autoimmunogenic chemicals streptozotocin (STZ) and HgCls that induce distinct adverse immune effects in the same mouse strain, were w-injected with the reporter antigen TNP-OVA into the footpad of BALB/c mice. Next, the type of immune response elicited to the reporter antigen was assessed by (intracellular) flowcytometry, TNP-specific ELISPOT assays, and immuno histochemistry. ResuL: A single injection of STZ induced an increase of IFN-y producing Th (CD4+) and Tc (CD8+) cells in the draining lymph node (data on IL-4 and IL-10 producing cells are pending). Moreover, the total number of CD4+ cells was decreased, whereas CD8+ cells, macroohaoes and aooototic cells were increased in conjunction with enhanced IgG2,‘andlgGsr, production to TNP-OVA. Injection of HgC12on the other hand, reduced the number of IFN-Y producing cells, inducedaccumulation of B cells and of CD4+ and CD8+ T cells, enhanced IgGr and IgE production to TNP-OVA, and primed for secondary IgG, and IgE production as well as for DTH reactions. Conclusion: Together these results indicate that STZ stimulates type-l responses, whereas HgCls enhances mixed type-1 and 2 responses in BALB/c mice. Importantly, these responses stimulated by a single injection of chemical match the (auto)immune effects elicited to unknown (auto) antigens following multiple injections of these compounds, suggesting that induction of selective costimulation by chemicals is an important factor provoking the onset of autoimmunity in susceptible individuals.
P.5.11 .l1
The use of reporter antigens In the popliteai lymph node assay to assess immunomoduiation by chemicals
Ft. Albers, E. van ‘t Erve, A. Broeden, A. van der Pijl, W. Seinen, R. Pieters. RITOX, lmmunotaxicoogy Section, Utrecht Univetsib PO Box 80.176, 3.506 TD Utrecht, The Netherlands Introduction: Various drugs and other low molecular weight chemicals can induce T cell-dependent B cell activation which may cause allergic or autoimmune-like diseases. Because the nature of the relevant (neo-) antigens or hapten-carder complexes are generally not (in case autoimmunity) or not alwavs (in case of allerov) known. we exolored the ootential use of well-defined reporter antigens to d&mine T cell-dependent B’cell activation by chemicals. TNP-ficoll and TNP-OVA were used as reporter antigens because they are recognized by the same TNP-specific B cells. For specific antibody production. however, distinct co-stimulation is required.