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Autoimmunity in a mouse model of type 1 diabetes mellitus is triggered by an endocrine abnormality
3 12
Pathogenesis of autoimmunity
Conclusion:Active vasculitis is associated with the presence of activated circulating neutrophils and monocytes. Furthermore, leukocyte activation correlated with disease activity as determined according to the BVAS scoring system. Upregulation of adhesion molecules on circulating leukocytes was only minimal. This may be due to increased adhesiveness of cells that demonstrate increased expression of adhesion molecules. 1P.5.11.25 1 Immunological, biochemical and other homeostasis disturbances in diabetes mellitus as a signs of total metabolic depressive syndrome M. Stenina, D. Voevodin, A. Skripnic, N. Snisar, G. Rosanova, N. Matveeva. Russian State Medical Univ, Moscow, Russia
Introduction: The present study was carded out to reveal immunosuppression as a pad of total metabolic depressive syndrome in type 1 diabetes mellitus (DM). Mat&al and Methods: Control tdals were performed by immunological, biochemical, microbiological, cytochemical monitoring of health status in 100 children with DM. The patient’s age ranged from 4 to 11years. Reeults: Evidence has been received, that changes in the above mentioned homeostasis monitored parameters were systemic that reflected signs of total metabolic depressive syndrome. So it was shown, that in DM the suppression of high affinity IgG-antibody response occured. Indeed, in diabetic population cholesterol blood level rose in parallel with reduction of IgG-quota in total blood protein. The normal population (50 children) showed a strong positive correlation between the rises of cholesterol and IgM-part in blood protein. Patients with visible hyperfipidsmia (up to 5 mkM&l or more) and low IgG-quota (12.3 f 1%) showed a significantly lesser level of antibodies to peptidoglycan than those in whom the above mentioned parameters remained 3.9 f 0.2 mkMolfl Ed 15.2 f
1.3. Conclusion: Our data may imply that the formation of low affinity antibody response in DM may be more relevant to the suppression of immune system than direct autoimmunity activation. The role of intestinal-derived antigenic/toxic factor(s) seems to be important in maintaining the reversible subversion of metabolic and immune processes in patients with DM.
P.5.11.26
Autolmmunity In a mouse model of type 1 diabetes mellitus is triggered by an endocrine abnormality
J.G.M. Rosmalen ‘, F. Homo-Delarche 2, M. Dardenne *, H.A. Drexhage ‘. ’ Dept.of Immunology Erasmus University Rotterdam, Rofterdam, The Nethenands, ‘CNRS
URA 1416, H6pitaf Necker, Paris, France
Introduction: lnsulitis in the NOD mouse-model for type 1 diabetes mellitus starts with an accumulation of macrophages and dendrftlc cells around the pancreatic islets of Langerhans. The mason for this accumulation of antigen presenting cells (APC) is unknown. In other endocrine organs, e.g. the thyroid, pituitary and ovary, it is known that dendritic cells play a role in the regulation of endocrine responses. Therefore, we hypothesized that the initial accumulation of APC was caused by an abberent endocrine function of the islets. Indeed, an endocrine overactivity proceeds the islet-infiltration in NOD mice. Histologically, this is reflected in the development of a cohort of abnormally large islets (mega-islets, defined as larger than 1OOOO pixels) as compared to control mice. These mega-islets also develop in NODscicVscid mice which lack functional B and T lymphocytes, indicating that the hyperprolfferation of the islets is not caused by infiltrating lymphocytes. MaterlaIr and Methods: By means of immunohistology with various markers for APC and lymphocytes, we correlated islet sizes to infiltration during the onset of the insulitis. Results: It was found that APC were predominantly situated near the megaislets in early phases of the insulitis in NOD and, although present in lower numbers, in NODscWscid mice (56/42% of normal islets vs. 72/56% of mega-islets are infiltrated with APC for NOD/NODsci&ciid). In addition, at the timepoint of lymphocyte infiltration into the islets, we were able to show that these cells as well were attracted to the mega-islets (55% of mega-islets are infiltrated with lymphocytes vs. 7% of normal islets). Furthermore, we found that downregulating endocrine activity by means of insulin administration from 3 weeks of age onwards results in a smaller cohort of mega-islets on 6 weeks of age (28&O% of islets vs. 11112%after insulin administration for NODn\lODscid/scid).This reduction in islet size is accompanied by a reduced infiltration with APC and lymphocytes. Such prophylactic insulin treatment is also known to delay diabetes incidence in animal models as well as human diabetes. Conclusion: In conclusion, we have shown that abnonal mega-islets are the initial target for infiltration with APC and subsequently lymphocytes. Furthermore, it is likely that the protective effect of prophylactic insulin treatment, which is currently being tested in clinical trials, is due to a downregulation of endocrine function.
24 June 1997 - Poster presentations
-P.5. 11.27 1 Bullous pemphigoid associated with chronic B-lymphocytlc leukemia: The antibody against the 230 kD-protein Is not produced by leukemic cells L. Misery ‘,*, F. Cambazard 3, J.P. Magaud 4, R. Ghohestani I, A. Gaudillsre’, J.L. Perrot 3, A. Claudy‘, D. Schmitt*, C. Vincent *. ‘Department of Dermatofogy Edouard Herriot Hospital, Lyon, France, * INSERM W46, Edouard Hemot Hospital, Lyon, France, 3Department of Dennatofcgy Saint Etienne, France, 4 Cytogenetic and Molecular Biology Laboratory Edouard Hemot Hospital, Lyon, France
A patient presented with chronic B-lymphoid leukemia and bullous pemphigoid associated with circulating antibodies against 180 and 230 kd epidennal antigens. We search for production of these antibodies by leukemic cells. B-lymphocytes were isolated from peripheral blood mononuclear cells, obtained after centrlfugatlon on Ficoll-Paque medium, by depletion of adherent cells and T-lymphocytes. They were cultivated in different conditions. There was no spontaneous lineage. The addition of IL-2 (IO U/ml) increased transiently the cell count but the survival does not exceed 22 days. A sample was immortalized with Epstein-Barr virus (EBV). The cell phenotype was studied in the course of the 21 passages. 23 mg of immunoglobulines (fg) were produced between the 19th and the 21th passage. Ig were isolated then analyzed by immunofluorescence and immunoblot. A cytogenetic and genetic study was performed on leukemic and immortalized cells. Before and after culture, cells were CDl9+, CD21+, CD25+, CD40+, y+, HLA-DR+. Dermo-epidermal junction and basal keratinccytes were stained by antibodies conjugated with FITC. These antibodies were directed against the 230 kD epidermal antigen as detected by immunoblot assay. There was no caryotypic abnormality. Leukemic and immortalized cells were clonal but the immunoglobulin gene rearrangements were not identical. Hence, immortalized B-cells produced anti-230 kd hemidesmosomal protein but are probably not of a leukemic origin.
P.5.11.26
identity of immunologlcal disorders in children with bronchial asthma and obstructlve bronchitis
V.A. Lopatina ‘, A.A. Akatova*, V.A. Chereshnev ‘, 1.1.Balabolkin 3. l/nstitute of Ecology and Genetics of Microorganisms, Perm, Russia, * Pem, Medical academy, Perm, Russia, 3Scientiffc Researching fnstitute of pediatrics RAMS, Moscow, Russia
Bronchial Asthma (BA) is one of the causes of invalidism in children with lung diseases. Late diagnosing and beginning of treatment are the reasons for this. The aim of the work is the exposing of peculiarities of immune status in children with recurring obstructive bronchitis (ROB) as a phase of development of BA. 30 children aged 7 months to 10 years old were examined. 10 children formed the control group with BA. Leukogram, level of CD3, CD4, CD6 cells, CD4/CD8 ratio, lg A, lg G, Ig M, Ig E, circulating immune complexes (CC) were estimated. The initial immunological status was exposed. In 2/3 of cases the allergic process developed on bases of leukopenia, lymphopenia, in l/2-with abstemious eosinophilia. The level of CD3 was increased in 79%, in 57% because of increasing of CD4 cells. More deep decreasing of CD6 per 50% was exposed in 70% of children. CD4/CD8 ratio disordered towards increased in 39%. In more than I/2 of children the ClCs were increased and lg A, lg G were decreased. In 2/3 of children the allergic process took place with increasing of Ig E level. Changes of CD6, lg E and ClCs were more expressed in the control group. Thus, the identity of immune deviations between BA and ROB reveals the commonality of their genesis and permits to consider children with ROB, especially in presence of dermal allergic manifestations and burdened heredity, as sick with light form of BA.
1P.5.11.29 1 “Bovine spongiform encephalopathy” (BSE) could be an autoimmune disease produced by bacteria showing “molecular mimicry” with bovine/ovine brain antigens A. Ebringer, J. Pin, C. Wilson, P. Cunningham, C. Thorpe, C. Ettelaie. Division of LL Sciences and Department of Computing, King’s College London, UK, Department of &chemistry Royal Free Hospital, London, UK
Bovine spongifon encephalopathy (BSE) could be an autoimmune disease produced following exposure of cattle to “green offal” feedstuffs containing bacteria showing molecular mimicry between bacterial components and bovine nervous tissues. Analysis of molecular sequence databases (Genbank and SwissProt) shows that 3 bacteria: Acinetobacter cakoaceticus, Ruminococcus a/bus and Agmbacter tumefaciens share sequences with the encephalitogenic peptide FSWGAEGQK of bovine myelin. These bacteria are present in environmental material to which cows, sheep and goats are frequently exposed.
Pathogenesis of autoimmunity
Conclusion:Active vasculitis is associated with the presence of activated circulating neutrophils and monocytes. Furthermore, leukocyte activation correlated with disease activity as determined according to the BVAS scoring system. Upregulation of adhesion molecules on circulating leukocytes was only minimal. This may be due to increased adhesiveness of cells that demonstrate increased expression of adhesion molecules. 1P.5.11.25 1 Immunological, biochemical and other homeostasis disturbances in diabetes mellitus as a signs of total metabolic depressive syndrome M. Stenina, D. Voevodin, A. Skripnic, N. Snisar, G. Rosanova, N. Matveeva. Russian State Medical Univ, Moscow, Russia
Introduction: The present study was carded out to reveal immunosuppression as a pad of total metabolic depressive syndrome in type 1 diabetes mellitus (DM). Mat&al and Methods: Control tdals were performed by immunological, biochemical, microbiological, cytochemical monitoring of health status in 100 children with DM. The patient’s age ranged from 4 to 11years. Reeults: Evidence has been received, that changes in the above mentioned homeostasis monitored parameters were systemic that reflected signs of total metabolic depressive syndrome. So it was shown, that in DM the suppression of high affinity IgG-antibody response occured. Indeed, in diabetic population cholesterol blood level rose in parallel with reduction of IgG-quota in total blood protein. The normal population (50 children) showed a strong positive correlation between the rises of cholesterol and IgM-part in blood protein. Patients with visible hyperfipidsmia (up to 5 mkM&l or more) and low IgG-quota (12.3 f 1%) showed a significantly lesser level of antibodies to peptidoglycan than those in whom the above mentioned parameters remained 3.9 f 0.2 mkMolfl Ed 15.2 f
1.3. Conclusion: Our data may imply that the formation of low affinity antibody response in DM may be more relevant to the suppression of immune system than direct autoimmunity activation. The role of intestinal-derived antigenic/toxic factor(s) seems to be important in maintaining the reversible subversion of metabolic and immune processes in patients with DM.
P.5.11.26
Autolmmunity In a mouse model of type 1 diabetes mellitus is triggered by an endocrine abnormality
J.G.M. Rosmalen ‘, F. Homo-Delarche 2, M. Dardenne *, H.A. Drexhage ‘. ’ Dept.of Immunology Erasmus University Rotterdam, Rofterdam, The Nethenands, ‘CNRS
URA 1416, H6pitaf Necker, Paris, France
Introduction: lnsulitis in the NOD mouse-model for type 1 diabetes mellitus starts with an accumulation of macrophages and dendrftlc cells around the pancreatic islets of Langerhans. The mason for this accumulation of antigen presenting cells (APC) is unknown. In other endocrine organs, e.g. the thyroid, pituitary and ovary, it is known that dendritic cells play a role in the regulation of endocrine responses. Therefore, we hypothesized that the initial accumulation of APC was caused by an abberent endocrine function of the islets. Indeed, an endocrine overactivity proceeds the islet-infiltration in NOD mice. Histologically, this is reflected in the development of a cohort of abnormally large islets (mega-islets, defined as larger than 1OOOO pixels) as compared to control mice. These mega-islets also develop in NODscicVscid mice which lack functional B and T lymphocytes, indicating that the hyperprolfferation of the islets is not caused by infiltrating lymphocytes. MaterlaIr and Methods: By means of immunohistology with various markers for APC and lymphocytes, we correlated islet sizes to infiltration during the onset of the insulitis. Results: It was found that APC were predominantly situated near the megaislets in early phases of the insulitis in NOD and, although present in lower numbers, in NODscWscid mice (56/42% of normal islets vs. 72/56% of mega-islets are infiltrated with APC for NOD/NODsci&ciid). In addition, at the timepoint of lymphocyte infiltration into the islets, we were able to show that these cells as well were attracted to the mega-islets (55% of mega-islets are infiltrated with lymphocytes vs. 7% of normal islets). Furthermore, we found that downregulating endocrine activity by means of insulin administration from 3 weeks of age onwards results in a smaller cohort of mega-islets on 6 weeks of age (28&O% of islets vs. 11112%after insulin administration for NODn\lODscid/scid).This reduction in islet size is accompanied by a reduced infiltration with APC and lymphocytes. Such prophylactic insulin treatment is also known to delay diabetes incidence in animal models as well as human diabetes. Conclusion: In conclusion, we have shown that abnonal mega-islets are the initial target for infiltration with APC and subsequently lymphocytes. Furthermore, it is likely that the protective effect of prophylactic insulin treatment, which is currently being tested in clinical trials, is due to a downregulation of endocrine function.
24 June 1997 - Poster presentations
-P.5. 11.27 1 Bullous pemphigoid associated with chronic B-lymphocytlc leukemia: The antibody against the 230 kD-protein Is not produced by leukemic cells L. Misery ‘,*, F. Cambazard 3, J.P. Magaud 4, R. Ghohestani I, A. Gaudillsre’, J.L. Perrot 3, A. Claudy‘, D. Schmitt*, C. Vincent *. ‘Department of Dermatofogy Edouard Herriot Hospital, Lyon, France, * INSERM W46, Edouard Hemot Hospital, Lyon, France, 3Department of Dennatofcgy Saint Etienne, France, 4 Cytogenetic and Molecular Biology Laboratory Edouard Hemot Hospital, Lyon, France
A patient presented with chronic B-lymphoid leukemia and bullous pemphigoid associated with circulating antibodies against 180 and 230 kd epidennal antigens. We search for production of these antibodies by leukemic cells. B-lymphocytes were isolated from peripheral blood mononuclear cells, obtained after centrlfugatlon on Ficoll-Paque medium, by depletion of adherent cells and T-lymphocytes. They were cultivated in different conditions. There was no spontaneous lineage. The addition of IL-2 (IO U/ml) increased transiently the cell count but the survival does not exceed 22 days. A sample was immortalized with Epstein-Barr virus (EBV). The cell phenotype was studied in the course of the 21 passages. 23 mg of immunoglobulines (fg) were produced between the 19th and the 21th passage. Ig were isolated then analyzed by immunofluorescence and immunoblot. A cytogenetic and genetic study was performed on leukemic and immortalized cells. Before and after culture, cells were CDl9+, CD21+, CD25+, CD40+, y+, HLA-DR+. Dermo-epidermal junction and basal keratinccytes were stained by antibodies conjugated with FITC. These antibodies were directed against the 230 kD epidermal antigen as detected by immunoblot assay. There was no caryotypic abnormality. Leukemic and immortalized cells were clonal but the immunoglobulin gene rearrangements were not identical. Hence, immortalized B-cells produced anti-230 kd hemidesmosomal protein but are probably not of a leukemic origin.
P.5.11.26
identity of immunologlcal disorders in children with bronchial asthma and obstructlve bronchitis
V.A. Lopatina ‘, A.A. Akatova*, V.A. Chereshnev ‘, 1.1.Balabolkin 3. l/nstitute of Ecology and Genetics of Microorganisms, Perm, Russia, * Pem, Medical academy, Perm, Russia, 3Scientiffc Researching fnstitute of pediatrics RAMS, Moscow, Russia
Bronchial Asthma (BA) is one of the causes of invalidism in children with lung diseases. Late diagnosing and beginning of treatment are the reasons for this. The aim of the work is the exposing of peculiarities of immune status in children with recurring obstructive bronchitis (ROB) as a phase of development of BA. 30 children aged 7 months to 10 years old were examined. 10 children formed the control group with BA. Leukogram, level of CD3, CD4, CD6 cells, CD4/CD8 ratio, lg A, lg G, Ig M, Ig E, circulating immune complexes (CC) were estimated. The initial immunological status was exposed. In 2/3 of cases the allergic process developed on bases of leukopenia, lymphopenia, in l/2-with abstemious eosinophilia. The level of CD3 was increased in 79%, in 57% because of increasing of CD4 cells. More deep decreasing of CD6 per 50% was exposed in 70% of children. CD4/CD8 ratio disordered towards increased in 39%. In more than I/2 of children the ClCs were increased and lg A, lg G were decreased. In 2/3 of children the allergic process took place with increasing of Ig E level. Changes of CD6, lg E and ClCs were more expressed in the control group. Thus, the identity of immune deviations between BA and ROB reveals the commonality of their genesis and permits to consider children with ROB, especially in presence of dermal allergic manifestations and burdened heredity, as sick with light form of BA.
1P.5.11.29 1 “Bovine spongiform encephalopathy” (BSE) could be an autoimmune disease produced by bacteria showing “molecular mimicry” with bovine/ovine brain antigens A. Ebringer, J. Pin, C. Wilson, P. Cunningham, C. Thorpe, C. Ettelaie. Division of LL Sciences and Department of Computing, King’s College London, UK, Department of &chemistry Royal Free Hospital, London, UK
Bovine spongifon encephalopathy (BSE) could be an autoimmune disease produced following exposure of cattle to “green offal” feedstuffs containing bacteria showing molecular mimicry between bacterial components and bovine nervous tissues. Analysis of molecular sequence databases (Genbank and SwissProt) shows that 3 bacteria: Acinetobacter cakoaceticus, Ruminococcus a/bus and Agmbacter tumefaciens share sequences with the encephalitogenic peptide FSWGAEGQK of bovine myelin. These bacteria are present in environmental material to which cows, sheep and goats are frequently exposed.