Biochemical studies of RT6 alloantigens in BB/Wor and normal rats: Evidence for intact unexpressed RT6a structural gene in diabetes-prone BB rats

L. Crisa, D. L. Greiner, J. P. Mordes, R. G. MacDonald, E. S. Handler, M. P. Czech, A. A. Rossini

Research output: Contribution to journalArticle

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Abstract

Lymphocytes bearing the T-lymphocyte differentiation antigen RT6 play an important immunoregulatory role in the development of autoimmune diabetes in BB rats. Immunofluorescence studies suggest that diabetes-prone (DP)- but not diabetes-resistant (DR)-BB rat lymphocytes fail to express RT6 antigen during ontogeny. Two alloantigenic forms of the molecule exist, i.e., RT6.1 and RT6.2; both are linked to cell membranes by a phosphatidylinositol (Pl) linkage. In these studies, Pl-phospholipase C (PLC) treatment of lymphocytes from BB and normal rats followed by immunoabsorption and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of released proteins with anti-RT6 allotype-specific monoclonal antibodies was performed. RT6.1 in several nondiabetic rat strains was found to consist of a family of nonglycosylated and variably glycosylated molecules: an N-Glycanase-resistant 24,000- to 26,000-M(r) peptide and four N-Glycanase-sensitive peptides of 29,000, 31,000, 33,000, and 34,000 M(r). In contrast, RT6.2 was found to be a 24,000- to 26,000-M(r) nonglycosylated polypeptide. The electrophoretic pattern of RT6.1 was observed to be the same when the antigen was extracted from W3/25+ (CD4+) versus W3/25- T lymphocytes or from resting versus mitogen-activated cells. A pattern of bands characteristic of the RT6.1 antigen found in normal rat strains was detected after PLC treatment or detergent solubilization of lymphocytes obtained from DR rats. In contrast, no evidence of either RT6 species was found after PLC or detergent treatment of comparable numbers of T lymphocytes from DP-BB rats. Interestingly, T lymphocytes from Wistar-Furth (RT6.2+) x DP (RT6-) F1 crosses were observed to coexpress both RT6.2 and RT6.1 molecules, with the electrophoretic pattern of RT6.1 being similar to that obtained in DR and other rat strains. This study provides biochemical evidence that DP rats may have an intact RT6a structural gene.

Original languageEnglish (US)
Pages (from-to)1279-1288
Number of pages10
JournalDiabetes
Volume39
Issue number10
DOIs
StatePublished - Jan 1 1990

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Inbred BB Rats
Isoantigens
Type C Phospholipases
Lymphocytes
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Genes
Phosphatidylinositols
T-Lymphocytes
Antigens
Detergents
T Lymphocyte Differentiation Antigens
Peptides
Type 1 Diabetes Mellitus
Mitogens
Sodium Dodecyl Sulfate
Fluorescent Antibody Technique
Polyacrylamide Gel Electrophoresis
Therapeutics
Monoclonal Antibodies
Cell Membrane

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism

Cite this

Biochemical studies of RT6 alloantigens in BB/Wor and normal rats : Evidence for intact unexpressed RT6a structural gene in diabetes-prone BB rats. / Crisa, L.; Greiner, D. L.; Mordes, J. P.; MacDonald, R. G.; Handler, E. S.; Czech, M. P.; Rossini, A. A.

In: Diabetes, Vol. 39, No. 10, 01.01.1990, p. 1279-1288.

Research output: Contribution to journalArticle

Crisa, L. ; Greiner, D. L. ; Mordes, J. P. ; MacDonald, R. G. ; Handler, E. S. ; Czech, M. P. ; Rossini, A. A. / Biochemical studies of RT6 alloantigens in BB/Wor and normal rats : Evidence for intact unexpressed RT6a structural gene in diabetes-prone BB rats. In: Diabetes. 1990 ; Vol. 39, No. 10. pp. 1279-1288.
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abstract = "Lymphocytes bearing the T-lymphocyte differentiation antigen RT6 play an important immunoregulatory role in the development of autoimmune diabetes in BB rats. Immunofluorescence studies suggest that diabetes-prone (DP)- but not diabetes-resistant (DR)-BB rat lymphocytes fail to express RT6 antigen during ontogeny. Two alloantigenic forms of the molecule exist, i.e., RT6.1 and RT6.2; both are linked to cell membranes by a phosphatidylinositol (Pl) linkage. In these studies, Pl-phospholipase C (PLC) treatment of lymphocytes from BB and normal rats followed by immunoabsorption and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of released proteins with anti-RT6 allotype-specific monoclonal antibodies was performed. RT6.1 in several nondiabetic rat strains was found to consist of a family of nonglycosylated and variably glycosylated molecules: an N-Glycanase-resistant 24,000- to 26,000-M(r) peptide and four N-Glycanase-sensitive peptides of 29,000, 31,000, 33,000, and 34,000 M(r). In contrast, RT6.2 was found to be a 24,000- to 26,000-M(r) nonglycosylated polypeptide. The electrophoretic pattern of RT6.1 was observed to be the same when the antigen was extracted from W3/25+ (CD4+) versus W3/25- T lymphocytes or from resting versus mitogen-activated cells. A pattern of bands characteristic of the RT6.1 antigen found in normal rat strains was detected after PLC treatment or detergent solubilization of lymphocytes obtained from DR rats. In contrast, no evidence of either RT6 species was found after PLC or detergent treatment of comparable numbers of T lymphocytes from DP-BB rats. Interestingly, T lymphocytes from Wistar-Furth (RT6.2+) x DP (RT6-) F1 crosses were observed to coexpress both RT6.2 and RT6.1 molecules, with the electrophoretic pattern of RT6.1 being similar to that obtained in DR and other rat strains. This study provides biochemical evidence that DP rats may have an intact RT6a structural gene.",
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AU - Mordes, J. P.

AU - MacDonald, R. G.

AU - Handler, E. S.

AU - Czech, M. P.

AU - Rossini, A. A.

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N2 - Lymphocytes bearing the T-lymphocyte differentiation antigen RT6 play an important immunoregulatory role in the development of autoimmune diabetes in BB rats. Immunofluorescence studies suggest that diabetes-prone (DP)- but not diabetes-resistant (DR)-BB rat lymphocytes fail to express RT6 antigen during ontogeny. Two alloantigenic forms of the molecule exist, i.e., RT6.1 and RT6.2; both are linked to cell membranes by a phosphatidylinositol (Pl) linkage. In these studies, Pl-phospholipase C (PLC) treatment of lymphocytes from BB and normal rats followed by immunoabsorption and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of released proteins with anti-RT6 allotype-specific monoclonal antibodies was performed. RT6.1 in several nondiabetic rat strains was found to consist of a family of nonglycosylated and variably glycosylated molecules: an N-Glycanase-resistant 24,000- to 26,000-M(r) peptide and four N-Glycanase-sensitive peptides of 29,000, 31,000, 33,000, and 34,000 M(r). In contrast, RT6.2 was found to be a 24,000- to 26,000-M(r) nonglycosylated polypeptide. The electrophoretic pattern of RT6.1 was observed to be the same when the antigen was extracted from W3/25+ (CD4+) versus W3/25- T lymphocytes or from resting versus mitogen-activated cells. A pattern of bands characteristic of the RT6.1 antigen found in normal rat strains was detected after PLC treatment or detergent solubilization of lymphocytes obtained from DR rats. In contrast, no evidence of either RT6 species was found after PLC or detergent treatment of comparable numbers of T lymphocytes from DP-BB rats. Interestingly, T lymphocytes from Wistar-Furth (RT6.2+) x DP (RT6-) F1 crosses were observed to coexpress both RT6.2 and RT6.1 molecules, with the electrophoretic pattern of RT6.1 being similar to that obtained in DR and other rat strains. This study provides biochemical evidence that DP rats may have an intact RT6a structural gene.

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