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In vivo evidence for the contribution of human histocompatibility leukocyte antigen (HLA)-DQ molecules to the development of diabetes.

Wen L, Wong FS, Tang J, Chen NY, Altieri M, David C, Flavell R, Sherwin R - J. Exp. Med. (2000)

Bottom Line: Although DQA1*0301/DQB1*0302 is the human histocompatibility leukocyte antigen (HLA) class II gene most commonly associated with human type 1 diabetes, direct in vivo experimental evidence for its diabetogenic role is lacking.In conclusion, substitution of HLA-DQA1*0301/DQB1*0302, but not HLA-DQA1*0103/DQB1*0601, for murine MHC class II provokes autoimmune diabetes in non-diabetes-prone rat insulin promoter (RIP).B7-1 C57BL/6 mice.Our data provide direct in vivo evidence for the diabetogenic effect of this human MHC class II molecule and a unique "humanized" animal model of spontaneous diabetes.

View Article: PubMed Central - PubMed

Affiliation: Section of Endocrinology, Department of Internal Medicine, the. Yale University Shool of Medicine, New Haven, CT, USA. li.wen.@yale.edu

ABSTRACT
Although DQA1*0301/DQB1*0302 is the human histocompatibility leukocyte antigen (HLA) class II gene most commonly associated with human type 1 diabetes, direct in vivo experimental evidence for its diabetogenic role is lacking. Therefore, we generated C57BL/6 transgenic mice that bear this molecule and do not express mouse major histocompatibility complex (MHC) class II molecules (DQ8(+)/mII(-)). They did not develop insulitis or spontaneous diabetes. However, when DQ8(+)/mII(-) mice were bred with C57BL/6 mice expressing costimulatory molecule B7-1 on beta cells (which normally do not develop diabetes), 81% of the DQ8(+)/mII(-)/B7-1(+) mice developed spontaneous diabetes. The diabetes was accompanied by severe insulitis composed of both T cells (CD4(+) and CD8(+)) and B cells. T cells from the diabetic mice secreted large amounts of interferon gamma, but not interleukin 4, in response to DQ8(+) islets and the putative islet autoantigens, insulin and glutamic acid decarboxylase (GAD). Diabetes could also be adoptively transferred to irradiated nondiabetic DQ8(+)/mII(-)/B7-1(+) mice. In striking contrast, none of the transgenic mice in which the diabetes protective allele (DQA1*0103/DQB1*0601, DQ6 for short) was substituted for mouse MHC class II molecules but remained for the expression of B7-1 on pancreatic beta cells (DQ6(+)/mII(-)/B7-1(+)) developed diabetes. Only 7% of DQ(-)/mII(-)/B7-1(+) mice developed diabetes at an older age, and none of the DQ(-)/mII(+)/B7-1(+) mice or DQ8(+)/mII(+)/B7-1(+) mice developed diabetes. In conclusion, substitution of HLA-DQA1*0301/DQB1*0302, but not HLA-DQA1*0103/DQB1*0601, for murine MHC class II provokes autoimmune diabetes in non-diabetes-prone rat insulin promoter (RIP).B7-1 C57BL/6 mice. Our data provide direct in vivo evidence for the diabetogenic effect of this human MHC class II molecule and a unique "humanized" animal model of spontaneous diabetes.

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Adoptively transferred diabetes. 10 × 107 diabetic DQ8+/mII−/B7+ splenocytes (depleted red blood cells) were transferred intravenously into irradiated recipients (as indicated). Diabetes was determined as above. Number of mice per group was as follows: n = 9 (4 female, 5 male) DQ8+/mII−/B7+; n = 8 (4 female, 4 male) DQ−/mII−/B7+; n = 3 (male) DQ8+/mII−/B7−; n = 3 (male) DQ8−/mII−/B7−; and n = 3 (male) DQ8+/mII−/B7+* (these recipients were adoptively transferred with purified CD8+ splenocytes only from diabetic DQ8+/mII−/B7+ mice).
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Figure 2: Adoptively transferred diabetes. 10 × 107 diabetic DQ8+/mII−/B7+ splenocytes (depleted red blood cells) were transferred intravenously into irradiated recipients (as indicated). Diabetes was determined as above. Number of mice per group was as follows: n = 9 (4 female, 5 male) DQ8+/mII−/B7+; n = 8 (4 female, 4 male) DQ−/mII−/B7+; n = 3 (male) DQ8+/mII−/B7−; n = 3 (male) DQ8−/mII−/B7−; and n = 3 (male) DQ8+/mII−/B7+* (these recipients were adoptively transferred with purified CD8+ splenocytes only from diabetic DQ8+/mII−/B7+ mice).

Mentions: Adoptive transfer experiments showed that diabetes in DQ8+/mII−/RIP.B7-1 mice was autoimmune mediated. Splenocytes (depleted of erythrocytes) from diabetic DQ8+/mII−/RIP.B7-1 mice were adoptively transferred into either DQ8+ or DQ8− irradiated recipients (8–10 wk of age; Fig. 2). Diabetes appeared in 78% of the DQ8+/mII−/RIP.B7-1 recipients (7/9) beginning at 3 wk after cell transfer, but in only 2 of 8 DQ8−/mII−/RIP.B7-1 recipients between 6 and 8 wk after adoptive transfer. In contrast, none of the RIP.B7-1 transgene-negative recipients, either DQ8+/mII− or DQ8−/mII−, developed diabetes 8 wk after the cell transfer ( Fig. 2). To ascertain that the diabetes, developed either spontaneously or after adoptive transfer, was not primarily CD8 mediated, since two “odd” mice became diabetic in both experiments, we performed another group of adoptive transfers using purified CD8 T cells derived from diabetic DQ8+/mII−/RIP.B7-1 splenocytes. None of the recipients (DQ8+/mII−/RIP.B7-1; n = 3) developed diabetes 8 wk after the transfer when the experiment was terminated ( Fig. 2), supporting a key role for the HLA-DQA1*0301/DQB1*0302 molecules and CD4 T cells selected by the molecules in this diabetic model.


In vivo evidence for the contribution of human histocompatibility leukocyte antigen (HLA)-DQ molecules to the development of diabetes.

Wen L, Wong FS, Tang J, Chen NY, Altieri M, David C, Flavell R, Sherwin R - J. Exp. Med. (2000)

Adoptively transferred diabetes. 10 × 107 diabetic DQ8+/mII−/B7+ splenocytes (depleted red blood cells) were transferred intravenously into irradiated recipients (as indicated). Diabetes was determined as above. Number of mice per group was as follows: n = 9 (4 female, 5 male) DQ8+/mII−/B7+; n = 8 (4 female, 4 male) DQ−/mII−/B7+; n = 3 (male) DQ8+/mII−/B7−; n = 3 (male) DQ8−/mII−/B7−; and n = 3 (male) DQ8+/mII−/B7+* (these recipients were adoptively transferred with purified CD8+ splenocytes only from diabetic DQ8+/mII−/B7+ mice).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2195792&req=5

Figure 2: Adoptively transferred diabetes. 10 × 107 diabetic DQ8+/mII−/B7+ splenocytes (depleted red blood cells) were transferred intravenously into irradiated recipients (as indicated). Diabetes was determined as above. Number of mice per group was as follows: n = 9 (4 female, 5 male) DQ8+/mII−/B7+; n = 8 (4 female, 4 male) DQ−/mII−/B7+; n = 3 (male) DQ8+/mII−/B7−; n = 3 (male) DQ8−/mII−/B7−; and n = 3 (male) DQ8+/mII−/B7+* (these recipients were adoptively transferred with purified CD8+ splenocytes only from diabetic DQ8+/mII−/B7+ mice).
Mentions: Adoptive transfer experiments showed that diabetes in DQ8+/mII−/RIP.B7-1 mice was autoimmune mediated. Splenocytes (depleted of erythrocytes) from diabetic DQ8+/mII−/RIP.B7-1 mice were adoptively transferred into either DQ8+ or DQ8− irradiated recipients (8–10 wk of age; Fig. 2). Diabetes appeared in 78% of the DQ8+/mII−/RIP.B7-1 recipients (7/9) beginning at 3 wk after cell transfer, but in only 2 of 8 DQ8−/mII−/RIP.B7-1 recipients between 6 and 8 wk after adoptive transfer. In contrast, none of the RIP.B7-1 transgene-negative recipients, either DQ8+/mII− or DQ8−/mII−, developed diabetes 8 wk after the cell transfer ( Fig. 2). To ascertain that the diabetes, developed either spontaneously or after adoptive transfer, was not primarily CD8 mediated, since two “odd” mice became diabetic in both experiments, we performed another group of adoptive transfers using purified CD8 T cells derived from diabetic DQ8+/mII−/RIP.B7-1 splenocytes. None of the recipients (DQ8+/mII−/RIP.B7-1; n = 3) developed diabetes 8 wk after the transfer when the experiment was terminated ( Fig. 2), supporting a key role for the HLA-DQA1*0301/DQB1*0302 molecules and CD4 T cells selected by the molecules in this diabetic model.

Bottom Line: Although DQA1*0301/DQB1*0302 is the human histocompatibility leukocyte antigen (HLA) class II gene most commonly associated with human type 1 diabetes, direct in vivo experimental evidence for its diabetogenic role is lacking.In conclusion, substitution of HLA-DQA1*0301/DQB1*0302, but not HLA-DQA1*0103/DQB1*0601, for murine MHC class II provokes autoimmune diabetes in non-diabetes-prone rat insulin promoter (RIP).B7-1 C57BL/6 mice.Our data provide direct in vivo evidence for the diabetogenic effect of this human MHC class II molecule and a unique "humanized" animal model of spontaneous diabetes.

View Article: PubMed Central - PubMed

Affiliation: Section of Endocrinology, Department of Internal Medicine, the. Yale University Shool of Medicine, New Haven, CT, USA. li.wen.@yale.edu

ABSTRACT
Although DQA1*0301/DQB1*0302 is the human histocompatibility leukocyte antigen (HLA) class II gene most commonly associated with human type 1 diabetes, direct in vivo experimental evidence for its diabetogenic role is lacking. Therefore, we generated C57BL/6 transgenic mice that bear this molecule and do not express mouse major histocompatibility complex (MHC) class II molecules (DQ8(+)/mII(-)). They did not develop insulitis or spontaneous diabetes. However, when DQ8(+)/mII(-) mice were bred with C57BL/6 mice expressing costimulatory molecule B7-1 on beta cells (which normally do not develop diabetes), 81% of the DQ8(+)/mII(-)/B7-1(+) mice developed spontaneous diabetes. The diabetes was accompanied by severe insulitis composed of both T cells (CD4(+) and CD8(+)) and B cells. T cells from the diabetic mice secreted large amounts of interferon gamma, but not interleukin 4, in response to DQ8(+) islets and the putative islet autoantigens, insulin and glutamic acid decarboxylase (GAD). Diabetes could also be adoptively transferred to irradiated nondiabetic DQ8(+)/mII(-)/B7-1(+) mice. In striking contrast, none of the transgenic mice in which the diabetes protective allele (DQA1*0103/DQB1*0601, DQ6 for short) was substituted for mouse MHC class II molecules but remained for the expression of B7-1 on pancreatic beta cells (DQ6(+)/mII(-)/B7-1(+)) developed diabetes. Only 7% of DQ(-)/mII(-)/B7-1(+) mice developed diabetes at an older age, and none of the DQ(-)/mII(+)/B7-1(+) mice or DQ8(+)/mII(+)/B7-1(+) mice developed diabetes. In conclusion, substitution of HLA-DQA1*0301/DQB1*0302, but not HLA-DQA1*0103/DQB1*0601, for murine MHC class II provokes autoimmune diabetes in non-diabetes-prone rat insulin promoter (RIP).B7-1 C57BL/6 mice. Our data provide direct in vivo evidence for the diabetogenic effect of this human MHC class II molecule and a unique "humanized" animal model of spontaneous diabetes.

Show MeSH
Related in: MedlinePlus