Sanjeevi C B, Lybrand T P, Landin-Olsson M, Kockum I, Dahlquist G, Hagopian W A, Palmer J P, Lernmark A
Karolinska Institute, Department of Endocrinology, Karolinska Hospital, Stockholm, Sweden.
Tissue Antigens. 1994 Aug;44(2):110-9. doi: 10.1111/j.1399-0039.1994.tb02366.x.
HLA-DR2 is negatively associated with insulin-dependent diabetes mellitus (IDDM). The aim of the present study was to analyze DR2-positive patients among 425 consecutively diagnosed unrelated Swedish children with IDDM and in 367 matched controls. HLA-DRB, -DQA and -DQB were determined by Taq I restriction fragment length polymorphism analysis. Amplification by polymerase chain reaction (PCR) and hybridization with sequence-specific oligonucleotide probes was done for DQA1, DQB1 and DRB1 and DRB5. DR2 was positive in 11/425 patients (3%) and 101/367 (28%) controls (OR 0.07, p < 0.0001). Of the 11 DR2-positive patients, PCR was done in 10, of whom 8 were positive for DRB11601-DRB50201 compared to 4/96 (4%) controls (OR 92.0: p < 0.001) while the remaining 2 were positive for DRB11501-DRB50101 compared to 92/96 (96%, OR 0.01; p < 0.001). In 2 patients, a recombination between the haplotypes DQB10502-DQA10102 (DQ5)-DRB11601-DRB50201 (DR16 Dw21) and DQB10301-DQA10501 (DQ7)-DRB11602-DRB50202 (DR16 Dw22) was observed resulting in the DQB10301-DQA10501 (DQ7) DRB11601-DRB50201 (DR16 Dw22) haplotypes. The second haplotype was DR3 DQ2 in 6/11 and DR4 DQ8 in 2/11 DR2-positive patients. In all 3 DQB10602-DQA10102-DR15-positive patients the second haplotype was DR4-positive. In order to test whether physicochemical properties of the DR2 molecules were associated with IDDM, we constructed three-dimensional models of the peptide binding and T-cell recognition sites (alpha 1 and beta 1 domains) of five subtypes of DR2-DRB1, based on the published DR1 crystal structure. No correlations were observed for DR molecule physicochemical properties and diabetes susceptibility. Islet cell antibodies, insulin autoantibodies and GAD65 antibodies, were measured in DR2-positive patients (n = 11) and controls (n = 101). Despite the presence of the DR2 haplotype the antibody markers were significantly elevated in the patients compared to the controls (GAD65 3/10 patients and 2/101 controls; ICA 7/11 patients and 1/101 controls and IAA 3/11 patients and 0/101 controls). In conclusion, of the five subtypes of DR2, only one, the DRB11501, DRB50101, DQB10602-DQA10102 haplotype, was negatively associated with IDDM. DQ may therefore confer more protection from the disease than DR.
HLA - DR2与胰岛素依赖型糖尿病（IDDM）呈负相关。本研究的目的是分析425例连续诊断的无亲缘关系的瑞典IDDM儿童及367例匹配对照中的DR2阳性患者。通过Taq I限制性片段长度多态性分析来确定HLA - DRB、- DQA和 - DQB。对DQA1、DQB1、DRB1和DRB5进行聚合酶链反应（PCR）扩增并与序列特异性寡核苷酸探针杂交。11/425例患者（3%）的DR2呈阳性，101/367例对照（28%）的DR2呈阳性（比值比0.07，p < 0.0001）。在11例DR2阳性患者中，对10例进行了PCR检测，其中8例DRB11601 - DRB50201呈阳性，与之相比，96例对照中有4例（4%）呈阳性（比值比92.0：p < 0.001），而其余2例DRB11501 - DRB50101呈阳性，与之相比，96例对照中有92例（96%）呈阳性（比值比0.01；p < 0.001）。在2例患者中，观察到单倍型DQB10502 - DQA10102（DQ5）- DRB11601 - DRB50201（DR16 Dw21）与DQB10301 - DQA10501（DQ7）- DRB11602 - DRB50202（DR16 Dw22）之间发生重组，产生了DQB10301 - DQA10501（DQ7）DRB11601 - DRB50201（DR16 Dw22）单倍型。在11例DR2阳性患者中，6例的第二种单倍型为DR3 DQ2，2例为DR4 DQ8。在所有3例DQB10602 - DQA10102 - DR15阳性患者中，第二种单倍型为DR4阳性。为了测试DR2分子的物理化学性质是否与IDDM相关，我们基于已发表的DR1晶体结构构建了DR2 - DRB1五种亚型的肽结合和T细胞识别位点（α1和β1结构域）的三维模型。未观察到DR分子物理化学性质与糖尿病易感性之间的相关性。对DR2阳性患者（n = 11）和对照（n = 101）检测了胰岛细胞抗体、胰岛素自身抗体和GAD65抗体。尽管存在DR2单倍型，但与对照相比，患者中的抗体标志物显著升高（GAD65：10例患者中有3例，101例对照中有2例；ICA：11例患者中有7例，101例对照中有1例；IAA：11例患者中有3例，101例对照中有0例）。总之，在DR2的五种亚型中，只有一种，即DRB11501、DRB50101、DQB10602 - DQA10102单倍型与IDDM呈负相关。因此，DQ可能比DR赋予更多对该疾病的保护作用。
