广西壮族系统性红斑狼疮与HLA-DQA1等位基因的相关性研究
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摘要
目的：探讨广西壮族系统性红斑狼疮（SLE）与HLA-DQA1等位基因相关性。方法：患者组为广西壮族SLE病人，男5例，女46例。正常对照组为无血缘关系的壮族健康者，男32例，女38例。SLE组与正常对照组双亲三代内均为壮族。应用经典的酚／氯仿抽提法提取外周血基因组DNA，用聚合酶链反应—序列特异性引物（PCR-SSP）技术，对51例SLE壮族患者的HLA-DQA1基因进行研究,以70例壮族健康者作对照。同时，采用免疫印迹技术测定SLE壮族患者自身抗体谱。结果：12对引物检出HLA-DQA1 8个亚型，HLA-DQA1*0201和HLA-DQA1*0302等位基因在两组中均未检测到。SLE组DQA1*0101频率显著高于对照组(RR=3.2727，χ2=7.321，P=0.009)，而DQA1*0104和DQA1*0301频率均显著低于对照组（RR=0.4561，χ2=3.885，P=0.049 和 RR=0.4317，χ2=4.843，P=0.028）。DQA1*0101与抗dsDNA正相关 （χ2=4.488，P=0.034），DQA1*0301与抗dsDNA及抗SSA/SSB负相关 （χ2=5.017，P=0.025及χ2=3.869，P=0.049）。结论：HLA-DQA1*0201和HLA-DQA1*0302等位基因空白构成了广西壮族人群的特殊性。HLA-DQA1*0101可能是广西壮族SLE易感基因，其易感性可能通过诱导抗dsDNA抗体产生实现。DQA1*0104和DQA1*0301可能通过抑制自身抗体的产生为保护基因。
Objective: To explore the association of HLA-DQA1 alleles with systemic lupus erythematosus of Zhuang nationality in Guangxi. Methods: The patients group with SLE of Guangxi Zhuang nationnality included 5 males and 46 females . The health control group of Guangxi Zhuang nationnality were non blood relationship . The parents and the grandparents of everyone in two groups were all Zhuang people. Classical phenol /chloroform technique was used to extracted the DNA of white blood cells. Then polymerase chain reaction-sequence specific primers technique(PCR-SSP)was used to comparatively study the allelic genes in HLA-DQA1 locus in 51 patients with SLE and 70 selected , unrelated healthy controls of Zhuang subjects. Meanwhile, the autoantibodies of SLE patients were detected by immunoblot technique. Results: Twelve pairs primers amplified out 8 isoforms within HLA-DQA1 region. The HLA-DQA1*0201 and HLA-DQA1*0302 alleles were not found. The allelic frequencies of HLA-DQA1*0101 was significantly increased in SLE patients than those of the controls (RR=3.2727，χ2=7.321，P=0.009). But the allelic frequencies of HLA-DQA1*0104 and HLA-DQA1*0301 were significantly less in SLE
patients than those of the controls（RR=0.4561，χ2=3.885，P=0.049 and RR=0.4317，χ2=4.843，P=0.028）. Furthermore, the anti-dsDNA associated with DQA1*0101 positively （χ2=4.488，P=0.034）but associated with DQA1*0301 negatively（χ2=5.017，P=0.025）. The anti-SSA/SSB associated with DQA1*0301 negatively (χ2=3.869，P=0.049). Conclusion: Our findings suggested that the deficiency of HLA- DQA1 *0201 and DQA1*0302 alleles is one of particularities in Guangxi Zhuang people. The HLA-DQA1*0101 allele may be susceptible gene for SLE in Guangxi Zhuang nationality and act by inducting the production of anti-dsDNA. Yet the HLA-DQA1*0104 and HLA-DQA1*0301 alleles may involve in inhibiting the production of autoantibodies and may be protecting genes for SLE in Guangxi Zhuang nationality.
Objective: To explore the association of HLA-DQA1 alleles with systemic lupus erythematosus of Zhuang nationality in Guangxi. Methods: The patients group with SLE of Guangxi Zhuang nationnality included 5 males and 46 females . The health control group of Guangxi Zhuang nationnality were non blood relationship . The parents and the grandparents of everyone in two groups were all Zhuang people. Classical phenol /chloroform technique was used to extracted the DNA of white blood cells. Then polymerase chain reaction-sequence specific primers technique(PCR-SSP)was used to comparatively study the allelic genes in HLA-DQA1 locus in 51 patients with SLE and 70 selected , unrelated healthy controls of Zhuang subjects. Meanwhile, the autoantibodies of SLE patients were detected by immunoblot technique. Results: Twelve pairs primers amplified out 8 isoforms within HLA-DQA1 region. The HLA-DQA1*0201 and HLA-DQA1*0302 alleles were not found. The allelic frequencies of HLA-DQA1*0101 was significantly increased in SLE patients than those of the controls (RR=3.2727，χ2=7.321，P=0.009). But the allelic frequencies of HLA-DQA1*0104 and HLA-DQA1*0301 were significantly less in SLE
patients than those of the controls（RR=0.4561，χ2=3.885，P=0.049 and RR=0.4317，χ2=4.843，P=0.028）. Furthermore, the anti-dsDNA associated with DQA1*0101 positively （χ2=4.488，P=0.034）but associated with DQA1*0301 negatively（χ2=5.017，P=0.025）. The anti-SSA/SSB associated with DQA1*0301 negatively (χ2=3.869，P=0.049). Conclusion: Our findings suggested that the deficiency of HLA- DQA1 *0201 and DQA1*0302 alleles is one of particularities in Guangxi Zhuang people. The HLA-DQA1*0101 allele may be susceptible gene for SLE in Guangxi Zhuang nationality and act by inducting the production of anti-dsDNA. Yet the HLA-DQA1*0104 and HLA-DQA1*0301 alleles may involve in inhibiting the production of autoantibodies and may be protecting genes for SLE in Guangxi Zhuang nationality.
引文
Tebbe B,Orfannos CE.Epidemiology and socioeconomic impact of skin disease in lupus erythematosus. Lupus,1997;6(2):96-104
Morrow WJW, Nelson L, Watts R, Isenberg DA. Autoimmuune rheumatic disease. 2nd ed.Oxford :Oxford University Press,1999；328-346
Trowsdale J. Major histocopatibility complex .Res-Immunol. 1997；148（6）：425-426
广西年鉴社编辑.广西年鉴.广西：广西年鉴出版社,1995，127.
Marsh SGE, Bodmer JG. HLA class region nucleotide sequences. Tissue Antigens，1995，46：258-280.
龙桂芳，阿卜迪，李卫等.应用序列特异引物PCR法对广西壮族HLA-DQA1进行基因分型.中华血液学杂志，1995；16（10）：532-534
江三多，吕宝忠.医学遗传数理统计方法，北京：科学技术出版社.1998；8-9
Yao Z,Kimura a,Hartung K et al. Polomorphism of HLA-DQA1 promoter region(QAP)and DRB1,QAP,DQA1,DQB1 haplotypes in SLE, SLE Study Group Members.Immunogenetics ,1993,38(6):421-429
Skarsvag S,Hansen KE,Holst, et al.Distribution of HLA class Ⅱ alleles among Scandinavian patients with systemic lupus erythematosus(SLE):an increased risk of SLE among non DRB1*03,DQA1*0501,DQB1*0201 class Ⅱ homozygotes? Tissue Antigen,1992,40(3):128-133
Cowland JB, Andersen V, Halberg P, Morling N. DNA polymorphism
of HLA class II genes in systemic lupus erythematosus. Tissue Antigens. 1994 Jan;43(1):34-37.
Reveille JD, Schrohenloher RE, Acton RT, et al. DNA analysis of HLA-DR and DQ genes in American black with systemic lupus erythematosus. Arthritis Rheum. 1989,32(10):1243-1251
Barron, KS, Silverman, ED, Gonzalesj, et al. Clinical, serologic and immunogenetic analysis. Arthritis Rheum. 1993,36:348-352
Dong RP, Kimura A, Hashimoto H, et al. Difference in HLA linked genetic background between MCTD and SLE. Tissue Antigens, 1993,41:20-25
Azizah MR,Ainoi SS,Kuak SH et. The association of the HLA class Ⅱ antigen with clinical and autoantibody expression in Malaysian Chinese patients with systemic lupus erythematosus. Asian Pac J Allergy Immunol,2001 Jun;19(2):93-100.
彭学标,徐文严,岳晓玉,等. 汉族系统性红斑狼疮患者HLA－DQA1位点多态性分析. 临床皮肤科杂志，1996；4：196-198
施为，刘稚然，谢红付. 系统性红斑狼疮患者HLA-DQ 基因与自身抗体及肾脏受累的研究. 中华皮肤科杂志，2001；34（3）：222-223
叶冬青，曹秀菁，李向培. HLA- DQA1等位基回与系统性红斑狼疮及自身抗体相关性. 中华流行病学杂志， 2001；22（5）：392-393
李晓岚，冒长峙，洪坤学等. 云南汉族系统性红斑狼疮与HLA-DRB1、DQA1、
DQB1相关性研究. 中华医学遗传学杂志，2001；18（5）：408-409
潘星华,陆建荣, 猪子英俊等. 中国汉人HLA-DQAl基因对系统性红斑狼疮的遗传易感性研究. 中国免疫学杂志. 1995，11（1）：19-22
李峰，冒长峙，邵剑春等. 云南白族系统性红斑狼疮与HLA-DR、DQ相关性的研究. 中华遗传学杂志. 1999；16（6）：612-413
李峰，冒长峙，邵剑春等.云南白族系统性红斑狠疮患者 HLA－DR、DQ多态性研究. 中华皮肤科杂志. 2000；33（1）：33
Steven G.E. Marsh, Julia G. Bodmer, HLA Class Ⅱ region nucleotide sequences, 1995. Tissue Antigen. 1995;45:258-280
古洁若,余步云,周俊宜等. 系统性红斑狼疮与HLA-DQA1基因的相关性. 中山医科大学学报. 1996；19（4）：286-289
Galeazzi M, Sebastiani GD, Morozzi G et al. HLA Class II DNA Typing in a Large Series of European Patients with Systemic Lupus Erythematosus: Correlations with Clinical and Autoantibody Subsets.Medicine,2002;81(3)：169-178
Olsen ML,Arnett FC,Reveille JD, et al . Contrasting molecular patterns of MHC class Ⅱ alleles associated with the anti-Sm and anti-RNP precipitin autoantibodies in systemic lupus erythematosus . Arthritis Rheum ，1993，36(1):94-104
彭学标,徐文严,费虹明,等.系统性红斑狼疮患者自身抗体与HLA-DR、DQ基因的相关性研究.中华皮肤科杂志，1998；31（1）：9-11
施为，刘稚然，谢红付. 湖南地区系统性红斑狠疮患者 HLA-DQ等位基因
相关性的研究. 中华皮肤科杂志，2000；33（1）：34
李晓岚，冒长峙，李学平，杨昭庆，李梅章，褚嘉. 云南汉族系统性红斑狼疮患者抗dsDNA抗体与HLA-Ⅱ类基因的相关性研究.中华皮肤科杂志，2001；34（4）：288-289
Morrow WJW, Nelson L, Watts R, Isenberg DA. Autoimmuune rheumatic disease. 2nd ed.Oxford :Oxford University Press,1999；328-346
Trowsdale J. Major histocopatibility complex .Res-Immunol. 1997；148（6）：425-426
广西年鉴社编辑.广西年鉴.广西：广西年鉴出版社,1995，127.
Marsh SGE, Bodmer JG. HLA class region nucleotide sequences. Tissue Antigens，1995，46：258-280.
龙桂芳，阿卜迪，李卫等.应用序列特异引物PCR法对广西壮族HLA-DQA1进行基因分型.中华血液学杂志，1995；16（10）：532-534
江三多，吕宝忠.医学遗传数理统计方法，北京：科学技术出版社.1998；8-9
Yao Z,Kimura a,Hartung K et al. Polomorphism of HLA-DQA1 promoter region(QAP)and DRB1,QAP,DQA1,DQB1 haplotypes in SLE, SLE Study Group Members.Immunogenetics ,1993,38(6):421-429
Skarsvag S,Hansen KE,Holst, et al.Distribution of HLA class Ⅱ alleles among Scandinavian patients with systemic lupus erythematosus(SLE):an increased risk of SLE among non DRB1*03,DQA1*0501,DQB1*0201 class Ⅱ homozygotes? Tissue Antigen,1992,40(3):128-133
Cowland JB, Andersen V, Halberg P, Morling N. DNA polymorphism
of HLA class II genes in systemic lupus erythematosus. Tissue Antigens. 1994 Jan;43(1):34-37.
Reveille JD, Schrohenloher RE, Acton RT, et al. DNA analysis of HLA-DR and DQ genes in American black with systemic lupus erythematosus. Arthritis Rheum. 1989,32(10):1243-1251
Barron, KS, Silverman, ED, Gonzalesj, et al. Clinical, serologic and immunogenetic analysis. Arthritis Rheum. 1993,36:348-352
Dong RP, Kimura A, Hashimoto H, et al. Difference in HLA linked genetic background between MCTD and SLE. Tissue Antigens, 1993,41:20-25
Azizah MR,Ainoi SS,Kuak SH et. The association of the HLA class Ⅱ antigen with clinical and autoantibody expression in Malaysian Chinese patients with systemic lupus erythematosus. Asian Pac J Allergy Immunol,2001 Jun;19(2):93-100.
彭学标,徐文严,岳晓玉,等. 汉族系统性红斑狼疮患者HLA－DQA1位点多态性分析. 临床皮肤科杂志，1996；4：196-198
施为，刘稚然，谢红付. 系统性红斑狼疮患者HLA-DQ 基因与自身抗体及肾脏受累的研究. 中华皮肤科杂志，2001；34（3）：222-223
叶冬青，曹秀菁，李向培. HLA- DQA1等位基回与系统性红斑狼疮及自身抗体相关性. 中华流行病学杂志， 2001；22（5）：392-393
李晓岚，冒长峙，洪坤学等. 云南汉族系统性红斑狼疮与HLA-DRB1、DQA1、
DQB1相关性研究. 中华医学遗传学杂志，2001；18（5）：408-409
潘星华,陆建荣, 猪子英俊等. 中国汉人HLA-DQAl基因对系统性红斑狼疮的遗传易感性研究. 中国免疫学杂志. 1995，11（1）：19-22
李峰，冒长峙，邵剑春等. 云南白族系统性红斑狼疮与HLA-DR、DQ相关性的研究. 中华遗传学杂志. 1999；16（6）：612-413
李峰，冒长峙，邵剑春等.云南白族系统性红斑狠疮患者 HLA－DR、DQ多态性研究. 中华皮肤科杂志. 2000；33（1）：33
Steven G.E. Marsh, Julia G. Bodmer, HLA Class Ⅱ region nucleotide sequences, 1995. Tissue Antigen. 1995;45:258-280
古洁若,余步云,周俊宜等. 系统性红斑狼疮与HLA-DQA1基因的相关性. 中山医科大学学报. 1996；19（4）：286-289
Galeazzi M, Sebastiani GD, Morozzi G et al. HLA Class II DNA Typing in a Large Series of European Patients with Systemic Lupus Erythematosus: Correlations with Clinical and Autoantibody Subsets.Medicine,2002;81(3)：169-178
Olsen ML,Arnett FC,Reveille JD, et al . Contrasting molecular patterns of MHC class Ⅱ alleles associated with the anti-Sm and anti-RNP precipitin autoantibodies in systemic lupus erythematosus . Arthritis Rheum ，1993，36(1):94-104
彭学标,徐文严,费虹明,等.系统性红斑狼疮患者自身抗体与HLA-DR、DQ基因的相关性研究.中华皮肤科杂志，1998；31（1）：9-11
施为，刘稚然，谢红付. 湖南地区系统性红斑狠疮患者 HLA-DQ等位基因
相关性的研究. 中华皮肤科杂志，2000；33（1）：34
李晓岚，冒长峙，李学平，杨昭庆，李梅章，褚嘉. 云南汉族系统性红斑狼疮患者抗dsDNA抗体与HLA-Ⅱ类基因的相关性研究.中华皮肤科杂志，2001；34（4）：288-289