Toll样受体基因多态性与福建籍非人类免疫缺陷病毒相关隐球菌性脑膜炎易感性的关联分析

Association between Toll-like receptor gene polymorphism and susceptibility to non-human immunodeficiency virus-related cryptococcal meningitis in patients from Fujian Province

目的:探讨Toll样受体(Toll-like receptor,TLR)基因多态性与福建籍非人类免疫缺陷病毒(human immunodeficiency virus,HIV)相关隐球菌性脑膜炎易感性的关联。方法:纳入2016年10月至2019年4月在上海市复旦大学附属华山医院和福建省福州市联勤保障部队第九〇〇医院仓山院区住院治疗的101例福建籍非HIV相关隐球菌性脑膜炎患者为病例组,270名福建籍门诊健康体格检查者为健康对照组。提取患者基因组DNA,采用多重SNaPshot单核苷酸多态性(single nucleotide polymorphism,SNP)分型技术,对既往文献报道的与疾病相关但未得到充分验证的8个TLR SNP位点进行基因分型。分别比较病例组和健康对照组、无易感因素患者组和健康对照组TLR基因多态性的分布差异。统计学分析采用χ^(2)检验或Fisher确切概率法。结果:除TLR1 rs5743563外,TLR1 rs5743604、TLR2 rs3804099、TLR4 rs1927907、TLR6 rs3796508、TLR6 rs5743794、TLR9 rs164637、TLR9 rs3521407个TLR SNP检测位点的等位基因频率分布均符合哈迪-温伯格平衡。病例组与健康对照组比较发现,TLR2 rs3804099位点的T/T基因型[52.5%(53/101)比40.4%(109/270),比值比(odds ratio,OR)=1.63,χ^(2)=4.378,P=0.036]和TLR6 rs5743794位点的G/G基因型[44.6%(45/101)比32.2%(87/270),OR=1.69,χ^(2)=4.877,P=0.027]为隐球菌性脑膜炎的风险基因型;而TLR6 rs3796508的G/G基因型[83.2%(84/101)比92.6%(250/270),OR=0.40,χ^(2)=7.271,P=0.007]和TLR9 rs164637的C/C基因型[96.0%(97/101)比100.0%(270/270),Fisher确切概率法,P=0.005]为隐球菌性脑膜炎的保护性因素。101例患者中有70例无易感因素。无易感因素患者组与健康对照组比较发现,TLR6 rs5743794位点的G/G基因型[52.9%(37/70)比32.2%(87/270),OR=2.36,χ^(2)=10.216,P=0.001]为隐球菌性脑膜炎的风险基因型,TLR6 rs3796508的G/G基因型[81.4%(57/70)比92.6%(250/270),OR=0.35,χ^(2)=7.906,P=0.005]和TLR9 rs164637的C/C基因型[97.1%(68/70)比100.0%(270/270),Fisher确切概率法,P=0.042]为隐球菌性脑膜炎的保护性因素。结论:TLR基因多态性与福建籍非HIV相关隐球菌性脑膜炎的易感性密切关联,提示TLR在隐球菌性脑膜炎的发病过程中可能起到重要作用。

Objective To explore the association between Toll-like receptor(TLR)gene polymorphism and susceptibility to non-human immunodeficiency virus(HIV)-related cryptococcal meningitis in patients from Fujian Province.Methods A total of 101 non-HIV patients with cryptococcal meningitis and 270 healthy controls in Huashan Hospital,Fudan University and Cangshan Hospital District,The 900th Hospital of the Joint Logistics Support Force from Fujian Province were prospectively enrolled in this case-control study.Genomic DNA was extracted.Genotyping was performed by multiplex SNaPshot technology on eight TLR single nucleotide polymorphism(SNP)which were reported to be related to cryptococcal meningitis but still lacking validations.The differences of gene polymorphism distributions were compared between all patients and healthy controls,and between patients without predisposing factors and healthy controls.Data were analyzed by chi-square test or Fisher exact test.Results Except for TLR1 rs5743563,the distributions of allele frequency in seven tested TLR SNPs(TLR1 rs5743604,TLR2 rs3804099,TLR4 rs1927907,TLR6 rs3796508,TLR6 rs5743794,TLR9 rs164637 and TLR9 rs352140)were in Hardy-Weinberg equilibrium.Comparisons between cases and controls found that TLR2 rs3804099 T/T genotype(52.5%(53/101)vs 40.4%(109/270),odds ratio(OR)=1.63,χ^(2)=4.378,P=0.036)and TLR6 rs5743794 G/G genotype(44.6%(45/101)vs 32.2%(87/270),OR=1.69,χ^(2)=4.877,P=0.027)were correlated with high risks of cryptococcal meningitis,while TLR6 rs3796508 G/G genotype((83.2%(84/101)vs 92.6%(250/270),OR=0.40,χ^(2)=7.271,P=0.007)and TLR9 rs164637 C/C genotype(96.0%(97/101)vs 100.0%(270/270),Fisher exact test,P=0.005)were found to be protective factors.Seventy out of 101 patients had no predisposing factors.Comparison between patients without predisposing factors and healthy controls also found similar results.TLR6 rs5743794 G/G genotype(52.9%(37/70)vs 32.2%(87/270),OR=2.36,χ^(2)=10.216,P=0.001)was risk factor,while TLR6 rs3796508 G/G genotype(81.4%(57/70)vs 92.6%(250/270),OR=0.35,χ^(2)=7.906,P=0.005)and TLR9 rs164637 C/C genotype(97.1%(68/70)vs 100.0%(270/270),Fisher exact test,P=0.042)were protective factors.Conclusion TLR gene polymorphism is significantly associated with non-HIV-related cryptococcal meningitis,which indicates that TLR might play an important role in the pathogenesis of cryptococcal meningitis.