幼年型与成年型强直性脊柱炎HLA*B等位基因高分辨分型研究

High-resolution typing of HLA*B allele in Juvenile-onset and Adult-onset Ankylosing Spondylitis

目的 通过对幼年型强直性脊柱炎(JAS)和成年型强直性脊柱炎(AAS)患者HLA*B等位基因进行高分辨测序分型,了解HLA-B基因多态性与JAS是否有相关性。方法 收集2019年8月—2021年12月就诊于本院的符合JAS和AAS患者各50例,采用PCR-SBT法对HLA*B位点特异性扩增,再使用ABI 3100毛细管测序仪对外显子1正向测序,外显子2、3、4、5同时进行正反向测序,采用uTYPETMHLA测序分析软件处理测序结果。结果 JAS组中男41例,女9例,AAS组中男43例,女7例,两组男女性别比例差异无统计学意义(P=0.585);JAS组发病年龄(12.5±2.9)岁,显著低于AAS组的(29.7±6.8)岁(P<0.001)。JAS组检出HLA-B*27纯合子5例占10.0%,AAS组检出HLA-B*27纯合子4例占8.0%,JAS组中45例杂合子中B*27:04、B*27:05、B*27:15、B*27:86分别为34例、6例、3例、2例,AAS组46例杂合子中B*27:04、B*27:02分别为45例、1例;HLA-B*27阳性的JAS和AAS各检出15例HLA-B*40,高分辨分型中以B*40:01:02为主要亚型。结论 HLA-B*27纯合子与等位基因HLA-B*40对JAS的早发病并无直接相关性。HLA-B*27:04是JAS和AAS主要基因亚型,HLA-B*27:05:02:01、HLA-B*27:15,在JAS中多于AAS组,携带该等位基因的患者倾向于早发。

Objective In this study,HLA*B alleles of Juvenile-onset ankylosing spondylitis(JAS)and Adult-onset ankylosing spondylitis(AAS)patients were genotyped by high-resolution sequencing to understand the correlation between HLA-B gene polymorphism and JAS.Methods 50 cases of JAS and 50 cases of AAS who attended our hospital from August 2019 to December 2021 were enrolled in the study.The HLA*B allele was specifically amplified by PCR-SBT method,and then the ABI 3100 capillary sequencer was used for forward sequencing of exon 1,and forward and reverse sequencing was performed for exons 2,3,4,and 5 at the same time.Sequencing results were processed using uTYPE TM HLA software.Results There were 41 males and 9 females in the JAS group,and 43 males and 7 females in the AAS group,there was no significant difference in the sex ratio between the two groups(P=0.585).The age of onset in the JAS group(12.5±2.9)was significantly lower than that(29.7±6.8)in the AAS group(P<0.001).There were 5 cases(10.0%)of HLA-B*27 homozygotes in the JAS group,and 4 cases(8.0%)of HLA-B*27 homozygotes in the AAS group.Among 45 heterozygotes in JAS group,34 cases were B*27:04,6 cases were B*27:05,3 cases were B*27:15,and 2 cases were B*27:86,while among 46 heterozygotes in AAS group,45 cases were B*27:04,and 1 case was B*27:02.15 cases of HLA-B*40 were detected in JAS and AAS respectively,and the main subtype of high resolution was B*40:01:02.Conclusions HLA-B*27 homozygotes and HLA-B*40 alleles are not directly associated with early onset of JAS.HLA-B*2704 is the major gene subtype of JAS and AAS.HLA-B*27:05:02:01 and HLA-B*2715 are more common in JAS than in AAS,patients carrying these alleles tend to have early onset.