HLA-G1及 Th17／Tregs 轴相关免疫调节分子在 B-ALL 中的表达变化及其相关性分析

摘要

目的：观察急性B淋巴细胞白血病（B-ALL）患者外周血中人白细胞抗原G1（HLA-G1）及Th17／Tregs轴相关免疫调节分子（FOXP3、RORα、t-bet、ROR-γ、GATA-3、IL-17A、IL-23和 CD25）的基因表达变化，并分析其相关性。方法收集经 MICM分型确诊的29例 B-ALL 患者外周血样本，初发 B-ALL 14例（初发组），复发难治的 B-ALL 8例（复发组），完全缓解的 B-ALL 7例（缓解组）；同时收集10例健康志愿者外周血样本作为对照组。采用实时荧光定量 PCR 法检测各组外周血中膜结合型 HLA-G1及 Th17／Tregs 轴相关免疫调节分子（FOXP3、RORα、t-bet、ROR-γ、GATA-3、IL-17A、IL-23和 CD25）的 mRNA 相对表达水平。结果初发组、复发组、缓解组、对照组膜结合型HLA-G1 mRNA 相对表达水平分别为0．415％、0．565％、0．108％、0．110％，初发组、复发组与缓解组及对照组比较， P 均＜0．05。初发组 IL-17A、IL-23、t-bet 和 CD25基因表达水平显著高于复发组（P 均＜0．05）。初发组的 FOXP3、IL-17A、IL-23、t-bet、GATA3和 CD25基因表达水平亦显著高于缓解组（P 均＜0．05）；复发组、初发组、缓解组 ROR-γmRNA 相对表达水平分别为2．777％、1．425％、0．089％，复发组与初发组、缓解组比较，P 均＜0．05。初发组 HLA-G1 mRNA 表达水平与 IL-17A 和 FOXP3相对表达水平呈正相关（r 分别为0．635、0．647；P 均＜0．05），且 HLA-G1 mRNA 表达水平与 IL-17A 呈正相关（r ＝0．573，P ＜0．05）。初发组 HLA-G1 mRNA 表达水平与 IL-23、ROR-γ、RORα、t-bet、GATA-3、CD25表达水平无明显相关关系（P 均＞0．05）。复发组 HLA-G1表达水平与各免疫相关调节分子均未发现明显相关性（P 均＞0．05）。缓解组 HLA-G1表达水平与 IL-17A 和 FOXP3表达水平呈负相关（r 分别为－0．857、－0．785，P 均＜0．05），而与其他免疫相关调节分子表达水平无显著相关关系（P 均＞0．05）。对照组HLA-G1表达水平与 t-bet 表达水平呈负相关（r ＝－0．665；P ＜0．05），与其他免疫相关调节分子均未见显著相关关系（P 均＞0．05）。结论初发及复发 B-ALL 患者外周血中膜结合型 HLA-G1基因表达水平显著升高，缓解后HLA-G1基因水平下降；且初发患者 HLA-G1表达水平与 IL-17A 和 FOXP3存在正相关关系，ROR-γ基因与复发 B-ALL 关系密切。HLA-G1和 ROR-γ可能参与调节 Th17／Tregs 轴的平衡，影响 B-ALL 预后。%Objective To investigate the expression changes of human leukocyte antigen-G1 (HLA-G1)and Th17 /Tregs axis associated regulatory factors (FOXP3,IL-17A,IL-23,ROR-γ,RORα,t-bet,GATA-3 and CD25)in periph-eral blood from adult patients with B-cell acute lymphocytic leukemia (B-ALL)and its correlation.Methods The mRNA expression levels of HLA-G and Th17 /Tregs axis associated regulatory factors (FOXP3,IL-17A,IL-23,ROR-γ,RORα, t-bet,GATA-3 and CD25)were detected in peripheral blood from 29 cases of MICMtype-confirmed patients with B-ALL, including 14 cases of primary patients (primary group),8 cases of patients with refractory relapse B-ALL (recurrence group)and 7 cases of patients with complete remission (CR)(remission group)by real-time quantification RT-PCR. Meanwhile,10 healthy individuals (HI)were selected as the control group.Results The expression levels of HLA-G1 gene in the primary group,recurrence group,remission group and control group were 0.415%,0.565%,0.108% and 0.110%,respectively.Significant differences were found among the primary group,recurrence group,remission group and control group (all P <0.05).The expression levels of IL-17A,IL-23,t-bet and CD25 genes in the primary group were significantly higher than those of the recurrence group (all P <0.05).The expression levels of FOXP3,IL-17A,IL-23,t-bet,GATA-3 and CD25 genes in the primary group were also significantly higher than those of the remission group (all P <0.05).The expression levels of ROR-γmRNA in the recurrence group,primary group and remission group were 2.777%, 1.425% and 0.089%,respectively (all P <0.05).The mRNA expression of HLA-G1 was positively correlated with IL-17A and FOXP3 in the primary group (r =0.635,0.647,respectively;all P <0.05),and meanwhile,the mRNA expres-sion of HLA-G1 was positively correlated with IL-17A (rs =0.573,P <0.05).No statistically significant correlation was found between the expression levels of HLA-G1 and IL-23,ROR-γ,RORα,t-bet,GATA-3 and CD25 in the primary group (all P >0.05).In the recurrence group,no statistically significant correlation was found between the HLA-G1 ex-pression and Th17 /Tregs axis associated regulatory factors (FOXP3,IL-17A,IL-23,ROR-γ,RORα,t-bet,GATA-3 and CD25)(all P >0.05).In the remission group,a negative correlation was found between the HLA-G1 expression and IL-17A (r =-0.857,P <0.05)or HLA-G1 and FOXP3 (r =-0.785,P <0.05).In the control group,the HLA-G1 ex-pression was negatively correlated with t-bet (r =-0.665,P =0.035),but was not related with the other regulatory mole-cules (all P >0.05).Conclusions HLA-G expression increases in untreated and refractory relapse B-ALL patients and decreases after CR.A positive correlation is found between the HLA-G1 expression and IL-17A,HLA-G1 and FOXP3 in the primary B-ALL patients.In the Th17 /Tregs axis associated regulatory factors,ROR-γgene is closely related with the recurrence of B-ALL.HLA-G1 and ROR-γgene may influence the balance of Th17 /Tregs and further affect the prognosis of B-ALL.