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

Expression and correlation of HLA-G1 and Th17/Tregs axis associated regulatory factors in B-ALL

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BALL 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HLAG1 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-G1mRNA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BALL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-17 A,IL-23,ROR-γ,RORα,t-bet,GATA-3 and CD25） in peripheral 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-17 A,IL-23,ROR-γ,RORα,t-bet,GATA-3 and CD25） were detected in peripheral blood from 29 cases of MICM type-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% and0. 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-17 A,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-17 A,IL-23,tbet,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-γ mRNA 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-17 A and FOXP3 in the primary group（ r = 0. 635,0. 647,respectively; all P〈0. 05）,and meanwhile,the mRNA expression 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 expression and Th17/Tregs axis associated regulatory factors（ FOXP3,IL-17 A,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 expression was negatively correlated with t-bet（ r =- 0. 665,P = 0. 035）,but was not related with the other regulatory molecules（ 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-17 A,HLA-G1 and FOXP3 in the primary B-ALL patients. In the Th17/Tregs axis associated regulatory factors,ROR-γ gene is closely related with the recurrence of B-ALL. HLA-G1 and ROR-γ gene may influence the balance of Th17/Tregs and further affect the prognosis of B-ALL.