干扰素诱导蛋白16-干扰素基因刺激因子通路在柯萨奇病毒A6型感染手足口病患儿的表达及其临床意义

Expression and clinical significance of interferon gamma inducible protein 16-stimulator of interferon genes pathway in children with hand,foot and mouth disease infected by Coxsackie virus A6

目的探索干扰素诱导蛋白16(IFI16)和干扰素基因刺激因子(STING)在柯萨奇病毒A6型(CV-A6)感染手足口病(HFMD)患儿的表达及其临床意义。方法收集2023年5月至8月西安交通大学第二附属医院、西安市儿童医院和西安市中心医院收治的CV-A6感染HFMD患儿外周血标本55例,同时匹配收集同期健康体检儿童血清20例作为对照。入组HMFD患儿根据疾病轻重程度分为轻症、重症患儿。采用酶联免疫吸附法(ELISA)检测不同严重程度、急性期与恢复期患儿IFI16、环化GMP-AMP合酶(cGAS)、STING、干扰素调节因子3(IRF-3)和α干扰素(IFN-α)的表达水平。结果与对照组相比,轻症CV-A6 HFMD患儿IFI16和STING的表达均升高,而在重症CV-A6 HFMD患儿中则表达下降[15.92(13.34,19.13)ng/ml vs.13.66(11.91,14.83)ng/ml:Z=-2.200、P=0.028;1345.45(991.55,1843.63)pg/ml vs.1072.26(947.25,1180.97)pg/ml:Z=-2.000、P=0.046],仅STING在重症CV-A6型HFMD患儿急性期与恢复期间表达差异具有统计学意义[1072.26(947.25,1180.97)vs.1665.29(1341.62,1961.83):Z=-3.237、P=0.001];与对照组相比,IFN-α在轻症CV-A6 HFMD患儿表达降低[864.47(721.41,952.89)pg/ml vs.715.08(575.41,896.69)pg/ml,Z=-2.054、P=0.040];但cGAS、IRF3在对照组、轻症和重症HFMD患儿间表达差异均无统计学意义(P均>0.05)。Sperman相关性分析显示,IFI16和STING表达具有正相关性(r=0.286、P=0.013)。临床特征方面,神经受累(病理反射阳性)患儿IFI16(Z=-3.307、P=0.001)和STING(Z=-2.702、P=0.007)水平均低于病理反射阴性患儿,有肢体抽动(Z=-2.489、P=0.013)、精神差(Z=2.542、P=0.011)和高血糖水平(Z=-2.828、P=0.005)患儿的IFI16水平也低于无以上特征的患儿,IFI16与血糖水平也具有负相关性(Sperman相关性分析:r=-0.427、P=0.001)。结论CV-A6感染可激活IFI16-STING通路。IFI16和STING的表达与CV-A6型HFMD的重症化相关联,其较高的表达水平可能是机体抵御重症化的保护因素。

Objective To investigate the expression and clinical significance of interferon gamma inducible protein 16(IFI16)and stimulator of interferon genes(STING)of children with hand,foot and mouth disease(HFMD)infected by Coxsackie virus A6(CV-A6).Methods The peripheral blood samples of 55 children with CV-A6 HFMD admitted to the Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an Children's Hospital and Xi'an Central Hospital from May to August 2023 were collected,and 20 samples were collected from healthy children as controls.According to the severity of the disease,the children were divided into mild and severe cases.Enzyme-linked immunosorbent(ELISA)assay was used to detect the expression levels of IFI16,Cyclic GMP-AMP Synthase(cGAS),STING,interferon regulatory factor 3(IRF-3)and interferon-α(IFN-α)in different severity,acute phase and recovery phase.Results Compared with control group,the expression of IFI16 and STING increased in mild CV-A6 HFMD children,and decreased in severe CV-A6 HFMD children[15.92(13.34,19.13)ng/ml vs.13.66(11.91,14.83)ng/ml:Z=-2.200,P=0.028;1345.45(991.55,1843.63)pg/ml vs.1072.26(947.25,1180.97)pg/ml:Z=-2.000,P=0.046].Only the expression of STING in children with severe CV-A6 HFMD was significantly different between the acute stage and the recovery stage[1072.26(947.25,1180.97)vs.1665.29(1341.62,1961.83):Z=-3.237,P=0.001].Compared with the control group,the expression of IFN-α in the mild CV-A6 HFMD group decreased[864.47(721.41,952.89)pg/ml vs.715.08(575.41,896.69)pg/ml:Z=-2.054,P=0.040].There was no significant difference in the expression of cGAS and IRF3 among the control group,mild HFMD group and severe HFMD group(all P>0.05).Sperman correlation analysis showed that IFI16 expression was positively correlated with STING expression(r=0.286,P=0.013).The expression of IFI16(Z=-3.307,P=0.001)and STING(Z=-2.702,P=0.007)in children with nerve involvement(positive pathological reflex)were lower than those in children with negative pathological reflex.Children with limb twitching(Z=-2.489,P=0.013),poor mental state(Z=-2.542,P=0.011),and high blood glucose level(Z=-2.828,P=0.005)also had lower IF116 levels than those without such characteristics,and IFI16 was negatively correlated with blood glucose levels(Sperman correlation analysis:r=-0.427,P=0.001).Conclusions CV-A6 infection can activate the IFI16-STING pathway.The expression of IFI16 and STING are associated with the severity of CV-A6-induced HFMD,and their higher expression levels may be a protective factor of the body against the progression to severe disease.