【摘要】 目的探讨HCMVgB抗原血症在HCMV感染中的诊断价值以及gB诱导的特异性CD8+T细胞在控制病毒激活中的作用。方法根据移植术后时间进行分组,免疫组化法检测92份异基因造血干细胞移植术后白细胞样本中的gB、IE和pp65抗原,分析gB抗原检测在监测HCMV感染中的意义。应用SYFPEITHI和REVEALTM主要组织相容性抗原-肽结合力测定筛选出潜在T细胞表位,进而合成MHC-肽五聚体,应用基于流式细胞术的五聚体染色分析筛选出gB相对优势表位。结果 HCMV gB抗原与pp65和IE抗原检测均呈正相关,其阳性率(82.6%)和pp65抗原阳性率(90.2%)差异无显著意义(P=0.065),但比IE抗原阳性率(92.4%)低(P=0.035)。三种抗原检测阳性率在移植术后1年内无显著差异;而在移植术1年后,gB阳性率低于IE和pp65阳性率(P=0.005),并且在gB阴性而IE或pp65阳性的标本中,IE或pp65阳性值处于一个较低的范围(1~6/5×104WBC)。gB332-340可能是CD8+T细胞的免疫优势靶位,其诱导的gB特异性CD8+T细胞与gB抗原血症的发生呈现负相关。结论 HCMV gB对移植术后早期诊断感染有潜在的价值,并可能是一个提示病毒感染能力的指标;在对长期随访病人HCMV感染的监测中,gB与pp65、IE正相关,或许成为诊断HCMV感染的重要补充;gB特异性CD8+T细胞的免疫应答在控制HCMV感染中可能起到重要作用。更多还原

【Abstract】 Objective To investigate the diagnostic value of human cytomegalovirus(HCMV) glycoprotein B(gB) antigenaemia in HCMV infection and study the role of gB-specifc CD8+T cells in the control of HCMV activation.Methods According to the time of posttransplantation,the positive rates of gB,immediate-early(IE) and phosphoprotein 65(pp65) antigen in 92 leukocyte samples from 64 allogeneic hematopoietic stem cell transplantation recipients were grouped and determined by immunohistochemical staining to analyze the signifcance of gB antigen detection in diagnosis of HCMV infection.Potential gB T-cell epitopes were screened by the SYFPEITHI algorithm and REVEALTMMHC-peptide binding assay to identify the relative dominant epitope(s) by fow cytometry-pentamer staining.Results The positive numbers of gB,IE and pp65 antigens were in a positive correlation comparing between any two of them.The positive rate of HCMV gB antigen(82.6%) was lower than that of HCMV IE antigen(92.4%,P=0.035),but there was no signifcant difference between the positive rate of gB and pp65 antigens(90.2%,P=0.065).Within 1 year after transplantation,the positive rate among the gB,IE and pp65 antigens was the same,while in the group of longer than 1 year after transplantation,gB positive rate was slightly lower than both IE and pp65(P=0.005).Besides,in the gB negative but IE or pp65 positive samples,the positive rate of IE or pp65 antigens ranged in a low level(1~6/5×104WBC).The gB332-340 peptide may be identifed as an immunodominant target recognized by HCMVspecifc CD8+T cells.There was a negative correlation between the frequency of gB332-340-specifc CD8+T cells and the level of gB antigenaemia.Conclusion HCMV gB had a potential application of early diagnosis in patients of allogeneic hematopoietic stem cell transplantation and maybe indicate the infectious ability of HCMV.Probably,gB was a candidate for diagnosing HCMV infection in the long term follow-up monitoring since there was a positive correlation between gB and pp65 or IE.In addition,gB-specifc CD8+ T-cell immunity may play an important role in the control of HCMV infection.更多还原