用显微切割技术和催化信号扩增法检测何杰金病瘤细胞(H/RS)中的人巨细胞病毒

为明确何杰金病 (Hodgkin’sdisease ,HD)瘤细胞中是否存在人巨细胞病毒 (humancytomegalovirus,HCMV)感染 ,采用显微切割技术结合PCR方法检测HD组织及其瘤细胞Hodgkin/Reed Sternberg(H/RS)中的HCMV核酸 ;采用免疫组织化学催化信号扩增 (catalysedsignalamplification ,CSA)法检测HD中HCMV的立即早期抗原、早期抗原和基质蛋白。结果在 5 4例HD中选取 13例HD的H/RS细胞 ,经显微切割分离后有 4例 (30 8% )经PCR扩增出HCMV的核酸 ;5 4例HD组织中 ,经PCR检测有 10例 (18 5 % )扩增出HCMV的核酸 ;CSA染色显示有 6例(11 1% )HCMV立即早期抗原和早期抗原 (DDG9/CCH2 )阳性 ,5例 (9 3 % )基质蛋白 (AAC10 )阳性。对照的 17例反应性增生淋巴结中HCMV核酸的PCR检测 ,以及三种HCMV抗原的CSA检测 ,均为阴性。表明HD组织的H/RS细胞中存在HCMV核酸和抗原 ,而反应性增生淋巴结中不存在 ,提示HCMV可能参与了何杰金病的发病过程。

Detection of Human Cytomegalovirus in Neoplastic Cells of Hodgkin's Disease by Microdissection Technique and Catalyzed Signal Amplification Method

In order to clearify whether human cytomegalovirus (HCMV)exists in neoplastic cells of Hodgkin's disease(HD), the microdissection technique, together with polymerase chain reaction, were employed for detection of HCMV DNA. The catalyzed signal amplification (CSA) method was used to detect immediate early antigen, early antigen and matrix protein of HCMV in HD tissues. Out of 54 cases of HD, 13 cases were selected to separate H /RS cells from their surroundings by microdissection technique. The 149bp fragment of HCMV was amplified from 4 of 13 cases (30.8%) in microdissected H/RS cells and from 10 of 54 cases(18.5%) in tissues of HD by PCR. With immunohistochemistry stain of CSA, immediate early antigen and early antigen of HCMV (DDG9/CCH2) were detected in 6 of 54 cases (11.1%), matrix protein of HCMV (AAC10) was detected in 5 of 54 cases (9.3%). In contrast, there were no detectable DNA and the three antigens of HCMV in 17 cases of reactive lymph nodes. The results indicate that human cytomegalovirus infection exists in H/RS cells in some cases of HD, and HCMV might be an agent for development of HD.