Colina F, Jucá N T, Moreno E, Ballestín C, Fariña J, Nevado M, Lumbreras C, Gómez-Sanz R
Department of Pathological Anatomy, Hospital Universitario 12 de Octubre, Madrid, Spain.
J Clin Pathol. 1995 Apr;48(4):351-7. doi: 10.1136/jcp.48.4.351.
AIMS--To determine the incidence of histologically documented cytomegalovirus (CMV) hepatitis following orthotopic liver transplantation (OLT) and to assess the effectiveness of immunohistochemistry and in situ hybridisation (ISH) in detecting CMV. To describe the histological pattern most frequently associated with CMV hepatitis in order to select the biopsy group in which these modern techniques are most effective. METHODS--A prospective histological study was carried out on 853 biopsy specimens, obtained from 191 liver allografts (160 patients). Specimens were stained with haematoxylin and eosin and immunohistochemically (avidin-biotin complex) using monoclonal antibodies directed against early and late CMV antigens. A retrospective selection was made of 23 specimens with viral inclusion bodies in cytomegalic cells (group A) to characterise the most frequently associated histological pattern, and of 34 other specimens without viral inclusion bodies (group B) but with the same microscopic features as group A. Re-cuts from both specimen groups were studied using immunohistochemistry and ISH with a CMV specific complementary DNA probe. RESULTS--CMV infection was confirmed in 35 specimens (29 by immunohistochemistry, 23 by presence of inclusion bodies in haematoxylin and eosin stained sections, 16 by ISH) from 27 patients (incidence 16.9%). CMV hepatitis was diagnosed within 46 +/- 19 (range 21-114) days posttransplant. Twenty on (91.3%) of the 23 biopsy specimens with inclusion bodies (group A) displayed heterogeneous inflammatory foci disseminated throughout the hepatic lobule. Nineteen specimens (82.6%) were positive by immunohistochemistry and 14 (60.9%) by ISH. In eight (23.5%) of the 34 group B specimens CMV infection was confirmed by immunohistochemistry (n = 6) or ISH (n = 2). Another 12 (35.3%) of the group B specimens negative on staining with haematoxylin and eosin, immunohistochemistry and ISH came from allografts in which previous or subsequent biopsy specimens were CMV positive. CONCLUSIONS--Demonstration of cytomegalic inclusion bodies in haematoxylin and eosin sections is sufficient for a diagnosis of CMV hepatitis. The routine use of immunohistochemistry in all allograft biopsy specimens in more sensitive than demonstration of inclusion bodies by staining with haematoxylin and eosin but may yield false negative results because of the focal distribution of positive cells. ISH was less sensitive than staining with haematoxylin and eosin and/or immunohistochemistry. A histological picture of "disseminated focal hepatitis" without viral inclusion bodies selects a group of allograft biopsy specimens in which immunohistochemistry and/or ISH may improve detection of CMV.
目的——确定原位肝移植（OLT）后组织学确诊的巨细胞病毒（CMV）肝炎的发病率，并评估免疫组织化学和原位杂交（ISH）检测CMV的有效性。描述与CMV肝炎最常相关的组织学模式，以便选择这些现代技术最有效的活检组。方法——对取自191例肝移植（160例患者）的853份活检标本进行前瞻性组织学研究。标本用苏木精和伊红染色，并使用针对CMV早期和晚期抗原的单克隆抗体进行免疫组织化学（抗生物素蛋白-生物素复合物）染色。回顾性选择23例在巨细胞内有病毒包涵体的标本（A组）以确定最常相关的组织学模式，以及另外34例无病毒包涵体但具有与A组相同显微镜特征的标本（B组）。使用免疫组织化学和ISH以及CMV特异性互补DNA探针研究两组标本的再次切片。结果——在27例患者的35份标本中确诊为CMV感染（29份通过免疫组织化学确诊，23份通过苏木精和伊红染色切片中存在包涵体确诊，16份通过ISH确诊）（发病率16.9%）。CMV肝炎在移植后46±19（范围21 - 114）天内被诊断。23份有包涵体的活检标本（A组）中有20份（91.3%）显示在肝小叶内散在分布的异质性炎性病灶。19份标本（82.6%）免疫组织化学呈阳性，14份（60.9%）ISH呈阳性。在34份B组标本中，8份（23.5%）通过免疫组织化学（n = 6）或ISH（n = 2）确诊为CMV感染。另外12份（35.3%）B组标本苏木精和伊红染色、免疫组织化学和ISH均为阴性，这些标本来自之前或之后的活检标本为CMV阳性的移植肝。结论——苏木精和伊红切片中巨细胞包涵体的显示足以诊断CMV肝炎。在所有移植肝活检标本中常规使用免疫组织化学比苏木精和伊红染色显示包涵体更敏感，但由于阳性细胞的局灶性分布可能产生假阴性结果。ISH比苏木精和伊红染色和/或免疫组织化学敏感性低。没有病毒包涵体的“弥漫性局灶性肝炎”组织学表现选择了一组移植肝活检标本，在其中免疫组织化学和/或ISH可能提高CMV的检测率。
