Hendrix R M, Wagenaar M, Slobbe R L, Bruggeman C A
Department of Medical Microbiology, University of Maastricht, The Netherlands.
J Clin Pathol. 1997 Jan;50(1):59-63. doi: 10.1136/jcp.50.1.59.
To determine the localisation of human cytomegalovirus (CMV) DNA in abdominal aorta, spleen, and transplantable organs, such as kidney, pancreas, and liver, obtained from healthy individuals; to characterise the cell type(s) in these tissues that serve as a reservoir for latent CMV.
CMV DNA was detected by dot blot DNA hybridisation and in situ DNA hybridisation with a probe for CMV major immediate early sequences (UL123) and nested PCR with primers derived from the CMV major immediate early (IE) gene exon 4 (UL123ex4). Samples of liver, abdominal aorta, spleen, kidney, and pancreas were obtained at necropsy or from donor kidneys from healthy subjects.
CMV DNA was detected in most tissue samples using dot blot hybridisation and nested PCR. In situ hybridisation demonstrated that, in addition to smooth muscle cells in the arterial wall, hepatocytes, tubular and glomerular kidney cells, splenic red pulp cells, and pancreatic acinar cells also harboured CMV DNA. CMV DNA was detected in seropositive and in some seronegative subjects.
CMV DNA is widely distributed in organs of healthy subjects. CMV DNA was found in various cell types in several organs, suggesting that during latency, CMV DNA is present thoughout the body.
确定从健康个体获取的腹主动脉、脾脏以及可移植器官(如肾脏、胰腺和肝脏)中人类巨细胞病毒(CMV)DNA的定位;鉴定这些组织中作为CMV潜伏库的细胞类型。
采用斑点印迹DNA杂交和原位DNA杂交技术,用针对CMV主要立即早期序列(UL123)的探针进行检测,并用源自CMV主要立即早期(IE)基因外显子4(UL123ex4)的引物进行巢式PCR。在尸检时或从健康受试者的供体肾脏获取肝脏、腹主动脉、脾脏、肾脏和胰腺的样本。
使用斑点印迹杂交和巢式PCR在大多数组织样本中检测到CMV DNA。原位杂交表明,除动脉壁平滑肌细胞外,肝细胞、肾小管和肾小球细胞、脾红髓细胞以及胰腺腺泡细胞也含有CMV DNA。在血清阳性和一些血清阴性受试者中均检测到CMV DNA。
CMV DNA广泛分布于健康受试者的器官中。在多个器官的多种细胞类型中发现了CMV DNA,这表明在潜伏期间,CMV DNA遍布全身。