新生乳鼠大脑神经胶质细胞对人巨细胞病毒易患性研究

目的探讨人巨细胞病毒（HCMV）体外感染新生Balb／c乳鼠原代培养大脑神经胶质细胞的特征及其敏感性。方法建立了新生乳鼠大脑皮质细胞原代培养方法并进行病毒感染。在感染后的不同时间，分别在光镜下连续观察摄影记录，并取样42份，进行抗HCMV单抗的免疫组化染色和地高辛标记的HCMV寡核苷酸探针检测病毒核酸及其胞内定位分布的原位杂交试验。同时，同样取正常对照组培养片36份进行试验。结果HCMV能感染新生乳鼠大脑皮质细胞，受感染的神经胶质细胞，在感染后第4天即可观察到细胞病变，表现为细胞肿胀变圆。测定感染细胞上清中病毒半数组织培养感染量，表明病毒在细胞内复制。免疫组化和原位杂交结果均证实，HCMV感染后48小时，即可分别在受感染细胞内检出病毒早期抗原和病毒核酸；且上述阳性结果的出现均比在同期受感染的神经细胞早，易观察。结论新生乳鼠大脑皮质神经胶质细胞较神经细胞对HCMV更为易感染，表明前者在HCMV致中枢神经系统感染的建立及病理的发生发展中具有重要作用。

Susceptibility of primary cultured central neurogliocytes cells from the newborn mice to human cytomegalovirus

Objective To investigate the suscptibility and feature of the central neurogliocytes fromBalb/c newborn mice by primary culture in vitro to human cytomegalovirus (HCMV). Methods The centralneurons and neurogliocytes were prepared and cultured, and the model for HCMV infection was made. The cellswere observed and recorded with photography under the microscope, and the 42 neuron cell cultures were taken forthe viral specific antigen, the viral DNA and its intracellular location were detected and analyzed with immunohistochemical stainning using HCMV specific McAbs and in situ hybridization utilizing digoxigenin labelled HCMVDNA probes, respectively, at 24 h,48 h, 72 h......1,2,3 and 4 weeks after infection. Results The cytopathiceffects of the central neurogliocytes were exhibited 4 days after HCMV postintection, that is, the cell werebeginning to enlarge and become round. The virus titer measured by TCID50 assay was proved to be on the increasein the supernatan of the infected cells. Viral eary antigens and its specific DNA sequences in the cells weredetectable at 48 hours after the infection. These observed results occurred earlier than those of the neurons.However, nothing was found in the negative controls. Conclusion The central neurogliocytes were moresusceptible than the neurons to HCMV infection. This finding in regard to the central neurogliocytes suggested thatthey might play important roles for the establishment of the HCMV infection and its pathological development in thecentral nervous system.