Lidin B, Gartland L, Marshall G, Sanchez V, Lamon E W
Birmingham Veterans Administration Medical Center, Department of Surgery, Alabama.
Viral Immunol. 1993 Summer;6(2):97-107. doi: 10.1089/vim.1993.6.97.
The intracellular expression of two early gene products of Epstein-Barr virus (EBV) was evaluated by fluorescence flow cytometry. Two Burkitt's lymphoma cell lines were superinfected by EBV from the P3HR-1 cell line. Twelve to 96 hr after superinfection the cells were fixed with paraformaldehyde and made permeable by saponin treatment. Monoclonal antibodies to the diffuse (D) and restricted (R) components of the early antigen (EA) complex were applied to the cells followed by fluorochrome-conjugated goat antibodies specific for the heavy chain isotypes of the monoclonal antibodies. Fluorescence flow cytometry revealed a clear separation in fluorescence intensity between cells containing EA-R or EA-D and negative cells. A major proportion of EA-positive cells displayed both antigens. In addition, a significant fraction expressed either EA-R or EA-D but not both. Expression of EA occurred more rapidly and peaked earlier in Daudi cells than in Raji cells. This apparent difference in EA expression between the two cell lines, however, was much less pronounced when one considered the proportion of cells expressing EA-R only, EA-D only, or both EA-R and EA-D, as a percentage of the total EA expression. Parallel fluorescence microscopy experiments revealed that the variation of the ratio of EA-R to EA-D expression with time correlated well between the two methods.
通过荧光流式细胞术评估了爱泼斯坦 - 巴尔病毒（EBV）两种早期基因产物的细胞内表达。两种伯基特淋巴瘤细胞系被来自P3HR - 1细胞系的EBV超感染。超感染后12至96小时，用多聚甲醛固定细胞，并通过皂角苷处理使其通透。将针对早期抗原（EA）复合物的弥散（D）和局限（R）成分的单克隆抗体应用于细胞，随后是针对单克隆抗体重链同种型的荧光染料偶联山羊抗体。荧光流式细胞术显示，含有EA - R或EA - D的细胞与阴性细胞之间在荧光强度上有明显分离。大部分EA阳性细胞同时显示两种抗原。此外，相当一部分细胞仅表达EA - R或EA - D中的一种，而非两种都表达。EA的表达在Daudi细胞中比在Raji细胞中更快出现且峰值更早。然而，当将仅表达EA - R、仅表达EA - D或同时表达EA - R和EA - D的细胞比例作为总EA表达的百分比来考虑时，这两种细胞系在EA表达上的明显差异就不那么显著了。平行的荧光显微镜实验表明，两种方法中EA - R与EA - D表达比例随时间的变化具有良好的相关性。
