Stoerker J, Glaser R
Proc Natl Acad Sci U S A. 1983 Mar;80(6):1726-9. doi: 10.1073/pnas.80.6.1726.
Transfection experiments using subgenomic fragments of the B95-8 strain of Epstein-Barr virus (EBV) DNA and EBV genome (HR-1)-positive epithelial/Burkitt hybrid cells (D98/HR-1) were carried out to determine whether an interaction between the transfecting virus fragment(s) and the endogenous HR-1 EBV genome could take place. Expression of EBV-specific antigens, including early antigen and virus capsid antigen, was examined in transfected cells by immunofluorescence. Attempts were also made to recover biologically active EBV from the D98/HR-1 cells after transfection with cloned fragments of B95-8 DNA. We found that D98/HR-1 cells transfected with the BamHI H or H, F, and X fragments were positive for early antigen 3 days after transfection. Spent media from transfected D98/HR-1 cells maintained for 20-30 days in culture were pooled, filtered, concentrated, and used as a potential source of virus to inoculate human umbilical cord blood lymphocytes. No evidence of transformation was observed with such preparations. However, if spent medium from D98/HR-1 cell cultures was first treated with iododeoxyuridine (to induce EBV DNA synthesis and replicative cycle) and then transfected with the BamHI H, F, and X fragments of B95-8 DNA and used to infect cord blood lymphocytes, transformation was obtained. A lymphoblastoid cell line derived in this manner, designated HI-HFX, is an EBV nuclear antigen-positive nonproducer cell line. Similar results were obtained with preparations from iododeoxyuridine-treated D98/HR-1 cells transfected with the EB 26-36 fragment of B95-8 DNA cloned in a Charon 4A vector. The EB 26-36 fragment contains the BamHI H, F, and X regions.
利用爱泼斯坦 - 巴尔病毒（EBV）B95 - 8株DNA的亚基因组片段和EBV基因组（HR - 1）阳性上皮/伯基特杂交细胞（D98/HR - 1）进行转染实验，以确定转染的病毒片段与内源性HR - 1 EBV基因组之间是否会发生相互作用。通过免疫荧光检测转染细胞中EBV特异性抗原的表达，包括早期抗原和病毒衣壳抗原。在用B95 - 8 DNA克隆片段转染D98/HR - 1细胞后，还尝试从这些细胞中回收具有生物活性的EBV。我们发现，用BamHI H或H、F和X片段转染的D98/HR - 1细胞在转染后3天早期抗原呈阳性。将在培养中维持20 - 30天的转染D98/HR - 1细胞的用过的培养基收集、过滤、浓缩，并用作接种人脐带血淋巴细胞的潜在病毒来源。用这种制剂未观察到转化的证据。然而，如果先用碘脱氧尿苷处理D98/HR - 1细胞培养物的用过的培养基（以诱导EBV DNA合成和复制周期），然后用B95 - 8 DNA的BamHI H、F和X片段转染并用于感染脐带血淋巴细胞，就会获得转化。以这种方式衍生的一个淋巴母细胞系命名为HI - HFX，是一个EBV核抗原阳性的非生产性细胞系。用克隆在Charon 4A载体中的B95 - 8 DNA的EB 26 - 36片段转染经碘脱氧尿苷处理的D98/HR - 1细胞制备物也得到了类似结果。EB 26 - 36片段包含BamHI H、F和X区域。
