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仙台病毒包膜介导的基因转移后人淋巴细胞中EB病毒（EBV）DNA和克隆DNA片段的表达

Expression of Epstein-Barr virus (EBV) DNA and cloned DNA fragments in human lymphocytes following Sendai virus envelope-mediated gene transfer.

作者信息

Volsky D J, Gross T, Sinangil F, Kuszynski C, Bartzatt R, Dambaugh T, Kieff E

出版信息

Proc Natl Acad Sci U S A. 1984 Oct;81(19):5926-30. doi: 10.1073/pnas.81.19.5926.

DOI:10.1073/pnas.81.19.5926
PMID:6091119
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC391831/
Abstract

Purified EBV DNA and cloned DNA fragments were trapped in Sendai virus (SV) envelopes during envelope reconstitution. The DNA-loaded reconstituted envelopes (RSVE/DNA) served as gene-transfer vehicles using the capability of RSVE to fuse with normal and tumor cells. The efficiency of RSVE-mediated EBV DNA transfer into lymphoid tumor cells and fresh human lymphocytes was 5-10% of the enveloped 3H-labeled EcoRI fragment B of EBV DNA. Purified intracellular EBV (B95-8 strain) DNA induced EBV nuclear antigen (EBNA) in 0.2-1% of human lymphocytes, transiently stimulated cellular DNA synthesis, but did not fully transform cells. Cloned Sal I F1 fragment [approximately equal to 9 kilobase pairs (kbp)] and a smaller BamHI K (5.2 kbp) fragment from the same region of B95-8 EBV DNA induced EBNA in 2-4% of human lymphocytes but did not stimulate DNA synthesis nor transform cells. Cloned BamHI D1 fragment (approximately equal to 9 kbp) from AG-876 virus DNA, or a combination of cloned BamHI X and H fragments (approximately equal to 2 and 7 kbp, respectively) from the similar region of B95-8 virus DNA, significantly stimulated lymphocyte DNA synthesis, but EBNA could not be detected and transformation was not achieved. Early antigen and viral capsid antigen were not observed with any of the fragments tested. Our results suggest that the induction of EBNA and stimulation of lymphocyte proliferation are not controlled by the same region of EBV DNA.

摘要

在包膜重构过程中，纯化的EBV DNA和克隆的DNA片段被捕获在仙台病毒（SV）包膜中。负载DNA的重构包膜（RSVE/DNA）利用RSVE与正常细胞和肿瘤细胞融合的能力作为基因传递载体。RSVE介导的EBV DNA转移到淋巴瘤细胞和新鲜人淋巴细胞中的效率为包膜化的3H标记的EBV DNA EcoRI片段B的5-10%。纯化的细胞内EBV（B95-8株）DNA在0.2-1%的人淋巴细胞中诱导EBV核抗原（EBNA），短暂刺激细胞DNA合成，但未完全转化细胞。来自B95-8 EBV DNA同一区域的克隆Sal I F1片段[约9千碱基对（kbp）]和较小的BamHI K（5.2 kbp）片段在2-4%的人淋巴细胞中诱导EBNA，但不刺激DNA合成也不转化细胞。来自AG-876病毒DNA的克隆BamHI D1片段（约9 kbp），或来自B95-8病毒DNA相似区域的克隆BamHI X和H片段（分别约2和7 kbp）的组合，显著刺激淋巴细胞DNA合成，但未检测到EBNA且未实现转化。在所测试的任何片段中均未观察到早期抗原和病毒衣壳抗原。我们的结果表明，EBNA的诱导和淋巴细胞增殖的刺激不受EBV DNA同一区域的控制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/501a/391831/d5832967688c/pnas00620-0025-e.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/501a/391831/2ff46761ccd7/pnas00620-0025-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/501a/391831/77039a4b189b/pnas00620-0025-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/501a/391831/fb16ed7fb6e3/pnas00620-0025-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/501a/391831/63d4bec28965/pnas00620-0025-d.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/501a/391831/d5832967688c/pnas00620-0025-e.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/501a/391831/2ff46761ccd7/pnas00620-0025-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/501a/391831/77039a4b189b/pnas00620-0025-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/501a/391831/fb16ed7fb6e3/pnas00620-0025-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/501a/391831/63d4bec28965/pnas00620-0025-d.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/501a/391831/d5832967688c/pnas00620-0025-e.jpg

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