Further characterization of marek's disease virus‐infected lymphocytes. I. In vivo infection
Further characterization of marek's disease virus‐infected lymphocytes. I. In vivo infection
复制标题
马立克氏病病毒感染淋巴细胞的进一步表征 I. 体内感染。
作者：
B. Calnek;K. Schat;L. Ross;W. Shek;C.
本实验室以前的报道表明，法氏囊来源的淋巴细胞(B细胞)和非B细胞分别是鸡马立克氏病病毒(MDV)早期溶细胞和随后潜伏感染的主要细胞类型。目前尚不清楚这些差异是定性的还是定量的，也不知道检测潜伏感染的方法(体外培养48小时后产生病毒抗原)是否足够敏感。为了进一步确定参与MDV感染不同阶段的细胞，我们使用了与B细胞、T细胞或携带1a抗原的细胞发生特异性反应的单抗。对MDV感染鸡新鲜感染的脾细胞或体外培养后的脾细胞进行双重荧光检测，以检测表面标志和病毒内部抗原(VIA)。同样的抗体也被用于玫瑰花环程序，以产生富含或耗尽T细胞、B细胞或含La细胞的部分。通过原位杂交或斑点杂交直接检测这些细胞的病毒DNA，并进行通孔培养。我们了解到，感染的T细胞也包括MDV感染的早期细胞溶解阶段的一部分，但在感染后3或4天，与B细胞(83-92%)相比，感染的T细胞构成少数群体(约2-3%)。虽然少数(2-4%)是B细胞，但潜伏感染的细胞被确定为主要是携带la的T细胞。在体外培养前，潜伏感染细胞明显缺乏用于原位杂交检测的病毒DNA，但更灵敏的斑点杂交程序显示，体外培养后VIA表达阳性的部分细胞中病毒DNA呈阳性。培养48h后，由于原位杂交检测到感染细胞，病毒DNA在潜伏感染细胞中明显复制。用碘脱氧尿嘧啶核苷、12-O-十四酰佛波醇-13-乙酸酯或正丁酸处理细胞，均不能增加脾细胞VIA的表达。
Previous reports from this laboratory identified bursa‐derived lymphocytes (B cells) and non‐B cells as the predominant cell types respectively involved in the early cytolytic and subsequent latent infection of chickens with Marek's disease virus (MDV). It was not known whether these differences were qualitative or quantitative or if the method for detection of latent infection (viral antigen production after 48 h of in vitro cultivation) was sensitive enough. To further define the cells involved in the various phases of MDV infection, we used monoclonal antibodies which specifically react with B cells, or T cells, or la‐antigen‐bearing cells. Dual fluorescence tests to detect surface markers and viral internal antigen (VIA) were conducted with infected spleen cell freshly collected from MDV‐infected chickens or after in vitro cultivation of those cells. The same antibodies were also used for a rosetting procedure to yield fractions enriched or depleted of T cells, B cells or la‐bearing cells. These were examined directly for viral DNA by in situ hybridization or dot blot DNA hybridization and for VIA cultivation. We learned that infected T cells also comprise part of the early cytolytic phase of MDV infection but constitute a minority population (approximately 2–3%) compared to B cells (83–92%) at 3 or 4 days post infection. Latently infected cells were definitively identified as mostly la‐bearing T cells, although a few (2–4%) were B cells. Prior to in vitro cultivation, latently infected cells apparently had insufficient viral DNA for detection by in situ hybridization, but the more sensitive dot blot procedure revealed viral DNA in fractions later found positive by VIA expression after in vitro cultivation. Viral DNA replication in latently infected cells apparently had occured after 48 h cultivation because in situ hybridization detected infected cells at that time. Treatment of cell cultures with iodo‐deoxyuridine, 12‐O‐tetradecanoyl phorbol‐13‐acetate or n‐butyrate failed to increase the number of spleen cells which expressed VIA.
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马立克氏病病毒和火鸡疱疹病毒的潜伏感染。
DOI：
--
发表时间：
1981
期刊：
Journal of the National Cancer Institute
影响因子：
--
作者：
Calnek,BW;Shek,WR;Schat,KA
通讯作者：
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DOI：
--
发表时间：
1982
期刊：
Journal of the National Cancer Institute
影响因子：
--
作者：
Schat,KA;Chen,CL;Shek,WR;Calnek,BW
通讯作者：
Calnek,BW
影响因子：
4.4
作者：
K. Schat;B. Calnek;J. Fabricant;H. Abplanalp
通讯作者：
K. Schat;B. Calnek;J. Fabricant;H. Abplanalp
马立克氏病病毒体外感染淋巴细胞。
DOI：
--
发表时间：
1982
期刊：
Journal of the National Cancer Institute
影响因子：
--
作者：
Calnek,BW;Schat,KA;Shek,WR;Chen,CL
通讯作者：
Chen,CL
DOI：
--
发表时间：
1983-03
期刊：
Journal of the National Cancer Institute
影响因子：
--
作者：
W. Shek;Calnek Bw;Schat Ka;Chen-lo H. Chen-Chen-lo-H.-Chen-2109482134
通讯作者：
W. Shek;Calnek Bw;Schat Ka;Chen-lo H. Chen-Chen-lo-H.-Chen-2109482134