Dienglewicz R L, Parcells M S
Department of Poultry Science, University of Arkansas, Fayetteville 72701, USA.
Acta Virol. 1999 Apr-Jun;43(2-3):106-12.
We have previously described the construction and characterization of mutant Marek's disease viruses (MDVs) having mutations within the unique-short (US) region of the genome that have retained oncogenicity (Anderson et al., 1998; Parcells et al., 1995). We have also reported the characterization of lymphoblastoid cell lines (LBCLs) derived using these mutant viruses (Parcells et al., 1998). These mutant MDVs were constructed using a lacZ expression cassette. Expression of lacZ was found to be constitutive during lytic infection but was found to be tightly repressed in tumors and the derived LBCLs. The construction of these viruses and the analysis of lacZ induction required the use of toxic substrates or antibody staining to detect lacZ expression. We now report the establishment of an MDV lymphoblastoid cell line, MDCC-UA04, that was derived from a tumor induced by an MDV having an insertion of a green fluorescent protein expression cassette into the US2 gene. Like previous mutant-derived LBCLs, expression of the marker cassette is constitutive in lytic infection, but repressed in tumors and in the UA04 cells. UA04 cells express CD3low, CD4, TCR-2low, MHC class II, and CD28 antigens on their surface. The percentage of UA04 cells expressing GFP is generally low (5-7%), but increases markedly within 48 hrs of 5'-iododeoxyuridine (IUdR) treatment (20-30%) in a manner similar to many MDV lytic antigens. Thus, induction of GFP expression in UA04 cells can serve as a non-invasive marker for MDV reactivation from latency.
我们之前描述了在基因组独特短区域（US）内发生突变但仍保留致癌性的马立克氏病病毒（MDV）突变体的构建和特性（Anderson等人，1998年；Parcells等人，1995年）。我们还报道了使用这些突变病毒衍生的淋巴母细胞系（LBCL）的特性（Parcells等人，1998年）。这些突变MDV是使用lacZ表达盒构建的。发现lacZ在裂解感染期间组成性表达，但在肿瘤和衍生的LBCL中被紧密抑制。这些病毒的构建以及lacZ诱导的分析需要使用有毒底物或抗体染色来检测lacZ表达。我们现在报告建立了一个MDV淋巴母细胞系MDCC-UA04，它源自由一种MDV诱导的肿瘤，该MDV在US2基因中插入了绿色荧光蛋白表达盒。与之前源自突变体的LBCL一样，标记盒在裂解感染中组成性表达，但在肿瘤和UA04细胞中受到抑制。UA04细胞在其表面表达CD3low、CD4、TCR-2low、MHC II类和CD28抗原。表达GFP的UA04细胞百分比通常较低（5-7%），但在5'-碘脱氧尿苷（IUdR）处理后48小时内显著增加（20-30%），其方式类似于许多MDV裂解抗原。因此，UA04细胞中GFP表达的诱导可作为MDV从潜伏状态重新激活的非侵入性标记。
