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网状内皮增生症病毒转化的淋巴母细胞系的稳定转染

Stable transfection of reticuloendotheliosis virus-transformed lymphoblastoid cell lines.

作者信息

Schat K A, Pratt W D, Morgan R, Weinstock D, Calnek B W

机构信息

Department of Avian and Aquatic Animal Medicine, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853.

出版信息

Avian Dis. 1992 Apr-Jun;36(2):432-9.

PMID:1320872
Abstract

Lymphoblastoid T cell lines were established by infection of chicken splenocytes with reticuloendotheliosis virus (REV). The target cells first were cultured in interleukin-containing conditioned medium or were stimulated by concanavalin A, or both. Most cell lines were T cells expressing CD3 and one of the T cell receptors, and all cell lines were positive for major histocompatibility complex (MHC) class II antigens. Several REV-transformed cell lines were stably transfected using electroporation with a selectable plasmid, pNL1, containing the neor gene. Transfected cell lines were selected using G418 and were maintained for periods up to 137 days. Transfected cell lines were susceptible to MHC class-I restricted lysis by cytotoxic T lymphocytes from REV-infected chickens.

摘要

通过用网状内皮组织增殖病病毒(REV)感染鸡脾细胞建立了淋巴母细胞样T细胞系。靶细胞首先在含白细胞介素的条件培养基中培养,或用刀豆球蛋白A刺激,或两者兼用。大多数细胞系是表达CD3和一种T细胞受体的T细胞,并且所有细胞系对主要组织相容性复合体(MHC)II类抗原呈阳性。使用含有新霉素抗性基因(neor基因)的可选择质粒pNL1通过电穿孔对几种REV转化的细胞系进行稳定转染。使用G418选择转染的细胞系,并维持长达137天。转染的细胞系易受来自REV感染鸡的细胞毒性T淋巴细胞的MHC I类限制性裂解作用。

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