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用于研究鼠巨细胞病毒的小鼠唾液腺衍生间充质细胞系的建立

Development of a mouse salivary gland-derived mesenchymal cell line for immunological studies of murine cytomegalovirus.

机构信息

Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, Louisiana, United States of America.

出版信息

PLoS One. 2022 Aug 17;17(8):e0265479. doi: 10.1371/journal.pone.0265479. eCollection 2022.

Abstract

The salivary glands are a crucial site of replication for human cytomegalovirus (HCMV) and its murine counterpart, murine cytomegalovirus (MCMV). Studies of MCMV often involve the use of BALB/c strain mice, but most in vitro assays are carried out in the NIH 3T3 cell line, which is derived from Swiss Albino mice. This report describes a BALB/c-derived mouse salivary gland cell line immortalized using the SV40 large T antigen. Cells stained positive for PDGFR1 and negative for E-cadherin and PECAM-1, indicating mesenchymal origin. This cell line, which has been named murine salivary gland mesenchymal (mSGM), shows promise as a tool for ex vivo immunological assays due to its MHC haplotype match with the BALB/c mouse strain. In addition, plaque assays using mSGM rather than NIH 3T3 cells are significantly more sensitive for detecting low concentrations of MCMV particles. Finally, it is demonstrated that mSGM cells express all 3 BALB/c MHC class I isotypes and are susceptible to T cell-mediated ex vivo cytotoxicity assays, leading to many possible uses in immunological studies of MCMV.

摘要

唾液腺是人巨细胞病毒(HCMV)及其鼠类对应物鼠巨细胞病毒(MCMV)的复制关键部位。MCMV 的研究通常涉及 BALB/c 品系小鼠的使用,但大多数体外检测都是在源自瑞士白化小鼠的 NIH 3T3 细胞系中进行的。本报告描述了一种使用 SV40 大 T 抗原永生化的 BALB/c 衍生的鼠唾液腺间质细胞系。细胞 PDGFR1 染色阳性,E-钙黏蛋白和 PECAM-1 染色阴性,表明其起源于间充质。该细胞系已被命名为鼠唾液腺间充质细胞(mSGM),由于其与 BALB/c 小鼠品系的 MHC 单倍型匹配,有望成为体外免疫检测的工具。此外,使用 mSGM 而不是 NIH 3T3 细胞进行的噬斑检测对于检测低浓度的 MCMV 颗粒更加敏感。最后,证明 mSGM 细胞表达所有 3 种 BALB/c MHC Ⅰ类同种型,并且容易受到 T 细胞介导的体外细胞毒性检测,从而在 MCMV 的免疫学研究中有许多可能的用途。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/46da/9385033/f11a6b108756/pone.0265479.g001.jpg

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