Up-regulation of the vascular cell adhesion molecule-1 (VCAM-1) induced by Theiler's murine encephalomyelitis virus infection of murine brain astrocytes.

Q2 Biochemistry, Genetics and Molecular Biology Cell Communication and Adhesion Pub Date : 2010-06-01 DOI:10.3109/15419061.2010.507827
Nazario Rubio, Francisco Sanz-Rodriguez, Maria-Angeles Arevalo
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引用次数: 13

Abstract

The present article reports the up-regulation of the expression of the vascular cell adhesion molecule-1 (VCAM-1) by SJL/J mouse brain astrocytes infected with Theiler's murine encephalomyelitis virus (TMEV). Complementary RNA (cRNA) from mock- and TMEV-infected cells was hybridized to the Affymetrix whole murine genome U74v2 DNA microarray. Hybridization data analysis revealed background expression in untreated cells and the up-regulation of three sequences coding for VCAM-1, as described by the SCOP (Structural Classification Of Proteins) database. The authors further studied its regulation, confirming and validating their mRNA increase by reverse transcriptase-polymerase chain reaction (RT-PCR) and quantitative real-time RT-PCR. The presence of the 100-kDa VCAM-1 protein in mock- and TMEV-infected cells was demonstrated in the cell membrane by a specific cell-based enzyme-linked immunosorbent assay (ELISA), in addition to flow cytometry and confocal immunohistochemistry. Further, Western blots were used to quantify the amount of VCAM-1 molecules in cell extracts. All these data demonstrated a mean 75% increase in the expression of VCAM-1 on the surface of TMEV-infected cells. Three inflammatory cytokines, interleukin-1alpha (IL-1alpha), interferon gamma (IFNgumma), and specially tumor necrosis factor alpha (TNF-α), some of which are also induced by TMEV in astrocytes (IL-1alpha and TNF-alpha), were potent inducers of VCAM-1 expression. To demonstrate whether the VCAM-1 molecules were biologically active, mediating adhesion to other cells as the integrin alpha4-expressing CD4+ T lymphocytes, the authors used a cell adhesion test. It was also demonstrated by immunohistochemistry that in vivo VCAM-1 expression is enhanced after TMEV intracraneal infection. The present data show a small but statistically significant overexpression of VCAM-1 after astrocyte infection with TMEV that could play a significant role in vivo.

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Theiler's小鼠脑脊髓炎病毒感染小鼠脑星形胶质细胞诱导血管细胞粘附分子-1 (VCAM-1)的上调
本文报道了血管细胞粘附分子-1 (vascular cell adhesion molecule-1, VCAM-1)在感染Theiler's鼠脑脊髓炎病毒(TMEV)的SJL/J小鼠脑星形胶质细胞中的表达上调。将模拟和tmev感染细胞的互补RNA (cRNA)杂交到Affymetrix全鼠基因组U74v2 DNA微阵列中。杂交数据分析显示,未经处理的细胞中有背景表达,并且三个编码VCAM-1的序列上调,如SCOP(蛋白质结构分类)数据库所描述的那样。作者进一步研究了其调控作用,通过逆转录聚合酶链反应(RT-PCR)和实时定量RT-PCR证实了其mRNA的增加。除了流式细胞术和共聚焦免疫组织化学外,通过特异性细胞酶联免疫吸附试验(ELISA)在模拟和tmev感染细胞的细胞膜中证实了100 kda的VCAM-1蛋白的存在。此外,Western blots用于定量细胞提取物中VCAM-1分子的数量。所有这些数据表明,在感染tmev的细胞表面,VCAM-1的表达平均增加75%。三种炎症因子,白细胞介素-1 α (il -1 α),干扰素γ (IFNgumma),特别是肿瘤坏死因子α (TNF-α),其中一些也被TMEV诱导在星形细胞中(il -1 α和TNF-α),是VCAM-1表达的有效诱导剂。为了证明VCAM-1分子是否具有生物活性,作为表达整合素α 4的CD4+ T淋巴细胞介导与其他细胞的粘附,作者使用了细胞粘附试验。免疫组化也证实了TMEV颅内感染后体内VCAM-1表达增强。目前的数据显示,星形胶质细胞感染TMEV后,VCAM-1的过表达量虽小,但具有统计学意义,可能在体内发挥重要作用。
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来源期刊
Cell Communication and Adhesion
Cell Communication and Adhesion 生物-生化与分子生物学
CiteScore
2.50
自引率
0.00%
发文量
0
审稿时长
>12 weeks
期刊介绍: Cessation Cell Communication and Adhesion is an international Open Access journal which provides a central forum for research on mechanisms underlying cellular signalling and adhesion. The journal provides a single source of information concerning all forms of cellular communication, cell junctions, adhesion molecules and families of receptors from diverse biological systems. The journal welcomes submission of original research articles, reviews, short communications and conference reports.
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