Expression of vascular cell adhesion molecule-1 mRNA and protein in rheumatoid synovium demonstrated by in situ hybridization and immunohistochemistry.
Expression of vascular cell adhesion molecule-1 mRNA and protein in rheumatoid synovium demonstrated by in situ hybridization and immunohistochemistry.
复制标题
原位杂交和免疫组化显示类风湿滑膜中血管细胞粘附分子1 mRNA和蛋白的表达。
DOI：
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发表时间：
1995
期刊：
影响因子：
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通讯作者：
S. Gay
中科院分区：
文献类型：
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作者：
J. Kriegsmann;G. Keyszer;T. Geiler;R. Bräuer;R. Gay;S. Gay
背景
血管细胞粘附分子-1（VCAM-1）在类风湿关节炎患者的滑膜组织中表达。VCAM-1-蛋白已被证明在非血管细胞中除了该分子的血管表达之外。关于表达VCAM-1的非血管细胞类型存在相互矛盾的结果。
实验设计
为了评价VCAM-1在类风湿性滑膜中的表达，已经通过碱性磷酸酶抗碱性磷酸酶（APAAP）技术证实了该分子。此外，VCAM-1 mRNA已被证明通过原位杂交来评估该分子在体内的从头合成。为了阐明表达VCAM-1 mRNA的细胞类型的性质，已经通过在同一组织切片中结合VCAM-1的原位杂交和免疫组织化学显示了该分子。已经用抗IV型胶原单克隆抗体进行双标记以描绘内皮细胞和周细胞，并用抗CD 68抗体进行双标记以阐明成纤维细胞样（B型）或巨噬细胞样（A型）滑膜细胞中VCAM-1 mRNA的表达。
结果
虽然已经报道了VCAM-1发生在细胞因子刺激后的内皮细胞上，但我们发现VCAM-1 mRNA和蛋白的血管表达是最小的，并且仅限于衬里细胞层下的小血管。VCAM-1 mRNA的进一步表达可以证明在含有血管基底膜的IV型胶原外的周细胞中。关于滑膜衬里层中VCAM-1的表达，我们可以通过原位杂交和免疫组织化学的结合清楚地证明衬里层中单核细胞/巨噬细胞谱系的CD 68阳性细胞（A型细胞）不表达VCAM-1 mRNA，衬里层中VCAM-1 mRNA的表达仅限于成纤维细胞样滑膜细胞（B型细胞）。显示VCAM-1 mRNA的散在基质细胞也为CD 68阴性。
结论
VCAM-1在RA滑膜的成纤维细胞样细胞中的强表达和在血管内皮中表达的缺乏表明VCAM-1的主要作用似乎与易于附着并随后侵入关节软骨的增殖滑膜细胞相关。
BACKGROUND
Vascular cell adhesion molecule-1 (VCAM-1) is expressed in synovial tissue of patients with rheumatoid arthritis. VCAM-1-protein has been demonstrated in nonvascular cells beside a vascular expression of this molecule. There are conflicting results about the nonvascular cell types expressing VCAM-1.
EXPERIMENTAL DESIGN
For the evaluation of VCAM-1 expression in rheumatoid synovium, this molecule has been demonstrated by alkaline phosphatase anti-alkaline phosphatase (APAAP) technique. Furthermore, VCAM-1 mRNA has been demonstrated by in situ hybridization to evaluate de novo synthesis of this molecule in vivo. To elucidate the nature of the cell types expressing VCAM-1 mRNA, this molecule has been shown by combined in situ hybridization for VCAM-1 and immunohistochemistry in the same tissue section. Double labeling has been performed with anti-collagen type IV monoclonal antibodies to delineate endothelial cells and pericytes and with anti-CD68 antibodies to elucidate the expression of VCAM-1 mRNA in fibroblast-like (type B) or macrophage-like (type A) synoviocytes.
RESULTS
Although it has been reported that VCAM-1 occurs on endothelial cells after cytokine stimulation, we show that vascular expression of VCAM-1 mRNA and protein was minimal and restricted to small vessels beneath the lining cell layer. Further expression of VCAM-1 mRNA could be demonstrated in pericytes outside the collagen type IV containing vascular basement membrane. With respect to the expression of VCAM-1 in the synovial lining layer, we could clearly demonstrate by combined in situ hybridization and immunohistochemistry that CD68 positive cells of the monocyte/macrophage lineage in the lining layer (type A cells) do not express VCAM-1 mRNA and that the expression of VCAM-1 mRNA in the lining layer was restricted to fibroblast-like synoviocytes (type B cells). Scattered stromal cells revealing VCAM-1 mRNA were also CD68 negative.
CONCLUSIONS
The strong expression of VCAM-1 in the fibroblast-like cells of RA synovium and the lack of expression in the vascular endothelium suggest that the major role of VCAM-1 appears to be associated with the proliferating synovial cells prone to attach and subsequently invade articular cartilage.