Characterization of a renal tubular epithelial cell line which secretes the autologous target antigen of autoimmune experimental interstitial nephritis.
Characterization of a renal tubular epithelial cell line which secretes the autologous target antigen of autoimmune experimental interstitial nephritis.
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DOI：
10.1083/jcb.107.4.1359
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发表时间：
1988-10
期刊：
影响因子：
--
通讯作者：
Neilson EG
中科院分区：
文献类型：
--
作者：
Haverty TP;Kelly CJ;Hines WH;Amenta PS;Watanabe M;Harper RA;Kefalides NA;Neilson EG
发现来自发展自身免疫性间质性肾炎的小鼠的近端肾小管上皮细胞表达致肾炎靶抗原3 M-1。抗3 M-1 mAb（α 3 M-1-Ab）用于阳性选择3 M-1分泌性肾小管上皮细胞，在培养中稳定后，检查该新细胞系（MCT）是否产生对免疫相互作用或细胞外基质发育重要的几个部分。碱性磷酸酶染色MCT细胞也表达上皮生长因子受体，Kd为0.87 nM，上皮生长因子受体常数（Ro）为2.1 × 10（4）受体/细胞。与I型和III型前胶原相比，MCT培养上清液含有更大量的层粘连蛋白以及IV型和V型前胶原，并且在I型胶原基质上生长的MCT细胞导致它们优先分泌甚至更多的IV型和V型前胶原。30，000-Mr 3 M-1抗原可从具有α 3 M-1-Ab的生物合成标记的MCT细胞上清液中免疫沉淀。通过免疫亲和层析从胶原酶溶解的小鼠肾小管基底膜中分离出相同大小的部分。3 M-1抗原可通过放射免疫测定法在MCT细胞表面上发现，或通过免疫荧光法在培养物中以线性阵列沉积在MCT细胞周围的细胞外基质中。通过北方杂交检测I类和II类主要组织相容性复合体（MHC）分子的成熟信使RNA种类，并通过单倍型特异性抗体染色的MCT细胞的细胞荧光照相检测其相应的细胞表面基因产物。细胞表面3 M-1和少量检测到的II类MHC分子似乎都具有生物学功能，因为MCT细胞可以支持培养物中3 M-1特异性II类MHC限制性辅助T细胞的增殖。这些发现表明，MCT细胞提供了所有必要的生物学参数，用于作为致肾炎免疫应答的靶点和作为新细胞外基质的潜在来源，所述新细胞外基质作为间质性肾炎的纤维化应答而发展。
Proximal tubular epithelial cells from mice which develop autoimmune interstitial nephritis were found to express the nephritogenic target antigen, 3M-1. Anti-3M-1 mAbs (alpha 3M-1-Ab) were used to positively select for 3M-1-secreting tubular epithelium and, after stabilization in culture, this new cell line (MCT) was examined for the production of several moieties important to either immune interactions or to the development of extracellular matrix. Alkaline phosphatase-staining MCT cells also express epithelial growth factor receptors with a Kd of 0.87 nM and an epithelial growth factor receptor constant (Ro) of 2.1 X 10(4) receptors/cell. MCT culture supernatants contain greater amounts of laminin, and types IV and V procollagens compared to types I and III procollagens, and growing MCT cells on type I collagen matrix causes them to preferentially secrete even more type IV and V procollagen. The 30,000-Mr 3M-1 antigen could be immunoprecipitated from biosynthetically labeled MCT cell supernatants with alpha 3M-1-Ab. An identical-sized moiety was isolated by immunoaffinity chromatography from collagenase-solubilized mouse kidney tubular basement membranes. The 3M-1 antigen can be found on the MCT cell surface by radioimmunoassay, or deposited in a linear array in the extracellular matrix surrounding the MCT cells in culture by immunofluorescence. Mature messenger RNA species for both class I and class II major histocompatibility complex (MHC) molecules were detected by Northern hybridization, and their corresponding cell surface gene products were detected by cytofluorography of MCT cells stained with haplotype- specific antibodies. Both the cell surface 3M-1 and the small amounts of detected class II MHC molecules appear to be biologically functional, as MCT cells can support the proliferation of 3M-1- specific, class II MHC-restricted helper T cells in culture. These findings suggest that MCT cells provide all the necessary biological parameters for interfacing both as the target of a nephritogenic immune response, and as a potential source for new extracellular matrix which develops as a fibrogenic response to interstitial nephritis.