Hori J, Shiro U, Anzai N, Itoh K, Ogawa M, Ohto M, Wakashin Y, Wakashin M
First Department of Internal Medicine, Chiba University, Japan.
Nihon Jinzo Gakkai Shi. 1995 Mar;37(3):157-64.
Cultured PTC clearly expressed intercellular adhesion molecule-1 (ICAM-1) under the influence of tubular basement membrane antigen (TBM)-primed lymphocytes. These TBM-primed lymphocytes also demonstrated a high cytotoxic activity against cultured PTC. A pure preparation of isolated PTC from BALB/c mouse kidney was brought into primary culture. PTC was prepared by the method of Boogaard PJ et al, and our modification. Briefly, kidney was perfused with buffer containing 0.08% (w/v) collagenase. The cortical tissue was then filtered through nylon-gauze. Viable PTC were separated from other materials by isopycnic centrifugation on a discontinuous Nycodenz gradient. The confluent monolayer of PTC showed a typical epithelial morphology with cobblestone-like cells in the center of the cell-islands. Typical dome formation was observed in PTC cultures. These cells also strongly expressed gamma-glutamyl transpeptitase activity. Coculture of PTC with syngeneic lymphocytes primed with TBM antigen induced ICAM-1 expression in PTC. The TBM-primed lymphocytes had a cytotoxic activity without complement. However, neither virgin lymphocytes nor liver antigen-primed lymphocytes had cytotoxic activity. This simple syngeneic experimental model may allow us further molecular biological examination of renal tubulointerstitial diseases.
在肾小管基底膜抗原（TBM）致敏淋巴细胞的影响下，培养的甲状旁腺细胞（PTC）明显表达细胞间黏附分子-1（ICAM-1）。这些TBM致敏淋巴细胞对培养的PTC也表现出高细胞毒性活性。从BALB/c小鼠肾脏分离得到的纯PTC制剂进行原代培养。PTC采用Boogaard PJ等人的方法并经我们改良制备。简要地说，用含0.08%（w/v）胶原酶的缓冲液灌注肾脏。然后将皮质组织通过尼龙纱布过滤。通过在不连续的Nycodenz梯度上进行等密度离心，将活的PTC与其他物质分离。PTC的汇合单层显示出典型的上皮形态，细胞岛中心为鹅卵石样细胞。在PTC培养物中观察到典型的穹顶形成。这些细胞还强烈表达γ-谷氨酰转肽酶活性。PTC与用TBM抗原致敏的同基因淋巴细胞共培养可诱导PTC中ICAM-1的表达。TBM致敏淋巴细胞具有无补体的细胞毒性活性。然而，未致敏淋巴细胞和肝抗原致敏淋巴细胞均无细胞毒性活性。这个简单的同基因实验模型可能使我们能够进一步对肾小管间质性疾病进行分子生物学研究。
