Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
Arthritis Rheumatol. 2015 Apr;67(4):1096-106. doi: 10.1002/art.39009.
To investigate whether human T lymphotropic virus type I (HTLV-I) directly infects salivary gland epithelial cells (SGECs) and induces the niche of the salivary glands in patients with Sjögren's syndrome (SS).
SGECs were cultured with the HTLV-I-producing CD4+ T cell line HCT-5 or with Jurkat cells. Antibody arrays, immunofluorescence analysis, and enzyme-linked immunosorbent assay (ELISA) were used to determine the profiles of inflammation-related molecules, and the profiles of apoptosis-related molecules were determined by antibody array and immunofluorescence analysis. The presence of HTLV-I-related molecules was assessed by immunofluorescence analysis and in situ polymerase chain reaction. Apoptosis of SGECs was evaluated by TUNEL staining.
Among the SGECs, 7.8 ± 1.3% (mean ± SD) were positive for HTLV-I-related proteins after 96-hour coculture with HCT-5 cells. Nuclear NF-κB p65 was also detected in 10% of the SGECs. The presence of HTLV-I proviral DNA in SGECs after coculture with HCT-5 cells was detected by in situ polymerase chain reaction. After coculture of SGECs with HCT-5, the expression of cytokines and chemokines, including soluble intercellular adhesion molecule 1, RANTES, and interferon γ-induced protein 10 kd (IP-10/CXCL10) was increased in a time-dependent manner. The expression of proapoptotic molecules (e.g., cytochrome c and Fas) and antiapoptotic molecules (e.g., Bcl-2, Heme oxygenase 2, and Hsp27) was increased in the SGECs cocultured with HCT-5, showing that apoptosis of SGECs was not detected after coculture with HCT-5 or Jurkat cells.
HTLV-I is thought to infect SGECs and alter their cellular functions. These changes may induce the niche of SS and contribute to the development of SS in anti-HTLV-I antibody-positive individuals.
研究人 T 淋巴细胞白血病病毒 I 型（HTLV-I）是否直接感染唾液腺上皮细胞（SGEC）并诱导干燥综合征（SS）患者的唾液腺龛位。
用 HTLV-I 产生的 CD4+T 细胞系 HCT-5 或 Jurkat 细胞培养 SGEC。采用抗体芯片、免疫荧光分析和酶联免疫吸附试验（ELISA）检测炎症相关分子谱，采用抗体芯片和免疫荧光分析检测凋亡相关分子谱。免疫荧光分析和原位聚合酶链反应检测 HTLV-I 相关分子的存在。通过 TUNEL 染色评估 SGEC 的凋亡。
在与 HCT-5 细胞共培养 96 小时后，SGEC 中 7.8±1.3%（平均值±标准差）为 HTLV-I 相关蛋白阳性。在 10%的 SGEC 中也检测到核 NF-κB p65。用原位聚合酶链反应检测到与 HCT-5 细胞共培养后的 SGEC 中存在 HTLV-I 前病毒 DNA。与 HCT-5 细胞共培养后，SGEC 中细胞因子和趋化因子的表达（包括可溶性细胞间黏附分子 1、RANTES 和干扰素 γ诱导蛋白 10 kd（IP-10/CXCL10））呈时间依赖性增加。与 HCT-5 共培养的 SGEC 中促凋亡分子（如细胞色素 c 和 Fas）和抗凋亡分子（如 Bcl-2、血红素加氧酶 2 和 Hsp27）的表达增加，表明与 HCT-5 或 Jurkat 细胞共培养后未检测到 SGEC 凋亡。
HTLV-I 被认为感染 SGEC 并改变其细胞功能。这些变化可能诱导 SS 的龛位，并有助于抗 HTLV-I 抗体阳性个体 SS 的发展。
