Vinay Dass S, Kim Jung D, Asai Tatsuhiko, Choi Beom K, Kwon Byoung S
LSU Eye Center, LSU Health Sciences Center School of Medicine, New Orleans, Louisiana, USA.
Invest Ophthalmol Vis Sci. 2007 Oct;48(10):4608-15. doi: 10.1167/iovs.07-0153.
To define the role of endogenous 4-1BB (an important T-cell costimulatory molecule) in the regulation of ocular disease, MRL-Fas(lpr) mice deficient in 4-1BB were generated, and their lacrimal gland function was studied.
4-1BB(-/-)MRL/MpJ-Tnfrs(lpr)/Tnfrs(lpr) (lpr/4-1BB(-/-)) mice were generated and used at the ninth backcross. Mice were killed at various times, and lacrimal gland cellularity was analyzed by flow cytometry. Tear and tissue samples were analyzed by Western blotting for the presence of aquaporin 5 (AQP5) and 120-kDa fragments of alpha-fodrin. Cytokine expression of lacrimal glands was assessed by flow cytometry and RT-PCR analysis.
Absence of the 4-1BB gene function in lpr mice resulted in early and increased infiltration of mononuclear cells into lacrimal glands compared with 4-1BB intact lpr mice. The severity of lesions in lpr/4-1BB(-/-) mice was closely associated with enhanced accumulation of primarily CD4(+) T cells within the lacrimal glands and with increased expression of IL-4. Elevated levels of AQP5 and cleaved 120-kDa fragments of alpha-fodrin were found in tears and lacrimal gland lysates, respectively, of lpr/4-1BB(-/-) but not lpr/4-1BB(+/+) mice.
Deletion of 4-1BB in lpr mice accelerates lacrimal gland lesions through increased CD4(+) T-cell infiltration and their production of immune modulators.
为了确定内源性4-1BB（一种重要的T细胞共刺激分子）在眼部疾病调节中的作用，我们培育了缺乏4-1BB的MRL-Fas(lpr)小鼠，并研究了它们的泪腺功能。
培育4-1BB(-/-)MRL/MpJ-Tnfrs(lpr)/Tnfrs(lpr)（lpr/4-1BB(-/-)）小鼠，并在第九代回交时使用。在不同时间点处死小鼠，通过流式细胞术分析泪腺细胞组成。通过蛋白质免疫印迹法分析泪液和组织样本中是否存在水通道蛋白5（AQP5）和α- fodrin的120-kDa片段。通过流式细胞术和逆转录-聚合酶链反应（RT-PCR）分析评估泪腺细胞因子的表达。
与4-1BB完整的lpr小鼠相比，lpr小鼠中4-1BB基因功能的缺失导致单核细胞早期且增加地浸润到泪腺中。lpr/4-1BB(-/-)小鼠病变的严重程度与泪腺内主要是CD4(+) T细胞的积累增加以及IL-4表达增加密切相关。在lpr/4-1BB(-/-)小鼠的泪液和泪腺裂解物中分别发现了升高水平的AQP5和裂解的α- fodrin的120-kDa片段，但在lpr/4-1BB(+/+)小鼠中未发现。
lpr小鼠中4-1BB的缺失通过增加CD4(+) T细胞浸润及其免疫调节剂的产生加速了泪腺病变。
