Ocular Surface Center, Doheny Eye Institute, Los Angeles, CA 90033, USA.
Cornea. 2012 Jun;31(6):693-701. doi: 10.1097/ICO.0b013e31823f8e47.
Autologous peripheral blood lymphocytes, activated in a mixed cell reaction when cocultured with purified rabbit lacrimal epithelial cells, are known to induce a severe autoimmune dacryoadenitis when injected directly into the donor animal's remaining inferior lacrimal gland (LG) or subcutaneously at a site remote from the LG. The purpose of the present study was to determine the ability of intravenously (IV) injected autologous stimulated lymphocytes to home to the LG and salivary gland (SG) and induce disease.
One inferior LG was surgically excised from each rabbit. Acinar epithelial cells were purified, cultured for 2 days, gamma-irradiated, and cocultured for 5 days with purified autologous peripheral blood lymphocytes. The activated lymphocytes were used for autoadoptive transfer.
Tear production was reduced 50% by 4 weeks and tear breakup time was 70% less than normal. Ocular surface defects assessed by rose bengal staining were present but not as pronounced as after direct injection. Four weeks after IV injection, as after direct injection, glands contained large infiltrates composed of predominantly CD4(+) T cells close to interlobular and intralobular ducts; however, they also contained unique areas of streaming lymphocytes. Histopathology at 8 weeks was more severe than at 4 weeks, and SG also showed clusters of abnormal epithelial cells and streaming lymphocytes.
Lymphocytes activated against lacrimal antigens and injected IV can home to the LG and SG and initiate autoimmune processes, suggesting that these sites constitutively contain not only antigen-presenting cells displaying potentially pathogenic autoantigen epitopes but also chemokines and homing molecules that recruit CD4(+) T cells. This new rabbit model more closely mimics Sjögren syndrome, in that SG manifestations accompany the LG disease. It should be well suited to elucidating Sjögren pathogenesis and pathophysiology and to evaluating experimental therapies.
当与纯化的兔泪腺上皮细胞共培养时，自体外周血淋巴细胞在混合细胞反应中被激活，当直接注射到供体动物剩余的下泪腺（LG）或远离 LG 的皮下部位时，已知会引起严重的自身免疫性泪腺炎。本研究的目的是确定静脉内（IV）注射的自体刺激淋巴细胞归巢到 LG 和唾液腺（SG）并诱导疾病的能力。
从每只兔子中手术切除一个下 LG。将腺泡上皮细胞纯化、培养 2 天、γ 射线照射并与纯化的自体外周血淋巴细胞共培养 5 天。激活的淋巴细胞用于自体过继转移。
泪液产生在 4 周时减少了 50%，泪膜破裂时间比正常情况少了 70%。通过玫瑰红染色评估的眼表面缺陷存在，但不如直接注射后明显。IV 注射后 4 周，与直接注射后一样，腺体含有主要由 CD4(+) T 细胞组成的大浸润物，靠近小叶间和小叶内导管；然而，它们还包含独特的淋巴细胞流区。8 周时的组织病理学比 4 周时更严重，SG 也显示出异常上皮细胞和淋巴细胞流的簇。
针对泪腺抗原激活并静脉内注射的淋巴细胞可以归巢到 LG 和 SG 并引发自身免疫过程，表明这些部位不仅含有呈现潜在致病自身抗原表位的抗原呈递细胞，还含有趋化因子和归巢分子，可招募 CD4(+) T 细胞。这种新的兔模型更类似于干燥综合征，因为 SG 表现伴随着 LG 疾病。它应该非常适合阐明干燥综合征的发病机制和病理生理学，并评估实验性治疗方法。
