Zhu Zejin, Stevenson Douglas, Schechter Joel E, Mircheff Austin K, Atkinson Roscoe, Trousdale Melvin D
Doheny Eye Institute, Los Angeles, CA 90033, USA.
Cornea. 2003 Jan;22(1):25-32. doi: 10.1097/00003226-200301000-00007.
To study the effects of induced autoimmune dacryoadenitis on lacrimal gland function, histopathology, and ocular surface disease in a rabbit model.
One lacrimal gland was surgically excised from each experimental rabbit, and epithelial cells were purified, cultured, irradiated, and then cocultured with autologous peripheral blood lymphocytes (PBLs) for 5 days. Autoimmune dacryoadenitis was induced by injecting the autologous mixed cell reactions (AMCRs) into the rabbit's remaining lacrimal gland. Normal rabbits and rabbits with both lacrimal glands injected with nonstimulated PBLs were examined as controls. Eyes were evaluated biweekly for 8 weeks by slit-lamp biomicroscopy, Schirmer testing, tear break-up time measurement, and rose bengal examination. Sections of lacrimal glands removed at 8 weeks post-operation were immunostained using antibodies against rabbit class II major histocompatibility complex molecule (MHC-II), CD4, CD8, CD18, and rabbit thymic lymphocyte antigen (RTLA). Relative numbers of positively stained cells were quantified with a ChromaVision image analysis system.
During an 8-week period, a continuous decrease in tear production and stability, accompanied by a continuous increase in rose bengal staining, occurred in eyes in which AMCR-PBL had been injected into the ipsilateral lacrimal glands. Similar, though generally less severe, changes occurred in eyes contralateral to the AMCR-PBL-injected eyes. No obvious changes by 8 weeks in these parameters were found in eyes in which the lacrimal glands had been injected with nonstimulated PBLs or in the lacrimal gland-excised eyes contralateral to normal eyes. Interstitial cells in normal lacrimal glands expressed CD18 and RTLA antigens, but few expressed CD4, CD8, or MHC-II. Focal mononuclear cell infiltrates were only found in lacrimal glands from animals with induced autoimmune dacryoadenitis. These cells were predominantly positive for CD4 (7.3-fold increase), RTLA (7.8-fold increase), or CD18 (42-fold increase). MHC-II expression in interstitial and ductal epithelial cells was also significantly greater in these animals than in control animals. The mononuclear cell infiltrates were frequently found enveloping venules, some of which appeared to be high endothelial cell venules. The ductal epithelium also contained CD4 and CD8 immunopositivity, within the epithelium, at the lumenal surface, or surrounding the ducts. Occasionally CD4 and CD8 immunopositive cells could be identified within the acinar lumens.
Injection of activated PBLs (i.e., AMCR-PBLs) in the lacrimal gland induces autoimmune dacryoadenitis with immunopathologic features similar to those of Sjögren's syndrome. The lacrimal immunopathology is accompanied by typical clinical manifestations of dry eye syndrome. The persistent significant dry eye does not appear to result just from failure of the diseased gland but from a more general dysfunction of the surface secretory tissues.
在兔模型中研究诱导性自身免疫性泪腺炎对泪腺功能、组织病理学及眼表疾病的影响。
从每只实验兔手术切除一个泪腺，纯化、培养、照射上皮细胞，然后与自体外周血淋巴细胞（PBL）共培养5天。通过将自体混合细胞反应（AMCR）注射到兔剩余的泪腺中诱导自身免疫性泪腺炎。以正常兔和双侧泪腺注射未刺激PBL的兔作为对照。通过裂隙灯生物显微镜检查、泪液分泌试验、泪膜破裂时间测量和孟加拉玫瑰红染色检查，每两周对眼睛评估一次，持续8周。术后8周切除的泪腺切片用抗兔II类主要组织相容性复合体分子（MHC-II）、CD4、CD8、CD18和兔胸腺淋巴细胞抗原（RTLA）的抗体进行免疫染色。用ChromaVision图像分析系统对阳性染色细胞的相对数量进行定量。
在8周期间，同侧泪腺注射AMCR-PBL的眼睛中，泪液分泌和稳定性持续下降，同时孟加拉玫瑰红染色持续增加。在注射AMCR-PBL眼睛的对侧眼睛中也出现了类似但通常较轻的变化。泪腺注射未刺激PBL的眼睛或正常眼睛对侧的泪腺切除眼睛在8周时这些参数未发现明显变化。正常泪腺中的间质细胞表达CD18和RTLA抗原，但很少表达CD-4、CD8或MHC-II。仅在诱导性自身免疫性泪腺炎动物的泪腺中发现局灶性单核细胞浸润。这些细胞主要CD4（增加7.3倍）、RTLA（增加7.8倍）或CD18（增加42倍）呈阳性。这些动物间质和导管上皮细胞中的MHC-II表达也明显高于对照动物。单核细胞浸润经常围绕小静脉，其中一些似乎是高内皮细胞小静脉。导管上皮在管腔内表面或导管周围的上皮内也含有CD4和CD8免疫阳性。偶尔在腺泡腔内可识别出CD4和CD8免疫阳性细胞。
在泪腺中注射活化的PBL（即AMCR-PBL）可诱导具有类似于干燥综合征免疫病理特征的自身免疫性泪腺炎。泪腺免疫病理学伴有干眼综合征的典型临床表现。持续性严重干眼似乎不仅仅是患病腺体功能衰竭的结果，而是表面分泌组织更普遍功能障碍的结果。
