多种自然和实验性炎性兔泪腺表型

Multiple Natural and Experimental Inflammatory Rabbit Lacrimal Gland Phenotypes.

作者信息

Mircheff Austin K, Wang Yanru, Schechter Joel E, Li Meng, Tong Warren, Attar Mayssa, Chengalvala Murty, Harmuth Joe, Prusakiewicz Jeffery J

机构信息

Department of Physiology & Biophysics, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA; Department of Ophthalmology, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.

Department of Physiology & Biophysics, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.

出版信息

Ocul Surf. 2016 Oct;14(4):460-483.e3. doi: 10.1016/j.jtos.2016.07.001. Epub 2016 Jul 15.

DOI:10.1016/j.jtos.2016.07.001

PMID:27423911

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5065763/

Abstract

PURPOSE

To investigate lacrimal gland (LG) immunophysiological and immune-mediated inflammatory process (IMIP) phenotype diversity.

METHODS

Ex vivo matured dendritic cells (mDC) were loaded with acinar cell microparticles (M). Peripheral blood lymphocytes (PBL) were activated in mixed cell reactions with mDC and injected directly into autologous, unilateral LG (1°ATD-LG) of two rabbit cohorts, one naïve, one immunized with a LG lysate membrane fraction (P). Autoimmune IgG titers were assayed by ELISA, MCR PBL stimulation indices (SI) by [H]-thymidine incorporation. Schirmer tests without and with topical anesthetic (STT-I, STT-I) and rose Bengal (RB) staining tests were performed. H&E and immunohistochemically stained sections were examined. RNA yields and selected transcript abundances were measured. Immune cell number and transcript abundance data were submitted to Principal Component Analysis (PCA).

RESULTS

Immunizing P dose influenced SI but not IgG titers. STT scores were decreased, and rose Bengal scores increased, by day 118 after immunization. Previous immunization exacerbated scores in 1°ATD-eyes and exacerbated 1°ATD-LG atrophy. IMIP were evident in 2°ATD-LG as well as 1°ATD-LG. PCA described diverse immunophysiological phenotypes in control LG and diverse IMIP phenotypes in ATD-LG. IgG titers and SI pre-adoptive transfer were significantly associated with certain post-adoptive transfer IMIP phenotype features, and certain LG IMIP features were significantly associated with RB and STT I scores.

CONCLUSIONS

The underlying variability of normal states may contribute to the diversity of experimental IMIP phenotypes. The ability to generate and characterize diverse phenotypes may lead to phenotype-specific diagnostic and therapeutic paradigms.

摘要

目的

研究泪腺（LG）免疫生理学及免疫介导的炎症过程（IMIP）的表型多样性。

方法

用腺泡细胞微粒（M）加载体外成熟的树突状细胞（mDC）。外周血淋巴细胞（PBL）在与mDC的混合细胞反应中被激活，并直接注射到两组兔的自体单侧LG（1°ATD-LG）中，一组为未免疫组，另一组用LG裂解物膜组分（P）免疫。通过ELISA检测自身免疫性IgG滴度，通过[H]-胸腺嘧啶核苷掺入法检测MCR PBL刺激指数（SI）。进行了无局部麻醉和有局部麻醉的Schirmer试验（STT-I、STT-II）以及孟加拉玫瑰红（RB）染色试验。检查了苏木精-伊红（H&E）染色和免疫组化染色切片。测量RNA产量和选定转录本丰度。将免疫细胞数量和转录本丰度数据进行主成分分析（PCA）。

结果

免疫P剂量影响SI，但不影响IgG滴度。免疫后第118天，STT评分降低，孟加拉玫瑰红评分升高。先前的免疫加剧了1°ATD眼的评分，并加剧了1°ATD-LG萎缩。IMIP在2°ATD-LG以及1°ATD-LG中均很明显。PCA描述了对照LG中不同的免疫生理表型以及ATD-LG中不同的IMIP表型。过继转移前的IgG滴度和SI与过继转移后的某些IMIP表型特征显著相关，并且某些LG IMIP特征与RB和STT I评分显著相关。