Lin X P, Almqvist N, Telemo E
Department of Rheumatology and Inflammation Research, Sahlgrenska Academy, University of Göteborg, Guldhedsg. 10, S-41346 Göteborg, Sweden.
Blood Cells Mol Dis. 2005 Sep-Oct;35(2):122-8. doi: 10.1016/j.bcmd.2005.05.011.
In humans, the small intestinal epithelial cells (IEC) have a high constitutive expression of MHC class II (MHC II), and contains lysosomes. The IEC also contains MHC II rich multivesicular compartments and has been shown to produce exosomes. This suggests a role for the IEC in antigen processing and presentation either directly or indirectly by the production of exosomes. However, the presence and localisation in the IEC of other key molecules involved in this process has not been studied previously. In the present work, we have investigated small intestinal biopsies from healthy adults and the HT29 IEC cell line with monoclonal antibodies against molecules involved in the antigen processing/presenting systems and molecules typically found on exosomes derived from professional APCs and IECs. Immunohistology was performed to study the expression and localisation of MHC II (HLA-DR), HLA-DM, MHC I (HLA-ABC), CD1d, Invariant chain, Lamp-1, CD68, CD63, B7.1, B7.2, ICAM-1, Cathepsin D/S/L and the IEC specific marker A33 in the IECs. We found that the IECs from the biopsies constitutively express MHC II, HLA-DM, MHC I, Invariant chain, Lamp-1, CD 68, CD63 and A33, and these markers were also found in the IFN-g treated HT-29 cells. All these molecules were found apically in the IECs of the biopsies, localised mainly in vesicular structures. Interestingly, in the baso-latereral area of the IEC, only MHC II, MHC I, Lamp 1, CD68, CD63 and A33 were found and also here with vesicular staining pattern which matches the molecules previously found on exosomes derived professional APCs and human IEC lines. CD1d, B7, ICAM-1, CD9 and cathepsin D and L were absent in the IEC compartment, but cathepsin S showed a relatively weak staining in the apical part of the IEC. The staining pattern and the morphological localisation of these markers suggest a prominent antigen processing/loading and trafficking compartment, and a possible baso-lateral release of exosomes in the normal human IEC.
在人类中,小肠上皮细胞(IEC)组成性高表达主要组织相容性复合体II类分子(MHC II),并含有溶酶体。IEC还含有富含MHC II的多囊泡小室,且已被证明能产生外泌体。这表明IEC在抗原加工和呈递过程中可能直接或通过产生外泌体间接发挥作用。然而,此前尚未研究参与该过程的其他关键分子在IEC中的存在情况和定位。在本研究中,我们使用针对参与抗原加工/呈递系统的分子以及通常在源自专职抗原呈递细胞(APC)和IEC的外泌体上发现的分子的单克隆抗体,对健康成年人的小肠活检组织和HT29 IEC细胞系进行了研究。通过免疫组织学方法研究了MHC II(HLA - DR)、HLA - DM、MHC I(HLA - ABC)、CD1d、恒定链、Lamp - 1、CD68、CD63、B7.1、B7.2、ICAM - 1、组织蛋白酶D/S/L以及IEC特异性标志物A33在IEC中的表达和定位。我们发现活检组织中的IEC组成性表达MHC II、HLA - DM、MHC I、恒定链、Lamp - 1、CD68、CD63和A33,并且在经干扰素 - γ处理的HT - 29细胞中也发现了这些标志物。所有这些分子在活检组织的IEC顶端均有发现,主要定位于囊泡结构中。有趣的是,在IEC的基底外侧区域,仅发现了MHC II、MHC I、Lamp 1、CD68、CD63和A33,并且此处也是囊泡染色模式,这与先前在源自专职APC和人IEC系的外泌体上发现的分子相匹配。IEC区室中不存在CD1d、B7、ICAM - 1、CD9以及组织蛋白酶D和L,但组织蛋白酶S在IEC顶端部分显示出相对较弱的染色。这些标志物的染色模式和形态学定位表明存在一个突出的抗原加工/装载和运输区室,并且在正常人类IEC中可能存在外泌体的基底外侧释放。