Expression of “non-classical” molecules of the main histocompatibility complex in rheumatoid arthritis and bronchial asthma

Olga S. Boeva, V. I. Borisevich, V. A. Kozlov, D. V. Vladimirovna, A. E. Sizikov, E. A. Pashkina
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Abstract

HLA-E is a minor understudied non-classical HLA genes. HLA-E transcription is revealed in many cell types, especially, in immune cells, e.g., T and B cells monocytes, macrophages. In this work, we evaluated expression of HLA-E on CD8+, CD4+ and CD14+ cells in conditionallу healthy donors and in the patients with bronchial asthma (BA) and rheumatoid arthritis (RA). Peripheral blood mononuclears (PBMNC) were used as initial biomaterial. PBMNC from RA patients (n = 15), BA (n = 11) and healthy donors were separated from peripheral blood in Ficoll-Urographin density gradient (1.077 g/mL). The cells were then stained with fluorochrome-conjugated monoclonal antibodies: anti-CD3-APC, anti-CD4-APC-Cy7, anti-CD-14-FITC, and anti-HLA-E-PerCP/Cy5. The cell phenotype was analyzed by flow cytometry with FACS Canto II (BD Biosciences, USA). We have found an increased expression of HLA-E on CD8+, CD4+Т cells, like as on CD14+ cells (monocytes) in the RA patients, when compared with BA patients. We have also shown significant differences of HLA-E expression on CD8+Т cells between the conditionally healthy donors and RA patients.
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主要组织相容性复合体“非经典”分子在类风湿关节炎和支气管哮喘中的表达
HLA- e是一个次要的未被充分研究的非经典HLA基因。HLA-E转录在许多细胞类型中发现,特别是在免疫细胞中,如T细胞和B细胞、单核细胞、巨噬细胞。在这项工作中,我们评估了HLA-E在条件健康供体和支气管哮喘(BA)和类风湿性关节炎(RA)患者的CD8+、CD4+和CD14+细胞上的表达。外周血单核细胞(PBMNC)作为初始生物材料。以Ficoll-Urographin密度梯度(1.077 g/mL)从外周血中分离RA患者(15例)、BA患者(11例)和健康献血者的PBMNC。然后用荧光染料偶联的单克隆抗体:抗cd3 - apc、抗cd4 - apc - cy7、抗cd -14- fitc和抗hla - e- percp /Cy5对细胞进行染色。流式细胞术分析细胞表型,使用FACS Canto II (BD Biosciences, USA)。我们发现,与BA患者相比,RA患者的CD8+、CD4+Т细胞上的HLA-E表达增加,与CD14+细胞(单核细胞)上的表达一样。我们还发现,在条件健康供者和RA患者之间,CD8+Т细胞上HLA-E的表达有显著差异。
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