Objective:To study the expression of human leukocyte antigen G (HLA-G) during human T lymphocytic leukemia virus type 1 (HTLV-1) infection, and to investigate the function and mechanism of HLA-G in HTLV-1 immune escape.Methods:HeLa and THP-1 cells were infected by MT2, a stable HTLV-1 infected cell line. Expression of HLA-G isomers at mRNA and protein levels was detected by qRT-PCR and Western blot, respectively. Flow cytometry was used to detect the expression of HLA-G1. Moreover, transfection and siRNA gene were respectively used to up-regulate and silence HLA-G expression in HeLa cells to observe the changes in the expression of HTLV-1-featured protein Tax on protein level after HTLV-1 infection.Results:HTLV-1 infection could induce differential expression of HLA-G isomers, mainly HLA-G1 and HLA-G5, in HeLa and THP-1 cells. HLA-G expression at both mRNA and protein levels was significantly up-regulated 24 h after HTLV-1 infection. Moreover, the expression of HTLV-1 protein Tax was significantly enhanced in HTLV-1-infected HeLa cells overexpressing HLA-G. An opposite result was obtained when the HLA-G gene in HeLa cells was silenced by siRNA.Conclusions:The immune-tolerant molecule HLA-G was significantly increased in HTLV-1-infected cells, thereby promoting the expression of HTLV-1 viral protein, which led to persistent viral infection.

Objective:To develop multiplex cytokine detection reagents to analyze expression levels of cytokines,angiogene-sis and bone remodeling proteins in relapse/refractory multiple myeloma(RRMM).Methods:Multiplex bead-based immunoassay by flow cytometry was used to develop quantitative detection reagents of multiplex cytokines,which were applied to detect serum samples from 55 RRMM patients and 22 healthy controls.Expression levels of cytokines,angiogenesis,and bone remodeling proteins in pa-tients,and their correlation with clinical characteristics were analyzed.Results:Detection reagents of 10-plex cytokine immunoassay were successfully developed in this study,with average sensitivity of 7.1 pg/ml,average recovery rate of 97.4%,average intra-assay CV of 4.8%,and average inter-assay CV of 9.0%.In addition,results of RRMM samples found that levels of IL-2,IL-17,DKK1,RANKL and OPG were positively correlated with the level of IgG monoclonal protein,and TIMP1 was positively correlated with levels of IgG and IgA monoclonal protein.Conclusion:In this study,ten kinds of cytokine detection reagents with high sensitivity and speci-ficity are developed,and we found that IL-2,IL-17,DKK1,RANKL,OPG and TIMP1 have potential value in tracking disease pro-gression in RRMM.The established development process of multiplex cytokine reagents has important reference significance for ex-panding the development and application of multiplex detection reagents for protein markers in the future.