| Literature DB >> 26155421 |
Simon Jasinski-Bergner1, Christine Stoehr2, Juergen Bukur1, Chiara Massa1, Juliane Braun3, Stefan Hüttelmaier3, Verena Spath2, Roland Wartenberg2, Wolfgang Legal4, Helge Taubert4, Sven Wach4, Bernd Wullich4, Arndt Hartmann2, Barbara Seliger1.
Abstract
In human tumors of distinct origin including renal cell carcinoma (RCC), the non-classical human leukocyte antigen G (HLA-G) is frequently expressed, thereby inhibiting the cytotoxic activity of T and natural killer (NK) cells. Recent studies demonstrated a strong post-transcriptional gene regulation of the HLA-G by miR-152, -148A, -148B and -133A. Standard methods were applied to characterize the expression and function of HLA-G, HLA-G-regulatory microRNAs (miRs) and the immune cell infiltration in 453 RCC lesions using a tissue microarray and five RCC cell lines linking these results to clinical parameters. Direct interactions with HLA-G regulatory miRs and the HLA-G 3' untranslated region (UTR) were detected and the affinities of these different miRs to the HLA-G 3'-UTR compared. qPCR analyses and immunohistochemical staining revealed an inverse expression of miR-148A and -133A with the HLA-G protein in situ and in vitro. Stable miR overexpression caused a downregulation of HLA-G protein enhancing the NK and LAK cell-mediated cytotoxicity in in vitro CD107a activation assays revealing a HLA-G-dependent cytotoxic activity of immune effector cells. A significant higher frequency of CD3+/CD8+ T cell lymphocytes, but no differences in the activation markers CD69, CD25 or in the presence of CD56+, FoxP3+ and CD4+ immune cells were detected in HLA-G+ compared to HLA-G- RCC lesions. This could be associated with higher WHO grade, but not with a disease-specific survival. These data suggest a miR-mediated control of HLA-G expression in RCC, which is associated with a distinct pattern of immune cell infiltration.Entities:
Keywords: ACTB, β-actin; APM, antigen processing machinery; B7-H1, B7 homolog 1; CDS, coding sequence; Cr, chromium; COPZ2, coatomer protein complex, subunit zeta 2; DAC, 5′-aza-2′-desoxycytidine, GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HLA-G, human leukocyte antigen G; HRP, horseradish peroxidase; IFNγ, interferon gamma; IHC, immunohistochemistry; IL, interleukin; ILT, immunoglobulin-like transcript; LAK, lymphokine-activated killer cell; MDSC, myeloid-derived suppressor cells; MFI, mean-specific fluorescence intensity; NK, natural killer cell; RCC, renal cell carcinoma; SNP, single nucleotide polymorphism; TGF-β, transforming growth factor β; TIL, tumor infiltrating lymphocyte; TMA, tissue microarray; Treg, regulatory T cell; UTR, untranslated region; WB, Western blot analysis; WT, wild type; immune escape; luc, luciferase; mAb, monoclonal antibody; miR, microRNA; miTRAP, miRNA trapping by RNA in vitro affinity purification; microRNA; n.d., not determined; n.o.s., not otherwise specified; ntc., non-template control; non-classical HLA class I molecules; renal cell carcinoma; sHLA-G, soluble HLA-G; tumor-infiltrating lymphocytes; β-gal, β-galactosidase; β2-m, β-2-microglobulin
Year: 2015 PMID： 26155421 PMCID： PMC4485830 DOI： 10.1080/2162402X.2015.1008805
Source DB: PubMed Journal: Oncoimmunology ISSN： 2162-4011 Impact factor: 8.110