Human papillomavirus promotes esophageal squamous cell carcinoma by regulating DNA methylation and expression of HLA-DQB1.

AIMS: Esophageal cancer (EC) is one of the most prevalent and deadly cancers worldwide. Along with nutrition, smoking and alcohol consumption, human papillomavirus (HPV) infection is one of the major risk factors, which is modulated by host immune response. This study is aimed at elucidating how HPV modifies host immune system in the EC pathogenesis. METHODS: The HPV and HLA-DQB1 levels in primary esophageal squamous cell (ESC) cancer cells from Han, Khazak and Uygur patients were analyzed by quantitative real-time PCR and immunoblotting. The ability of HPV16 E6/E7 to transform normal primary ESCs was investigated by infecting ESC with pMSCVpuro-carried E6 or E7. The shRNA against HPV16 E6 or E7 was delivered by adenovirus into esophageal squamous cell carcinoma (ESCC) cells with high HPV content. The DNA methylation level of HLA-DQB1 was measured by methylation-specific PCR. RESULTS: The HLA-DQB1 expression level was correlated with the levels of HPV and inversely related to DNA methylation level of HLA-DQB1. Overexpressing HPV16 E6 or E7 alone was enough to transform normal primary ESCs. However, single knockdown of either E6 or E7 in ESC cancer cells did not reduce HLA-DQB1 expression. CONCLUSIONS: Oncogenic HPV E6 and E7 genes promoted ESCC pathogenesis by upregulating susceptible HLA-DQB1 via DNA demethylation.

Follicular lymphoma-protective HLA class II variants correlate with increased HLA-DQB1 protein expression.

Multiple follicular lymphoma (FL) susceptibility single-nucleotide polymorphisms in the human leukocyte antigen (HLA) class I and II regions have been identified, including rs6457327, rs3117222, rs2647012, rs10484561, rs9268853 and rs2621416. Here we validated previous expression quantitative trait loci results with real-time reverse transcription quantitative PCR and investigated protein expression in B-lymphoblastoid cell lines and primary dendritic cells using flow cytometry, cell-based enzyme-linked immunosorbent assay and western blotting. We confirmed that FL-protective rs2647012-linked variants, in high linkage disequilibrium with the extended haplotype DRB1*15:01-DQA1*01:02-DQB1*06:02, correlate with increased HLA-DQB1 expression. This association remained significant at the protein level and was reproducible across different cell types. We also found that differences in HLA-DQB1 expression were not related to changes in activation markers or class II, major histocompatibility complex, transactivator expression, suggesting the role of an alternative regulatory mechanism. However, functional analysis using RegulomeDB did not reveal any relevant regulatory candidates. Future studies should focus on the clinical relevance of increased HLA-DQB1 protein expression facilitating tumor cell removal through increased immune surveillance.

Profiles of epigenetic histone post-translational modifications at type 1 diabetes susceptible genes.

Both genetic and environmental factors are implicated in type 1 diabetes (T1D). Because environmental factors can trigger epigenetic changes, we hypothesized that variations in histone post-translational modifications (PTMs) at the promoter/enhancer regions of T1D susceptible genes may be associated with T1D. We therefore evaluated histone PTM variations at known T1D susceptible genes in blood cells from T1D patients versus healthy nondiabetic controls, and explored their connections to T1D. We used the chromatin immunoprecipitation-linked to microarray approach to profile key histone PTMs, including H3-lysine 4 trimethylation (H3K4me3), H3K27me3, H3K9me3, H3K9 acetylation (H3K9Ac), and H4K16Ac at genes within the T1D susceptible loci in lymphocytes, and H3K4me3, H3K9me2, H3K9Ac, and H4K16Ac at the insulin-dependent diabetes mellitus 1 region in monocytes of T1D patients and healthy controls separately. We screened for potential variations in histone PTMs using computational methods to compare datasets from T1D and controls. Interestingly, we observed marked variations in H3K9Ac levels at the upstream regions of HLA-DRB1 and HLA-DQB1 within the insulin-dependent diabetes mellitus 1 locus in T1D monocytes relative to controls. Additional experiments with THP-1 monocytes demonstrated increased expression of HLA-DRB1 and HLA-DQB1 in response to interferon-γ and TNF-α treatment that were accompanied by changes in H3K9Ac at the same promoter regions as that seen in the patient monocytes. These results suggest that the H3K9Ac status of HLA-DRB1 and HLA-DQB1, two genes highly associated with T1D, may be relevant to their regulation and transcriptional response toward external stimuli. Thus, the promoter/enhancer architecture and chromatin status of key susceptible loci could be important determinants in their functional association to T1D susceptibility.

Differential expressions of MHC-DQB1 mRNA in Chinese merino sheep infected with Echinococosus granuclosus.

The purpose of the present study was to investigate the dynamic changes of MHC-DQB1 mRNA expression in sheep infected with Echinococosus granuclosus. A total of 14 healthy Chinese merino sheep were experimentally infected with E. granuclosus. The blood samples were collected on days 0 (initiation of the infection), 7, 21, 30, and 60 post-infection, respectively. On day 60 post-infection, when the experiment was terminated, all sheep were euthanized to make a diagnosis of cystic echinococcosis (CE) using routine meat inspection and microscopical examination, respectively. The sheep were then divided into two groups according to the diagnostic results: group A (n = 8) consisted of sheep which were diagnosed as CE infection, while group B (n = 6) comprised sheep diagnosed as self-cured or healthy controls. Blood samples obtained during the period of the study were correspondingly divided into groups A and B. The mRNA expression levels of DQB1 revealed significant alterations detected at different stages of E. granuclosus infection in the two groups. Results showed that in group A, DQB1 mRNA expression underwent a progressive increase from day 0 to day 21 post-infection (P = 0.073), and suddenly, suffered from a dramatic drop until day 30 post-infection, and then jumped rapidly and peaked on day 60 post-infection (P = 0.004). Meanwhile, in group B, DQB1 mRNA expression displayed a sharp increase from day 0 to day 7 post-infection (P = 0.000), which thereafter showed a marked decrease until day 30 post-infection, and experienced a plateau from day 30 to day 60 post-infection, remaining at or above that on day 0. It is concluded that DQB1 mRNA expression levels varied in different stages of E. granuclosus infection in sheep. In addition, it appears that the ability to eliminate the parasites possibly depends, at least in part, on the DQB1 expression in the early stage of infection, especially in the first week post-infection.