Different expression pattern of human cytomegalovirus-encoded microRNAs in circulation from virus latency to reactivatio
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Journal of Translational Medicine Open Access
RESEARCH
Different expression pattern of human cytomegalovirus‑encoded microRNAs in circulation from virus latency to reactivation Wanqing Zhou1,2,3†, Cheng Wang2,3†, Meng Ding4, Yuying Bian2,3, Yujie Zhong2,3, Han Shen1, Junjun Wang2, Chen‑Yu Zhang3* and Chunni Zhang2,3*
Abstract Background: Human cytomegalovirus (HCMV) is a beta-hersvirinae that has a high latent infection rate worldwide and can cause serious consequences in immunocompromised patients when reactivation; however, the mechanism of how HCMV convert from latent to reactivation has rarely been investigated. In the present study, we aimed to perform a comprehensive analysis of the HCMV-encoded microRNA (miRNA) profile in serum of patients upon HCMV reactivation from latency and to further evaluate its clinical significance for the disease monitoring and preventing usefulness. Methods: Serum samples from 59 viremia patients and 60 age-gender matched controls were enrolled in this study for screening and validation of different expression of HCMV miRNAs. Serum concentrations of 22 known HCMV miRNAs were determined by a hydrolysis probe-based stem-loop quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay. HCMV DNA was measured by quantitative real-time PCR (qPCR) with the whole blood sam‑ ple. Serum HCMV IgG and IgM were assessed using enzyme linked immunosorbent assay (ELISA). Another 47 samples from 5 patients at different time points were collected to evaluate the monitoring effectiveness and disease predic‑ tion ability of differential expression HCMV-miRNAs during the antiviral treatment. Results: The RT-qPCR analysis revealed that the serum levels of 16 of the 22 examined HCMV miRNAs were elevated in HCMV viremia patients compared with controls, and a profile of 8 HCMV miRNAs including hcmv-miR-US25-2-3p, hcmv-miR-US4-5p, hcmv-miR-US25-2-5p, hcmv-miR-US25-1-3p, hcmv-miR-US25-1, hcmv-miR-UL36, hcmv-miRUL148D, hcmv-miR-US29-3p were markedly elevated (fold change > 2, P 500 IU/mL), HCMV IgG seropositive and HCMV IgM seronegative as the case set (defined as reactivation infectious), and another 24 patients with HCMV DNA levels less than 500 IU/ mL, HCMV IgG seropositive and HCMV IgM seronegative as the control set (defined as latency infectious) was used for screening the differential expression pattern of HCMV miRNAs. A validation cohort that containing 36 patients for the case set and 36 patients for the control set with the same above criterial was used to confirm the results of the training cohort. An additional independent cohort including 47 samples from 5 patients with leukemia (2 severe aplastic anemia patients, 1 myelodysplastic syndromes patient, 1 acute myeloid leukemia M2a
Zhou et al. J Transl Med
(2020) 18:469
patient and 1 acute lymphoblastic leukemia patient) who underwent bone marrow transplantation (the samples were collected at different time points during the antiviral therapy with ganciclovir) were also collected. The overall study design is shown in F
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