Complexity of Host Micro‐RNA Response to Cytomegalovirus Reactivation After Organ Transplantation

Human (Homo sapiens) micro‐RNAs (hsa‐miRNAs) regulate virus and host‐gene translation, but the biological impact in patients with human cytomegalovirus (hCMV) infection is not well defined in a clinically relevant model. First, we compared hsa‐miRNA expression profiles in peripheral blood mononuclear cells from 35 transplant recipients with and without CMV viremia by using a microarray chip covering 847 hsa‐miRNAs. This approach demonstrated a set of 142 differentially expressed hsa‐miRNAs. Next, we examined the effect of each of these miRNAs on viral growth by using human fibroblasts (human foreskin fibroblast‐1) infected with the hCMV Towne strain, identifying a subset of proviral and antiviral hsa‐miRNAs. miRNA‐target prediction software indicated potential binding sites within the hCMV genome (e.g., hCMV‐UL52 and ‐UL100 [UL = unique long]) and host‐genes (e.g., interleukin‐1 receptor, IRF1). Luciferase‐expressing plasmid constructs and immunoblotting confirmed several predicted miRNA targets. Finally, we determined the expression of selected proviral and antiviral hsa‐miRNAs in 242 transplant recipients with hCMV‐viremia. We measured hsa‐miRNAs before and after antiviral therapy and correlated hsa‐miRNA expression levels to hCMV‐replication dynamics. One of six antiviral hsa‐miRNAs showed a significant increase during treatment, concurrent with viral decline. In contrast, six of eight proviral hsa‐miRNAs showed a decrease during viral decline. Our results indicate that a complex and multitargeted hsa‐miRNA response occurs during CMV replication in immunosuppressed patients. This study provides mechanistic insight and potential novel biomarkers for CMV replication.     

A built-in co-carcinogenic effect due to viruses involved in latent or persistent infections

A new hypothesis for some cancers, which combines the chromosomal instability theories with a co-carcinogenic effect of viruses causing latent or persistent infection, is presented. The hypothesis incorporates the multi-step model of cancer and that pre-cancerous cells reach a state of chromosomal instability. Because of chromosomal instability, the genome of these cell lines will lead to changes from generation to generation and will face a remarkable selection pressure both from lost traits, apoptosis, and from the immune system. Viruses causing latent or persistent infections have evolved many different genes capable to evade the immune system. If these viruses are harboured in the genome of pre-cancerous cells they could provide them with "superpowers" and with genes that may assist the cells to elude the immune system. The theory explains why cancer predominantly is a disease of old age. Upon aging, the immune system becomes reduced including the ability to control and suppress the viruses that cause latent or persistent infections. The risk of cancer could thereby increase as the immune functions decrease. The theory provides new insights to the genesis of cancers.     

Analysis of relationships between polymorphisms in the genes encoding the pentameric complex and neutralization of clinical cytomegalovirus isolates

IntroductionAs congenital cytomegalovirus (CMV) infection is one of the major causes of birth defects and developmental abnormalities, it is essential to develop vaccines and therapeutic antibodies against CMV. Clinical trials demonstrated that the subunit vaccine based on glycoprotein B, which had been believed to be the major target for neutralization, did not induce sufficient protective immunity. On the other hand, it has been reported that the immunization of animals with the Pentamer, the pentameric complex of gH/gL/UL128/UL130/UL131A, induced strong neutralizing antibodies. Here, we sought to clarify whether any polymorphic alterations present in the Pentamer of clinical isolates affect neutralization by anti-Pentamer antibodies.MethodsSequences of the genes encoding the Pentamer components of 25 Japanese clinical isolates were determined. Neutralization of infection by two seropositive sera and by anti-Pentamer serum was measured using a CMV reporter cell line based on ARPE-19.ResultsPolymorphisms of the amino acid sequence of UL128, UL130, and UL131A ORFs were limited and clustered into two major groups. The identified alterations, except UL128 I140T, were mapped outside of the reported regions recognized by neutralizing antibodies. Anti-Pentamer serum neutralized infection with all isolates to a similar degree and had no correlation with the polymorphic groups.ConclusionsOur findings indicate that Pentamer antigens prepared from Merlin Fix strain induce antibodies that neutralize infection with all isolates to a similar level and that anti-Pentamer antibodies neutralize CMV infection better than do human sera, suggesting that vaccines and therapeutic antibodies based on Pentamer as an antigen have some promise.     

Cytomegalovirus shedding in the oral cavity of allogeneic haematopoietic stem cell transplant patients

OBJECTIVE: This study was designed to investigate the effect of allogeneic haematopoietic stem cell transplantation (HSCT) on cytomegalovirus (CMV) shedding in the saliva by nested polymerase chain reaction (nested PCR) and its impact on patient survival. PATIENTS AND METHODS: One hundred and twenty-four HSCT patients and 124 healthy volunteers were included in the study. Oral swabs were taken before, after 100 days and 1 year of HSCT at the buccal mucosa. Nested PCR was used to detect CMV in the saliva. Time of death after HSCT was displayed, by means of the Kaplan-Meier method, for the following parameters: age and gender of the patient, donor gender, primary disease, stem cell source, platelet number, chronic graft vs host disease (cGVHD) of salivary glands and oral mucosa, and oral CMV shedding. Cox proportional hazards model was used for multivariate survival analysis. RESULTS: While none of the individuals in the control group showed positive swabs for CMV, the frequency of positive CMV oral swabs in patients at day + 100 after HSCT (45.2%) was statistically higher than before (7.2%) and 1 year after HSCT (17.5%). The presence of CMV was not associated with cGVHD and did not have any impact on post-transplant survival. CONCLUSIONS: The present study shows that oral CMV shedding occurs after HSCT, especially at day +100 post-transplant. Identification of CMV in the saliva might be important for the early diagnosis of CMV infection in allo-HSTC.     

HIV/AIDS患者CD4+T淋巴细胞数水平与巨细胞病毒感染分析

目的 分析住院及门诊人类免疫缺陷病毒（human immunodeficiency virus, HIV）感染者/获得性免疫缺陷综合征（acquired immunodeficiency syndrome, AIDS）患者（简称HIV感染者/AIDS患者）不同免疫状态下合并巨细胞病毒（human cytomegalovirus, HCMV）感染率情况,进一步了解HIV/AIDS合并HCMV感染的相关影响因素。方法 用流式细胞术进行HIV/AIDS体内CD4⁺T淋巴细胞亚群的计数,采用聚合酶链-荧光法进行HIV/AIDS尿液中HCMV-DNA检测,采用χ²检验分析HIV/AIDS不同来源、不同免疫状态下合并HCMV感染率差异。采用Logistic回归分析HIV/AIDS患者合并HCMV感染的相关因素。结果 817例HIV/AIDS患者合并HCMV感染阳性率21.5%（147/817）。HIV/AIDS患者的年龄、性别在是否合并HCMV感染中差异无统计学意义。CD4⁺T细胞≤50个/μL、合并梅毒感染为HIV/AIDS患者合并HCMV感染危险因素（P<0.001,OR=6.410,95%CI=4.141~9.922;P<0.05,OR=1.790,95%CI=1.206~2.657）,门诊和住院HIV/AIDS患者合并HCMV感染率差异有统计学意义（χ²=36.042,P<0.001）。以患者来源为分层因素进行CD4⁺T淋巴细胞计数与HCMV感染率分析,住院HIV/AIDS患者CD4⁺T细胞≤50个/μL时为合并HCMV危险因素（P<0.001,OR=4.796,95%CI=2.998~7.668）;门诊HIV/AIDS患者,CD4⁺T细胞≤50个/μL时为HIV/AIDS患者合并HCMV危险因素（P<0.001,OR=18.468,95%CI=6.668~51.154）。结论 AIDS期患者应及时筛查有无巨细胞病毒合并感染, AIDS CD4⁺T细胞数≤50个/μL、合并梅毒感染为HIV/AIDS患者合并HCMV感染的危险因素,门诊AIDS患者CD4⁺T细胞≤50个/μL其合并HCMV感染可能性为CD4⁺T细胞>50个/μL的18.468倍,应予以重视,尽早治疗,获得更好预后。     

抗病毒药物滴眼对巨细胞病毒阳性青光眼睫状体炎综合征患者炎症反应及眼压的影响

目的　探讨抗病毒药物滴眼对巨细胞病毒（cytomegalovirus,CMV）阳性青光眼睫状体炎综合征患者炎症反应水平及眼压的影响。方法　选取我院2015年4月至2018年10月收治的CMV阳性青光眼睫状体炎综合征患者共73例73眼,均给予20 g·L^-1更昔洛韦滴眼液治疗,比较患者治疗前后角膜后沉着物（keratic precipitates,KP）分布情况、房水闪辉分布情况、激素使用率、激素用量、眼压、降眼压药物使用率及降眼压药物使用种类。结果　患者治疗后羊脂状KP、钱币状KP及房水闪辉比例分别为52.05%、2.74%、0.00%,均显著低于治疗前的87.67%、26.03%、12.33%（均为P<0.05）。患者治疗后激素使用率和激素用量分别为47.95%,每天0（0~3）次,均显著低于治疗前的90.41%,每天3（0~4）次（均为P<0.05）。患者治疗后眼压、降眼压药物使用率及使用种类分别为（15.97±4.23）mmHg（1 kPa=7.5 mmHg）、41.10%、每天0（0~3）种,均显著低于治疗前的（26.34±5.10）mmHg、72.60%、每天2（0~3）种（均为P<0.05）。结论　抗病毒药物滴眼用于CMV阳性青光眼睫状体炎综合征患者可显著提高炎症反应控制效果,降低眼压,减少激素和降眼压药物使用,具有良好的临床应用价值。