Coinfection of individual leukocytes with human cytomegalovirus and human immunodeficiency virus is a rare event in vivo

Infection with the human cytomegalovirus (HCMV) accelerates disease progression in human immunodeficiency virus 1 (HIV-1)-infected individuals. This has been attributed to the transaction of HIV-1 gene expression by HCMV gene products. For transactivation to be effective in vivo both viruses must be present in the same cell. We therefore examined blood samples from 13 HIV-1-infected patients with HCMV viremia for coinfection of individual leukocytes. In four of the patients lymph nodes were also examined. Multiple samples contained defined numbers (between 10 and 1000) of CD4⁺ lymphocytes or CD14⁺ monocytes were sorted by a FACS-based automated cell deposition unit. Samples were then analysed by a multiplex nested polymerase chain reaction, which can detect simultaneously HCMV and HIV DNA. The percentage of infected cells was calculated for each virus using the Poisson distribution. Between 0.43% and 6.2% of the CD4⁺ lymphocytes were infected with HIV and less than 0.15% with HCMV. The level of infection in CD14⁺ monocytes was always ≤0.11% for HIV and ranged between <0.05% and 0.58% for HCMV. Only seven of 1030 sorted samples from blood were positive for both viruses. In lymph nodes, none of the 144 samples tested were double-positive. This clearly shows that coinfection of individual human leukocytes with HIV and HCMV is a very rare event in vivo. Therefore, direct transactivation of HIV by HCMV in coinfected cells obtained from blood and lymph nodes may not explain the effect of HCMV on the prognosis of HIV-infected individuals. © 1996 Wiley-Liss, Inc.     

人巨细胞病毒立即早期蛋白功能的研究进展

人巨细胞病毒基因表达分为立即早期、早期和晚期三个时相。立即早期基因的表达在病毒基因和宿主细胞基因的表达和调控方面有重要作用,并能够影响宿主细胞周期进程。本文对立即早期蛋白调节病毒复制以及影响细胞周期进程方面的功能作一综述。     

套式聚合酶链反应及限制酶分析对孕早期孕妇巨细胞病毒感染的研究

在人巨细胞病毒（ＨＣＭＶ）－ＡＤ１６９株直接早期基因的ＥｃｏＲＩＪ片段内，自行设计二对引物，建立套式聚合酶链反应（ＰＣＲ）加限制酶分析检测ＨＣＭＶＤＮＡ，经实验证明有高度的特异性与敏感性，对其它疱疹类病毒及正常人基因组ＤＮＡ无交叉反应，一次性ＰＣＲ检测ＨＣＭＶ－ＡＤ１６９株基因组的最小检出量为１００ｆｇ，而套式ＰＣＲ可达１０ｆｇ，经ＰｓｔＩ酶切后，得到的片段与设计相符，对５８名孕早期孕妇外周静脉血     

Use of flow cytometry for characterization of human cytomegalovirus vaccine particles

Despite a 40-year effort, an effective vaccine against human cytomegalovirus (HCMV) remains an unmet medical need. The discovery of potent neutralizing epitopes on the pentameric gH complex (gH/gL/UL128/130/131) has reenergized HCMV vaccine development. Our whole-virus vaccine candidate, currently in a Phase I clinical trial, is based on the attenuated AD169 strain with restored expression of the pentameric gH complex. Given the complexity of a whole-virus vaccine, improved analytical methods have been developed to better characterize heterogeneous viral particles released from infected cells during vaccine production. Here we show the utility of a commercial flow cytometer for the detection of individual HCMV particles, either via light scattering or using fluorescence after labeling of specific antigens. Rapid measurements requiring minimal material provide near real-time information on particle concentration, distributions of different particle types, and product purity. Additionally, utilizing immunoreagents has allowed us to characterize the distribution of key antigens across individual particles and particle types.     

Diagnosis and counseling of fetal and neonatal HCMV infection

Fetal HCMV infection is investigated by amniocentesis when a maternal primary infection is diagnosed or ultrasound (US/MRI) abnormalities are observed. In fetal blood, prognostic markers of symptomatic congenital infection may be evaluated for parental counseling. At birth, viral load measurement in body fluids may correlate with long-term sequelae, but the prognostic accuracy of symptomatic infection increases when maternal, fetal, and neonatal parameters are combined.     

CD45RA expression on HCMV-specific effector memory CD8+ T cells is associated with the duration and intensity of HCMV replication after transplantation

In this cross-sectional study on 42 solid organ transplant recipients, the association of kinetics of human cytomegalovirus (HCMV) replication and EMRA HCMV-specific CD8+ T cells was investigated. Correlation was observed between the duration of HCMV replication after transplantation and CD45RA+CD27− (r = 0.609; p = 0.004), CD45RA+ CD28− (r = 0.579; p = 0.008) or CD45RA+CCR7− (r = 0.488; p = 0.029) HCMV-specific CD8+ T cells percentages. In the multivariate regression analyses, CD45RA+CD27−, CD45RA+CD28− or CD45RA+CCR7− HCMV-specific CD8+ T cells percentages increased 5.58% (p = 0.001), 5.35% (p = 0.001) or 4.49% (p = 0.012), respectively, with every 10-day increase in the duration of HCMV replication. Moreover, CD45RA+CD27− or CD45RA+CD28− frequencies increased 4.16% (p = 0.024) or 3.58% (p = 0.049), respectively, with every unity increase in log₁₀ genomes/mL. These observations support the major association between the frequency of EMRA HCMV-specific CD8+ T cells and the duration of post-transplant HCMV replication episodes in solid organ transplantation recipients.     

人巨细胞病毒感染对非特异性下腰痛患者内皮功能的影响

目的探讨内皮功能异常与人巨细胞病毒(HCMV)感染致非特异性下腰痛(NLBP)的关系,及其在NLBP发病中的作用。方法NLBP组50例,对照组36例,检测尿沉渣HCMV-DNA和包涵体,外周血HCMV-IgM、IgG和内皮素水平。结果NLBP组尿HCMV-DNA和包涵体,血HCMV-IgM、IgG阳性率高于正常对照组(P<0·05)。NLBP组内皮素水平增高(P<0·05)。结论NLBP常伴有HCMV感染及内皮功能异常。