Human Cytomegalovirus Infection in Lung Transplant Recipients Triggers a CXCL‐10 Response

Human cytomegalovirus (HCMV) causes significant morbidity in lung transplant recipients (LTRs). The clinical effects of HCMV replication are determined partly by a type 1 T‐helper cell (Th1) response. Because the chemokine interferon‐inducible protein of 10 kilodaltons (IP‐10, CXCL‐10) induces a Th1 response, we investigated whether HCMV triggers IP‐10 in LTRs. The IP‐10 concentration and HCMV DNA load were determined in 107 plasma and 46 bronchoalveolar lavage fluid (BALF) samples from 36 LTRs. Initial HCMV detection posttransplantation was significantly associated with increased plasma IP‐10, regardless of whether the patients showed HCMV DNAemia (p = 0.001) or HCMV replication only in the allograft (p < 0.0001). In subsequent episodes of HCMV detection, plasma IP‐10 increased regardless of whether HCMV was detected in blood (p = 0.0078) or only in BALF (p < 0.0001) and decreased after successful antiviral therapy (p = 0.0005). Furthermore, levels of HCMV DNA and IP‐10 correlated statistically (p = 0.0033). Increased IP‐10 levels in HCMV‐positive BALF samples were significantly associated with severe airflow obstruction, as indicated by a decrease in forced expiratory volume in one second (FEV1). Our data indicate that HCMV replication in LTRs evokes a plasma IP‐10 response and that, when an IP‐10 response is observed in BALF, it is associated with inflammatory airway obstruction in the allograft.     

肝炎综合征婴儿HCMV-IgM定量检测的临床价值

目的：探讨检测肝炎综合征婴儿血清中人巨细胞病毒（human cytomegalovirus,HCMV）IgM含量在诊断和动态监测婴儿HCMV感染中的临床意义。方法：ELISA检测了97例肝炎综合征婴儿（婴肝患儿）血清HCMV-IgM含量,并对其中48例HCMV-IgM阳性患儿在治疗过程中进行了血清HCMV-IgM定量和肝功能的动态监测。结果：97例婴肝患儿血清HCMV-IgM阳性率为49.48%（48/97）,HCMV肝炎患儿治疗前后血清HCMV-IgM定量检测均值分别为（27.7±3.92）、（12.84±2.87）IU/ml,HCMV肝炎患儿治疗后血清HCMV-IgM定量检测值显著下降（t=19.35,P〈0.05）,并与肝脏转氨酶及总胆红素的下降同步。结论：HCMV感染为婴肝患儿的主要病因,血清HCMV-IgM定量检测有助于婴肝患儿的诊断和婴儿巨细胞病毒性肝炎疗效观察。     

HCMV感染患儿血浆可溶性HLA-G及IL-10检测的意义

目的探讨血浆可溶性人类白细胞抗原-G(soluble human leukocyte antigen-G,sHLA-G)及白细胞介素-10(in-terleukin-10,IL-10)在儿童巨细胞病毒(human cytomegalovirus,HCMV)感染中的意义。方法分别收集75例HCMV活动性感染患儿的外周血及尿液标本,采用酶联免疫吸附试验(ELISA)测定血浆sHLA-G和IL-10水平;采用实时荧光定量PCR(FQ-PCR)检测尿液HCMV DNA载量。结果 HCMV活动性感染患儿血浆sHLA-G水平[54.91(6.75～282.72)U/ml]明显高于对照组[21.32(1.07～260.35)U/ml],差异有统计学意义(P<0.001);血浆IL-10的水平[9.24(1.61～41.77)pg/ml]明显高于对照组[1.82(1.03～3.98)pg/ml],差异有统计学意义(P<0.001);血浆sHLA-G和IL-10水平与尿液HCMV DNA载量之间无明显相关性(P>0.05)。结论 HCMV活动性感染患儿血浆sHLA-G和IL-10水平显著升高,可作为HCMV活动性感染诊断的指标。     

Comparative study of three PCR assays with antigenaemia and serology for the diagnosis of HCMV infection in thoracic transplant recipients

Three PCR assays were evaluated for the detection of human cytomegalovirus (HCMV) infection in heart and lung transplant recipients in comparison with HCMV antigenaemia and serology assay. Polymorphonuclear leucocyte (PMNL) samples taken at regular intervals after transplantation were tested for HCMV DNA using primer sets homologous to the glycoprotein B (gp58), major immediate early (IE1), and structural phosphoprotein (pp150) regions. The detection of HCMV infection at various times after transplantation showed all three primer sets to have a sensitivity of 100% and a specificity of 92.3% for the detection of HCMV infection although overall the gp58 primer set was found to be significantly more frequently associated with a positive PCR result than the IE1 (P = 0.0228) and pp150 (P = 0.0015) primer sets. The positive PCR result had a positive predictive value of 27.8% for HCMV disease. Detection of HCMV infection was first by the PCR assay, and significantly before the HCMV antigenaemia assay. Of nine patients who received antiviral therapy while PCR positive, only one patient cleared HCMV DNA from PMNLs during treatment but became positive again 17 days later. Quantitative PCR methodologies may improve the predictive value of PCR for HCMV disease and its value for monitoring antiviral therapy. © 1996 Wiley-Liss, Inc.     

Monitoring for cytomegalovirus infection in organ transplant recipients: Analysis of pp65 antigen,DNA and late mRNA in peripheral blood leukocytes

The use of sensitive and specific methods for rapid and reliable diagnosis is required due to the considerable impact of human cytomegalovirus (HCMV) in organ transplant recipients. For this purpose the demonstration of the presence of viral antigens in peripheral blood leukocytes (PMNLs) and of viral nucleic acids in the same cells or in sera would seem to be of valid support. The present study was designed to test pp65 antigen, HCMV DNA and HCMV late mRNA in order to provide clinical information for the management of the infection. Fifty solid organ recipients were monitored for six months after transplant. The data obtained from the various tests were analysed from the first evidence of HCMV infection revealed by positive antigenaemia and/or DNA-polymerase chain reaction (PCR). In 3 asymptomatic and in 7 symptomatic patients, PCR became positive 1–2 weeks before antigenaemia but PCR did not discriminate the clinical evolution of HCMV infection. The antigenaemia test well correlated to the development of viral infection being positive in all symptomatics and in 31, 2% of asymptomatics. The antigenic load >100/2 × 10⁵ positive cells was always associated with clinical signs of illness. The detection of late mRNA was more indicative of the virus replicative status in the follow-up of patients treated with ganciclovir. In some cases there was evidence, prior to the other two tests, the block of viral replication due to the antiviral therapy and in others the onset of HCMV infection relapse. J. Med. Virol. 53:189–195, 1997. © 1997 Wiley-Liss, Inc.     

抗HCMV不同多肽的6株McAb的研究

目有：为了制备人巨细胞病毒（HCMV）单特异性可溶性多肽的单克隆抗体。方法：采用细胞杂交技术建立了分泌单克隆抗体（McAb）的13株杂交瘤细胞,本文选取6株杂交癌细胞作了进一步研究。结果：6株杂交瘤细胞分泌的McAb均为IgGl,重链、轻链分子量分别为50KD、25KD。间接免疫荧光试验表明：McAb8B8相应的多肽为即刻早期抗原;而McAb7B4、7D7、7E7、7E11、8D6的相应病毒多肽为晚期抗原。免疫印迹试验表明：McAb7B4、7D7、7E7、7E11、8B8、8D6相应的病毒多肽分子量分别为46、150、38、51、72、65KDO结论：6株McAb可用作HCMV感染的诊断。     

抗HCMV不同多肽McAb的鉴定

目的为了将单克隆抗体 (Mc Ab)用于诊断技术 ,我们首先建立了 13株抗人巨细胞病毒 (HCMV)的 Mc Ab,选取 6株 Mc Ab作进一步鉴定。方法这 6株 Mc Ab的鉴定主要采用间接免疫荧光试验、免疫印迹试验等方法。结果间接免疫荧光试验表明 :Mc Ab8B8相应的多肽为早期抗原 ;而 Mc Ab7B4、7D7、7E7、7E11、8D6的相应病毒多肽为晚期抗原。免疫印迹试验的结果表明 :Mc Ab7B4、7D7、7E7、7E11、8B8、8D6相应的病毒多肽分子量分别为 46、15 0、38、5 1、72、6 5 kd。结论 Mc Ab8B8可不用于 HCMV的快速诊断。     

Circulating cytomegalic endothelial cells are associated with high human cytomegalovirus (HCMV) load in AIDS patients with late-stage disseminated HCMV disease

The prevalence of circulating cytomegalic endothelial cells, detected currently by the pp65-antigenemia assay and described previously in blood of transplanted and AIDS patients with disseminated human cytomegalovirus (HCMV) infection, was found to be 2.9% in the AIDS population and 6.5% in the fraction of the AIDS population with HCMV in blood. Cytomegalic endothelial cells increased to 39.7% and 48.4%, respectively, in AIDS patients with very high levels of antigenemia and viremia, while an end organ disease reached an incidence of 76.4%. Positive and negative predictive values of cytomegalic endothelial cell detection for diagnosis of HCMV end organ disease were 73.1% and 21.4% with antigenemia levels >1,000, respectively. On the other hand, in a selected group of 38 cytomegalic endothelial cell-positive AIDS patients with <50 CD4+ T cells/μl and late-stage HCMV disease, who were followed-up for variable periods of time, the prevalence of high level antigenemia was 95.3%, that of viremia 86.0% and that of L-DNAemia 92.7%, while the incidence of HCMV end organ disease was 84.2%. In this population, it was shown that cytomegalic endothelial cell presence was associated with lack of (56.0% of episodes) or insufficient (4.0%) anti-HCMV treatment or emergence of HCMV drug-resistant strains (17.3%) or short-term response to antiviral treatment (22.7%); was determined in the same patient by different conditions during follow-up. Longitudinal observations indicated that cytomegalic endothelial cells were detected often in blood at least 3 months later than end organ disease suggesting that the duration of end organ disease was a cofactor associated with the appearance of cytomegalic endothelial cells. J. Med. Virol. 55:64–74, 1998. © 1998 Wiley-Liss, Inc.     

Human cytomegalovirus glycoprotein B genotypes in blood of AIDS patients: Lack of association with either the viral DNA load in leukocytes or presence of retinitis

It has been suggested that human cytomegalovirus (HCMV) glycoprotein B (gB) genotypes could be used as a marker for viral virulence in patients with AIDS. The present study was designed to evaluate a possible association between specific gB genotypes, the presence of HCMV retinitis, and the HCMV viral load. Fifty-four blood samples were obtained from 54 HIV- and HCMV-infected patients. Twenty-seven of these patients were asymptomatic for HCMV, whereas the other 27 patients had been diagnosed recently with HCMV retinitis. HCMV gB genotyping was carried out by using restriction enzyme analysis of PCR-amplified PMNL extracts. Determination of the HCMV viral load in the same specimens was carried out using a quantitative-PCR. HCMV gB genotype 2 was found more frequently than other genotypes in PCR-amplified polymorphonuclear leukocytes (PMNL) of patients with AIDS (P < 0.05) but not more frequently in samples from patients with HCMV retinitis. No significant association was found between any HCMV gB genotypes and the viral load in blood. In conclusion, the actual HCMV gB genotyping system using PMNL provides no additional benefit over the viral load in blood for identification of HIV-infected subjects at risk of HCMV disease. J. Med. Virol. 59:98–103, 1999. © 1999 Wiley-Liss, Inc.