Human cytomegalovirus load in plasma and bronchoalveolar lavage fluid: a longitudinal study of lung transplant recipients.

BACKGROUND: In lung transplant recipients (LTRs), the measurement of human cytomegalovirus (HCMV) load dynamics in the local environment of the lung allograft may offer distinct advantages over their assessment in the peripheral blood compartment. METHODS: We compared HCMV load in paired bronchoalveolar lavage (BAL) and plasma samples in a prospective cohort of LTRs. RESULTS: In all, 182 paired samples were collected from 41 LTRs. Qualitative findings were concordant for 79.7% of the paired samples (virus load detected or absent in 8.1% and 71.5% of paired samples, respectively). In 35 paired samples (20.3%), HCMV was detected in the BAL but not the plasma sample; the converse was not found for any of the paired samples. HCMV inclusions were histologically detected in the lungs of 8 patients on 9 occasions. HCMV was detected in both BAL and plasma samples on 7 of the 9 occasions and in only the BAL sample on 2 occasions. Application of a threshold virus load in BAL samples, to predict histologically proven HCMV infection, was associated with improved diagnostic sensitivity and specificity. CONCLUSIONS: Quantification of HCMV in the lung allograft correlates with histological detection and, in addition, offers the potential to monitor subclinical viral replication in the lung allograft.

HCMV DNA detection and quantitation in the plasma and PBL of lung transplant recipients: COBAS Amplicor HCMV monitor test versus in-house quantitative HCMV PCR.

BACKGROUND: Human cytomegalovirus (HCMV) reactivation may cause severe disease in immunosuppressed patients. Quantitation of HCMV viral load in the blood has been shown to be important in predicting for HCMV disease, however the particular blood compartment that should be tested, plasma versus peripheral blood leukocytes (PBL), has been subject to debate. OBJECTIVES: To simultaneously compare HCMV viral loads in the PBL using an in-house quantitative HCMV polymerase chain reaction (PCR) assay and in the plasma using the commercially available COBAS Amplicor HCMV monitor test, in a cohort of lung transplant recipients (LTR). STUDY DESIGN: Sequential paired plasma and PBL samples were collected (n=98) from a total of 21 LTR during the first 6 months post lung transplantation. HCMV viral loads were assessed in the PBL using an in-house quantitative HCMV PCR assay and the plasma using the COBAS Amplicor HCMV monitor test. HCMV disease in LTR was defined as histopathologically proven HCMV pneumonitis. RESULTS: HCMV deoxyribonucleic acid (DNA) was detected in 39/98 (40%) of total samples with excellent agreement between the two strategies. HCMV was detected in both the plasma and PBL in 38/39 (97%) of HCMV positive samples. HCMV viral loads were higher in patients with HCMV pneumonitis in both the PBL and plasma compared to patients without HCMV pneumonitis. Greater than 10-fold increases in HCMV DNA levels had a sensitivity of 88% and a specificity of 73% in the plasma for HCMV pneumonitis and a positive and negative predictive value of 70 and 89%, respectively. Greater that 10-fold increases in HCMV DNA levels in the PBL had a sensitivity of 88% and a specificity of 64%, for HCMV pneumonitis and a positive and negative predictive value of 64 and 88%, respectively. CONCLUSIONS: Acknowledging the difference in assay methods used for HCMV DNA quantitation, the in-house PCR PBL assay and the COBAS Amplicor HCMV monitor plasma assay predicted equally well for HCMV pneumonitis in LTR.

Relation of local platelet glycoprotein IIb/IIIa independent activation during coronary angioplasty in acute myocardial infarction to recovery of left ventricular function.

We assessed glycoprotein (GP) IIb/IIIa independent platelet activation in coronary sinus and peripheral blood from patients who underwent angioplasty for acute myocardial infarction and stable angina. Despite complete blockade of the activated GP IIb/IIIa receptor with abciximab in patients with acute myocardial infarction, unsuppressed local GP IIb/IIIa independent activation was associated with a lack of recovery of left ventricular function.

Bronchiolitis obliterans syndrome and early human cytomegalovirus DNAaemia dynamics after lung transplantation.

BACKGROUND: Bronchiolitis obliterans syndrome (BOS) remains a major cause of morbidity and mortality after lung transplantation. The major identified risk factors for BOS are acute rejection and human cytomegalovirus (HCMV) infection, the latter despite the use of relatively insensitive and nonspecific measures such as HCMV pneumonitis and HCMV serostatus, respectively. We hypothesized that a more accurate prospective analysis of HCMV reactivation in lung transplant recipients (LTRs) would improve our understanding of the association between HCMV and BOS development. METHODS: In 26 LTRs, HCMV DNAaemia was measured using quantitative polymerase chain reaction at monthly intervals during the initial 6 months posttransplantation. BOS was defined as a sustained irreversible 20% decrease in FEV1 compared with the best baseline FEV1 posttransplantation in the absence of any other cause. RESULTS: Of the 26 LTRs, 23 were assessable with regard to the BOS outcome variable. At a median follow-up of 37 months, 10 patients had developed BOS. During the first 6-month monitoring period, HCMV DNAaemia was detected in 15 of the 23 patients on at least one occasion, and there were 12 episodes of HCMV pneumonitis in eight patients. Episodes of grade A3 or greater acute rejection occurred in eight LTRs, six of whom had been HCMV DNAaemia positive at least once and four of whom also demonstrated HCMV pneumonitis. Our results revealed a strong association between BOS and early HCMV DNAaemia detection (univariate analysis [P=0.002] and freedom from BOS analysis [P=0.006]). CONCLUSION: Early HCMV DNAaemia in LTRs is associated with the development of BOS despite routine ganciclovir prophylaxis.

beta-Herpesvirus (human cytomegalovirus and human herpesvirus 6) reactivation in at-risk lung transplant recipients and in human immunodeficiency virus-infected patients.

Human herpesvirus (HHV)-6 is a beta-herpesvirus-like human cytomegalovirus (HCMV) with the potential to reactivate in immunocompromised persons. HHV-6 and HCMV were assessed in the peripheral blood leukocytes of 26 lung transplant recipients and of 37 human immunodeficiency virus (HIV)-infected patients receiving highly active antiretroviral therapy, to determine the degree of concordance between HHV-6 and HCMV reactivation in different biologic settings. In the lung transplant recipients (145 samples), HHV-6 was not detected, even though 44 (30%) of 145 samples were from 9 HCMV DNA-positive patients (13 episodes of HCMV pneumonitis). Among the HIV-infected patients (172 samples), HCMV DNA was detected in 29 (17%) of 172 samples from 10 patients (4 episodes of HCMV disease). HHV-6 DNA was detected in 2 HIV-infected patients who did not have HCMV detected at that time. These findings suggest that the pathobiologic control mechanisms for these 2 beta-herpesviruses may be significantly different.