Detection of cytomegalovirus proteins by flow cytometry in the blood of patients undergoing hematopoietic stem cell transplantation

Cytomegalovirus (CMV) infection and associated diseases continue to be a major complication encountered by patients undergoing high-dose chemoradiotherapy and hematopoietic stem cell transplantation (HSCT), A number of studies revealed that identification of CMV in the blood of HSCT patients was a predictor of future CMV disease. The purpose of this study was to determine if CMV proteins detected by flow cytometry could be a rapid and more quantitative way to monitor CMV infections and CMV antigenemia in HSCT patients. Preliminary studies showed that CMV immediate early (IE), early (E), and late (L) tegument proteins were specifically identified in CMV-infected cell lines and not in uninfected cells, We evaluated CMV antigen detection by flow cytometry in blood samples collected before and after transplantation in 56 serially collected blood samples from 17 HSCT patients and CMV protein expression was compared to CMV isolation, CMV IE and E proteins were not detected in any of the samples analyzed. However, CMV L protein detection by flow cytometry correlated with virus isolation in serially collected blood samples, Samples from 14 patients were evaluated by both techniques, at the same time intervals, There was a 100% correlation (8/8) between the lack of CMV antigen detection by flow cytometry and the failure to isolate infectious virus. Moreover, 5 of 6 patients who were positive for CMV L antigen by flow cytometry also were positive by virus isolation techniques. When flow cytometry and virus isolation did not detect CMV antigen on the same day, CMV positivity was first detected by flow cytometry, Then, 1-2 weeks later, positive virus isolation was documented. This study indicates that flow cytometric identification of CMV antigenemia correlates with isolation of CMV in HSCT patients and may be a predictive test for the rapid detection of CMV in the blood. (C) 1999 International Society for Experimental Hematology, Published by Elsevier Science Inc.