Abstract
To establish a practical monitoring system of human herpesviruses reactivation in patients undergoing stem cell transplantation, we developed a new, very rapid, highly sensitive, and quantitative PCR assay for accurate measurement of human cytomegalovirus (CMV), human herpesvirus 6 (HHV-6) and Epstein–Barr virus (EBV) DNA using LightCycler. The LightCycler system revealed that there was a linear correlation in the wide range of viral template DNA at the indicated number of PCR cycles. Peripheral blood cells were collected from 16 patients undergoing stem cell transplantation. The cut-off level of CMV and HHV-6 was assessed as 102 copies/μg and that of EBV as 103. High numbers of CMV genomes were detected in 3/13 patients after transplant, and reactivation of HHV-6 was frequently seen, whereas none of the patient showed an elevation of EBV genome copies until the end of the observation period. In the present study, the reactivation of β herpesviruses is associated with the occurrence of thrombotic microangiopathy (TMA) in two patients undergoing allogeneic BMT. Therefore, it may contribute in clarifying the pathological potential of human herpesviruses using a large number of clinical samples. Our results suggest that this system may be useful for monitoring viral reactivation. Bone Marrow Transplantation (2001) 28, 975–980.
Similar content being viewed by others
References
Ljungman P . Immune reconstitution and viral infections after stem cell transplantation Bone Marrow Transplant 1998 21 (Suppl. 2): S72–74
Wang F-Z, Dahl H, Linde A et al. Lymphotropic herpesvirus in allogeneic bone marrow transplantation Blood 1996 88: 3615–3620
Broers AE, van Der Holt R, van Esser JW et al. Increased transplant-related morbidity and mortality in CMV-seropositive patients despite highly effective prevention of CMV disease after allogeneic T-cell-depleted stem cell transplantation Blood 2000 95: 2240–2245
Wang F-Z, Linde A, Dahl H, Ljungman P . Human herpesvirus 6 infection inhibits specific lymphocyte proliferation and is related to lymphocytopenia after allogeneic bone marrow transplantation Bone Marrow Transplant 1999 24: 1201–1206
Bethge W, Beck R, Jahn G et al. Successful treatment of human herpesvirus-6 encephalitis after bone marrow transplantation Bone Marrow Transplant 1999 24: 1245–1248
Cone RW, Hackman R, Huang M-LW et al. Human herpesvirus 6 in lung tissue from patients with pneumonitis after bone marrow transplantation New Engl J Med 1993 329: 156–161
Cone RW, Huang ML, Grey L et al. Human herpesvirus 6 infections after bone marrow transplantation: clinical and virologic manifestations J Infect Dis 1999 179: 311–318
Lei KI, Chan LY, Chan WY et al. Quantitative analysis of circulating cell-free Epstein–Barr virus (EBV) DNA levels in patients with EBV-associated lymphoid malignancies Br J Haematol 2000 111: 239–246
Ohyashiki JH, Suzuki A, Aritaki K et al. Use of real-time PCR to monitor human herpesvirus 6 reactivation after allogeneic bone marrow transplantation Int J Mol Med 2000 6: 427–432
Ohyashiki JH, Abe K, Ojima T et al. Quantification of human herpesvirus 6 in healthy volunteers and patients with lymphoproliferative disorders by PCR-ELISA Leukemia Res 1999 23: 625–630
Ohyashiki JH, Yamamoto K, Suzuki A, Ohyashiki K . use of real-time PCR to monitor human β herpesviruses infection in hematologic disorders Jpn J Clin Hematol 2001 42: 468–471
Nagata H, Konno A, Kimura N et al. Characterization of novel natural killer (NK)-cell and gammadelta T-cell lines established from primary lesions of nasal T/NK-cell lymphomas associated with the Epstein–Barr virus Blood 2001 97: 708–713
Maeda Y, Teshima T, Yamada M et al. Monitoring of human herpesvirus after allogeneic peripheral blood stem cell transplantation and bone marrow transplantation Br J Haematol 1999 105: 295–302
Tanaka N, Kimura H, Hoshino Y et al. Monitoring for herpesviruses in unrelated cord blood transplantation Bone Marrow Transplant 2000 26: 1193–1197
Takemoto Y, Takatsuka H, Wada H et al. Evaluation of CMV/human herpes virus-6 positivity in bronchoalveolar lavage fluids as early detection of acute GVHD following BMT: evidence of a significant relationship Bone Marrow Transplant 2000 26: 77–81
Vogelsang GB . How I treat chronic graft-versus-host disease Blood 2001 97: 1196–1201
Zeigler Z, Kelton J, Moore J et al. Calpain activity in bone marrow transplant-associated thrombotic thrombocytopenic purpura Bone Marrow Transplant 1999 24: 641–645
Matsuda Y, Hara J, Miyoshi H et al. Thrombotic microangiopathy associated with reactivation of human herpesvirus 6 following high-dose chemotherapy with autologous bone marrow transplantation in young children Bone Marrow Transplant 1999 24: 919–923
Visseren FL, Verkerk MS, Bouter KP et al. Interleukin-6 production by endothelial cells after infection with influenza virus and cytomegalovirus J Lab Clin Med 1999 134: 623–630
Wu CA, Shanley JD . Chronic infection of human umbilical vein endothelial cells by human herpesvirus-6 J Gen Virol 1998 79: 1247–1256
Waldman WJ, Knight DA, Adams PW . Cytolytic activity against allogeneic human endothelia: resistance of cytomegalovirus-infected cells and virally activated lysis of uninfected cells Transplantation 1998 66: 67–77
Fish KN, Soderberg-Naucler C, Mills LK et al. Human cytomegalovirus persistently infects aortic endothelial cells J Virol 1998 72: 5661–5668
Kahn M, Siegel-Acel D, Stenglein S et al. Efficient lytic infection of human arterial endothelial cells by human cytomegalovirus strains J Virol 2000 74: 7628–7635
Ljungman P, Wang FZ, Clark DA et al. High levels of human herpesvirus 6 DNA in peripheral leukocytes are correlated to platelet engraftment and disease in allogeneic stem cell transplantation Br J Haematol 2000 111: 774–781
Isomura H, Yoshida M, Namba H et al. Suppressive effects on human herpesvirus-6 on thrombopoietin-inducible megakaryocyte colony formation in vitro J Gen Virol 2000 81: 3663–3667
Imbert-Marcille BM, Tang XW, Lepelletier D et al. Human herpesvirus 6 infection after autologous or allogeneic stem cell transplantation: a single-center prospective longitudinal study of 92 patients Clin Infect Dis 2000 31: 881–886
Nitsche A, Muller CW, Radonic A et al. Human herpesvirus 6A DNA is detected frequently in plasma but rarely in peripheral blood leukocytes of patients after bone marrow transplantation J Infect Dis 2001 183: 130–133
Ihira M, Yoshikawa T, Suzuki K et al. Correlation between human herpesvirus 6 and 7 infections after living related liver transplantation Microbiol Immunol 2001 45: 225–232
Lo YM, Chan LY, Chan AT et al. Quantitative and temporal correlation between circulating cell-free Epstein–Barr virus DNA and tumor recurrence in nasopharyngeal carcinoma Cancer Res 1999 59: 5452–5455
Orii T, Ohkohchi N, Kikuchi H et al. Usefulness of quantitative real-time polymerase chain reaction in following up patients with Epstein-Barr virus infection after liver transplantation Clin Transplant 2000 4: 308–317
Zhou CF, Abe K, Wang P et al. Efficient propagation of human herpesvirus 6B in a T cell line, derived from a patient with adult T cell leukemia/lymphoma Microbiol Immunol 1999 43: 425–436
Acknowledgements
The authors are indebted to Prof J Patrick Barron of the International Medical Communications Center of Tokyo Medical University for his review of this manuscript. The authors wish to thank Dr Naoko Shoji, Ms Ayako Hirota and Ms Michiko Kimura for their help.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Aritaki, K., Ohyashiki, J., Suzuki, A. et al. A rapid monitoring system of human herpesviruses reactivation by LightCycler in stem cell transplantation. Bone Marrow Transplant 28, 975–980 (2001). https://doi.org/10.1038/sj.bmt.1703266
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.bmt.1703266
- Springer Nature Limited