Lymphoproliferative disease after allogeneic stem cell transplantation--pre-emptive diagnosis by quantification of Epstein-Barr virus DNA in serum.
BACKGROUND
Lymphoproliferative disease (PTLD) is a life-threatening complication of organ transplantation. In matched, allogeneic, non-T-cell-depleted stem-cell transplantations (SCT) the disease develops early but has been thought to be rare.


OBJECTIVE
We determined by strict histopathological criteria the incidence of fatal Epstein-Barr-virus (EBV)-related PTLD in a large number of SCT, and assessed the diagnostic value of a real-time quantitative polymerase chain reaction (qPCR) for EBV-DNA in serum specimens.


METHODS
Of the 257 SCT performed in Helsinki during 1994-1999, 132 (51%) recipients were alive and 125 (49%) had succumbed by June 2001. The necropsies were analyzed for EBV-related PTLD as evidenced by disseminated lymphocytic infiltrates labeled histochemically for antigens and RNA (EBER 1 and 2) detectable by in situ technology. From a subset of the PTLD cases (N=12) and a series of corresponding stem-cell recipient controls (N=36), consecutive samples of serum (N=103 and 364, respectively) were studied by qPCR for EBV-DNA, and the clinical data were reviewed.


RESULTS
The post-mortem analysis revealed 18 cases of PTLD (14% of the deceased), all of whom had received intensive immunosuppressive treatment including anti-thymocyte globulin for treatment or prophylaxis of graft versus host disease (GVHD). By using qPCR all the PTLD patients became EBV-DNA positive, in progressively rising copy numbers. EBV-DNA was first detectable 70 (median; range 24-154) days after SCT or 23 (4-86) days before death; i.e. earlier than the symptoms which appeared 15 (2-85) days before death. Among the SCT controls, EBV-DNA occurred sporadically (in only 3.9% sera).


CONCLUSIONS
qPCR for EBV-DNA in serum is a highly sensitive (100%) and specific (96%) diagnostic approach. Intensely immunosuppressed stem-cell recipients are at a great risk of developing PTLD, and should be carefully monitored for EBV-DNA, for pre-emptive treatment of this life-threatening disorder.
Aalto
Sanna M
SM
Department of Virology, Haartman Institute, University of Helsinki and Helsinki University Central Hospital, FIN-00290 Helsinki, Finland.
Juvonen
Eeva
E
Tarkkanen
Jussi
J
Volin
Liisa
L
Ruutu
Tapani
T
Mattila
Petri S
PS
Piiparinen
Heli
H
Knuutila
Sakari
S
Hedman
Klaus
K
D004279:DNA, Viral / Q000097:blood*; D020031:Epstein-Barr Virus Infections / Q000821:virology; D004854:Herpesvirus 4, Human / Q000302:isolation & purification*; D006801:Humans; D015994:Incidence; D008232:Lymphoproliferative Disorders / Q000453:epidemiology / Q000401:mortality*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D033581:Stem Cell Transplantation / Q000009:adverse effects*; D014184:Transplantation, Homologous / Q000009:adverse effects*; D019562:Viral Load
D004279:DNA, Viral