|
|
 Previous Article | Table of Contents | Next Article 
Blood, 1 November 2000, Vol. 96, No. 9, pp. 3279-3281
BRIEF REPORT
Molecular evidence of organ-related transmission of Kaposi
sarcoma-associated herpesvirus or human herpesvirus-8 in
transplant patients
Mario Luppi,
Patrizia Barozzi,
Gaia Santagostino,
Raffaella Trovato,
Thomas F. Schulz,
Roberto Marasca,
Davide Bottalico,
Lucia Bignardi, and
Giuseppe Torelli
From the Department of Medical Sciences, Section of
Hematology, University of Modena and Division of Nephrology, University
of Parma, Italy; and Department of Medical Microbiology and
Genitourinary Medicine, University of Liverpool, UK.
 |
Abstract |
In transplant patients, Kaposi sarcoma (KS)-associated herpesvirus
or human herpesvirus-8 (HHV-8) infection is associated with the
development of KS, primary effusion lymphoma and Castleman disease.
Whether HHV-8 is either reactivated in the recipient or transmitted by
the donor has been investigated so far only by serologic studies. Thus,
we addressed the issue of HHV-8 transmission in the transplantation
setting by molecular methods. We exploited the high level variability
of the orf-K1 gene and the polymorphism of the
orf-73 gene of the HHV-8 genome to assess the genetic
relatedness of the HHV-8 strains identified in the posttransplant KS
lesions that developed, simultaneously, 20 months after
transplantation, in 2 recipients of twin kidneys from the same cadaver
donor. The 100% identity of nucleotide sequence of the most
variable viral region and the presence of the same, single
orf-73 type in both patients provides strong molecular
evidence of organ-related transmission of HHV-8 in the setting of transplantation.
(Blood. 2000;96:3279-3281)
© 2000 by The American Society of Hematology.
| |
Introduction |
Kaposi sarcoma-associated herpesvirus (KSHV) or
human herpesvirus-8 (HHV-8) has been implicated in the pathogenesis of
Kaposi sarcoma (KS), primary effusion lymphoma (PEL), and multicentric Castleman disease (CD) of plasma cell type.1,2 HHV-8 is
not ubiquitous in the general population, with seroprevalence rates being very low in the UK and US and higher only in certain geographic areas of the world (the Mediterranean area and Africa) with a known
high incidence of classic/endemic KS.1,2 HHV-8 is spread sexually, but nonsexual routes of transmission are likely to occur in
HHV-8 endemic areas, where this herpesviral infection is acquired early
in childhood.1,2
In the setting of transplantation, HHV-8 infection has been associated
with an increased risk of developing posttransplant KS,3-10 and few cases of HHV-8-associated PEL, CD, and
plasmacytic proliferations have been also described in recipients of
solid organs.11-13 It has been recently proposed that
posttransplant KS is primarily due to HHV-8 reactivation in endemic
areas and to primary infection in nonendemic areas.6
However, all previous reports have focused only on the use of serologic
assays to investigate whether HHV-8 infection is either preexisting in
the recipient or transmitted by the donor.3-10
In this report, we used molecular methods to address the issue of HHV-8
transmission in the unique clinical setting of 2 renal transplant
patients, who had cutaneous KS develop, simultaneously, 20 months after
receiving twin kidneys from the same cadaver donor. HHV-8 genome
contains, at the left end, the orf-K1 gene, encoding a
transmembrane protein that exhibits much more sequence variability than
the rest of the viral genome.14,15 The variability in this
gene among different isolates has so far been exploited to study the
molecular epidemiology of HHV-8 obtained from individuals in different
parts of the world.14,15 Moreover, molecular polymorphism has been recently found within the internal repeat domain of the orf-73 gene of the HHV-8 genome.16 The size of
the orf-73 internal repeat domain, which is dependent of the
variable number of repeats, is characteristic of each individual HHV-8
genotype or isolate, and has been used to develop a new genotyping
technique.16 Thus, we exploited the high-level variability
in the orf-K1 gene and the polymorphism of the
orf-73 gene to assess the degree of genetic relatedness of
the HHV-8 strains detected in the posttransplant KS lesions of the 2 renal transplant patients.
| |
Study design |
In August 1994, 2 Italian women, 46 and 45 years old,
respectively, living in different cities, received renal grafts from the same cadaver donor, a 53-year-old Italian man. Both patients received immunosuppression with cyclosporin A (CsA) and
methylprednisolone (MP), but, during the posttransplantation course,
both were also treated with MP pulses for an episode of acute
rejection. Azathioprine was also added to the immunosuppressive regimen
in one case. Twenty months after transplantation, cutaneous KS of the
lower extremities developed simultaneously in both patients. A detailed
description of the clinical histories of the 2 transplant patients has
been reported elsewhere.17 Reduction of immunosuppressive
therapy has led to KS regression in both patients, which still persists.
Frozen biopsy specimens from the cutaneous KS lesions of both patients,
as well as the Ficoll separated peripheral blood mononuclear cells
(PBMCs) collected about 4 years after the initial diagnosis of KS, were
available for molecular analysis. DNA was purified after proteinase
digestion and phenol chloroform extraction. HHV-8 DNA was amplified on
the same material by polymerase chain reaction (PCR) with primers
specific for the orf-K1 gene, as we previously described.15 The PCR products were subjected to direct
automated sequence analysis, and phylogenetic analysis of the
K1 gene was performed as we previously
reported.15 PCR amplification of the fragment of the
orf-73 internal repeat domain was performed as described by
Gao et al.16 To avoid false-positive results, all
procedures were performed in strict adherence to the recommendations of
Kwok and Higuchi.18 Negative controls consisting of all
reagents, except sample DNA, were also present during the DNA
extraction and equalled or exceeded the number of samples assayed.
| |
Results and discussion |
HHV-8 DNA was detected by PCR in the cutaneous posttransplant KS
lesion from both renal recipients. Sequence analysis of the 2 highly
variable regions of the orf-K1 gene from the 2 patients showed 100% identity (Figure 1A) and
phylogenetic analysis showed that the infecting strain belonged to
clade C, which is rather common in Italy.15 The
orf-K1 gene sequence results were also the same in the PBMCs
collected from the 2 patients about 4 years after the initial diagnosis
of KS. PCR of the orf-73 internal repeat domain showed a
single band of the same size in both patients (Figure 1B). In the same
assay, a single band of different size was detected in one classic KS
and in one PEL specimen, indicating the occurrence of different
isolates in these latter cases (Figure 1B).
View larger version (45K):
[in this window]
[in a new window]
| Figure 1.
Alignment of the nucleotide sequences and genotyping of
the internal repeat domain detected in the KS lesions.
(A) Alignment of the nucleotide sequences of the 2 highly
variable regions (VR1 and VR2) of the orf-K1 gene. The
alignment includes a prototype K1 sequence (clade A) from the BC-1 PEL
cell line (GeneBank accession number U75698), and the K1 sequences
(clade C) detected in the KS lesions from each of the 2 renal
recipients (Pt 1 and Pt 2), after transplantation. Dots indicate
identity with the first sequence in the alignment. (B) Genotyping of
the polymorphic internal repeat domain of orf-73 gene
detected in the KS lesions from each of the 2 renal recipients (Pt 1 and Pt 2), after transplantation, in the cutaneous KS lesion from an
elderly Italian patient with classic KS (KS), and in the pleural PEL
specimen from an elderly HIV negative Italian patient (PEL). nc,
negative control, represented by the DNA from HHV-8 negative Raji cell
line; mk VII, molecular weight marker (Boehringer Mannheim,
Germany).
|
|
Sera collected before and after transplantation from 28 recipients who
had posttransplant KS develop, have so far been examined for
anti-HHV-8 antibodies in 8 independent studies.3-10
Twenty-three patients, most of whom originated from endemic areas, were
infected with HHV-8 before the graft, suggesting that KS was mainly due to virus reactivation.3,5-10 In the remaining 5 patients
who seroconverted after transplantation, transmission of HHV-8 from the
organ donor to the recipient was suggested.3,4 However, for 4 of these patients, originating from nonendemic areas, the source
of infection was not determined, as serum samples from their paired
donors were not available.4 For one patient, originating from an endemic area (southern Italy), serum from the living-related donor was seropositive for HHV-8, providing the only clear available example of organ-related transmission of HHV-8, although the serum was
tested more than 2 years after transplantation (Table
1).3
View this table:
[in this window]
[in a new window]
|
Table 1.
Updated review of serologic studies for anti-HHV-8
antibodies in patients with posttransplant Kaposi
sarcoma
|
|
In our study, the finding of a 100% identity of nucleotide sequence of
the most variable region of HHV-8 genome and the presence of the same,
single orf-73 type in the posttransplant KS lesions of the 2 patients receiving twin kidneys from the same cadaver donor, provide
strong molecular evidence of organ-related transmission of HHV-8.
Although among populations with high HHV-8 seroprevalence, the
development of posttransplant KS has been found to be mainly associated
with HHV-8 reactivation, the occurrence of HHV-8 transmission from the
donor should not be underestimated. Analysis of the genetic relatedness
in the highly variable orf-K1 gene and the polymorphic orf-73 gene of HHV-8 isolates from the transplant recipients
and their paired donors represents a useful tool to obtain a molecular tracing of HHV-8 transmission in this clinical setting.
| |
Footnotes |
Submitted March 6, 2000; accepted July 3, 2000.
M.L., P.B., and G.S. equally contributed to the study.
Supported by a grant from Associazione Italiana per la Ricerca sul
Cancro (A.I.R.C.), Milan, Italy, and the European Concerted Action on
HHV-8/KSHV (M.L.), a grant from M.U.R.S.T. (G.T.), and a grant from NHS
Biomedical R&D Fund (RDO/22/09) (T.F.S.).
The publication costs of this
article were defrayed in part by
page charge payment. Therefore,
and solely to indicate this fact,
this article is hereby marked
"advertisement"
in accordance with 18 U.S.C.
section 1734.
Reprints: Giuseppe Torelli, Department of Medical Sciences,
Section of Hematology, Policlinico, Via del Pozzo 71, 41100, Modena,
Italy; e-mail: gtorelli{at}unimo.it.
| |
References |
1.
Schulz TF.
Kaposi's sarcoma-associated herpesvirus (human herpesvirus-8).
J Gen Virol.
1998;79:1573-1591[Medline]
[Order article via Infotrieve].
2.
Moore PS, Chang Y.
Kaposi's sarcoma (KS), KS-associated herpesvirus, and the criteria for causality in the age of molecular biology.
Am J Epidemiol.
1998;147:217-221[Free Full Text].
3.
Parravicini C, Olsen SJ, Capra M, et al.
Risk of Kaposi's sarcoma-associated herpes virus transmission from donor allografts among Italian posttransplant Kaposi's sarcoma patients.
Blood.
1997;90:2826-2829[Abstract/Free Full Text].
4.
Regamey N, Tamm M, Wernli M, et al.
Transmission of human herpesvirus 8 infection from renal transplant donors to recipients.
N Engl J Med.
1998;339:1358-1363[Abstract/Free Full Text].
5.
Frances C, Mouquet C, Calvez V.
Human herpesvirus 8 and renal transplantation [Discussion].
N Engl J Med.
1999;340:1045[Free Full Text].
6.
Rabkin CS, Sheperd FA, Wade JA.
Human herpesvirus 8 and renal transplantation.
N Engl J Med.
1999;340:1045-1046.
7.
Farge D, Lebbe C, Marjanovic Z, et al.
Human herpes virus-8 and other risk factors for Kaposi's sarcoma in kidney transplant recipients. Groupe Cooperatif de Transplantation d' Ile de France.
Transplantation.
1999;67:1236-1242[Medline]
[Order article via Infotrieve].
8.
Lee P-C, Wang Y-W, Su I-J, Lin Y-J, Lei H-Y.
Immunosuppressive drugs and HHV-8 in a patient with a renal transplant and Kaposi's sarcoma.
Lancet.
1998;351:1175-1176[Medline]
[Order article via Infotrieve].
9.
Mendez JC, Procop GW, Espy MJ, Smith TF, McGregor CGA, Paya CV.
Relationship of HHV-8 replication and Kaposi's sarcoma after solid organ transplantation.
Transplantation.
1999;67:1200-1201[Medline]
[Order article via Infotrieve].
10.
Sachsenberg-Studer EM, Dobrynski N, Sheldon J, et al.
Human herpes-virus 8 seropositive patient with skin and graft Kaposi's sarcoma after lung transplantation.
J Am Acad Dermatol.
1999;40:308-311[Medline]
[Order article via Infotrieve].
11.
Jones D, Ballestas ME, Kaye KM, et al.
Primary effusion lymphoma and Kaposi's sarcoma in a cardiac transplant recipient.
N Engl J Med.
1998;339:444-449[Free Full Text].
12.
Dotti G, Fiocchi R, Motta T, et al.
Primary effusion lymphoma after heart transplantation: a new entity associated with human herpesvirus-8.
Leukemia.
1999;13:664-670[Medline]
[Order article via Infotrieve].
13.
Matsushima AY, Strauchen JA, Lee G, et al.
Posttransplantation plasmacytic proliferations related to Kaposi's sarcoma-associated herpesvirus.
Am J Surg Pathol.
1999;23:1393-1400[Medline]
[Order article via Infotrieve].
14.
Zong J-C, Ciufo DM, Alcendor DJ, et al.
High level variability in the ORF-K1 membrane protein gene at the left end of the Kaposi's sarcoma-associated herpesvirus genome defines four major virus subtypes and multiple variants or clades in different human populations.
J Virol.
1999;73:4156-4170[Abstract/Free Full Text].
15.
Cook P, Whitby D, Calabro ML, et al.
Variability and evolution of Kaposi's sarcoma-associated herpesvirus in different geographic regions of Europe and Africa.
AIDS.
1999;13:1165-1176[Medline]
[Order article via Infotrieve].
16.
Gao SJ, Zhang YJ, Deng JH, Rabkin CS, Flore O, Jenson HB.
Molecular polymorphism of Kaposi's sarcoma-associated herpesvirus (human herpesvirus 8) latent nuclear antigen: evidence for a large repertoire of viral genotypes and dual infection with different viral genotypes.
J Infect Dis.
1999;180:1466-1476[Medline]
[Order article via Infotrieve].
17.
Bottalico D, Santabosti Barbone G, Giancaspro V, Bignardi L, Arisi L, Cambi V.
Post-transplantation Kaposi's sarcoma appearing simultaneously in same cadaver renal transplant recipients.
Nephrol Dial Transplant.
1997;12:1055-1057[Abstract/Free Full Text].
18.
Kwok S, Higuchi R.
Avoiding false positives with PCR.
Nature.
1989;339:237-238[Medline]
[Order article via Infotrieve].
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
M. Bergallo, C. Costa, S. Margio, F. Sidoti, D. Re, G. P. Segoloni, and R. Cavallo
Human herpes virus 8 infection in kidney transplant patients from an area of northwestern Italy (Piemonte region)
Nephrol. Dial. Transplant.,
June 1, 2007;
22(6):
1757 - 1761.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. L. Morris, R. R. Arnold, and J. Webster-Cyriaque
Signaling Cascades Triggered by Bacterial Metabolic End Products during Reactivation of Kaposi's Sarcoma-Associated Herpesvirus
J. Virol.,
June 1, 2007;
81(11):
6032 - 6042.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. S. Laney, J. S. Peters, S. M. Manzi, L. A. Kingsley, Y. Chang, and P. S. Moore
Use of a Multiantigen Detection Algorithm for Diagnosis of Kaposi's Sarcoma-Associated Herpesvirus Infection.
J. Clin. Microbiol.,
October 1, 2006;
44(10):
3734 - 3741.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. Caselli, M. Galvan, E. Cassai, A. Caruso, L. Sighinolfi, and D. Di Luca
Human herpesvirus 8 enhances human immunodeficiency virus replication in acutely infected cells and induces reactivation in latently infected cells
Blood,
October 15, 2005;
106(8):
2790 - 2797.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. Gentile, A. Capobianchi, A. Volpi, G. Palu, F. Pica, A. Calistri, M. A. Biasolo, and P. Martino
Human Herpesvirus 8 DNA in Serum During Seroconversion in Allogeneic Bone Marrow Transplant Recipients
J Natl Cancer Inst,
July 6, 2005;
97(13):
1008 - 1011.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. P. Naranatt, H. H. Krishnan, S. R. Svojanovsky, C. Bloomer, S. Mathur, and B. Chandran
Host Gene Induction and Transcriptional Reprogramming in Kaposi's Sarcoma-Associated Herpesvirus (KSHV/HHV-8)-Infected Endothelial, Fibroblast, and B Cells: Insights into Modulation Events Early during Infection
Cancer Res.,
January 1, 2004;
64(1):
72 - 84.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Delorme, I. Houde, and L. Deschenes
Seroprevalence of Antibodies against Human Herpesvirus 8 in a Population of Renal Transplant Recipients at Hotel-Dieu de Quebec Hospital
J. Clin. Microbiol.,
November 1, 2003;
41(11):
5207 - 5208.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. C. Gartner, A. Kloss, H. Kaul, U. Sester, K. Roemer, H. Pees, H. Kohler, and N. Mueller-Lantzsch
Risk of Occupational Human Herpesvirus 8 Infection for Health Care Workers
J. Clin. Microbiol.,
May 1, 2003;
41(5):
2156 - 2157.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. J. Jenkins, D. T. Rowe, and C. R. Rinaldo Jr.
Herpesvirus Infections in Organ Transplant Recipients
Clin. Vaccine Immunol.,
January 1, 2003;
10(1):
1 - 7.
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. D. Fakhari and D. P. Dittmer
Charting Latency Transcripts in Kaposi's Sarcoma-Associated Herpesvirus by Whole-Genome Real-Time Quantitative PCR
J. Virol.,
May 13, 2002;
76(12):
6213 - 6223.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
Top
Abstract
Introduction