Department of Clinical Virology1 and Department of Pathology2, Huddinge University Hospital, F68, Karolinska Institutet, SE-141 86 Stockholm, Sweden
The Norwegian Cancer Registry, Oslo, Norway2
Clinical Epidemiology Unit, Department of Medicine, Karolinska Hospital, Karolinska Institutet, Stockholm, Sweden3
Author for correspondence: Thomas Tolfvenstam. Fax +46 8 585 879 33. e-mail thomas.tolfvenstam{at}impi.ki.se
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Abstract |
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Introduction |
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Human parvovirus B19 (B19) is a ubiquitous pathogen that causes the childhood disease erythema infectiosum (Anderson et al., 1984 ). It is highly prevalent in society at large, with a seroprevalence of 5070% among adults (Cohen & Buckley, 1988
). B19 is known to be a concern in pregnant women, in whom foetal infection may result in foetal death, and also among individuals with haematological disorders, in whom the infection may be fatal (Tolfvenstam et al., 2001
; Young, 1988
). The virus mediates its pathogenicity by infecting erythroid progenitor cells by means of its cellular receptor, the blood group P antigen (Brown et al., 1993
). Although the virus has been shown to persist in the bone marrow of some individuals over long periods of time, it has not been previously associated with carcinogenesis (Kurtzman et al., 1987
; Lundqvist et al., 1999
). Recently, Gray et al. (1998)
reported a high frequency of B19 DNA in testicular tissue from testicular cancer patients as compared to controls. Diss et al. (1999)
later confirmed these findings but also concluded that B19 DNA could be found in normal testicular tissue. With the study design used by these authors, it cannot be ruled out that infection occurred subsequent to the development of cancer. To overcome this problem of reversed causality, we studied seroreactivity to B19 among cases and controls using serum samples drawn before the onset of disease in addition to an elucidation of the frequency of viral DNAs in a retrospectively collected 2-year testicular carcinoma series in our area of referral.
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Methods |
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Since 1960, all inhabitants of Norway have been assigned a unique, 11-digit identification number. All newly diagnosed cases of cancer in Norway are registered in the Cancer Registry of Norway, with compulsory reporting by both hospital departments and histopathological laboratories. Patients are entered in the Cancer Registry under their individual identification number. On a yearly basis, the identification numbers of all blood donors contained in the JANUS serum bank are linked with the Cancer Registry; as of October 1993, a total of 14000 donors had developed some form of cancer.
Persons eligible as cases for this nested case-control analysis were male blood donors listed in the Cancer Registry through 1993 who had been diagnosed with invasive testicular cancer. There were 81 eligible cases but one case was excluded due to a lack of sample material. Three blood donors, matched with each index case by birth date (within 1 year), were randomly selected from the JANUS serum bank as potential controls. To verify that a potential control was alive and had not been diagnosed with testicular cancer at the time of diagnosis of the corresponding case, we examined the records, linked through the individual identification number, in the Death Registry and the Cancer Registry of Norway. One control was excluded on this basis and one case was excluded due to a lack of sample material, leaving 241 to be included in the serological analysis.
To confirm the overrepresentation of B19 DNA in germ cell cancer reported in the previous studies in a Scandinavian setting, testicular tissue samples were collected retrospectively from all cases of testicular carcinoma referred during the years 19981999 (n=24) to the Department of Pathology at Huddinge University Hospital (Stockholm, Sweden), which serves all three hospitals in the southern part of the greater Stockholm area. Histopathologically, 21 cases represented seminomas, two cases were mixed testicular carcinomas and one case was an embryonic carcinoma. Controls were included retrospectively and consisted of all cases of orchidectomy referred to the same unit during the years 19961999, in which the diagnosis was not testicular carcinoma and the tissue was histopathologically normal (n=11). In these cases, orchidectomy was performed owing to suspected malignancy (five cases), prostatic cancer (four cases) and complicated hydrocele (two cases).
Ethical approval.
This study was approved by the Ethics Committee of Huddinge University Hospital, Karolinska Institutet, Stockholm, Sweden.
Serum analyses.
B19-specific IgG class antibodies were measured by a sandwich ELISA (Biotrin). Antibody levels were expressed as IU/ml and determined by five-point co-titration of the WHO B19 standard in each analysis. Information on seroreactivity to cytomegalovirus, EpsteinBarr virus, Chlamydia trachomatis, human papillomavirus types 16 and 18, herpes simplex virus types 1 and 2 and human herpesvirus type 8 was used in the multivariate analysis to control for potential confounding. The methods used for detecting seroreactivity to these agents are described elsewhere (Akre et al., 1999 ).
Tissue analysis.
Tissue sections of 25 mg were cut from the paraffin-embedded material representing the tumour, as indicated by the corresponding haematoxylin and eosin (H&E)-stained slide. Xenol and ethanol washings subsequently removed the paraffin and DNA was extracted using QiaAmp DNA Mini kit (Qiagen), according to the manufacturer's instructions. The extraction procedure was verified by amplification of a conserved region in the chromosomal major histocompatibility complex class II gene (Ehrlich & Bugawan, 1989 ). B19-specific DNA was amplified by nested PCR, as described previously, with the exception of the use of a modified outer forward primer (5' GGCAGCATGTGTTAAAGTGG 3') (Broliden et al., 1998
). Amplification resulted in a 284 bp fragment originating from the B19 non-structural protein (NS1). Immunohistochemistry was performed on formalin-fixed microtome-cut tissue sections using a monoclonal antibody directed to the B19 structural protein 2 (VP2), visualized by a peroxidase system. The slides were reviewed, together with routinely stained H&E tissue sections, by a senior pathologist experienced in B19-induced histopathology.
Statistical analysis.
The prevalence of seropositivity, as well as mean titres, were determined. In addition to analyses of the risk of being seropositive, dose-response relationships were evaluated by dichotomizing seropositivity into high (above median) and low (below median) positive titres based on the distribution in the seropositive population. Data were modelled by means of conditional logistic regression using the SAS statistical package. The parameters and standard errors in the models were converted to odds ratios (ORs) with 95% confidence intervals (CIs). All reported P values are two-tailed. Separate analyses for the two main histological subgroups of testicular cancer, seminomas and non-seminomas, were performed to evaluate potential aetiological differences.
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Results |
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Discussion |
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In summary, in this study we found no indication of an aetiological role of B19 in the development of testicular carcinoma but confirmed the observation of an overrepresentation of B19 DNA-positivity in testicular tumours compared to controls. We speculate that this finding may be due to susceptibility of the carcinoma cells to B19 infection owing to high-level expression of the viral receptor and possibly other putative cellular factors. This infection would have occurred subsequent to the development of cancer and would have a defect, low-grade replication, possibly protected by the bloodtestis barrier.
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Acknowledgments |
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References |
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Akre, O., Lipworth, L., Tretli, S., Linde, A., Engstrand, L., Adami, H. O., Melbye, M., Andersen, A. & Ekbom, A. (1999). EpsteinBarr virus and cytomegalovirus in relation to testicular-cancer risk: a nested case-control study. International Journal of Cancer 82, 1-5.
Anderson, M. J., Lewis, E., Kidd, I. M., Hall, S. M. & Cohen, B. J. (1984). An outbreak of erythema infectiosum associated with human parvovirus infection. Journal of Hygiene 93, 85-93.[Medline]
Broliden, K., Tolfvenstam, T., Ohlsson, S. & Henter, J. I. (1998). Persistent B19 parvovirus infection in pediatric malignancies. Medical and Pediatric Oncology 31, 66-72.[Medline]
Brown, K. E., Anderson, S. M. & Young, N. S. (1993). Erythrocyte P antigen: cellular receptor for B19 parvovirus. Science 262, 114-117.[Medline]
Cohen, B. J. & Buckley, M. M. (1988). The prevalence of antibody to human parvovirus B19 in England and Wales. Journal of Medical Microbiology 25, 151-153.[Abstract]
Cooling, L. L., Koerner, T. A. & Naides, S. J. (1995). Multiple glycosphingolipids determine the tissue tropism of parvovirus B19. Journal of Infectious Diseases 172, 1198-1205.[Medline]
Diss, T. C., Pan, L. X., Du, M. Q., Peng, H. Z. & Kerr, J. R. (1999). Parvovirus B19 is associated with benign testes as well as testicular germ cell tumours. Molecular Pathology 52, 349-352.[Abstract]
Ehrlich, H. & Bugawan, T. (1989). HLA class II gene polymorphism: DNA typing, evolution and relationship to disease susceptibility. In PCR Technology: Principles and Applications for DNA amplification , pp. 193-208. Edited by H. A. Ehrlich. Oxford: Oxford University Press.
Gray, A., Guillou, L., Zufferey, J., Rey, F., Kurt, A. M., Jichlinski, P., Leisinger, H. J. & Benhattar, J. (1998). Persistence of parvovirus B19 DNA in testis of patients with testicular germ cell tumours. Journal of General Virology 79, 573-579.[Abstract]
Jellum, E., Andersen, A., Lund-Larsen, P., Theodorsen, L. & Orjasaeter, H. (1995). Experiences of the Janus Serum Bank in Norway. Environmental Health Perspectives 103 (Suppl. 3), 8588.[Medline]
Kurtzman, G. J., Ozawa, K., Cohen, B., Hanson, G., Oseas, R. & Young, N. S. (1987). Chronic bone marrow failure due to persistent B19 parvovirus infection. New England Journal of Medicine 317, 287-294.[Medline]
Lundqvist, A., Tolfvenstam, T., Bostic, J., Soderlund, M. & Broliden, K. (1999). Clinical and laboratory findings in immunocompetent patients with persistent parvovirus B19 DNA in bone marrow. Scandinavian Journal of Infectious Diseases 31, 11-16.[Medline]
Morey, A. L., Keeling, J. W., Porter, H. J. & Fleming, K. A. (1992). Clinical and histopathological features of parvovirus B19 infection in the human fetus. British Journal of Obstetrics and Gynaecology 99, 566-574.[Medline]
Newell, G. R., Mills, P. K. & Johnson, D. E. (1984). Epidemiologic comparison of cancer of the testis and Hodgkin's disease among young males. Cancer 54, 1117-1123.[Medline]
Ohyama, C., Fukushi, Y., Satoh, M., Saitoh, S., Orikasa, S., Nudelman, E., Straud, M. & Hakomori, S. (1990). Changes in glycolipid expression in human testicular tumor. International Journal of Cancer 45, 1040-1044.
Olie, R. A., Fenderson, B., Daley, K., Oosterhuis, J. W., Murphy, J. & Looijenga, L. H. (1996). Glycolipids of human primary testicular germ cell tumours. British Journal of Cancer 74, 133-140.[Medline]
Pallier, C., Greco, A., Le Junter, J., Saib, A., Vassias, I. & Morinet, F. (1997). The 3' untranslated region of the B19 parvovirus capsid protein mRNAs inhibits its own mRNA translation in nonpermissive cells. Journal of Virology 71, 9482-9489.[Abstract]
Tolfvenstam, T., Papadogiannakis, N., Norbeck, O., Petersson, K. & Broliden, K. (2001). Frequency of human parvovirus B19 infection in intrauterine fetal death. Lancet 357, 1494-1497.[Medline]
Weigel-Kelley, K. A., Yoder, M. C. & Srivastava, A. (2001). Recombinant human parvovirus B19 vectors: erythrocyte P antigen is necessary but not sufficient for successful transduction of human hematopoietic cells. Journal of Virology 75, 4110-4116.
Wenk, J., Andrews, P. W., Casper, J., Hata, J., Pera, M. F., von Keitz, A., Damjanov, I. & Fenderson, B. A. (1994). Glycolipids of germ cell tumors: extended globo-series glycolipids are a hallmark of human embryonal carcinoma cells. International Journal of Cancer 58, 108-115.
Young, N. (1988). Hematologic and hematopoietic consequences of B19 parvovirus infection. Seminars in Hematology 25, 159-172.[Medline]
Received 14 March 2002;
accepted 29 April 2002.
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