infection of peripheral blood mononuclear cells by hepatitis C virus Free

Abstract

To study the susceptibility of peripheral blood mononuclear cells (PBMC) to hepatitis C virus (HCV), we incubated cells from healthy donors with HCV-positive sera. Using RT-PCR and hybridization, the genomic viral RNA was detected in PBMC and in their supernatants until 25 days post-incubation. The PBMC of the different donors were not all permissive to HCV, but results were more constantly positive when cells from four donors were pooled. Quantification of the genomic viral RNA by the branched-DNA assay showed a decrease in the HCV RNA concentration during the first week of culture followed by a peak during the second or third week, and also an increase in the total amount of viral RNA in the inoculated cells. Although HCV RNA could be detected in the supernatants by RT-PCR, the concentration was very low. Using a sense-specific RT-PCR method, the HCV negative-strand was also detected in the cells but not in the supernatants. In two experiments PBMC were successfully infected using HCV-positive culture supernatants, therefore suggesting that infectious particles can be produced in this system. Our findings demonstrate that PBMC are permissive for HCV replication but the replication level is very low. The HCV RNA concentration measured in PBMC of 10 chronically infected patients was not significantly higher than the maximal concentration obtained in PBMC infected .

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1995-10-01
2024-03-29
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References

  1. Alter H., Purcell R., Shih J., Melpoder J., Houghton M., Choo Q., Kuo G. 1989; Detection of antibody to hepatitis C virus in prospectively followed transfusion recipients with acute and chronic non-A, non-B hepatitis. New England Journal of Medicine 321:1494–1500
    [Google Scholar]
  2. Bertolini L., Iovacci S., Ponzetto A., Gorini G., Battaglia M., Carloni G. 1993; The human bone-marrow B-cell line CE, susceptible to hepatitis C virus infection. Research in Virology 144:281–285
    [Google Scholar]
  3. Bouffard P., Hayashi P. H., Acevedo R., Levy N., Zeldis J. B. 1992; Hepatitis C virus is detected in a monocyte/macrophage subpopulation of peripheral blood mononuclear cells of infected patients. Journal of Infectious Diseases 166:1276–1280
    [Google Scholar]
  4. Choo Q. L., Richman K. H., Han J. H., Berger K., Lee C., Dong C., Gallegos C., Coit D., Median-Selby A., Barr P. J., Weiner A. J., Bradley D. W., Kuo G., Houghton M. 1991; Genetic organization and diversity of the hepatitis C virus. Proceedings of the National Academy of Sciences, USA 88:2451–2455
    [Google Scholar]
  5. Ferri C., Monti M., La Civita L., Longonbardo G., Greco F., Pasero G., Gentilini P., Bombardieri S., Zignego A. L. 1993; Infection of peripheral blood mononuclear cells by hepatitis C virus in mixed cryoglobulinemia. Blood 82:3701–3704
    [Google Scholar]
  6. Henin C., Makris M., Brown J., Peake I. R., Preston E. F. 1994; Peripheral mononuclear cells of haemophiliacs with chronic liver disease are infected with replicating hepatitis C virus. British Journal of Haematology 87:215–217
    [Google Scholar]
  7. Iovacci S., Sargiacomo M., Parolini I., Ponzetto A., Peschle C., Carloni G. 1993; Replication and multiplication of hepatitis C virus genome in human foetal liver cells. Research in Virology 144:275–279
    [Google Scholar]
  8. Kaito M., Watanabe S., Tsukiyama-Kohara K., Yamagushi K., Kobayashi Y., Koshini M., Yokoi M., Ishida S., Suzuki S., Kohara M. 1994; Hepatitis C virus particle detected by immuno-electron microscopic study. Journal of General Virology 75:1755–1760
    [Google Scholar]
  9. Kuo G., Choo Q. L., Alter H. J., Gitnick L., Redecker A. G., Purcell R. H., Miyamura T., Dienstag J. L., Alter M. J., Stevens C. E., Tegmeier G. E., Bonino F., Colombo M., Lee W. S., Kuo C., Berger K., Sauster J. K., Overby L. R., Bradley D. W., Houghton M. 1989; An assay for circulating antibodies to a major etiologic virus of non-A, non-B hepatitis. Science 244:362–364
    [Google Scholar]
  10. Lanford R. E., Sureau C., Jacob J. R., White R., Fuerst T. R. 1994; Demonstration of in vitro infection of chimpanzee hepatocytes with hepatitis C virus using strand-specific RT-PCR. Virology 202:606–614
    [Google Scholar]
  11. MacGuiness P. H., Bishop G. A., MacCaughan G. W., Trowbridge R., Gowans E. J. 1994; False detection of negative strand hepatitis C virus RNA. Lancet 343:551–552
    [Google Scholar]
  12. Miller R. H., Purcell R. H. 1990; Hepatitis C virus shares amino acid sequence similarity with pestiviruses and flaviviruses as well as members of two plant virus supergroups. Proceedings of the National Academy of Sciences, USA 87:2057–2061
    [Google Scholar]
  13. Moldvay J., Deny P., Pol S., Brechot C., Lamas E. 1994; Detection of hepatitis C virus RNA in peripheral blood mononuclear cells of infected patients by in situ hybridization. Blood 83:269–273
    [Google Scholar]
  14. Muller H. M., Pfaff E., Goeser T., Kallinowski B., Solbach C., Theilman L. 1993; Peripheral blood leukocytes serve as a possible extrahepatic site for hepatitis C virus replication. Journal of General Virology 74:669–676
    [Google Scholar]
  15. Nissen E., Hohne M., Schreier E. 1994; In vitro, replication of hepatitis C virus in a human lymphoid cell line (H9). Journal of Hepatology 20:437
    [Google Scholar]
  16. Shimizu Y. K., Iwamoto A., Hijikata M., Purcell R. H., Yoshikuba H. 1992; Evidence for in vitro replication of hepatitis C virus genome in a human T-cell line. Proceedings of the National Academy of Sciences, USA 89:5477–5481
    [Google Scholar]
  17. Shimizu Y. K., Purcell R. H., Yoshikura H. 1993; Correlation between the infectivity of hepatitis C virus in vitro and its infectivity in vitro. Proceedings of the National Academy of Sciences, USA 90:6037–6041
    [Google Scholar]
  18. Simmonds P., Holmes E. C., Cha T. A., Chan S. W., McOmish F., Irvine B., Beall E., Yap P. L., Kolberg J., Urdea M. S. 1993; Classification of hepatitis C virus into six major genotypes and a series of subtypes by phylogenetic analysis of the NS-5 region. Journal of General Virology 74:2391–2399
    [Google Scholar]
  19. Wang J. T., Sheu J. C., Lin J. T., Wang T. H., Chen D. S. 1992; Detection of replicative form of hepatitis C virus RNA in peripheral blood mononuclear cells. Journal of Infectious Diseases 166:1167–1169
    [Google Scholar]
  20. Willems M., Peerlinck K., Moshage H., Deleu I., van den Eynden C., Vermylen J., Yap S. H. 1994; Hepatitis C virus RNAs in plasma and in peripheral blood mononuclear cells of hemophiliacs with chronic hepatitis C: evidence for viral replication in peripheral blood mononuclear cells. Journal of Medical Virology 42:272–278
    [Google Scholar]
  21. Yoo B. J., Selby M. J., Choe J., Suh B. S., Choi S. H., Joh J. S., Nuovo G. J., Lee H. S., Houghton M., Han J. H. 1995; Transfection of a differentiated human hepatoma cell line (Huh7) with in vitro-transcribed hepatitis C virus (HCV) RNA and establishment of a long-term culture persistently infected with HCV. Journal of Virology 69:32–38
    [Google Scholar]
  22. Zignegno A. L., Macchia D., Monti M., Thiers V., Mazetti M., Foschi M., Maggi E., Romagnani S., Gentilini P., Brechot C. 1992; Infection of peripheral mononuclear cells by hepatitis C virus. Journal of Hepatology 15:382–386
    [Google Scholar]
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