Skip to main content
PMC home page
Clinical and Diagnostic Laboratory Immunology logoLink to Clinical and Diagnostic Laboratory Immunology
. 1994 Mar;1(2):160–163. doi: 10.1128/cdli.1.2.160-163.1994

Presence of hepatitis C virus (HCV) genomic RNA and viral replicative intermediates in bone marrow and peripheral blood mononuclear cells from HCV-infected patients.

A Manzin 1, M Candela 1, S Paolucci 1, M L Caniglia 1, A Gabrielli 1, M Clementi 1
PMCID: PMC368220  PMID: 7496938

Abstract

The cellular tropism of hepatitis C virus (HCV) was studied in vivo in samples from patients with persistent HCV infection. Plasma, liver, peripheral blood mononuclear cell (PBMC), and bone marrow cell (BMC) samples from 15 subjects positive for anti-HCV antibodies were tested for the presence of HCV RNA sequences by reverse transcription PCR. Virus-specific RNA sequences were found to be present in liver samples from all subjects (100%), in plasma samples from 13 of 15 patients (86.7%), in PBMC samples from 3 patients (20%), and in BMC samples from 9 (60%) of the 15 anti-HCV-positive patients enrolled in this study. The presence of the molecular intermediate of HCV replication (the negative-stranded HCV RNA) was evident in the two of the three PBMC and in five of the nine BMC HCV RNA-positive samples. Finally, we studied the nucleotide sequence of a large portion (-270 to -59) of the 5'untranslated region of HCV amplified from plasma samples of 12 of the 15 patients with and without HCV in BMCs; the degree of heterogeneity compared with the prototype HCV sequence was similar in both groups. The data principally indicate that HCV infection of PBMCs and BMCs is frequent in persistently infected patients, as shown by the occurrence of positive- and negative-stranded HCV RNA, thus suggesting the possibility of extrahepatic replication of HCV.

Full text

PDF
161

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Agnello V., Chung R. T., Kaplan L. M. A role for hepatitis C virus infection in type II cryoglobulinemia. N Engl J Med. 1992 Nov 19;327(21):1490–1495. doi: 10.1056/NEJM199211193272104. [DOI] [PubMed] [Google Scholar]
  2. Alter H. J., Purcell R. H., Shih J. W., Melpolder J. C., Houghton M., Choo Q. L., Kuo G. Detection of antibody to hepatitis C virus in prospectively followed transfusion recipients with acute and chronic non-A, non-B hepatitis. N Engl J Med. 1989 Nov 30;321(22):1494–1500. doi: 10.1056/NEJM198911303212202. [DOI] [PubMed] [Google Scholar]
  3. Bouffard P., Hayashi P. H., Acevedo R., Levy N., Zeldis J. B. Hepatitis C virus is detected in a monocyte/macrophage subpopulation of peripheral blood mononuclear cells of infected patients. J Infect Dis. 1992 Dec;166(6):1276–1280. doi: 10.1093/infdis/166.6.1276. [DOI] [PubMed] [Google Scholar]
  4. Bukh J., Purcell R. H., Miller R. H. Sequence analysis of the 5' noncoding region of hepatitis C virus. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):4942–4946. doi: 10.1073/pnas.89.11.4942. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chen P. J., Lin M. H., Tu S. J., Chen D. S. Isolation of a complementary DNA fragment of hepatitis C virus in Taiwan revealed significant sequence variations compared with other isolates. Hepatology. 1991 Jul;14(1):73–78. doi: 10.1002/hep.1840140113. [DOI] [PubMed] [Google Scholar]
  6. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  7. Choo Q. L., Richman K. H., Han J. H., Berger K., Lee C., Dong C., Gallegos C., Coit D., Medina-Selby R., Barr P. J. Genetic organization and diversity of the hepatitis C virus. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2451–2455. doi: 10.1073/pnas.88.6.2451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fong T. L., Shindo M., Feinstone S. M., Hoofnagle J. H., Di Bisceglie A. M. Detection of replicative intermediates of hepatitis C viral RNA in liver and serum of patients with chronic hepatitis C. J Clin Invest. 1991 Sep;88(3):1058–1060. doi: 10.1172/JCI115368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Han J. H., Shyamala V., Richman K. H., Brauer M. J., Irvine B., Urdea M. S., Tekamp-Olson P., Kuo G., Choo Q. L., Houghton M. Characterization of the terminal regions of hepatitis C viral RNA: identification of conserved sequences in the 5' untranslated region and poly(A) tails at the 3' end. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1711–1715. doi: 10.1073/pnas.88.5.1711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Houghton M., Weiner A., Han J., Kuo G., Choo Q. L. Molecular biology of the hepatitis C viruses: implications for diagnosis, development and control of viral disease. Hepatology. 1991 Aug;14(2):381–388. [PubMed] [Google Scholar]
  11. Johnson R. J., Gretch D. R., Yamabe H., Hart J., Bacchi C. E., Hartwell P., Couser W. G., Corey L., Wener M. H., Alpers C. E. Membranoproliferative glomerulonephritis associated with hepatitis C virus infection. N Engl J Med. 1993 Feb 18;328(7):465–470. doi: 10.1056/NEJM199302183280703. [DOI] [PubMed] [Google Scholar]
  12. Katkov W. N., Dienstag J. L., Cody H., Evans A. A., Choo Q. L., Houghton M., Kuo G. Role of hepatitis C virus in non-B chronic liver disease. Arch Intern Med. 1991 Aug;151(8):1548–1552. [PubMed] [Google Scholar]
  13. Lee C. H., Cheng C., Wang J., Lumeng L. Identification of hepatitis C viruses with a nonconserved sequence of the 5' untranslated region. J Clin Microbiol. 1992 Jun;30(6):1602–1604. doi: 10.1128/jcm.30.6.1602-1604.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lenzi M., Johnson P. J., McFarlane I. G., Ballardini G., Smith H. M., McFarlane B. M., Bridger C., Vergani D., Bianchi F. B., Williams R. Antibodies to hepatitis C virus in autoimmune liver disease: evidence for geographical heterogeneity. Lancet. 1991 Aug 3;338(8762):277–280. doi: 10.1016/0140-6736(91)90418-o. [DOI] [PubMed] [Google Scholar]
  15. Manzin A., Menzo S., Bagnarelli P., Varaldo P. E., Bearzi I., Carloni G., Galibert F., Clementi M. Sequence analysis of the hepatitis B virus pre-C region in hepatocellular carcinoma [HCC] and nontumoral liver tissues from HCC patients. Virology. 1992 Jun;188(2):890–895. doi: 10.1016/0042-6822(92)90548-4. [DOI] [PubMed] [Google Scholar]
  16. Marcellin P., Descamps V., Martinot-Peignoux M., Larzul D., Xu L., Boyer N., Pham B. N., Crickx B., Guillevin L., Belaich S. Cryoglobulinemia with vasculitis associated with hepatitis C virus infection. Gastroenterology. 1993 Jan;104(1):272–277. doi: 10.1016/0016-5085(93)90862-7. [DOI] [PubMed] [Google Scholar]
  17. Michel G., Ritter A., Gerken G., Meyer zum Büschenfelde K. H., Decker R., Manns M. P. Anti-GOR and hepatitis C virus in autoimmune liver diseases. Lancet. 1992 Feb 1;339(8788):267–269. doi: 10.1016/0140-6736(92)91332-3. [DOI] [PubMed] [Google Scholar]
  18. Miller R. H., Purcell R. H. Hepatitis C virus shares amino acid sequence similarity with pestiviruses and flaviviruses as well as members of two plant virus supergroups. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2057–2061. doi: 10.1073/pnas.87.6.2057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Misiani R., Bellavita P., Fenili D., Borelli G., Marchesi D., Massazza M., Vendramin G., Comotti B., Tanzi E., Scudeller G. Hepatitis C virus infection in patients with essential mixed cryoglobulinemia. Ann Intern Med. 1992 Oct 1;117(7):573–577. doi: 10.7326/0003-4819-117-7-573. [DOI] [PubMed] [Google Scholar]
  20. Pascual M., Perrin L., Giostra E., Schifferli J. A. Hepatitis C virus in patients with cryoglobulinemia type II. J Infect Dis. 1990 Aug;162(2):569–570. doi: 10.1093/infdis/162.2.569. [DOI] [PubMed] [Google Scholar]
  21. Pechère-Bertschi A., Perrin L., de Saussure P., Widmann J. J., Giostra E., Schifferli J. A. Hepatitis C: a possible etiology for cryoglobulinaemia type II. Clin Exp Immunol. 1992 Sep;89(3):419–422. doi: 10.1111/j.1365-2249.1992.tb06973.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Qian C., Camps J., Maluenda M. D., Civeira M. P., Prieto J. Replication of hepatitis C virus in peripheral blood mononuclear cells. Effect of alpha-interferon therapy. J Hepatol. 1992 Nov;16(3):380–383. doi: 10.1016/s0168-8278(05)80674-9. [DOI] [PubMed] [Google Scholar]
  23. Takehara T., Hayashi N., Mita E., Hagiwara H., Ueda K., Katayama K., Kasahara A., Fusamoto H., Kamada T. Detection of the minus strand of hepatitis C virus RNA by reverse transcription and polymerase chain reaction: implications for hepatitis C virus replication in infected tissue. Hepatology. 1992 Mar;15(3):387–390. doi: 10.1002/hep.1840150305. [DOI] [PubMed] [Google Scholar]
  24. Zignego A. L., Macchia D., Monti M., Thiers V., Mazzetti M., Foschi M., Maggi E., Romagnani S., Gentilini P., Bréchot C. Infection of peripheral mononuclear blood cells by hepatitis C virus. J Hepatol. 1992 Jul;15(3):382–386. doi: 10.1016/0168-8278(92)90073-x. [DOI] [PubMed] [Google Scholar]

Articles from Clinical and Diagnostic Laboratory Immunology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES