Binding of hepatitis C virus to CD81

Science

Volumn 282, Issue 5390, 1998, Pages 938-941

  Pileri, Piero ^a   Uematsu, Yasushi ^a   Campagnoli, Susanna ^a   Galli, Giuliano ^a   Falugi, Fabiana ^a   Petracca, Roberto ^a   Weiner, Amy J ^b   Houghton, Michael ^b   Rosa, Domenico ^a   Grandi, Guido ^a   Abrignani, Sergio ^a  

^a CHIRON VACCINES   (Italy)

^b NOVARTIS PHARMA AG   (Switzerland)

Author keywords

[No Author keywords available]

Indexed keywords

CD81 ANTIGEN; ENVELOPE PROTEIN; NEUTRALIZING ANTIBODY;

ANTIGEN BINDING; ANTIGEN EXPRESSION; ARTICLE; B LYMPHOCYTE; BINDING AFFINITY; CRYOGLOBULINEMIA; HEPATITIS C VIRUS; LIVER CELL; LYMPHOCYTE PROLIFERATION; PRIORITY JOURNAL; VIRUS INFECTION;

AMINO ACID SEQUENCE; ANIMALS; ANTIBODIES; ANTIBODIES, VIRAL; ANTIGENS, CD; CELL LINE; DNA, COMPLEMENTARY; GENE LIBRARY; HEPACIVIRUS; HEPATITIS C; HUMANS; LIVER; LYMPHOCYTES; MEMBRANE PROTEINS; MICE; MOLECULAR SEQUENCE DATA; PAN TROGLODYTES; RECOMBINANT FUSION PROTEINS; RECOMBINANT PROTEINS; SEQUENCE ALIGNMENT; TUMOR CELLS, CULTURED; VIRAL ENVELOPE PROTEINS;

EID: 0032582538     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.282.5390.938     Document Type: Article

References (36)

1. M. Houghton, in Virology, B. N. Fields, D. M. Knipe, P. M. Howley, Eds. (Lippincott-Raven, Philadelphia, PA, 1996), pp. 1035-1058.
2. World Health Organization, Weekly Epidemiol. Rec. 72, 65 (1997).
3. J. H. Hoofnagle, Hepatology 26 (suppl. 1), 15S (1997).
4. Cryoglobulinemia is a B lymphocyte proliferative disorder characterized by the presence of cryoglobulins (immunoglobulins insoluble at 4°C) in the serum. The disease is often associated with multiple organ involvement and is due to the deposition of immune complexes in small blood vessels. Three types of cryoglobulinemia can be distinguished. Type I is formed by monoclonal immunoglobulin M (IgM) and is also known as Waldenstrom's disease. Type II is formed by IgM-IgG complexes in the serum; the IgG fraction is polyclonal, whereas the IgM fraction is monoclonal with rheumatoid factor activity. Type III is formed by polyclonal immunoglobulins. More than 80% of the patients with type II and type III cryoglobulinemia are infected by HCV, often without liver disease [C. Ferri et al., Clin. Exp. Rheumatol. 9, 95 (1991); V. Agnello, R. T. Chung, L. M. Kaplan, N. Engl. J. Med. 327, 1490 (1992); F. Silvestri et al., Blood 87, 4296 (1996); M. Ivanovski et al., ibid. 91, 2433 (1998)].
5. Cryoglobulinemia is a B lymphocyte proliferative disorder characterized by the presence of cryoglobulins (immunoglobulins insoluble at 4°C) in the serum. The disease is often associated with multiple organ involvement and is due to the deposition of immune complexes in small blood vessels. Three types of cryoglobulinemia can be distinguished. Type I is formed by monoclonal immunoglobulin M (IgM) and is also known as Waldenstrom's disease. Type II is formed by IgM-IgG complexes in the serum; the IgG fraction is polyclonal, whereas the IgM fraction is monoclonal with rheumatoid factor activity. Type III is formed by polyclonal immunoglobulins. More than 80% of the patients with type II and type III cryoglobulinemia are infected by HCV, often without liver disease [C. Ferri et al., Clin. Exp. Rheumatol. 9, 95 (1991); V. Agnello, R. T. Chung, L. M. Kaplan, N. Engl. J. Med. 327, 1490 (1992); F. Silvestri et al., Blood 87, 4296 (1996); M. Ivanovski et al., ibid. 91, 2433 (1998)].
6. Cryoglobulinemia is a B lymphocyte proliferative disorder characterized by the presence of cryoglobulins (immunoglobulins insoluble at 4°C) in the serum. The disease is often associated with multiple organ involvement and is due to the deposition of immune complexes in small blood vessels. Three types of cryoglobulinemia can be distinguished. Type I is formed by monoclonal immunoglobulin M (IgM) and is also known as Waldenstrom's disease. Type II is formed by IgM-IgG complexes in the serum; the IgG fraction is polyclonal, whereas the IgM fraction is monoclonal with rheumatoid factor activity. Type III is formed by polyclonal immunoglobulins. More than 80% of the patients with type II and type III cryoglobulinemia are infected by HCV, often without liver disease [C. Ferri et al., Clin. Exp. Rheumatol. 9, 95 (1991); V. Agnello, R. T. Chung, L. M. Kaplan, N. Engl. J. Med. 327, 1490 (1992); F. Silvestri et al., Blood 87, 4296 (1996); M. Ivanovski et al., ibid. 91, 2433 (1998)].
7. Cryoglobulinemia is a B lymphocyte proliferative disorder characterized by the presence of cryoglobulins (immunoglobulins insoluble at 4°C) in the serum. The disease is often associated with multiple organ involvement and is due to the deposition of immune complexes in small blood vessels. Three types of cryoglobulinemia can be distinguished. Type I is formed by monoclonal immunoglobulin M (IgM) and is also known as Waldenstrom's disease. Type II is formed by IgM-IgG complexes in the serum; the IgG fraction is polyclonal, whereas the IgM fraction is monoclonal with rheumatoid factor activity. Type III is formed by polyclonal immunoglobulins. More than 80% of the patients with type II and type III cryoglobulinemia are infected by HCV, often without liver disease [C. Ferri et al., Clin. Exp. Rheumatol. 9, 95 (1991); V. Agnello, R. T. Chung, L. M. Kaplan, N. Engl. J. Med. 327, 1490 (1992); F. Silvestri et al., Blood 87, 4296 (1996); M. Ivanovski et al., ibid. 91, 2433 (1998)].
8. Q.-L. Choo et al., Science 244, 359 (1989).
9. D. Rosa et al., Proc. Natl. Acad. Sci. U.S.A. 93, 1759 (1996). E2 binds to chimpanzee mononuclear cells, whereas it does not bind to rat, rabbit, or green monkey mononuclear cells or hepatocytes (22).
10. Human cells (B and T lymphoma and hepatocarcinoma cell lines) were incubated with recombinant E2 expressed in mammalian cells (CHO) (6) and stained with biotin-labeled anti-E2 as described (6). Cells with the highest E2-binding capacity were sorted with a FacsVantage (Becton-Dickinson) and subcloned by limiting dilution. Growing clones were screened for E2 binding at the Facs, and clones with the highest mean fluorescence intensity were further expanded.
11. L. Dailey and C. Basilico, J. Virol. 54, 739 (1985).
12. 7 cells per milliliter) with recombinant E2 (10 μg/ml). After incubation of the cell suspension with a monoclonal antibody (mAb) to E2 (clone 291) (22), those cells that had acquired the capacity to bind E2 were isolated with Dynabeads magnetic beads (Dynal) coated with goat anti-mouse IgG. Plasmid DNA was recovered from bead-bound cells with the protocol described by Campbell et al. (24) and amplified in E. coli, and the purified DNA (150 μg) was used for a subsequent round of enrichment of the E2-binding cells. The enrichment step was repeated three times, and finally the transfected WOP cells were incubated with recombinant E2 and then transferred to a petri dish (90-mm diameter) coated with mAb to E2. From the bound cells, plasmid DNA was prepared, amplified in E. coli, and used for the last transfection experiment. The E2-binding capacity of WOP cells was detected by FACScan (Becton-Dickinson) with phycoerythrin-conjugated goat anti-mouse IgG.
13. S. Levy, S. C. Todd, H. T. Maecker, Annu. Rev. Immunol. 16, 89 (1998).
14. Fixed and embedded liver biopsy samples were used for immunohistochemical analyses. Briefly, air-dried, acetone-fixed cryostat sections were incubated with biotin-labeled recombinant E2 followed by incubation with peroxidase-labeled streptavidin.
15. 4, resuspended in 15 ml of 20 mM phosphate buffer and 500 mM NaCl (pH 6.0), and loaded onto a 2-ml nickel-activated chelating Sepharose Fast Flow (Pharmacia) column. Retained proteins were eluted with a 30-ml, 0 to 200 mM Imidazole gradient, and the fractions containing the TRX-EC2 fusion were pooled, dialyzed against PBS, and stored at -80°C
16. Purified TRX-EC2 recombinant proteins were subjected to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) in nonreducing conditions. After electroblotting, the PVDF membrane (Millipore) was probed overnight with recombinant E2 (1 μg/ml) at room temperature. Incubation with an mAb to E2 (clone 291A2) was followed by chemiluminescent detection with a peroxidase-conjugated polyclonal anti-mouse IgG (Amersham).
17. + mouse stable transfectants (NIH 3T3) that bound E2. However, we consistently failed to measure substantial virus attachment to the cell surface by PCR because of high background inherent to the technique.
18. In our assay, we captured only enveloped RNA molecules. The highest available concentration of human TRX-EC2 for coating beads was 100 μg/ml. At this concentration, about 7% of HCV input was bound by the beads. Using TRX-EC2 (100 μg/ml) and increasing numbers of beads, we captured about 10% of the HCV input, in terms of RNA molecules, further demonstrating that HCV binding is dependent on the CD81 concentration. Moreover, our experience with antibody (from mouse, chimp, or human)-coated beads has shown that the percentage of HCV that can be captured is negligible, further proving that CD81 is indeed a very effective binder of HCV particles.
19. Q.-L. Choo et al., Proc. Natl. Acad. Sci. U.S.A. 91, 1294 (1994).
20. K. Blight, R. R. Lesniewski, J. T. LaBrooy, E. J. Gowans, Hepatology 20, 553 (1994); P. Bouffard et al., J. Infect. Dis. 166, 1276 (1992); L. Zignego et al., J. Hepatol. 15, 382 (1992).
21. K. Blight, R. R. Lesniewski, J. T. LaBrooy, E. J. Gowans, Hepatology 20, 553 (1994); P. Bouffard et al., J. Infect. Dis. 166, 1276 (1992); L. Zignego et al., J. Hepatol. 15, 382 (1992).
22. K. Blight, R. R. Lesniewski, J. T. LaBrooy, E. J. Gowans, Hepatology 20, 553 (1994); P. Bouffard et al., J. Infect. Dis. 166, 1276 (1992); L. Zignego et al., J. Hepatol. 15, 382 (1992).
23. F. Berditchevski, M. Zutter, M. E. Hemler, Mol. Biol. Cell 7, 193 (1996); B. A. Mannion, F. Berditchevski, S.-K. Kraeft, L. B. Chen, M. E. Hemler, J. Immunol. 157, 2039 (1996).
24. F. Berditchevski, M. Zutter, M. E. Hemler, Mol. Biol. Cell 7, 193 (1996); B. A. Mannion, F. Berditchevski, S.-K. Kraeft, L. B. Chen, M. E. Hemler, J. Immunol. 157, 2039 (1996).
25. L. E. Bradbury, G. S. Kansas, S. Levy, R. L. Evans, T. F. Tedder, J. Immunol. 149, 2841 (1991).
26. D. T. Fearon and R. H. Carter, Annu. Rev. Immunol. 13, 127 (1995).
27. N. R. Cooper, M. D. Moore, G. R. Nemerow, ibid. 6, 85 (1988).
28. S. Abrignani, unpublished data.
29. P. Chomczinsky and N. Sacchi, Anal. Biochem. 162, 156 (1987).
30. I. G. Campbell, T. A. Jones, W. D. Foulkes, J. Trowsdale, J. Cancer Res. 51, 5329 (1991).
31. U. E. Gibson, C. A. Held, P. M. Williams, Genome Res. 6, 995 (1996).
32. P. Pileri, unpublished data.
33. E. E. Geisert, L. Yang, M. H. Irwin, J. Neurosci. 16, 5478 (1996).
34. M. L. Andrina, C.-L. Hsich, R. Oren, U. Francke, S. Levy, J. Immunol. 147, 1030 (1991).
35. P. Pileri et al., data not shown.
36. We thank C. Moretto for a contribution at the early stages of this project; S. Nuti and C. Satetti for help in flow cytometry; J. Kansopon, D. Chien, Q.-L. Choo, S. Coates, K. Crawford, K. Berger, C. Dong, D. Piccioli, R. La Gaetana, and F. Masciopinto for technical help and reagents; G. Corsi for artwork; and R. Rappuoli, G. Del Giudice, L. Galli-Stampino, and N. Valiante for critical reading of the manuscript. We also thank D. Slade (Pharmacia and Upjohn) for providing the fluorescent detection probe.