Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1995 Dec;69(12):7383–7390. doi: 10.1128/jvi.69.12.7383-7390.1995

Human immunodeficiency virus type 1 infection of SK-N-MC cells: domains of gp120 involved in entry into a CD4-negative, galactosyl ceramide/3' sulfo-galactosyl ceramide-positive cell line.

J M Harouse 1, R G Collman 1, F González-Scarano 1
PMCID: PMC189674  PMID: 7494242

Abstract

The primary receptor for human immunodeficiency virus (HIV) is the CD4 molecule; however, in vitro evidence suggests that a neutral glycolipid, galactosyl ceramide (GalCer) or a derivative molecule, 3' sulfogalactosyl ceramide (GalS), may serve as an alternative receptor for HIV type 1 (HIV-1) in cells of neural and colonic origin. Biochemical studies have demonstrated that recombinant gp120 envelope protein binds to GalCer/GalS in both solid-phase enzyme-linked immunosorbent assay and high-performance thin-layer chromatography overlays. We have used the SK-N-MC cell line, a CD4-negative, GalCer/GalS-positive cell line previously characterized as susceptible to HIV-1 infection, to identify virus isolates with either a positive infection phenotype, HIVHxB2, or a negative infection phenotype, HIV-1(89.6). Using a solid-phase virus binding assay, we determined the level of restriction in HIV-1(89.6) infection to be at the level of virus-glycolipid binding. Furthermore, using HIV-1HxB2-HIV-1(89.6) chimeras, we have identified a 193-amino-acid fragment from the envelope region of HIV-1HxB2 containing the V3, V4, and V5 regions which confers a positive infection phenotype on the HIV-1(89.6) background. Recombinant viruses which separate this 193-amino-acid fragment into two distinct chimeras are each able to confer a positive infection phenotype on the background of HIV89.6, suggesting that a stable GalCer/GalS-envelope interaction is dependent on the conformation of the envelope protein in the context of the viral membrane. Alternatively, the GalCer/GalS-gp120 bond may involve multiple sites on the oligomeric envelope protein.

Full Text

The Full Text of this article is available as a PDF (342.2 KB).

Selected References

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

  1. Alizon M., Dragic T. CD26 antigen and HIV fusion? Science. 1994 May 20;264(5162):1161–1165. doi: 10.1126/science.7909962. [DOI] [PubMed] [Google Scholar]
  2. Bagasra O., Farzadegan H., Seshamma T., Oakes J. W., Saah A., Pomerantz R. J. Detection of HIV-1 proviral DNA in sperm from HIV-1-infected men. AIDS. 1994 Dec;8(12):1669–1674. doi: 10.1097/00002030-199412000-00005. [DOI] [PubMed] [Google Scholar]
  3. Bansal R., Warrington A. E., Gard A. L., Ranscht B., Pfeiffer S. E. Multiple and novel specificities of monoclonal antibodies O1, O4, and R-mAb used in the analysis of oligodendrocyte development. J Neurosci Res. 1989 Dec;24(4):548–557. doi: 10.1002/jnr.490240413. [DOI] [PubMed] [Google Scholar]
  4. Batinić D., Robey F. A. The V3 region of the envelope glycoprotein of human immunodeficiency virus type 1 binds sulfated polysaccharides and CD4-derived synthetic peptides. J Biol Chem. 1992 Apr 5;267(10):6664–6671. [PubMed] [Google Scholar]
  5. Bhat S., Spitalnik S. L., Gonzalez-Scarano F., Silberberg D. H. Galactosyl ceramide or a derivative is an essential component of the neural receptor for human immunodeficiency virus type 1 envelope glycoprotein gp120. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7131–7134. doi: 10.1073/pnas.88.16.7131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Boyd M. T., Simpson G. R., Cann A. J., Johnson M. A., Weiss R. A. A single amino acid substitution in the V1 loop of human immunodeficiency virus type 1 gp120 alters cellular tropism. J Virol. 1993 Jun;67(6):3649–3652. doi: 10.1128/jvi.67.6.3649-3652.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Callahan L. N., Phelan M., Mallinson M., Norcross M. A. Dextran sulfate blocks antibody binding to the principal neutralizing domain of human immunodeficiency virus type 1 without interfering with gp120-CD4 interactions. J Virol. 1991 Mar;65(3):1543–1550. doi: 10.1128/jvi.65.3.1543-1550.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Callebaut C., Krust B., Jacotot E., Hovanessian A. G. T cell activation antigen, CD26, as a cofactor for entry of HIV in CD4+ cells. Science. 1993 Dec 24;262(5142):2045–2050. doi: 10.1126/science.7903479. [DOI] [PubMed] [Google Scholar]
  9. Cann A. J., Churcher M. J., Boyd M., O'Brien W., Zhao J. Q., Zack J., Chen I. S. The region of the envelope gene of human immunodeficiency virus type 1 responsible for determination of cell tropism. J Virol. 1992 Jan;66(1):305–309. doi: 10.1128/jvi.66.1.305-309.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cheng-Mayer C., Rutka J. T., Rosenblum M. L., McHugh T., Stites D. P., Levy J. A. Human immunodeficiency virus can productively infect cultured human glial cells. Proc Natl Acad Sci U S A. 1987 May;84(10):3526–3530. doi: 10.1073/pnas.84.10.3526. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chesebro B., Buller R., Portis J., Wehrly K. Failure of human immunodeficiency virus entry and infection in CD4-positive human brain and skin cells. J Virol. 1990 Jan;64(1):215–221. doi: 10.1128/jvi.64.1.215-221.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Chesebro B., Wehrly K., Nishio J., Perryman S. Macrophage-tropic human immunodeficiency virus isolates from different patients exhibit unusual V3 envelope sequence homogeneity in comparison with T-cell-tropic isolates: definition of critical amino acids involved in cell tropism. J Virol. 1992 Nov;66(11):6547–6554. doi: 10.1128/jvi.66.11.6547-6554.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Chiodi F., Valentin A., Keys B., Schwartz S., Asjö B., Gartner S., Popovic M., Albert J., Sundqvist V. A., Fenyö E. M. Biological characterization of paired human immunodeficiency virus type 1 isolates from blood and cerebrospinal fluid. Virology. 1989 Nov;173(1):178–187. doi: 10.1016/0042-6822(89)90233-x. [DOI] [PubMed] [Google Scholar]
  14. Clapham P. R., Blanc D., Weiss R. A. Specific cell surface requirements for the infection of CD4-positive cells by human immunodeficiency virus types 1 and 2 and by Simian immunodeficiency virus. Virology. 1991 Apr;181(2):703–715. doi: 10.1016/0042-6822(91)90904-P. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Clapham P. R., Weber J. N., Whitby D., McIntosh K., Dalgleish A. G., Maddon P. J., Deen K. C., Sweet R. W., Weiss R. A. Soluble CD4 blocks the infectivity of diverse strains of HIV and SIV for T cells and monocytes but not for brain and muscle cells. Nature. 1989 Jan 26;337(6205):368–370. doi: 10.1038/337368a0. [DOI] [PubMed] [Google Scholar]
  16. Collman R., Balliet J. W., Gregory S. A., Friedman H., Kolson D. L., Nathanson N., Srinivasan A. An infectious molecular clone of an unusual macrophage-tropic and highly cytopathic strain of human immunodeficiency virus type 1. J Virol. 1992 Dec;66(12):7517–7521. doi: 10.1128/jvi.66.12.7517-7521.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Collman R., Godfrey B., Cutilli J., Rhodes A., Hassan N. F., Sweet R., Douglas S. D., Friedman H., Nathanson N., Gonzalez-Scarano F. Macrophage-tropic strains of human immunodeficiency virus type 1 utilize the CD4 receptor. J Virol. 1990 Sep;64(9):4468–4476. doi: 10.1128/jvi.64.9.4468-4476.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Collman R., Hassan N. F., Walker R., Godfrey B., Cutilli J., Hastings J. C., Friedman H., Douglas S. D., Nathanson N. Infection of monocyte-derived macrophages with human immunodeficiency virus type 1 (HIV-1). Monocyte-tropic and lymphocyte-tropic strains of HIV-1 show distinctive patterns of replication in a panel of cell types. J Exp Med. 1989 Oct 1;170(4):1149–1163. doi: 10.1084/jem.170.4.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Cook D. G., Fantini J., Spitalnik S. L., Gonzalez-Scarano F. Binding of human immunodeficiency virus type I (HIV-1) gp120 to galactosylceramide (GalCer): relationship to the V3 loop. Virology. 1994 Jun;201(2):206–214. doi: 10.1006/viro.1994.1287. [DOI] [PubMed] [Google Scholar]
  20. Dragic T., Picard L., Alizon M. Proteinase-resistant factors in human erythrocyte membranes mediate CD4-dependent fusion with cells expressing human immunodeficiency virus type 1 envelope glycoproteins. J Virol. 1995 Feb;69(2):1013–1018. doi: 10.1128/jvi.69.2.1013-1018.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Fantini J., Cook D. G., Nathanson N., Spitalnik S. L., Gonzalez-Scarano F. Infection of colonic epithelial cell lines by type 1 human immunodeficiency virus is associated with cell surface expression of galactosylceramide, a potential alternative gp120 receptor. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2700–2704. doi: 10.1073/pnas.90.7.2700. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Furuta Y., Eriksson K., Svennerholm B., Fredman P., Horal P., Jeansson S., Vahlne A., Holmgren J., Czerkinsky C. Infection of vaginal and colonic epithelial cells by the human immunodeficiency virus type 1 is neutralized by antibodies raised against conserved epitopes in the envelope glycoprotein gp120. Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12559–12563. doi: 10.1073/pnas.91.26.12559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Harouse J. M., Bhat S., Spitalnik S. L., Laughlin M., Stefano K., Silberberg D. H., Gonzalez-Scarano F. Inhibition of entry of HIV-1 in neural cell lines by antibodies against galactosyl ceramide. Science. 1991 Jul 19;253(5017):320–323. doi: 10.1126/science.1857969. [DOI] [PubMed] [Google Scholar]
  24. Harouse J. M., Kunsch C., Hartle H. T., Laughlin M. A., Hoxie J. A., Wigdahl B., Gonzalez-Scarano F. CD4-independent infection of human neural cells by human immunodeficiency virus type 1. J Virol. 1989 Jun;63(6):2527–2533. doi: 10.1128/jvi.63.6.2527-2533.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Howell D. N., Berger A. E., Cresswell P. Human T-B lymphoblast hybrids express HLA-DR specificities not expressed by either parent. Immunogenetics. 1982;15(2):199–206. doi: 10.1007/BF00621951. [DOI] [PubMed] [Google Scholar]
  26. Hwang S. S., Boyle T. J., Lyerly H. K., Cullen B. R. Identification of the envelope V3 loop as the primary determinant of cell tropism in HIV-1. Science. 1991 Jul 5;253(5015):71–74. doi: 10.1126/science.1905842. [DOI] [PubMed] [Google Scholar]
  27. Kim F. M., Kolson D. L., Balliet J. W., Srinivasan A., Collman R. G. V3-independent determinants of macrophage tropism in a primary human immunodeficiency virus type 1 isolate. J Virol. 1995 Mar;69(3):1755–1761. doi: 10.1128/jvi.69.3.1755-1761.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Koito A., Stamatatos L., Cheng-Mayer C. Small amino acid sequence changes within the V2 domain can affect the function of a T-cell line-tropic human immunodeficiency virus type 1 envelope gp120. Virology. 1995 Feb 1;206(2):878–884. doi: 10.1006/viro.1995.1010. [DOI] [PubMed] [Google Scholar]
  29. Kunsch C., Hartle H. T., Wigdahl B. Infection of human fetal dorsal root ganglion glial cells with human immunodeficiency virus type 1 involves an entry mechanism independent of the CD4 T4A epitope. J Virol. 1989 Dec;63(12):5054–5061. doi: 10.1128/jvi.63.12.5054-5061.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Li X. L., Moudgil T., Vinters H. V., Ho D. D. CD4-independent, productive infection of a neuronal cell line by human immunodeficiency virus type 1. J Virol. 1990 Mar;64(3):1383–1387. doi: 10.1128/jvi.64.3.1383-1387.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Long D., Berson J. F., Cook D. G., Doms R. W. Characterization of human immunodeficiency virus type 1 gp120 binding to liposomes containing galactosylceramide. J Virol. 1994 Sep;68(9):5890–5898. doi: 10.1128/jvi.68.9.5890-5898.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Maddon P. J., Dalgleish A. G., McDougal J. S., Clapham P. R., Weiss R. A., Axel R. The T4 gene encodes the AIDS virus receptor and is expressed in the immune system and the brain. Cell. 1986 Nov 7;47(3):333–348. doi: 10.1016/0092-8674(86)90590-8. [DOI] [PubMed] [Google Scholar]
  33. Malykh A., Reitz M. S., Jr, Louie A., Hall L., Lori F. Multiple determinants for growth of human immunodeficiency virus type 1 in monocyte-macrophages. Virology. 1995 Jan 10;206(1):646–650. doi: 10.1016/s0042-6822(95)80082-4. [DOI] [PubMed] [Google Scholar]
  34. Moore J. P., Sattentau Q. J., Wyatt R., Sodroski J. Probing the structure of the human immunodeficiency virus surface glycoprotein gp120 with a panel of monoclonal antibodies. J Virol. 1994 Jan;68(1):469–484. doi: 10.1128/jvi.68.1.469-484.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Moore J. P., Thali M., Jameson B. A., Vignaux F., Lewis G. K., Poon S. W., Charles M., Fung M. S., Sun B., Durda P. J. Immunochemical analysis of the gp120 surface glycoprotein of human immunodeficiency virus type 1: probing the structure of the C4 and V4 domains and the interaction of the C4 domain with the V3 loop. J Virol. 1993 Aug;67(8):4785–4796. doi: 10.1128/jvi.67.8.4785-4796.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nuovo G. J., Gallery F., MacConnell P., Braun A. In situ detection of polymerase chain reaction-amplified HIV-1 nucleic acids and tumor necrosis factor-alpha RNA in the central nervous system. Am J Pathol. 1994 Apr;144(4):659–666. [PMC free article] [PubMed] [Google Scholar]
  37. O'Brien W. A., Koyanagi Y., Namazie A., Zhao J. Q., Diagne A., Idler K., Zack J. A., Chen I. S. HIV-1 tropism for mononuclear phagocytes can be determined by regions of gp120 outside the CD4-binding domain. Nature. 1990 Nov 1;348(6296):69–73. doi: 10.1038/348069a0. [DOI] [PubMed] [Google Scholar]
  38. Piatak M., Jr, Luk K. C., Williams B., Lifson J. D. Quantitative competitive polymerase chain reaction for accurate quantitation of HIV DNA and RNA species. Biotechniques. 1993 Jan;14(1):70–81. [PubMed] [Google Scholar]
  39. Piatak M., Jr, Saag M. S., Yang L. C., Clark S. J., Kappes J. C., Luk K. C., Hahn B. H., Shaw G. M., Lifson J. D. High levels of HIV-1 in plasma during all stages of infection determined by competitive PCR. Science. 1993 Mar 19;259(5102):1749–1754. doi: 10.1126/science.8096089. [DOI] [PubMed] [Google Scholar]
  40. Ranscht B., Clapshaw P. A., Price J., Noble M., Seifert W. Development of oligodendrocytes and Schwann cells studied with a monoclonal antibody against galactocerebroside. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2709–2713. doi: 10.1073/pnas.79.8.2709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Ratner L., Haseltine W., Patarca R., Livak K. J., Starcich B., Josephs S. F., Doran E. R., Rafalski J. A., Whitehorn E. A., Baumeister K. Complete nucleotide sequence of the AIDS virus, HTLV-III. Nature. 1985 Jan 24;313(6000):277–284. doi: 10.1038/313277a0. [DOI] [PubMed] [Google Scholar]
  42. Roderiquez G., Oravecz T., Yanagishita M., Bou-Habib D. C., Mostowski H., Norcross M. A. Mediation of human immunodeficiency virus type 1 binding by interaction of cell surface heparan sulfate proteoglycans with the V3 region of envelope gp120-gp41. J Virol. 1995 Apr;69(4):2233–2239. doi: 10.1128/jvi.69.4.2233-2239.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Stefano K. A., Collman R., Kolson D., Hoxie J., Nathanson N., Gonzalez-Scarano F. Replication of a macrophage-tropic strain of human immunodeficiency virus type 1 (HIV-1) in a hybrid cell line, CEMx174, suggests that cellular accessory molecules are required for HIV-1 entry. J Virol. 1993 Nov;67(11):6707–6715. doi: 10.1128/jvi.67.11.6707-6715.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Sweet R. W., Truneh A., Hendrickson W. A. CD4: its structure, role in immune function and AIDS pathogenesis, and potential as a pharmacological target. Curr Opin Biotechnol. 1991 Aug;2(4):622–633. doi: 10.1016/0958-1669(91)90089-n. [DOI] [PubMed] [Google Scholar]
  45. Takeuchi Y., Akutsu M., Murayama K., Shimizu N., Hoshino H. Host range mutant of human immunodeficiency virus type 1: modification of cell tropism by a single point mutation at the neutralization epitope in the env gene. J Virol. 1991 Apr;65(4):1710–1718. doi: 10.1128/jvi.65.4.1710-1718.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Tornatore C., Chandra R., Berger J. R., Major E. O. HIV-1 infection of subcortical astrocytes in the pediatric central nervous system. Neurology. 1994 Mar;44(3 Pt 1):481–487. doi: 10.1212/wnl.44.3_part_1.481. [DOI] [PubMed] [Google Scholar]
  47. Tornatore C., Nath A., Amemiya K., Major E. O. Persistent human immunodeficiency virus type 1 infection in human fetal glial cells reactivated by T-cell factor(s) or by the cytokines tumor necrosis factor alpha and interleukin-1 beta. J Virol. 1991 Nov;65(11):6094–6100. doi: 10.1128/jvi.65.11.6094-6100.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Weber J., Clapham P., McKeating J., Stratton M., Robey E., Weiss R. Infection of brain cells by diverse human immunodeficiency virus isolates: role of CD4 as receptor. J Gen Virol. 1989 Oct;70(Pt 10):2653–2660. doi: 10.1099/0022-1317-70-10-2653. [DOI] [PubMed] [Google Scholar]
  49. Westervelt P., Trowbridge D. B., Epstein L. G., Blumberg B. M., Li Y., Hahn B. H., Shaw G. M., Price R. W., Ratner L. Macrophage tropism determinants of human immunodeficiency virus type 1 in vivo. J Virol. 1992 Apr;66(4):2577–2582. doi: 10.1128/jvi.66.4.2577-2582.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Wyatt R., Thali M., Tilley S., Pinter A., Posner M., Ho D., Robinson J., Sodroski J. Relationship of the human immunodeficiency virus type 1 gp120 third variable loop to a component of the CD4 binding site in the fourth conserved region. J Virol. 1992 Dec;66(12):6997–7004. doi: 10.1128/jvi.66.12.6997-7004.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Yahi N., Baghdiguian S., Moreau H., Fantini J. Galactosyl ceramide (or a closely related molecule) is the receptor for human immunodeficiency virus type 1 on human colon epithelial HT29 cells. J Virol. 1992 Aug;66(8):4848–4854. doi: 10.1128/jvi.66.8.4848-4854.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Yahi N., Sabatier J. M., Baghdiguian S., Gonzalez-Scarano F., Fantini J. Synthetic multimeric peptides derived from the principal neutralization domain (V3 loop) of human immunodeficiency virus type 1 (HIV-1) gp120 bind to galactosylceramide and block HIV-1 infection in a human CD4-negative mucosal epithelial cell line. J Virol. 1995 Jan;69(1):320–325. doi: 10.1128/jvi.69.1.320-325.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. van den Berg L. H., Sadiq S. A., Lederman S., Latov N. The gp120 glycoprotein of HIV-1 binds to sulfatide and to the myelin associated glycoprotein. J Neurosci Res. 1992 Dec;33(4):513–518. doi: 10.1002/jnr.490330403. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES