Summary
It has been suggested that threonine or serine residues in the V3 loop of HIV-1 gp120 are glycosylated with the short-chain O-linked oligosaccharides Tn or sialosyl-Tn that function as epitopes for broadly neutralizing carbohydrate specific antibodies. In this study we examined whether mutation of such threonine or serine residues could decrease the sensitivity to infectivity inhibition by Tn or sialosyl-Tn specific antibodies. All potentially O-glycosylated threonine and serine residues in the V3 loop of cloned HIV-1BRU were mutagenized to alanine thus abrogating any O-glycosylation at these sites. Additionally, one of these T-A mutants (T308A) also abrogated the signal for N-glycosylation at N306 inside the V3-loop. The mutant clones were compared with the wild type virus as to sensitivity to neutralization with monoclonal and polyclonal antibodies specific for the tip of the V3 loop of BRU or for the O-linked oligosaccharides Tn or sialosyl-Tn. Deletion of the N-linked oligosaccharide at N306 increased the neutralization sensitivity to antibodies specific for the tip of the loop, which indicates that N-linked glycosylation modulates the accessibility to this immunodominant epitope. However, none of the mutants with deletions of O-glycosylation signals in the V3 loop displayed any decrease in sensitivity to anti-Tn or anti-sialosyl-Tn antibody. This indicates that these broadly specific neutralization epitopes are located outside the V3 loop of gp120.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Back NKT, Smit L, De Jong J-J, Keulen W, Schutten M, Goudsmit J, Tersmette M (1994) An N-glycan within the human immunodeficiency virus type 1 gp 120 V3 loop affects virus neutralization. Virology 199: 431–438
Bernstein HB, Tucker SP, Hunter E, Schutzbach JS, Compans RW (1994) Human immunodeficiency virus type 1 envelope glycoprotein is modified by O-linked oligosaccharides. J Virol 68: 463–468
Clausen H, Pallesen T, Wandall H, Dabelsteen E, Hansen J-ES (1994) Simple mucin type O-glycans of HIV-1: enzymatic prediction of glycosylation sites for vaccine construction. In: Bock K, Clausen H (eds) Complex carbohydrates in drug research. 36th Alfred Benzon Symposium, Copenhagen 1993, Munksgaard, pp 297–315
Hansen J-ES, Hofmann B, Pallesen T, Clausen H (1994) Rôle of carbohydrate on HIV and possibilities for anti-viral intervention. In: Bock K, Clausen H (eds) Complex carbohydrates in drug research. 36th Alfred Benzon Symposium, Copenhagen 1993, Munksgaard, pp 414–427
Hansen J-ES (1992) Carbohydrates of human immunodeficiency virus. APMIS 100: 96–108
Hansen J-ES, Clausen H, Hu S-L, Nielsen JO, Olofsson S (1992) An O-linked carbohydrate neutralization epitope of HIV gp120 is expressed by HIV env gene recombinant vaccinia virus without involvement of HIV regulatory genes. Arch Virol 126: 11–20
Hansen J-ES, Nielsen C, Arendrup M, Olofsson S, Mathiesen LR, Nielsen JO, Clausen H (1991) Broadly neutralizing antibodies targeted to mucin-type carbohydrate epitopes of human immunodeficiency virus. J Virol 65: 6461–6467
Harada S, Koyanagi Y, Yamamoto N (1985) Infection of HTLV-III/LAV in HTLV-I-carrying cells MT-2 and MT-4 and application in a plaque assay. Science 229: 563–566
LaRosa GJ, Davide JP, Weinhold K, Waterbury JA, Profy AT, Lewis JA, Langlois AJ, Dreesman GR, Boswell RN, Shadduck P, Holley LH, Karplus M, Bolognesi DP (1990) Conserved sequence and structural elements in the HIV-1 principal neutralizing determinant. Science 249: 932–935
McCutchan JH, Pagano JS (1968) Enhancement of the infecticity of simian virus 40 deoxyribonucleic acid with diethyl-aminoethyl-dextran. J Natl Cancer Inst 41: 351–357
Moore JP, Jarret RF (1988) Sensitive ELISA for the gp120 and gp160 surface glycoproteins of HIV-1. AIDS Res Hum Retroviruses 5: 369–379
Moore JP, Sattentau QJ, Wyatt R, Sodroski J (1994) Probing the structure of the human immunodeficiency virus surface glycoprotein gp120 with a panel of monoclonal antibodies. J Virol 68: 469–484
Moore JP, Wallace LA, Follett EA, McKeating JA (1988) An enzyme-linked immunosorbent assay for antibodies to the envelope glycoproteins of divergent strains of HIV-1. AIDS 3: 155–163
Popovic M, Sarngadharan MG, Read E, Gallo RC (1984) Detection, isolation and continous production of cytopathic retroviruses (HTLV-III) from patients with AIDS and pre-AIDS. Science 224: 497–500
Putney SD, McKeating JA (1990) Antigenic variation in HIV. AIDS 4 [Suppl 1]: 129–136
Reed LJ, Muench H (1938) A simple method of estimating fifty per cent endpoints. Am J Hyg 27: 493–497
Ricksteen A, Svenson C, Welinder C, Rymo L (1987) Identification of sequences in Epstein-Barr virus DNA required for the expression of the second Epstein-Barr virus-determined nuclear antigen in COS-1 cells. J Gen Virol 68: 2407–2418
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Veronese FM, Reitz MS, Gupta G, Robert-Guroff M, Boyer-Thompson C, Louie A, Gallo RC, Lusso P (1993) Loss of a neutralizing epitope by a spontaneous point mutation in the V3 loop of HIV-1 isolated from an infected laboratory worker. J Biol Chem 268: 25894–25901
Åkerblom L, Hinkula J, Broliden PA, Mäkitalo B, Fridberger T, Rosen J, Vilacres-Eriksson M, Morein B, Wahren B (1990) Neutralizing cross-reactive and non- neutralizing monoclonal antibodies to HIV-1 gp 120. AIDS 4: 953–960
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hansen, J.E.S., Jansson, B., Gram, G.J. et al. Sensitivity of HIV-1 to neutralization by antibodies against O-linked carbohydrate epitopes despite deletion of O-glycosylation signals in the V3 loop. Archives of Virology 141, 291–300 (1996). https://doi.org/10.1007/BF01718400
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01718400