The transmembrane envelope (TM) protein gp41 of HIV-1 is an attractive target when designing a vaccine to induce neutralizing antibodies. A few broadly neutralizing antibodies (2F5, 4E10, and 10E8) that target conserved epitopes in the membrane proximal external region (MPER) of gp41 have been isolated from infected individuals. However, attempts to induce such antibodies by immunizations with gp41 and Env derivatives containing the MPER were successful only to some extent. In contrast, immunizations with the ectodomain of the TM protein p15E of different gamma retroviruses resulted in the induction of neutralizing antibodies. These sera recognized epitopes located in the MPER and in the fusion peptide proximal region (FPPR) of p15E. Based on these results, both regions of p15E were substituted with the corresponding sequences derived from gp41 of HIV-1. Thus, four different hybrid antigens were produced. One of the inserted sequences contained the epitopes of 2F5 and 4E10 in the MPER; the other corresponded to the FPPR. Vaccination of rats, guinea pigs, and a goat induced binding antibodies directed against the FPPR of gp41 and the 2F5 epitope (ELDKWA) located in the MPER. Despite the exact recognition of the 2F5 epitope, no or very weak neutralization of HIV-1NL4-3 by the immune sera was demonstrated. Nonetheless, using the strategy of hybrid proteins, antibodies targeting the desired epitope were successfully induced.
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References
1.
Doria-RoseNA, KleinRM, ManionMM, et al.: Frequency and phenotype of human immunodeficiency virus envelope-specific B cells from patients with broadly cross-neutralizing antibodies. J Virol, 2009; 83(1):188–199.
2.
SatherDN, ArmannJ, ChingLK, et al.: Factors associated with the development of cross-reactive neutralizing antibodies during human immunodeficiency virus type 1 infection. J Virol, 2009; 83(2):757–769.
3.
WalkerLM and BurtonDR: Rational antibody-based HIV-1 vaccine design: Current approaches and future directions. Curr Opin Immunol, 2010; 22(3):358–366.
4.
DimitrovAS, LouisJM, BewleyCA, CloreGM, and BlumenthalR: Conformational changes in HIV-1 gp41 in the course of HIV-1 envelope glycoprotein-mediated fusion and inactivation. Biochemistry, 2005; 44(37):12471–12479.
5.
ChanDC and KimPS: HIV entry and its inhibition. Cell, 1998; 93(5):681–684.
6.
BuzonV, NatrajanG, SchibliD, CampeloF, KozlovMM, and WeissenhornW: Crystal structure of HIV-1 gp41 including both fusion peptide and membrane proximal external regions. PLoS Pathog, 2010; 6(5):e1000880.
7.
HeY, LiuS, LiJ, et al.: Conserved salt bridge between the N- and C-terminal heptad repeat regions of the human immunodeficiency virus type 1 gp41 core structure is critical for virus entry and inhibition. J Virol, 2008; 82(22):11129–11139.
8.
ShenX, ParksRJ, MontefioriDC, et al.: In vivo gp41 antibodies targeting the 2F5 monoclonal antibody epitope mediate human immunodeficiency virus type 1 neutralization breadth. J Virol, 2009; 83(8):3617–3625.
9.
HuangJ, OfekG, LaubL, et al.: Broad and potent neutralization of HIV-1 by a gp41-specific human antibody. Nature, 2012; 491(7424):406–412.
10.
BomselM, TudorD, DrilletAS, et al.: Immunization with HIV-1 gp41 subunit virosomes induces mucosal antibodies protecting nonhuman primates against vaginal SHIV challenges. Immunity, 2011; 34(2):269–280.
11.
WangJ, TongP, LuL, et al.: HIV-1 gp41 core with exposed membrane-proximal external region inducing broad HIV-1 neutralizing antibodies. PLoS One, 2011; 6(3):e18233.
12.
ArnoldGF, VelascoPK, HolmesAK, et al.: Broad neutralization of human immunodeficiency virus type 1 (HIV-1) elicited from human rhinoviruses that display the HIV-1 gp41 ELDKWA epitope. J Virol, 2009; 83(10):5087–5100.
13.
YeL, WenZ, DongK, et al.: Induction of HIV neutralizing antibodies against the MPER of the HIV envelope protein by HA/gp41 chimeric protein-based DNA and VLP vaccines. PLoS One, 2011; 6(5):e14813.
14.
MascolaJR and MontefioriDC: The role of antibodies in HIV vaccines. Annu Rev Immunol, 2010; 28:413–444.
15.
KwongPD, MascolaJR, and NabelGJ: Rational design of vaccines to elicit broadly neutralizing antibodies to HIV-1. Cold Spring Harb Perspect Med, 2011; 1(1):a007278.
16.
GuenagaJ, DosenovicP, OfekG, et al.: Heterologous epitope-scaffold prime: Boosting immuno-focuses B cell responses to the HIV-1 gp41 2F5 neutralization determinant. PLoS One, 2011; 6(1):e16074.
17.
MatyasGR, WieczorekL, BeckZ, et al.: Neutralizing antibodies induced by liposomal HIV-1 glycoprotein 41 peptide simultaneously bind to both the 2F5 or 4E10 epitope and lipid epitopes. AIDS, 2009; 23(16):2069–2077.
18.
WatsonDS and SzokaFCJr: Role of lipid structure in the humoral immune response in mice to covalent lipid-peptides from the membrane proximal region of HIV-1 gp41. Vaccine, 2009; 27(34):4672–4683.
19.
FiebigU, StephanO, KurthR, and DennerJ: Neutralizing antibodies against conserved domains of p15E of porcine endogenous retroviruses: Basis for a vaccine for xenotransplantation?. Virology, 2003; 307(2):406–413.
20.
KaulitzD, FiebigU, EschrichtM, WurzbacherC, KurthR, and DennerJ: Generation of neutralising antibodies against porcine endogenous retroviruses (PERVs). Virology, 2011; 411(1):78–86.
21.
FiebigU, HartmannMG, BannertN, KurthR, and DennerJ: Transspecies transmission of the endogenous koala retrovirus. J Virol, 2006; 80(11):5651–5654.
22.
LanghammerS, FiebigU, KurthR, and DennerJ: Neutralising antibodies against the transmembrane protein of feline leukaemia virus (FeLV). Vaccine, 2005; 23(25):3341–3348.
23.
LanghammerS, FiebigU, KurthR, and DennerJ: Increased neutralizing antibody response after simultaneous immunization with leucogen and the feline leukemia virus transmembrane protein. Intervirology, 2010; 54(2):78–86.
24.
LanghammerS, HubnerJ, KurthR, and DennerJ: Antibodies neutralizing feline leukaemia virus (FeLV) in cats immunized with the transmembrane envelope protein p15E. Immunology, 2006; 117(2):229–237.
25.
LanghammerS, HubnerJ, JarrettO, KurthR, and DennerJ: Immunization with the transmembrane protein of a retrovirus, feline leukemia virus: Absence of antigenemia following challenge. Antiviral Res, 2011; 89(1):119–123.
26.
FiebigU, SchmolkeM, EschrichtM, KurthR, and DennerJ: Mode of interaction between the HIV-1-neutralizing monoclonal antibody 2F5 and its epitope. AIDS, 2009; 23(8):887–895.
27.
Hager-BraunC, KatingerH, and TomerKB: The HIV-neutralizing monoclonal antibody 4E10 recognizes N-terminal sequences on the native antigen. J Immunol, 2006; 176(12):7471–7481.
28.
Bellamy-McIntyreAK, LayCS, BaarS, et al.: Functional links between the fusion peptide-proximal polar segment and membrane-proximal region of human immunodeficiency virus gp41 in distinct phases of membrane fusion. J Biol Chem, 2007; 282(32):23104–23116.
29.
NoahE, BironZ, NaiderF, ArshavaB, and AnglisterJ: The membrane proximal external region of the HIV-1 envelope glycoprotein gp41 contributes to the stabilization of the six-helix bundle formed with a matching N’ peptide. Biochemistry, 2008; 47(26):6782–6792.
30.
BussowK, CahillD, NietfeldW, et al.: A method for global protein expression and antibody screening on high-density filters of an arrayed cDNA library. Nucleic Acids Res, 1998; 26(21):5007–5008.
31.
MontefioriDC: Measuring HIV neutralization in a luciferase reporter gene assay. Methods Mol Biol, 2009; 485:395–405.
32.
BehrendtR, FiebigU, NorleyS, GurtlerL, KurthR, and DennerJ: A neutralization assay for HIV-2 based on measurement of provirus integration by duplex real-time PCR. J Virol Methods, 2009; 159(1):40–46.
33.
BehrendtR, FiebigU, SchmolkeM, KurthR, and DennerJ: Induction of antibodies specific for Gp41 of HIV-1 by gene gun DNA vaccination. J Vaccines Vaccin, 2012; 3(145):2.
34.
GeonnottiAR, BilskaM, YuanX, et al.: Differential inhibition of human immunodeficiency virus type 1 in peripheral blood mononuclear cells and TZM-bl cells by endotoxin-mediated chemokine and gamma interferon production. AIDS Res Hum Retroviruses, 2010; 26(3):279–291.
35.
DennisonSM, AnastiK, ScearceRM, et al.: Nonneutralizing HIV-1 gp41 envelope cluster II human monoclonal antibodies show polyreactivity for binding to phospholipids and protein autoantigens. J Virol, 2011; 85(3):1340–1347.
36.
ShibataR, IgarashiT, HaigwoodN, et al.: Neutralizing antibody directed against the HIV-1 envelope glycoprotein can completely block HIV-1/SIV chimeric virus infections of macaque monkeys. Nat Med, 1999; 5(2):204–210.
37.
MascolaJR, LewisMG, StieglerG, et al.: Protection of macaques against pathogenic simian/human immunodeficiency virus 89.6PD by passive transfer of neutralizing antibodies. J Virol, 1999; 73(5):4009–4018.
38.
MascolaJR, StieglerG, VanCottTC, et al.: Protection of macaques against vaginal transmission of a pathogenic HIV-1/SIV chimeric virus by passive infusion of neutralizing antibodies. Nat Med, 2000; 6(2):207–210.
39.
FerrantelliF, RasmussenRA, BuckleyKA, et al.: Complete protection of neonatal rhesus macaques against oral exposure to pathogenic simian-human immunodeficiency virus by human anti-HIV monoclonal antibodies. J Infect Dis, 2004; 189(12):2167–2173.
40.
BabaTW, LiskaV, Hofmann-LehmannR, et al.: Human neutralizing monoclonal antibodies of the IgG1 subtype protect against mucosal simian-human immunodeficiency virus infection. Nat Med, 2000; 6(2):200–206.
41.
HessellAJ, PoignardP, HunterM, et al.: Effective, low-titer antibody protection against low-dose repeated mucosal SHIV challenge in macaques. Nat Med, 2009; 15(8):951–954.
42.
LiJ, ChenX, JiangS, and ChenYH: Deletion of fusion peptide or destabilization of fusion core of HIV gp41 enhances antigenicity and immunogenicity of 4E10 epitope. Biochem Biophys Res Commun, 2008; 376(1):60–64.
43.
HaynesBF: Cardiolipin polyspecific autoreactivity in two broadly neutralizing HIV-1 antibodies. Science, 2005; 308(5730):1906–1908.
44.
SchererEM, ZwickMB, TeytonL, and BurtonDR: Difficulties in eliciting broadly neutralizing anti-HIV antibodies are not explained by cardiolipin autoreactivity. AIDS, 2007; 21(16):2131–2139.
45.
VcelarB, StieglerG, WolfHM, et al.: Reassessment of autoreactivity of the broadly neutralizing HIV antibodies 4E10 and 2F5 and retrospective analysis of clinical safety data. AIDS, 2007; 21(16):2161–2170.
46.
MatyasGR, BeckZ, KarasavvasN, and AlvingCR: Lipid binding properties of 4E10, 2F5, and WR304 monoclonal antibodies that neutralize HIV-1. Biochim Biophys Acta, 2009; 1788(3):660–665.
47.
PetrovasC, VlachoyiannopoulosPG, KordossisT, and MoutsopoulosHM: Anti-phospholipid antibodies in HIV infection and SLE with or without anti-phospholipid syndrome: Comparisons of phospholipid specificity, avidity and reactivity with beta2-GPI. J Autoimmun, 1999; 13(3):347–355.
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