Restricted accessResearch articleFirst published online 2012-11
Combinations of Griffithsin with Other Carbohydrate-Binding Agents Demonstrate Superior Activity Against HIV Type 1,HIV Type 2,and Selected Carbohydrate-Binding Agent-Resistant HIV Type 1 Strains
Carbohydrate-binding agents (CBAs) are potential HIV microbicidal agents with a high genetic barrier to resistance. We wanted to evaluate whether two mannose-specific CBAs, recognizing multiple and often distinct glycan structures on the HIV envelope gp120, can interact synergistically against HIV-1, HIV-2, and HIV-1 strains that were selected for resistance against particular CBAs [i.e., 2G12 mAb and microvirin (MVN)]. Paired CBA/CBA combinations mainly showed synergistic activity against both wild-type HIV-1 and HIV-2 but also 2G12 mAb- and MVN-resistant HIV-1 strains as based on the median effect principle with combination indices (CIs) ranging between 0.29 and 0.97. Upon combination, an increase in antiviral potency of griffithsin (GRFT) up to ∼12-fold (against HIV-1), ∼8-fold (against HIV-2), and ∼6-fold (against CBA-resistant HIV-1) was observed. In contrast, HHA/GNA combinations showed additive activity against wild-type HIV-1 and HIV-2 strains, but remarkable synergy with HHA and GNA was observed against 2G12 mAb- and MVN-resistant HIV-1 strains (CI, 0.64 and 0.49, respectively). Overall, combinations of GRFT and other CBAs showed synergistic activity against HIV-1, HIV-2, and even against certain CBA-resistant HIV-1 strains. The CBAs tested appear to have distinct binding patterns on the gp120 envelope and therefore do not necessarily compete with each other's glycan binding sites on gp120. As a result, there might be no steric hindrance between two different CBAs in their competition for glycan binding (except for the HHA/GNA combination). These data are encouraging for the use of paired CBA combinations in topical microbicide applications (e.g., creams, gels, or intravaginal rings) to prevent HIV transmission.
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References
1.
LeonardCK, SpellmanMW, RiddleLet al.Assignment of intrachain disulfide bonds and characterization of potential glycosylation sites of the type 1 recombinant human immunodeficiency virus envelope glycoprotein (gp120) expressed in Chinese hamster ovary cells. J Biol Chem, 1990; 265,18:10373–10382.
2.
ScanlanCN, OfferJ, ZitzmannNet al.Exploiting the defensive sugars of HIV-1 for drug and vaccine design. Nature, 2007; 446,7139:1038–1045.
3.
BalzariniJ. Targeting the glycans of glycoproteins: A novel paradigm for antiviral therapy. Nat Rev Microbiol, 2007; 5,8:583–597.
4.
BalzariniJ, ScholsD, NeytsJet al.Alpha-(1-3)- and alpha-(1-6)-D-mannose-specific plant lectins are markedly inhibitory to human immunodeficiency virus and cytomegalovirus infections in vitro. Antimicrob Agents Chemother, 1991; 35,3:410–416.
5.
SwansonMD, WinterHC, GoldsteinIJet al.A lectin isolated from bananas is a potent inhibitor of HIV replication. J Biol Chem, 2010; 285,12:8646–8655.
6.
EmauP, TianB, O'KeefeB Ret al.Griffithsin, a potent HIV entry inhibitor, is an excellent candidate for anti-HIV microbicide. J Med Primatol, 2007; 36,4–5:244–253.
7.
MoriT, O'KeefeBR, SowderRC2ndet al.Isolation and characterization of griffithsin, a novel HIV-inactivating protein, from the red alga Griffithsia sp. J Biol Chem, 2005; 280,10:9345–9353.
8.
KouokamJC, HuskensD, ScholsDet al.Investigation of griffithsin's interactions with human cells confirms its outstanding safety and efficacy profile as a microbicide candidate. PLoS One, 2011; 6,8:e22635.
9.
HuskensD, FérirG, VermeireKet al.Microvirin, a novel {alpha}(1,2)-mannose-specific lectin isolated from microcystis aeruginosa, has comparable anti-HIV-1 activity as cyanovirin-N, but a much higher safety profile. J Biol Chem, 2010; 285,32:24845–24854.
10.
BuchacherA, PredlR, StrutzenbergerKet al.Generation of human monoclonal antibodies against HIV-1 proteins; electrofusion and Epstein-Barr virus transformation for peripheral blood lymphocyte immortalization. AIDS Res Hum Retroviruses, 1994; 10,4:359–369.
11.
ScanlanCN, PantophletR, WormaldMRet al.The broadly neutralizing anti-human immunodeficiency virus type 1 antibody 2G12 recognizes a cluster of alpha1→2 mannose residues on the outer face of gp120. J Virol, 2002; 76,14:7306–7321.
BrouwersJ, VermeireK, GrammenCet al.Early identification of availability issues for poorly water-soluble microbicide candidates in biorelevant media: A case study with saquinavir. Antiviral Res, 2011; 91,2:217–223.
NelAM, CoplanP, van de WijgertJHet al.Safety, tolerability, and systemic absorption of dapivirine vaginal microbicide gel in healthy, HIV-negative women. AIDS, 2009; 23,12:1531–1538.
17.
NelAM, CoplanP, SmytheSCet al.Pharmacokinetic assessment of dapivirine vaginal microbicide gel in healthy, HIV-negative women. AIDS Res Hum Retroviruses, 2010; 26,11:1181–1190.
18.
RomanoJ, VarianoB, CoplanPet al.Safety and availability of dapivirine (TMC120) delivered from an intravaginal ring. AIDS Res Hum Retroviruses, 2009; 25,5:483–488.
19.
Campbell-YesufuOT, GandhiRT. Update on human immunodeficiency virus (HIV)-2 infection. Clin Infect Dis, 2011; 52,6:780–787.
20.
MacNeilA, SarrAD, SankaleJLet al.Direct evidence of lower viral replication rates in vivo in human immunodeficiency virus type 2 (HIV-2) infection than in HIV-1 infection. J Virol, 2007; 81,10:5325–5330.
21.
MarlinkR, KankiP, ThiorIet al.Reduced rate of disease development after HIV-2 infection as compared to HIV-1. Science, 1994; 265,5178:1587–1590.
22.
Schim van der LoeffMF, JaffarS, AveikaAAet al.Mortality of HIV-1, HIV-2 and HIV-1/HIV-2 dually infected patients in a clinic-based cohort in The Gambia. AIDS, 2002; 16,13:1775–1783.
23.
NtemgwaML, d'Aquin ToniT, BrennerBGet al.Antiretroviral drug resistance in human immunodeficiency virus type 2. Antimicrob Agents Chemother, 2009; 53,9:3611–3619.
24.
O'KeefeBR, VojdaniF, BuffaVet al.Scaleable manufacture of HIV-1 entry inhibitor griffithsin and validation of its safety and efficacy as a topical microbicide component. Proc Natl Acad Sci USA, 2009; 106,15:6099–6104.
25.
HuskensD, Van LaethemK, VermeireKet al.Resistance of HIV-1 to the broadly HIV-1-neutralizing, anti-carbohydrate antibody 2G12. Virology, 2007; 360,2:294–304.
26.
PauwelsR, BalzariniJ, BabaMet al.Rapid and automated tetrazolium-based colorimetric assay for the detection of anti-HIV compounds. J Virol Methods, 1988; 20,4:309–321.
27.
VermeireK, PrincenK, HatseSet al.CADA, a novel CD4-targeted HIV inhibitor, is synergistic with various anti-HIV drugs in vitro. AIDS, 2004; 18,16:2115–2125.
28.
ChouTC, TalalayP. Quantitative analysis of dose-effect relationships: the combined effects of multiple drug or enzyme inhibitors. Adv Enzyme Regul, 1984; 22:27–55.
29.
DoncelGF, ClarkMR. Preclinical evaluation of anti-HIV microbicide products: New models and biomarkers. Antiviral Res, 2010; 88,Suppl 1:S10–18.
30.
BaetenJM, BenkiS, ChohanVet al.Hormonal contraceptive use, herpes simplex virus infection, and risk of HIV-1 acquisition among Kenyan women. AIDS, 2007; 21,13:1771–1777.
31.
HerreraC, CranageM, McGowanIet al.Colorectal microbicide design: triple combinations of reverse transcriptase inhibitors are optimal against HIV-1 in tissue explants. AIDS, 2011; 25,16:1971–1979.
32.
GantlettKE, WeberJN, SattentauQJ. Synergistic inhibition of HIV-1 infection by combinations of soluble polyanions with other potential microbicides. Antiviral Res, 2007; 75,3:188–197.
33.
KetasTJ, HoluigueS, MatthewsKet al.Env-glycoprotein heterogeneity as a source of apparent synergy and enhanced cooperativity in inhibition of HIV-1 infection by neutralizing antibodies and entry inhibitors. Virology, 2012; 422,1:22–36.
34.
ShenL, PetersonS, SedaghatARet al.Dose-response curve slope sets class-specific limits on inhibitory potential of anti-HIV drugs. Nat Med, 2008; 14,7:762–766.
35.
FergusonNM, FraserC, AndersonRM. Viral dynamics and anti-viral pharmacodynamics: Rethinking in vitro measures of drug potency. Trends Pharmacol Sci, 2001; 22,2:97–100.
36.
BalzariniJ, Van LaethemK, PeumansWJet al.Mutational pathways, resistance profile, and side effects of cyanovirin relative to human immunodeficiency virus type 1 strains with N-glycan deletions in their gp120 envelopes. J Virol, 2006; 80,17:8411–8421.
37.
HuskensD, VermeireK, VandemeulebrouckeEet al.Safety concerns for the potential use of cyanovirin-N as a microbicidal anti-HIV agent. Int J Biochem Cell Biol, 2008; 40,12:2802–2814.
38.
Shahzad-ul-HussanS, GustchinaE, GhirlandoRet al.Solution structure of the monovalent lectin microvirin in complex with Man(alpha)(1-2)Man provides a basis for anti-HIV activity with low toxicity. J Biol Chem, 2011; 286,23:20788–20796.
39.
AlexandreKB, GrayES, PantophletRet al.Binding of the mannose-specific lectin, griffithsin, to HIV-1 gp120 exposes the CD4-binding site. J Virol, 2011; 85,17:9039–9050.
40.
HoorelbekeB, HuskensD, FérirGet al.Actinohivin, a broadly neutralizing prokaryotic lectin, inhibits HIV-1 infection by specifically targeting high-mannose-type glycans on the gp120 envelope. Antimicrob Agents Chemother, 2010; 54,8:3287–3301.
41.
Abdool KarimQ, Abdool KarimSS, FrohlichJAet al.Effectiveness and safety of tenofovir gel, an antiretroviral microbicide, for the prevention of HIV infection in women. Science, 2010; 329,5996:1168–1174.
42.
AndreiG, LiscoA, VanpouilleCet al.Topical tenofovir, a microbicide effective against HIV, inhibits herpes simplex virus-2 replication. Cell Host Microbe, 2011; 10,4:379–389.
43.
FérirG, VermeireK, HuskensDet al.Synergistic in vitro anti-HIV type 1 activity of tenofovir with carbohydrate-binding agents (CBAs)Antiviral Res, 2011; 90:200–204.
44.
FérirG, PalmerKE, ScholsD. Synergistic activity profile of griffithsin in combination with tenofovir, maraviroc and enfuvirtide against HIV-1 clade C. Virology, 2011; 417,2:253–258.
45.
BertauxC, DaelemansD, MeertensLet al.Entry of hepatitis C virus and human immunodeficiency virus is selectively inhibited by carbohydrate-binding agents but not by polyanions. Virology, 2007; 366,1:40–50.
46.
MeulemanP, AlbeckaA, BelouzardSet al.Griffithsin has antiviral activity against hepatitis C virus. Antimicrob Agents Chemother, 2011; 55,11:5159–5167.
47.
AlexandreKB, GrayES, MufhanduHet al.The lectins griffithsin, cyanovirin-N and scytovirin inhibit HIV-1 binding to the DC-SIGN receptor and transfer to CD4(+) cells. Virology, 2012; 423,2:175–186.
48.
TsaiCC, EmauP, JiangYet al.Cyanovirin-N inhibits AIDS virus infections in vaginal transmission models. AIDS Res Hum Retroviruses, 2004; 20,1:11–18.
49.
TsaiCC, EmauP, JiangYet al.Cyanovirin-N gel as a topical microbicide prevents rectal transmission of SHIV89.6P in macaques. AIDS Res Hum Retroviruses, 2003; 19,7:535–541.
50.
LagenaurLA, Sanders-BeerBE, BrichacekBet al.Prevention of vaginal SHIV transmission in macaques by a live recombinant Lactobacillus. Mucosal Immunol, 2011; 4,6:648–657.
51.
Van DammeL, RamjeeG, AlaryMet al.Effectiveness of COL-1492, a nonoxynol-9 vaginal gel, on HIV-1 transmission in female sex workers: A randomised controlled trial. Lancet, 2002; 360,9338:971–977.
52.
Abdool KarimS, RichardsonB, RamjeeGet al.Safety and effectiveness of BufferGel and 0.5% PRO2000 gel for the prevention of HIV infection in women. AIDS, 2011; 25,7:957–966.
