HIV-2 neutralization by intact V3-specific Fab fragments

Sourial, Samer; Nilsson, Charlotta

doi:10.1186/1743-422x-5-96

Cited by 3 publications

(8 citation statements)

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“…conformation-sensitive (3C4) and a linear site-specific (7C8) and their respective papain-generated Fab fragments suggested that whole 7C8 and 3C4 MAbs were sterically hindered from neutralizing HIV-2. Alternatively the smaller size of Fab fragments capably accessed the virion surface V3 region (Sourial & Nilsson 2008). Further proceeding in this direction, the crystal structure of the Fab fragment of 7C8 was presented for structural analysis which showed a deep and narrow highly hydrophobic antigen-binding cleft (Uchtenhagen et al 2011).…”

Section: Conventional Attemptsmentioning

confidence: 99%

HIV-2 and its role in conglutinated approach towards Acquired Immunodeficiency Syndrome (AIDS) Vaccine Development

2013

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Acquired Immunodeficiency Syndrome (AIDS) is one of the most critically acclaimed endemic diseases, caused by two lentiviruses HIV-1 and 2. HIV-2 displays intimate serological and antigenic resemblance to Simian Immunodeficiency Virus (SIV) along with less pathogenicity, lower infectivity and appreciable cross reactivity with HIV-1 antigens. The present era is confronted with the challenge to fabricate a vaccine effective against all clades of both the species of HIV. But vaccine development against HIV-1 has proven highly intricate, moreover the laborious and deficient conventional approaches has slackened the pace regarding the development of new vaccines. These concerns may be tackled with the development of HIV-2 vaccine as a natural control of HIV-1 that has been found in ancestors of HIV-2 i.e. African monkeys, mangabeys and macaques. Thereby, suggesting the notion of cross protection among HIV-2 and HIV-1. Assistance of bioinformatics along with vaccinomics strategy can bring about a quantum leap in this direction for surpassing the bottleneck in conventional approaches. These specifics together can add to our conception that HIV-2 vaccine design by in silico strategy will surely be a constructive approach for HIV-1 targeting.Electronic supplementary materialThe online version of this article (doi:10.1186/2193-1801-2-7) contains supplementary material, which is available to authorized users.

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Section: Conventional Attemptsmentioning

confidence: 99%

HIV-2 and its role in conglutinated approach towards Acquired Immunodeficiency Syndrome (AIDS) Vaccine Development

2013

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“…Fab fragments of the monoclonal antibody 7C8 were prepared through limited digestion with papain (Sigma; Sourial & Nilsson, 2008). The reaction was carried out in 20 mM sodium phosphate buffer pH 7.4 containing 150 mM NaCl, 10 mM EDTA and 10 mM cysteine (Sigma).…”

Section: Purification Of 7c8 Fab Fragmentmentioning

confidence: 99%

“…The hybridoma cell line that expresses 7C8, classified as being of the IgG 1 subtype (Sourial & Nilsson, 2008), was originally isolated from mice immunized with a 15-mer peptide derived from the HIV-2 V3-loop region that includes the conserved stretch of residues FHSQ Size-exclusion chromatography of the purified murine antibody 7C8 following papain digestion. Preparative chromatography was carried out in 20 mM Tris-HCl pH 7.4 containing 150 mM NaCl using a Superdex 75 10/300 GL size-exclusion column with a flow rate of 0.5 ml min À1 .…”

Section: Production and Purificationmentioning

confidence: 99%

“…A panel of HIV-2 V3 loop-specific murine monoclonal antibodies, some of which displayed neutralizing capacity, has previously been described in a functional setting (Bjö rling et al, 1994). The monoclonal antibody 7C8 binds with high specificity and efficiency to peptides containing the FHSQ epitope and its Fab fragments neutralize HIV-2 isolates (Bjö rling et al, 1994;Mö rner et al, 1999;Sourial et al, 2006;Sourial & Nilsson, 2008). Interestingly, the third CDR of the heavy chain (CDRH3) of 7C8 is elongated when compared with the average length distribution in mouse antibodies (13 amino-acid residues versus an average of eight to nine; Wu et al, 1993;Johnson & Wu, 1998;Sourial & Nilsson, 2008).…”

Section: Introductionmentioning

confidence: 99%

“…The monoclonal antibody 7C8 binds with high specificity and efficiency to peptides containing the FHSQ epitope and its Fab fragments neutralize HIV-2 isolates (Bjö rling et al, 1994;Mö rner et al, 1999;Sourial et al, 2006;Sourial & Nilsson, 2008). Interestingly, the third CDR of the heavy chain (CDRH3) of 7C8 is elongated when compared with the average length distribution in mouse antibodies (13 amino-acid residues versus an average of eight to nine; Wu et al, 1993;Johnson & Wu, 1998;Sourial & Nilsson, 2008). This concords with data from human neutralizing antibodies specific for gp120 and suggests an important role for the elongated CDRH3 loop, which is believed to be able to reach and overcome sterical hindrance, binding to potentially masked epitopes on the HIV envelope proteins (Johnson & Wu, 1998;Kwong et al, 1998;Saphire et al, 2001;Collis et al, 2003;Ofek et al, 2004;Stanfield et al, 2004;Burton et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

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Production, purification, crystallization and preliminary X-ray diffraction analysis of the HIV-2-neutralizing V3 loop-specific Fab fragment 7C8

Uchtenhagen¹,

Sourial²,

Friemann³

et al. 2009

Acta Cryst Sect F

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7C8 is a mouse monoclonal antibody that is specific for the third hypervariable loop (V3 loop) of the human immunodeficiency virus type 2 (HIV-2) associated protein gp125. Fab fragments of 7C8 effectively neutralize HIV-2. 7C8 was expressed and purified from a hybridoma cell line in order to establish the molecular basis underlying the specificity of the 7C8 antibody for the V3 loop as well as the specific role of the elongated third complementarity-determining region of the heavy chain (CDRH3). The antibody was digested with papain and Fab fragments were purified using size-exclusion chromatography. Hanging-drop vapour-diffusion crystallization techniques were employed and the protein was crystallized in 50 mM ammonium sulfate, 100 mM Tris-HCl pH 8.5, 25%(w/v) PEG 8000 and 2.5%(w/v) PEG 400 at 275 K. The analysed crystals belonged to the rhombohedral space group P3(2)21, with unit-cell parameters a = b = 100.1, c = 196.8 A, and diffracted to 2.7 A resolution.

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Crystal Structure of the HIV-2 Neutralizing Fab Fragment 7C8 with High Specificity to the V3 Region of gp125

et al. 2011

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7C8 is a mouse monoclonal antibody specific for the third hypervariable region (V3) of the human immunodeficiency virus type 2 (HIV-2)-associated protein gp125. The three-dimensional crystal structure of the Fab fragment of 7C8, determined to 2.7 Å resolution, reveals a deep and narrow antigen-binding cleft with architecture appropriate for an elongated epitope. The highly hydrophobic cleft is bordered on one side by the negatively charged second complementarity determining region (CDR2) and the unusually long positively charged CDR3 of the heavy chain and, on the other side, by the CDR1 of the light chain. Analysis of 7C8 in complex with molecular models of monomeric and trimeric gp125 highlights the importance of a conserved stretch of residues FHSQ that is localized centrally on the V3 region of gp125. Furthermore, modeling also indicates that the Fab fragment neutralizes the virus by sterically impairing subsequent engagement of the gp125 trimer with the co-receptor on the target cell.

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HIV-2 neutralization by intact V3-specific Fab fragments

Cited by 3 publications

References 28 publications

HIV-2 and its role in conglutinated approach towards Acquired Immunodeficiency Syndrome (AIDS) Vaccine Development

HIV-2 and its role in conglutinated approach towards Acquired Immunodeficiency Syndrome (AIDS) Vaccine Development

Production, purification, crystallization and preliminary X-ray diffraction analysis of the HIV-2-neutralizing V3 loop-specific Fab fragment 7C8

Crystal Structure of the HIV-2 Neutralizing Fab Fragment 7C8 with High Specificity to the V3 Region of gp125

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