Selection of human anti-human immunodeficiency virus type 1 envelope single-chain antibodies from a peripheral blood cell-based phage repertoire.

Haard, J.J.W. de; Kazemier, Bert; Oudshoorn, P.; Boender, Piet J.; Gemen, Bob van; Koolen, M. J. M.; Groen, G van der; Hoogenboom, Hennie R.; Arends, Jan–Willem

doi:10.1099/0022-1317-79-12-2883

Cited by 7 publications

(5 citation statements)

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“…To construct genes encoding scFvs (AB31, AB32, 4E10, X5 and 48d), overlapping primers for the recursive PCR were synthesized according to the published sequences [34,62]. The end primers of recursive PCR were designed in such a way that the Xba I and the Xma I sites were introduced at 5' and 3' ends of the scFv genes, respectively.…”

Section: Gene Constructionmentioning

confidence: 99%

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GPI-anchored single chain Fv - an effective way to capture transiently-exposed neutralization epitopes on HIV-1 envelope spike

et al. 2010

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BackgroundIdentification of broad neutralization epitopes in HIV-1 envelope spikes is paramount for HIV-1 vaccine development. A few broad neutralization epitopes identified so far are present on the surface of native HIV-1 envelope spikes whose recognition by antibodies does not depend on conformational changes of the envelope spikes. However, HIV-1 envelope spikes also contain transiently-exposed neutralization epitopes, which are more difficult to identify.ResultsIn this study, we constructed single chain Fvs (scFvs) derived from seven human monoclonal antibodies and genetically linked them with or without a glycosyl-phosphatidylinositol (GPI) attachment signal. We show that with a GPI attachment signal the scFvs are targeted to lipid rafts of plasma membranes. In addition, we demonstrate that four of the GPI-anchored scFvs, but not their secreted counterparts, neutralize HIV-1 with various degrees of breadth and potency. Among them, GPI-anchored scFv (X5) exhibits extremely potent and broad neutralization activity against multiple clades of HIV-1 strains tested. Moreover, we show that GPI-anchored scFv (4E10) also exhibited more potent neutralization activity than its secretory counterpart. Finally, we demonstrate that expression of GPI-anchored scFv (X5) in the lipid raft of plasma membrane of human CD4+ T cells confers long-term resistance to HIV-1 infection, HIV-1 envelope-mediated cell-cell fusion, and the infection of HIV-1 captured and transferred by human DCs.ConclusionsThus GPI-anchored scFv could be used as a general and effective way to identify antibodies that react with transiently-exposed neutralization epitopes in envelope proteins of HIV-1 and other enveloped viruses. The GPI-anchored scFv (X5), because of its breadth and potency, should have a great potential to be developed into anti-viral agent for HIV-1 prevention and therapy.

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Section: Gene Constructionmentioning

confidence: 99%

“…AB31 and AB32 are high affinity antibodies. AB31 recognizes cluster III determinant of gp41 and AB32 interacts with gp120, but its epitope is not well characterized [34]. Antibody (TG15) recognizes the cluster II determinant (amino acid residues 644-663) which resides within the second heptad repeat (HR2) of HIV-1 gp41 [23].…”

Section: Introductionmentioning

confidence: 99%

GPI-anchored single chain Fv - an effective way to capture transiently-exposed neutralization epitopes on HIV-1 envelope spike

et al. 2010

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“…The Ig gene usage was also analyzed for 44 human mAbs specific for epitopes in the envelope proteins of HIV-1 other than V3; these included 11 anti-CD4bd mAbs, 12 anti-CD4i mAbs and 21 anti-gp41 mAbs (Table 3). Thirty-four nucleotide sequences for the Ig variable fragment of the heavy chain were obtained from published data or GenBank (Andris et al, 1991;Bagley et al, 1994;Barbas et al, 1993;Cavacini et al, 1998;David et al, 1995b;de Haard et al, 1998;Felgenhauer et al, 1990;Huang et al, 2004;Kunert et al, 1998;Kunert et al, 2004;Marasco et al, 1992;Moran et al, 1993;Thali et al, 1993;van der Donk et al, 1994;Zhang et al, 2004;Zwick et al, 2001) while 10 human anti-gp41 mAbs, 50-69 (Gorny et al, 1989), 126-6, 167-7, 181-D, 240-D and 246-D (Xu et al, 1991), 1281, 1342, 1367and 1379(Gorny et al, 2000, were produced and sequenced in our laboratory (Table 3).…”

Section: Human Anti-gp120 and Gp41 Mabsmentioning

confidence: 99%

Preferential use of the VH5-51 gene segment by the human immune response to code for antibodies against the V3 domain of HIV-1

Gorny

Wang

Williams

et al. 2009

Molecular Immunology

107

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Human anti-V3 monoclonal antibodies (mAbs) generated from HIV-1 infected individuals display diversity in the range of their cross-neutralization that may be related to their immunogenetic background. The study of the immunoglobulin (Ig) variable region gene usage of heavy chains have shown a preferential usage of the VH5-51 gene segment which was detected in 35% of 51 human anti-V3 mAbs. In contrast, human mAbs against other envelope regions of HIV-1 (anti-Env), including the CD4-binding domain, the CD4-induced epitope, and gp41 preferentially used the VH1-69 gene segment, and none of them used the VH5-51 gene. Furthermore, the usage of the VH4 family by anti-V3 mAbs was restricted to only one gene segment, VH4-59, while the VH3 gene family was used at a significantly lower frequency by all of the analyzed anti-HIV-1 mAbs. Multivariate analysis showed that usage of VH gene segments was significantly different between anti-V3 and anti-Env mAbs, and compared to antibodies from healthy subjects. In addition, the anti-V3 mAbs preferentially used the JH3 and D2-15 gene segments. The preferential usage of selected Ig gene segments and the characteristic pattern of Ig gene usage by anti-V3 mAbs can be related to the conserved structure of the V3 region.

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“…13,14 Alternatively, it may also be exploited to study in vivo humoral immune responses to particular antigens. 5,15,16 We set out to study the humoral immune response in a colorectal cancer (CRC) patient by screening phage antibody repertoires made from patient material for the presence of antibodies reactive with colon tumors. Since lymph node tissue has proven to be a valuable source of variable (V)-genes for the construction of phage antibody repertoires 17 and since this tissue has also been shown to be a rich source of antibodies to known tumor antigens, 5 we decided to use patient-derived lymph nodes (LN) draining a CRC for the construction of "immune" phage antibody repertoires.…”

mentioning

confidence: 99%

Evidence for a bias toward intracellular antigens in the local humoral anti-tumor immune response of a colorectal cancer patient revealed by phage display

et al. 2001

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Many patients with colorectal carcinoma (CRC) mount a cellular as well as a humoral immune response to the tumor. To investigate the nature and specificity of the humoral immune response in a CRC patient, lymphocytes infiltrating the primary colorectal tumor and lymph nodes draining the tumor were used as antibody variable (V)-gene pools for the construction of phage antibody repertoires. These libraries were first validated by selection on the antigen tetanus toxoid and shown to contain antibodies that were probably derived from both naive and memory B cells. The repertoires were then screened for the presence of antibodies directed to CRC by selection on the cell line CaCo2. For comparison, the same selections were performed with a phage antibody repertoire made from B cells of healthy donors. Striking differences were observed in the panel of specificities selected from these different repertoires: although a large panel of antibodies reactive with patient-derived primary tumors was obtained from the immune repertoires, none of these discriminated between normal colonic epithelium and colon cancer and none were reactive with cell-surface antigens. However, selections using the non-immune library did result in numerous antibodies that recognized cell surface markers on CaCo2. These data suggest a bias in the local humoral immune response in this CRC patient, directed primarily toward intracellular epithelial-cell specific target antigens.

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Selection of human anti-human immunodeficiency virus type 1 envelope single-chain antibodies from a peripheral blood cell-based phage repertoire.

Cited by 7 publications

References 50 publications

GPI-anchored single chain Fv - an effective way to capture transiently-exposed neutralization epitopes on HIV-1 envelope spike

GPI-anchored single chain Fv - an effective way to capture transiently-exposed neutralization epitopes on HIV-1 envelope spike

Preferential use of the VH5-51 gene segment by the human immune response to code for antibodies against the V3 domain of HIV-1

Evidence for a bias toward intracellular antigens in the local humoral anti-tumor immune response of a colorectal cancer patient revealed by phage display

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