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Identification of sites within gp41 that serve as targets for antibody-dependent cellular cytotoxicity by using human monoclonal antibodies.

Publication ,  Journal Article
Tyler, DS; Stanley, SD; Zolla-Pazner, S; Gorny, MK; Shadduck, PP; Langlois, AJ; Matthews, TJ; Bolognesi, DP; Palker, TJ; Weinhold, KJ
Published in: J Immunol
November 15, 1990

In an effort to determine the functional activity of anti-HIV-1 human mAb and to define the epitopes against which they are directed, supernatants from 10 EBV-transformed lymphoblastoid cell lines producing mAb to HIV were tested. Five clones producing mAb to gp41 and five producing mAb to p24 were identified. The anti-HIV-1 human mAb were tested in neutralization and cell fusion assays in the form of cell culture supernatants at concentrations ranging from 1.7 to 22.0 micrograms/ml. None of the human mAb were found either to inhibit HIV-1-(IIIB or RF) associated cell fusion or to neutralize HIV-1 (IIIB) infection of AA5 cells. All human mAb were additionally tested in 6 h 51Cr release assays for their ability to direct HIV-1 specific antibody-dependent cellular cytotoxicity (ADCC). For ADCC assays, PBMC were isolated from healthy seronegative donors and used as effector cells. HIV-1 infected (IIIB, RF, and MN) CEM.NKR cells as well as CEM.NKR cells with purified gp120 adsorbed onto their surface served as targets. None of the anti-p24 mAb mediated ADCC. In contrast, three of the anti-gp41 mAb were able to direct a significant level of ADCC against each of the infected targets, but as expected, failed to lyse gp120 adsorbed cells. To define the specific epitopes against which the anti-gp41 mAb were directed, seven small peptides homologous to regions within the extracellular domain of gp41 were synthesized. Using RIA, two of the mAb could be mapped. The most effective ADCC-directing human mAb bound to a peptide comprising amino acids 644-663, whereas the least effective ADCC directing anti-gp41 human mAb bound to a region within the immunodominant portion of gp41 outlined by amino acids 579-604. Together, these results for the first time assign a functional activity to human mAb directed at specific regions within gp41 by demonstrating that areas within this molecule can serve as targets for ADCC.

Duke Scholars

Published In

J Immunol

ISSN

0022-1767

Publication Date

November 15, 1990

Volume

145

Issue

10

Start / End Page

3276 / 3282

Location

United States

Related Subject Headings

  • Neutralization Tests
  • Molecular Sequence Data
  • Immunology
  • Humans
  • HIV Envelope Protein gp41
  • Epitopes
  • Antibody-Dependent Cell Cytotoxicity
  • Antibodies, Monoclonal
  • Amino Acid Sequence
  • 3204 Immunology
 

Citation

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ICMJE
MLA
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Tyler, D. S., Stanley, S. D., Zolla-Pazner, S., Gorny, M. K., Shadduck, P. P., Langlois, A. J., … Weinhold, K. J. (1990). Identification of sites within gp41 that serve as targets for antibody-dependent cellular cytotoxicity by using human monoclonal antibodies. J Immunol, 145(10), 3276–3282.
Tyler, D. S., S. D. Stanley, S. Zolla-Pazner, M. K. Gorny, P. P. Shadduck, A. J. Langlois, T. J. Matthews, D. P. Bolognesi, T. J. Palker, and K. J. Weinhold. “Identification of sites within gp41 that serve as targets for antibody-dependent cellular cytotoxicity by using human monoclonal antibodies.J Immunol 145, no. 10 (November 15, 1990): 3276–82.
Tyler DS, Stanley SD, Zolla-Pazner S, Gorny MK, Shadduck PP, Langlois AJ, et al. Identification of sites within gp41 that serve as targets for antibody-dependent cellular cytotoxicity by using human monoclonal antibodies. J Immunol. 1990 Nov 15;145(10):3276–82.
Tyler DS, Stanley SD, Zolla-Pazner S, Gorny MK, Shadduck PP, Langlois AJ, Matthews TJ, Bolognesi DP, Palker TJ, Weinhold KJ. Identification of sites within gp41 that serve as targets for antibody-dependent cellular cytotoxicity by using human monoclonal antibodies. J Immunol. 1990 Nov 15;145(10):3276–3282.

Published In

J Immunol

ISSN

0022-1767

Publication Date

November 15, 1990

Volume

145

Issue

10

Start / End Page

3276 / 3282

Location

United States

Related Subject Headings

  • Neutralization Tests
  • Molecular Sequence Data
  • Immunology
  • Humans
  • HIV Envelope Protein gp41
  • Epitopes
  • Antibody-Dependent Cell Cytotoxicity
  • Antibodies, Monoclonal
  • Amino Acid Sequence
  • 3204 Immunology