Dataset Information

Neutralizing epitope mapping of six beta1-bungarotoxin monoclonal antibodies and its application in beta1-bungarotoxin peptide vaccine design.

ABSTRACT: Twenty three stable monoclonal antibodies (mAbs) against beta1-bungarotoxin (beta1-bgt) were prepared by the hybridoma technique. Seven of the 23 mAbs (mAbs 2, 6, 8, 11, 17, 21 and 22) could inhibit more than 70% of phospholipase A2 activity of beta1-bgt and neutralize the toxin. Six of these neutralizing mAbs (mAbs 2, 6, 8, 17, 21 and 22) recognized continuous epitopes on the A chain of beta1-bgt and the other one (mAb 11) recognized a conformational epitope on the toxin. The continuous epitopes of these six mAbs were mapped using synthetic peptide and proteolytic enzymes. Experimental results indicate that mAb 17 recognized the A-chain residues 31-37; mAbs 2 and 8 recognized residues 46-51; mAbs 21 and 22 recognized residues 91-98; and mAb 6 recognized residue 100-106. The competitive-antibody-binding inhibition experiments showed that the affinity of these neutralizing mAbs to the native beta1-bgt is compatible with synthetic peptides. Furthermore, mice immunized with BSA-conjugated A-chain-peptide sequences A(31-37), A(46-51), A(91-98) or A(100-106) were protected from a high-dose beta1-bgt challenge. Subsequently, the peptide-immunized sera were passively injected into Balb/c mice and a significantly protective effect was also observed. To our knowledge, this study is the first systematic demonstration of multiple neutralizing B-cell epitopes of beta1-bgt, and this study is also the first report of the protective synthetic-peptide vaccine against beta1-bgt challenge.

SUBMITTER: Yang CC

PROVIDER: S-EPMC1219165 | biostudies-other | 1998 Feb

REPOSITORIES: biostudies-other

ACCESS DATA

Similar Datasets

Project description:In an attempt to generate broadly cross-reactive, neutralizing monoclonal antibodies (MAbs) to simian immunodeficiency virus (SIV), we compared two immunization protocols using different preparations of oligomeric SIV envelope (Env) glycoproteins. In the first protocol, mice were immunized with soluble gp140 (sgp140) from CP-MAC, a laboratory-adapted variant of SIVmacBK28. Hybridomas were screened by enzyme-linked immunosorbent assay, and a panel of 65 MAbs that recognized epitopes throughout the Env protein was generated. In general, these MAbs detected Env by Western blotting, were at least weakly positive in fluorescence-activated cell sorting (FACS) analysis of Env-expressing cells, and preferentially recognized monomeric Env protein. A subset of these antibodies directed toward the V1/V2 loop, the V3 loop, or nonlinear epitopes were capable of neutralizing CP-MAC, a closely related isolate (SIVmac1A11), and/or two more divergent strains (SIVsmDeltaB670 CL3 and SIVsm543-3E). In the second protocol, mice were immunized with unfixed CP-MAC-infected cells and MAbs were screened for the ability to inhibit cell-cell fusion. In contrast to MAbs generated against sgp140, the seven MAbs produced using this protocol did not react with Env by Western blotting and were strongly positive by FACS analysis, and several reacted preferentially with oligomeric Env. All seven MAbs potently neutralized SIVmac1A11, and several neutralized SIVsmDeltaB670 CL3 and/or SIVsm543-3E. MAbs that inhibited gp120 binding to CD4, CCR5, or both were identified in both groups. MAbs to the V3 loop and one MAb reactive with the V1/V2 loop interfered with CCR5 binding, indicating that these regions of Env play similar roles for SIV and human immunodeficiency virus. Remarkably, several of the MAbs generated against infected cells blocked CCR5 binding in a V3-independent manner, suggesting that they may recognize a region analogous to the conserved coreceptor binding site in gp120. Finally, all neutralizing MAbs blocked infection through the alternate coreceptor STRL33 much more efficiently than infection through CCR5, a finding that has important implications for SIV neutralization assays using CCR5-negative human T-cell lines.

Dataset Information

Neutralizing epitope mapping of six beta1-bungarotoxin monoclonal antibodies and its application in beta1-bungarotoxin peptide vaccine design.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets