Production and characterization of monoclonal antibodies to simian immunodeficiency virus envelope glycoproteins.
Twelve monoclonal antibodies (MAbs), TB1 to TB12, were produced against a soluble vaccinia recombinant envelope glycoprotein (gp140) from simian immunodeficiency virus SIVmac251. These MAbs recognized SIV gp140 with a relatively high affinity (K0.5 from 6.7 x 10(-8) to 4 x 10(-9) M). All the MAbs except TB9, TB11, and TB12 cross-reacted with HIV-2 envelope glycoproteins, but none of the 12 MAbs recognized those from HIV-1. Using a panel of 87 overlapping synthetic peptides containing 20 amino acid residues, with an overlap of 10 amino acids and spanning the entire primary sequence of gp140, 3 linear epitopes were identified. The first mapped with a neutralizing MAb, TB12, which recognized a linear sequence around amino acids 28-31 within the N-terminal end of the external envelope glycoprotein. The two other new nonneutralizing MAbs recognized linear epitopes around amino acid sequence 380-381 by MAbs TB1, TB2, and TB3, and at the transmembrane glycoprotein amino acids 581-600 by MAb TB6. Seven of the 12 MAbs, TB4, TB5, TB7-9, TB10, and TB11, failed to bind the linear synthetic peptides in ELISA. Moreover, among these seven MAbs only MAbs TB4, TB5, TB9, and TB10 failed to recognize SIV envelope glycoproteins in Western blot (WB) or ELISA after reduction of disulfide bridges by dithiothreitol (DTT), suggesting that they are directed against conformational or discontinuous epitopes. It is of interest to note that MAb TB10 can block the binding of gp140 to the CD4 receptor when the MAb is previously incubated with gp140. Consistent with this result, MAb TB10 cannot bind to gp140 that has been previously complexed with the CD4 receptor. All these results suggest that MAb TB10 recognizes a conformational or discontinuous epitope overlapping or close to the CD4-binding site. These properties are probably implicated in the neutralizing activity observed with this MAb.
Babas
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T
Laboratoire d'Immunovirologie des Lentivirus des Primates, UFR SVT, Université Paul Sabatier, Toulouse, France.
Le Grand
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R
Dormont
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Bahraoui
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E
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000914:Antibodies, Viral / Q000096:biosynthesis*; D000915:Antibody Affinity; D000918:Antibody Specificity; D015153:Blotting, Western; D015704:CD4 Antigens / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D007963:Leukocytes, Mononuclear / Q000821:virology; D008253:Macaca mulatta; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology; D011863:Radioimmunoassay; D011994:Recombinant Proteins / Q000276:immunology; D015302:Simian Immunodeficiency Virus / Q000235:genetics / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D015704:CD4 Antigens; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins