Epitope mapping of the human immunodeficiency virus type 1 gp120 with monoclonal antibodies.
A soluble form of recombinant gp120 of human immunodeficiency virus type 1 was used as an immunogen for production of murine monoclonal antibodies. These monoclonal antibodies were characterized for their ability to block the interaction between gp120 and the acquired immunodeficiency syndrome virus receptor, CD4. Three of the monoclonal antibodies were found to inhibit this interaction, whereas the other antibodies were found to be ineffective at blocking binding. The gp120 epitopes which are recognized by these monoclonal antibodies were mapped by using a combination of Western blot (immunoblot) analysis of gp120 proteolytic fragments, immunoaffinity purification of fragments of gp120, and antibody screening of a random gp120 gene fragment expression library produced in the lambda gt11 expression system. Two monoclonal antibodies which blocked gp120-CD4 interaction were found to map to adjacent sites in the carboxy-terminal region of the glycoprotein, suggesting that this area is important in the interaction between gp120 and CD4. One nonblocking antibody was found to map to a position that was C terminal to this CD4 blocking region. Interestingly, the other nonblocking monoclonal antibodies were found to map either to a highly conserved region in the central part of the gp120 polypeptide or to a highly conserved region near the N terminus of the glycoprotein. N-terminal deletion mutants of the soluble envelope glycoprotein which lack these highly conserved domains but maintain the C-terminal CD4 interaction sites were unable to bind tightly to the CD4 receptor. These results suggest that although the N-terminal and central conserved domains of intact gp120 do not appear to be directly required for CD4 binding, they may contain information that allows other parts of the molecule to form the appropriate structure for CD4 interaction.
Dowbenko
D
D
Department of Molecular Immunology, Genentech, Inc., South San Francisco, California 94080.
Nakamura
G
G
Fennie
C
C
Shimasaki
C
C
Riddle
L
L
Harris
R
R
Gregory
T
T
Lasky
L
L
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000945:Antigens, Differentiation, T-Lymphocyte / Q000276:immunology*; D001667:Binding, Competitive; D015153:Blotting, Western; D002460:Cell Line; D002846:Chromatography, Affinity; D000939:Epitopes / Q000032:analysis*; D005786:Gene Expression Regulation; D015488:HIV Antigens / Q000032:analysis / Q000276:immunology; D015699:HIV Envelope Protein gp120; D015497:HIV-1 / Q000235:genetics / Q000276:immunology*; D006801:Humans; D009154:Mutation; D011991:Receptors, Virus / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D012191:Retroviridae Proteins / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000945:Antigens, Differentiation, T-Lymphocyte; D000939:Epitopes; D015488:HIV Antigens; D015699:HIV Envelope Protein gp120; D011991:Receptors, Virus; D011994:Recombinant Proteins; D012191:Retroviridae Proteins