Mode of interaction between the HIV-1-neutralizing monoclonal antibody 2F5 and its epitope.
OBJECTIVE
To determine the mechanism of interaction between the HIV-1 gp41-specific broadly neutralizing monoclonal antibody (mAb) 2F5, its epitope in the membrane proximal external region and a domain located in the fusion peptide proximal region in the N-terminal region of gp41. Knowledge of these interactions would be useful for the design of antigens used to induce 2F5-like antibodies.


METHODS
The binding and avidity of the mAb 2F5 were analyzed using enzyme-linked immunosorbent assays, epitope mapping and surface plasmon resonance analysis. Inhibition of virus neutralization by 2F5 was analyzed using peptides corresponding to the gp41 sequence.


RESULTS
Using transmembrane envelope proteins of gammaretroviruses, we had previously induced neutralizing antibodies that recognize two epitopes, one located in the N-terminal part of the transmembrane protein (designated E1) and the other in the C-terminal membrane proximal external region (E2). The E2 epitope corresponds to the mAb 2F5/4E10 epitope in the gp41 of HIV and we have now identified a corresponding E1 domain in gp41. Although 2F5 did not bind directly to E1, the presence of E1 peptides increased the binding of 2F5 to peptides carrying its epitope. Neutralization of HIV-1 by 2F5 was inhibited more effectively by both gp41-derived peptides E1 and E2 together than with the peptide E2 alone.


CONCLUSIONS
The interaction between the E1 and E2 domains of gp41 increased the efficacy of mAb 2F5 binding to its E2 epitope. Such an interaction may occur after gp41 folding into a six-helix bundle. Antigens containing both domains might be used to induce broadly neutralizing 2F5-like antibodies.
Robert Koch Institute, Nordufer 20, Berlin D-13353, Germany. FiebigU@rki.de
Uwe
U
Fiebig
Mirco
M
Schmolke
Magdalena
M
Eschricht
Reinhard
R
Kurth
Joachim
J
Denner
D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D000939:Epitopes / Q000187:drug effects*; D015497:HIV-1 / Q000187:drug effects*; D006801:Humans; D007155:Immunologic Factors / Q000494:pharmacology*; D009500:Neutralization Tests; D020349:Surface Plasmon Resonance; D014759:Viral Envelope Proteins / Q000494:pharmacology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007155:Immunologic Factors; D014759:Viral Envelope Proteins