Characterization of a novel human immunodeficiency virus type 1 neutralizable epitope within the immunodominant region of gp41.
Previously, we generated human monoclonal antibodies using peripheral blood mononuclear cells from an asymptomatic human immunodeficiency virus type 1 (HIV-1)-seropositive donor. One of these monoclonal antibodies (designated clone 3, CL3) recognized 10 amino acids (GCSGKLICTT) within the immunodominant region (cluster I) of the transmembrane envelope glycoprotein gp41 and neutralized infection of target cells with different laboratory isolates. Because the epitope recognized by CL3 has two cysteine residues that could potentially produce a disulfide loop in gp41, we analyzed binding of our monoclonal antibody to the cyclic and linear motif of the peptide sequence IWGCSGKLICTTAVP (residues 600-614). The CL3 antibody did not bind to the synthetic cyclic peptide but did recognize the linear form. Two polyclonal rabbit sera against both the linear and cyclic peptides were then generated. Both antisera bound to viral glycoproteins gp41 and gp160, but neither sera neutralized HIV-1 laboratory isolates. Using a set of alanine-substituted IWGCSGKLICTTAV peptides, we analyzed binding of polyclonal antisera and CL3. The profile of binding of polyclonal antisera to these peptides was different from that of CL3 to the same peptides. This suggests that CL3 recognized a unique neutralizable core epitope, which was not immunogenic in either the cyclic or the linear IWGCSGKLICTTAVP peptides used as immunogens in the rabbits.
Viveros
M
M
Instituto de Investigaciones Biomedicas, Universidad Nacional Autonoma de Mexico, Mexico City, Mexico 70228, USA.
Dickey
C
C
Cotropia
J P
JP
Gevorkian
G
G
Larralde
C
C
Broliden
K
K
Levi
M
M
Burgess
A
A
Cao
C
C
Weiner
D B
DB
Agadjanyan
M G
MG
Ugen
K E
KE
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D002460:Cell Line; D003545:Cysteine / Q000276:immunology / Q000378:metabolism; D004220:Disulfides / Q000276:immunology / Q000378:metabolism; D015488:HIV Antigens / Q000737:chemistry / Q000276:immunology*; D019302:HIV Envelope Protein gp160 / Q000276:immunology; D015700:HIV Envelope Protein gp41 / Q000737:chemistry / Q000276:immunology*; D006679:HIV Seropositivity / Q000276:immunology / Q000821:virology; D015497:HIV-1 / Q000235:genetics / Q000276:immunology* / Q000502:physiology; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D016056:Immunodominant Epitopes / Q000737:chemistry / Q000276:immunology*; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010456:Peptides, Cyclic / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D011817:Rabbits; D014779:Virus Replication
D000911:Antibodies, Monoclonal; D004220:Disulfides; D015488:HIV Antigens; D019302:HIV Envelope Protein gp160; D015700:HIV Envelope Protein gp41; D007106:Immune Sera; D016056:Immunodominant Epitopes; D010456:Peptides, Cyclic; D003545:Cysteine