TY - JOUR
T1 - Characterization of neutralization epitopes of simian immunodeficiency virus (SIV) recognized by rhesus monoclonal antibodies derived from monkeys infected with an attenuated SIV strain
AU - Stefano Cole, Kelly
AU - Alvarez, Martha
AU - Elliott, Debra H.
AU - Lam, Hoa
AU - Martin, Effie
AU - Chau, Thao
AU - Micken, Katie
AU - Rowles, Jennifer L.
AU - Clements, Janice E.
AU - Murphey-Corb, Michael
AU - Montelaro, Ronald C.
AU - Robinson, James E.
N1 - Funding Information:
We thank Dr. Edmundo Kraiselburd for kindly providing us with the B7 cell line and Dr. Jodi Craigo for assistance with the artwork in Fig. 4. This work was supported by NIH Grants AI28243, AI07487 (K.S.C.), NS38008 (J.E.C.), and AI32375 (J.E.R.) and amfAR Grant 02713 (K.S.C.).
Funding Information:
Synthetic overlapping peptides (EVA 774.1–774.49), 20 amino acids in length with 10-amino-acid overlaps covering the entire sequence of SIVmac251 gp120 (32H isolate), were obtained from the National Institute for Biological Standards and Control Centralised Facility for AIDS Reagents supported by EU Programme EVA (Contract BMH4 97/2515) and the UK Medical Research Council (South Mimms, Potters Bar, Hertsfordshire, UK). A similar set of peptides based on the sequence of SIVmac239 was obtained from the AIDS Research and Reagent Program. Peptides (10 µg/well diluted in 0.05 M sodium bicarbonate, pH 9.6) were fixed onto the wells of ELISA plates (Corning or Dynatech Corp.) by incubation overnight at room temperature. Rhesus MAbs (5 µg/ml) were tested for binding to solid-phase peptides by ELISA, and detection of bound MAbs was determined using peroxidase-conjugated goat anti-monkey or goat anti-human IgG (Sigma) as previously described (Cole et al., 2000).
PY - 2001/11/10
Y1 - 2001/11/10
N2 - A major limitation in the simian immunodeficiency virus (SIV) system has been the lack of reagents with which to identify the antigenic determinants that are responsible for eliciting neutralizing antibody responses in macaques infected with attenuated SIV. Most of our information on SIV neutralization determinants has come from studies with murine monoclonal antibodies (MAbs) produced in response to purified or recombinant SIV envelope proteins or intact SIV-infected cells for relatively short periods of time. While these studies provide some basic information on the potential immunogenic determinants of SIV envelope proteins, it is unclear whether these murine MAbs identify epitopes relevant to antibody responses elicited in monkeys during infection with either wild-type or attenuated SIV strains. To accomplish maximum biological relevance, we developed a reliable method for the production of rhesus monoclonal antibodies. In the present study, we report on the production and characterization of a unique panel of monoclonal antibodies derived from four individual monkeys inoculated with SIV/17E-CL as an attenuated virus strain at a time when protective immunity from pathogenic challenge was evident. Results from these studies identified at least nine binding domains on the surface envelope glycoprotein; these included linear determinants in the V1, V2, cysteine loop (analogous to the V3 loop in human immunodeficiency virus type 1), and C5 regions, as well as conformational epitopes represented by antibodies that bind the C-terminal half of gp120 and those sensitive to defined mutations in the V4 region. More importantly, three groups of antibodies that recognize closely related, conformational epitopes exhibited potent neutralizing activity against the vaccine strain. Identification of the epitopes recognized by these neutralizing antibodies will provide insight into the antigenic determinants responsible for eliciting neutralizing antibodies in vivo that can be used in the design of effective vaccine strategies.
AB - A major limitation in the simian immunodeficiency virus (SIV) system has been the lack of reagents with which to identify the antigenic determinants that are responsible for eliciting neutralizing antibody responses in macaques infected with attenuated SIV. Most of our information on SIV neutralization determinants has come from studies with murine monoclonal antibodies (MAbs) produced in response to purified or recombinant SIV envelope proteins or intact SIV-infected cells for relatively short periods of time. While these studies provide some basic information on the potential immunogenic determinants of SIV envelope proteins, it is unclear whether these murine MAbs identify epitopes relevant to antibody responses elicited in monkeys during infection with either wild-type or attenuated SIV strains. To accomplish maximum biological relevance, we developed a reliable method for the production of rhesus monoclonal antibodies. In the present study, we report on the production and characterization of a unique panel of monoclonal antibodies derived from four individual monkeys inoculated with SIV/17E-CL as an attenuated virus strain at a time when protective immunity from pathogenic challenge was evident. Results from these studies identified at least nine binding domains on the surface envelope glycoprotein; these included linear determinants in the V1, V2, cysteine loop (analogous to the V3 loop in human immunodeficiency virus type 1), and C5 regions, as well as conformational epitopes represented by antibodies that bind the C-terminal half of gp120 and those sensitive to defined mutations in the V4 region. More importantly, three groups of antibodies that recognize closely related, conformational epitopes exhibited potent neutralizing activity against the vaccine strain. Identification of the epitopes recognized by these neutralizing antibodies will provide insight into the antigenic determinants responsible for eliciting neutralizing antibodies in vivo that can be used in the design of effective vaccine strategies.
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U2 - 10.1006/viro.2001.1144
DO - 10.1006/viro.2001.1144
M3 - Article
C2 - 11883006
AN - SCOPUS:0035841649
SN - 0042-6822
VL - 290
SP - 59
EP - 73
JO - Virology
JF - Virology
IS - 1
ER -