TY - JOUR
T1 - Characterization of V3 loop-Pseudomonas exotoxin chimeras
T2 - Candidate vaccines for human immunodeficiency virus-1
AU - FitzGerald, David J.
AU - Fryling, Charlotte M.
AU - McKee, Marian L.
AU - Vennari, Jo Ann C.
AU - Wrin, Terri
AU - Cromwell, Mary E.M.
AU - Daugherty, Ann L.
AU - Mrsny, Randall J.
PY - 1998/4/17
Y1 - 1998/4/17
N2 - To develop a candidate vaccine for human immunodeficiency virus, type 1 (HIV-1), chimeric proteins were constructed by inserting sequences derived from the V3 loop of gp120 into a nontoxic form of Pseudomonas exotoxin (PE). Inserts of 14 or 26 amino acids, constrained by a disulfide bond, were introduced between domains II and III of PE. V3 loop-toxin proteins expressed in Escherichia coil and corresponding to either MN (subtype B) or Thai (subtype E) strains, were recognized by strain-specific monoclonal anti- gp120 antibodies. When loop sequences were in introduced into an enzymatically active form of the toxin, there was no loss of toxin-mediated cell killing, suggesting that these sequences were co-transported to the cytosol. Sera from rabbits injected with nontoxic PE-V3 loop chimeras were reactive for strain-specific gp120s in Western blots, immunocapture assays, enzyme-linked immunosorbent assays, and neutralized HIV-I infectivity. Since toxin vectors were designed to receive oligonucleotide duplexes encoding any V3 loop sequence, this approach should allow for the production of V3 loop- toxin chimeras corresponding to multiple HIV isolates.
AB - To develop a candidate vaccine for human immunodeficiency virus, type 1 (HIV-1), chimeric proteins were constructed by inserting sequences derived from the V3 loop of gp120 into a nontoxic form of Pseudomonas exotoxin (PE). Inserts of 14 or 26 amino acids, constrained by a disulfide bond, were introduced between domains II and III of PE. V3 loop-toxin proteins expressed in Escherichia coil and corresponding to either MN (subtype B) or Thai (subtype E) strains, were recognized by strain-specific monoclonal anti- gp120 antibodies. When loop sequences were in introduced into an enzymatically active form of the toxin, there was no loss of toxin-mediated cell killing, suggesting that these sequences were co-transported to the cytosol. Sera from rabbits injected with nontoxic PE-V3 loop chimeras were reactive for strain-specific gp120s in Western blots, immunocapture assays, enzyme-linked immunosorbent assays, and neutralized HIV-I infectivity. Since toxin vectors were designed to receive oligonucleotide duplexes encoding any V3 loop sequence, this approach should allow for the production of V3 loop- toxin chimeras corresponding to multiple HIV isolates.
UR - http://www.scopus.com/inward/record.url?scp=0032540369&partnerID=8YFLogxK
U2 - 10.1074/jbc.273.16.9951
DO - 10.1074/jbc.273.16.9951
M3 - Article
C2 - 9545339
AN - SCOPUS:0032540369
SN - 0021-9258
VL - 273
SP - 9951
EP - 9958
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 16
ER -