Generation of neutralizing antibody to the reverse transcriptase of human immunodeficiency virus type 1 by immunizing of mice with an infectious vaccinia virus recombinant.
Antibodies inhibiting the reverse transcriptase (RT) of human immunodeficiency virus type-1 (HIV-1) were found to be generated in the serum of mice repeatedly infected with a vaccinia virus recombinant, WRRT, expressing the enzyme. A monoclonal antibody (mAb), 7C4, which specifically and almost completely inhibits the RNA-dependent DNA polymerase activity of HIV-1 RT was produced from a mouse repeatedly immunized with WRRT. 7C4 seems to be specific for HIV-1 among retroviruses: 7C4 inhibited RT activity of three strains of HIV-1 (IIIB, Bru, and IMS-1) but not of two strains of HIV-2 (GH-1 and LAV-2) or two strains of SIV (MAC and MND). The immunoglobulin isotype of three out of four mAbs produced from spleen cells of the immunized mouse were IgG2a. This immunization method that avoids protein denaturation may preferentially induce a T helper type-1 immune response and increase the chances of producing the only occasionally obtainable mAb capable of recognizing a conformational epitope and completely inhibiting enzyme activity.
Chiba
J
J
Department of Biological Science and Technology, Science University of Tokyo, Chiba, Japan. chibaj@rs.noda.sut.ac.jp
Nakano
M
M
Suzuki
Y
Y
Aoyama
K
K
Ohba
H
H
Kobayashi
T
T
Yasuda
A
A
Kojima
A
A
Kurata
T
T
D000818:Animals; D019138:Antibodies, Blocking / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D003001:Cloning, Molecular; D015870:Gene Expression / Q000276:immunology; D015483:HIV Antibodies / Q000096:biosynthesis*; D054303:HIV Reverse Transcriptase / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics / Q000276:immunology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D011995:Recombination, Genetic; D014611:Vaccination; D014614:Vaccines, Synthetic / Q000276:immunology*; D014616:Vaccinia virus / Q000235:genetics / Q000276:immunology
D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; D015483:HIV Antibodies; D014614:Vaccines, Synthetic; D054303:HIV Reverse Transcriptase