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1996-02-26
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Document 0104
DOCN M9620104
TI Construction and characterization of a Salmonella typhi-based human
immunodeficiency virus type 1 vector vaccine.
DT 9602
AU Fouts TR; Lewis GK; Hone DM; Department of Geographic Medicine, School
of Medicine, University; of Maryland at Baltimore 21201, USA.
SO Vaccine. 1995 Apr;13(6):561-9. Unique Identifier : AIDSLINE MED/96065464
AB Since the human immunodeficiency virus type 1 (HIV-1) is transmitted
either parenterally or sexually, both systemic and mucosal immune
responses might be required to provide protective immunity. One option
is to express HIV proteins in attenuated Salmonella vectors that elicit
immune responses in both compartments. The first step to constructing
such a strain was achieved by integrating a gene expression cassette
encoding recombinant HIV-1 gp120 (rgp120) into the aroC locus of an
attenuated vaccine strain of S. typhi. This rgp120 expression cassette
utilizes the strong constitutive promoter, P1pp/lacUV5, and produces
rgp120 to 0.05-01% of the total bacterial cell protein. Immunoblot
analysis shows that the S. typhi strains containing the integrated
cassette express a protein that is both recognized by anti-gp120
monoclonal antibodies (mAbs) and is the appropriate size for
nonglycosylated full-length gp120 (52 kDa). Immunoblot analysis also
demonstrates that the recombinant S. typhi strains express the rgp120 as
monomers and multimers found predominantly in the insoluble fraction of
the bacteria. Antigen-capture ELISA, using antibodies specific for
continuous epitopes on gp120, revealed that the exposure of these
epitopes on S. typhi-expressed rgp120 differs from exposure of these
epitopes on baculovirus-expressed rgp120 that binds CD4. Epitopes in the
first conserved region (109-113) and the third conserved/fourth variable
regions (376-380, 382-384, 395-400) are more surface-exposed, while one
epitope in the third variable region (313-324) is more buried relative
to the corresponding epitopes of baculovirus expressed gp120. Antibodies
recognizing discontinuous epitopes of the CD4 binding domain do not
react with the S. typhi expressed rgp120.(ABSTRACT TRUNCATED AT 250
WORDS)
DE AIDS Vaccines/*GENETICS/IMMUNOLOGY B-Lymphocytes/IMMUNOLOGY Base
Sequence Cloning, Molecular Comparative Study Epitopes/IMMUNOLOGY
Genetic Vectors HIV Envelope Protein gp120/*GENETICS/*IMMUNOLOGY
HIV-1/*IMMUNOLOGY Molecular Sequence Data Mutagenesis, Insertional
Recombinant Proteins/GENETICS/IMMUNOLOGY Salmonella
typhi/*GENETICS/METABOLISM Support, Non-U.S. Gov't Support, U.S.
Gov't, P.H.S. Vaccines, Attenuated/GENETICS/IMMUNOLOGY JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).