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M9610170.TXT
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1996-01-30
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Document 0170
DOCN M9610170
TI Expression and immunogenicity of an 18-residue epitope of HIV1 gp41
inserted in the flagellar protein of a Salmonella live vaccine.
DT 9601
AU Newton SM; Joys TM; Anderson SA; Kennedy RC; Hovi ME; Stocker BA;
Department of Microbiology and Immunology, Stanford University; School
of Medicine, CA 94305-5402.
SO Res Microbiol. 1995 Mar-Apr;146(3):203-16. Unique Identifier : AIDSLINE
MED/96008859
AB A synthetic oligonucleotide specifying residues 735-752 of the product
of the env gene of HIV1 IIIB was inserted by blunt-end ligation at
restriction sites in the hypervariable, antigenically determinant region
IV of two flagellin genes. Its integration, in frame and correct
orientation, into gene fliC(d) in plasmid pLS408 allowed production of
functional flagella when the plasmid was placed in a flagellin-negative
aroA live-vaccine Salmonella dublin strain, SL5928. Bacteria thus made
motile were immobilized and agglutinated by anti-(735-752 peptide)
serum; expression was also shown by immunoelectron-microscopy and by
Western blot of whole-cell lysates. Enzyme immunoassay (EIA) of sera of
mice given three doses by intraperitoneal injection of the live-vaccine
strain producing chimeric flagellin, or of concentrated flagella from
it, showed production of antibody with affinity for the peptide, and in
some sera, also for r-gp160. Pooled serum from mice given five i.p.
doses of the live vaccine strain expressing the gp41 epitope at the
surface of its flagellar filaments had higher titres of anti-peptide and
anti-r-gp160 antibody and weak neutralizing activity on the IIIB isolate
(90% neutralization at 1/100). The sera of nine mice given two doses of
the live vaccine by the oral route had either no or only very low titres
of antibody to flagellar antigen d; they were therefore not tested for
anti-peptide activity.
DE Animal Antibody Formation/*IMMUNOLOGY AIDS Vaccines/IMMUNOLOGY
Bacterial Vaccines/IMMUNOLOGY Blotting, Western
Epitopes/GENETICS/*IMMUNOLOGY Flagella/ULTRASTRUCTURE
Flagellin/GENETICS/*IMMUNOLOGY HIV-1/GENETICS/*IMMUNOLOGY In Vitro
Membrane Glycoproteins/GENETICS/*IMMUNOLOGY Mice Mice, Inbred BALB C
Rabbits Salmonella/CHEMISTRY/GENETICS/IMMUNOLOGY Support, Non-U.S.
Gov't Support, U.S. Gov't, Non-P.H.S. Support, U.S. Gov't, P.H.S.
Vaccines, Attenuated/IMMUNOLOGY JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).