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New acellular anti-pertussis vaccine; pertussis toxin protein engineering; filamentous hemagglutinin and 69 kDa protein production by Bordetella pertussis deletion mutant
Patent Report to identify those who are virus-positive and symptom-free for longer than the normal time for appearance of disease symptoms (at least 8 yr); culturing virus from the individuals; screening the virus for properties (growth rate) of (I); and cloning virus having the properties of (I). HIV (I) (claimed) is cultured from IMM-I, IMM-29 and IMM-41 cells for use as a live or inactivated vaccine and in therapy. Highly adaptive nonpathogenic (I) is prepared by: identifying loci in the sequence of (I) where a base change correlates with a change in phenotype from pathogenicity to nonpathogenicity, and where base-pairing between the two loci participates in secondary structure formation of viral DNA or RNA ; mutating the 2nd locus so that interaction between the two loci preserves the secondary structure and reversion to a pathogenic sequence at the 1st locus fails to base-pair at the 2nd locus; and transforming a host cell with DNA or RNA of the virus. 061-92
cholerae toxin or its A1 subunit (ctx gene) and DNA (II) encoding zonula occludens toxin are deleted, thus making the strain avirulent while retaining its capacity to colonize the intestine of a host animal and reducing residual side effects in the host. The culture is 100% effective for vaccination against cholera. The new culture was isolated using a method (claimed) which involves : constructing e.g. plasmid pCVD51 containing (I), (II) and an ampicillin-resistance gene (III), where extrachromosomal replication in V. cholerae is not possible; mating a host containing the plasmid with virulent V. cholerae E7946 containing (I) and (II) inserted between flanking identical copies of a sequence long enough to promote detectable in vivo recombination; selecting V. cholerae expressing (III) ; growing selected V. cholerae in the absence of selective agent; and selecting for V. cholerae which no longer expresses (III) and therefore has (II) and (III) deleted. Plasmid pCVD622.2B encoding Hg-resistance may also be used. 063-92
New acellular anti-pertussis vaccine; pertussis toxin protein engineering; filamentous hemagglutinin and 6 9 k D a protein production by Bordetella pertussis deletion mutant Sclavo Eur 462 534; 27 December 1991 New acellular anti-pertussis vaccines comprise a non-toxic double mutant of pertussis toxin (PT), filamentous hemagglutinin (FHA), 69kDa protein and pharmaceutically acceptable vectors, optionally in association with diphtheria anatoxin and/or tetanus anatoxin. The FHA and 69 kDa protein are preferably from the deletion mutant Bordetella pertussis delta-Tox, which has a deleted pertussis toxin gene, or from a mutant which produces non-toxic PT. The PT non-toxic mutant may contain the following substitutions in the SI subunit: Glu-129 replaced by Gly; Arg-9 replaced by Lys; Trp-26 replaced by Ile ; Arg-13 replaced by Leu ; or Asp-11 and Trp-26 replaced by Ser and Ile, respectively. The mutants may be stabilized by treatment with 0.035-0.7 wt./vol.% formaldehyde. The antigens may be purified from culture of B. pertussis in a liquid culture medium. The proteins are particularly suitable for preparing safe aceUular antipertussis vaccines with improved immunogenicity. In an example, B. pertussis W28-9K/129G was cultured, and mutant PT was purified by filtration, acidification and hydroxyapatite chromatography. 062-92
New Vibrio cholerae strains; cholera toxin deletion mutant selection for use as whole cell vaccine Univ. Maryland World 9118 979; 12 December 1991 Vibrio cholerae CVD 110 is of the Ogawa or Inaba serotype. Restriction fragments of chromosomal DNA (I) encoding V.
New 16 kDa surface protein of Plasmodium falciparum; Pfsl6 gene cloning and expression in transformant; DNA sequence; potential malaria vaccine Univ. Nijmegen World 9118 922; 12 December 1991 A 16kDa protein (A) of Plasmodium falciparum (Pf) comprising a specific protein sequence and immunogenic derivatives of it (including mutants) are new. (A)is obtained from the membrane of gametes and sporozoites (SPs) and is capable of eliciting an immune response in mammals against the SP and exoerythrocyte stages of the parasite. Also claimed are: i. a fusion protein (FP) comprising (A); ii. (A) linked to a macromolecule; iii. a specified DNA sequence (1) encoding (A) or FP; iv. an expression vector comprising (1); v. a host, e.g. bacterium or virus transformed with (iv.); and vi. production of (A) by transforming a host with (I), culturing the host and isolating (A) from the medium. A cDNA gene bank from Pf NF54 was screened for the presence of sexual stage-specific sequences using cDNA constructed from RNA isolated from sexual and asexual blood stages of the parasite. By eliminating clones which hybridized with the asexual probe, a novel Pfsl6 gene was isolated. (A) may be used in a vaccine for the prevention or mitigation of malarial infections in man, and/or blocking of transmission of the malaria parasite. 064-92
Vaccine, Vol. 10, Issue 8. 1992
559
cholerae toxin or its A1 subunit (ctx gene) and DNA (II) encoding zonula occludens toxin are deleted, thus making the strain avirulent while retaining its capacity to colonize the intestine of a host animal and reducing residual side effects in the host. The culture is 100% effective for vaccination against cholera. The new culture was isolated using a method (claimed) which involves : constructing e.g. plasmid pCVD51 containing (I), (II) and an ampicillin-resistance gene (III), where extrachromosomal replication in V. cholerae is not possible; mating a host containing the plasmid with virulent V. cholerae E7946 containing (I) and (II) inserted between flanking identical copies of a sequence long enough to promote detectable in vivo recombination; selecting V. cholerae expressing (III) ; growing selected V. cholerae in the absence of selective agent; and selecting for V. cholerae which no longer expresses (III) and therefore has (II) and (III) deleted. Plasmid pCVD622.2B encoding Hg-resistance may also be used. 063-92
New acellular anti-pertussis vaccine; pertussis toxin protein engineering; filamentous hemagglutinin and 6 9 k D a protein production by Bordetella pertussis deletion mutant Sclavo Eur 462 534; 27 December 1991 New acellular anti-pertussis vaccines comprise a non-toxic double mutant of pertussis toxin (PT), filamentous hemagglutinin (FHA), 69kDa protein and pharmaceutically acceptable vectors, optionally in association with diphtheria anatoxin and/or tetanus anatoxin. The FHA and 69 kDa protein are preferably from the deletion mutant Bordetella pertussis delta-Tox, which has a deleted pertussis toxin gene, or from a mutant which produces non-toxic PT. The PT non-toxic mutant may contain the following substitutions in the SI subunit: Glu-129 replaced by Gly; Arg-9 replaced by Lys; Trp-26 replaced by Ile ; Arg-13 replaced by Leu ; or Asp-11 and Trp-26 replaced by Ser and Ile, respectively. The mutants may be stabilized by treatment with 0.035-0.7 wt./vol.% formaldehyde. The antigens may be purified from culture of B. pertussis in a liquid culture medium. The proteins are particularly suitable for preparing safe aceUular antipertussis vaccines with improved immunogenicity. In an example, B. pertussis W28-9K/129G was cultured, and mutant PT was purified by filtration, acidification and hydroxyapatite chromatography. 062-92
New Vibrio cholerae strains; cholera toxin deletion mutant selection for use as whole cell vaccine Univ. Maryland World 9118 979; 12 December 1991 Vibrio cholerae CVD 110 is of the Ogawa or Inaba serotype. Restriction fragments of chromosomal DNA (I) encoding V.
New 16 kDa surface protein of Plasmodium falciparum; Pfsl6 gene cloning and expression in transformant; DNA sequence; potential malaria vaccine Univ. Nijmegen World 9118 922; 12 December 1991 A 16kDa protein (A) of Plasmodium falciparum (Pf) comprising a specific protein sequence and immunogenic derivatives of it (including mutants) are new. (A)is obtained from the membrane of gametes and sporozoites (SPs) and is capable of eliciting an immune response in mammals against the SP and exoerythrocyte stages of the parasite. Also claimed are: i. a fusion protein (FP) comprising (A); ii. (A) linked to a macromolecule; iii. a specified DNA sequence (1) encoding (A) or FP; iv. an expression vector comprising (1); v. a host, e.g. bacterium or virus transformed with (iv.); and vi. production of (A) by transforming a host with (I), culturing the host and isolating (A) from the medium. A cDNA gene bank from Pf NF54 was screened for the presence of sexual stage-specific sequences using cDNA constructed from RNA isolated from sexual and asexual blood stages of the parasite. By eliminating clones which hybridized with the asexual probe, a novel Pfsl6 gene was isolated. (A) may be used in a vaccine for the prevention or mitigation of malarial infections in man, and/or blocking of transmission of the malaria parasite. 064-92
Vaccine, Vol. 10, Issue 8. 1992
559