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New non-infectious HIV virus particles produced in mammal cell culture used as vaccine against AIDS
Patent Report Production of vaccine for control of footrot in sheep comprising basic serine protease from virulent or intermediate strain of Bacteroides nodosus CSIRO World 8809 668; 15 December 1988 A new vaccine for protection against or treatment of footrot contains an immunologically effective amount of the basic serine protease of Bacteroides nodosus virulent strain 198 or virulent strain 334 as the active component. The enzyme can be replaced by a synthetic polypeptide with the same antigenicity. The synthetic polypeptide comprises the expression product of a host cell containing new recombinant DNA with a specified nucleotide sequence encoding all or part of the serine protease. Also new are polyclonal and monoclonal antibodies to the serine protease, and a diagnostic kit for the detection of virulent and/or intermediate strains ofB. nodosus. Gel filtration of culture media from B. nodosus virulent or intermediate strains produced two peaks of protease activity. The first peak was purified on sulphopropyl-Sephadex C-25 to give homogeneous basic serine protease. The enzyme has a mol.wt of 37000 and exists as a tetramer (mol. wt 150 000). The active component of the vaccine can be the purified enzyme, or a crude or partially purified supernatant of a B. nodosus culture. The vaccine may include an adjuvant e.g. AI(OH)3, etc. 059-89
New immunoreactive protein of human B-lymphotropic virus recombinant vaccine production and disease diagnosis Baylor Coll. Med., US Dept. Health Human Serv World 8809 814; 15 December 1988 An isolated immunoreactive human B-lymphotropic virus protein encoded by a portion of a 9 kb HindIII DNA fragment of the virus genome is new. Also new are: (1) a recombinant expression vector, preferably a plasmid, containing the DNA sequence; (2) bacteria and cells transformed with the expression vector; (3) a method for producing recombinant human B-lymphotropic virus immunoreactive protein which comprises transforming a host with a vector and isolating the protein from the host; (4) a method for diagnosing human B-lymphotropic virus diseases in a biological sample using an immunoadsorbent; (5) an immunoadsorbent; (6) a kit for immunoassay containing the immunoadsorbent; and (7) an immunogen comprising an immunogenic amount of recombinant protein and a physiologically acceptable carrier. The bacterial host is preferably Escherichia coli. The immunogens are useful in recombinant vaccine production. 060-89
Recombinant vaccinia virus expressing gB glycoprotein from herpes-simplex virus-1 useful for recombinant vaccine production City of Hope Eur. 297 924; 4 January 1989 A recombinant vaccinia virus which expresses gB glycoprotein identical to that expressed by herpes simplex virus-1 (HSV1) is new. Also new is the recombinant gB glycoprotein. The gB glycoprotein is an important inducer of protective immunity to HSV1 infection and plays an important role in cell penetration. It may be used in recombinant vaccine production. The gB gene is modified at the 5' end by in vitro oligonucleotide directed mutagenesis to conver the gB translation initiation codon (ATG) to an EcoRI site so that ligation to EcoRI-cleaved plasmid pKB results in an in-frame fusion to a vaccinia virus derived ATG codon. The modified gB sequence is inserted into the vaccinia virus genome under the control of the vaccinia virus promoter. The mature gB glycoprotein is glycosylated, expressed at the cell surface and is indistinguishable from authentic HSV1 gB. 061-89
370 Vaccine, Vol. 7, August 1989
New non-infectious HIV virus particles produced in mammal cell culture used as vaccine against AIDS Immune Response World 8809 670; 15 December 1988 An immunogen comprising non-infectious retroviral, specifically HIV virus, particles devoid of outer envelope proteins, specifically gp160 and gpl20, is new. Also new is a monoclonal antibody reactive with one or more retroviral proteins but not reactive with the outer envelope proteins, and procedures for stimulating the immune system of humans with retro virus infection, and for preventing HIV virus infection. The new procedures and reagents can be used in the prevention and therapy of retroviral, e.g. HIV virus, infections. In an example, cells infected with HIV virus were cultured in RPMI 1640 medium containing 10% fetal bovine serum, 25mM HEPES, 50 /~g m l - I gentamicin and 100 gg ml-1 streptomycin. The cultures were expanded and supernatant from 5-7-day-old suspensions of the HIV virus-infected cells were filtered. A fl propiolactone solution was added to the supernatant and incubated at 37°C and pH 7.2-7.4 for 5 h. The supernatant was frozen and exposed to 4.5 mR Co60 gamma-irradiation. The supernatant was then thawed and the new immunogen was isolated free of outer envelope proteins. 062-89
Immunogenic, lymphoproliferative fraction with interleukin stimulation activity isolation from Leishmania infantum useful in vaccine production and diagnosis lnst Pasteur; Uni. Compiegne Fr. 2615 103; 18 November 1988 Fractions extracted from Leishmania spp., e.g. Leishmania infantum, are new. The seven fractions are antigenic and can be used to produce vaccines and for diagnosis of Leishmaniosis. Some of the fractions also have lymphoproliferative activity and induce the release of interleukin-1 from human monocytes. Fractions F1 and F2 contain mannose, glucose, L-fucose and fl-galactose while F3 and F4 contain mannose, glucose, and L-fucose. The fractions are prepared without using toxic reagents. Leishmania spp. promastigotes are cultured at 26°C and pH 6-8 with a partial pressure of 0 2 of 10-80% and agitation. The culture is continued at a partial pressure of 0 2 of 50-100%. The culture is centrifuged to recover the parasites, which are suspended in buffer. The extract is centrifuged and sonicated repeatedly and subject to molecular sieve chromatography. The fractions are eluted with ammonium acetate. 063-89
Production of modified RNA-replicase for protection against viral infection of plant and animal: enzyme engineering by amino acid substitution Yahult Honsha Jpn 3287 483; 24 November 1988 A new process for producing modified RNA-replicase involves substituting amino acid residues for amino acid X in a common amino acid sequence, Tyr-X-Asp-Asp-, which exists in the RNA-replicase protein of RNA virus. More specifically, the RNA-replicase of Q-fl phase is modified by replacing Gly, in the sequence Tyr-Gly-Asp-Asp (amino acid 356-359) of RNA-replicase fl-subunit, with Ala, Ser, Pro, Met or Val. The modified RNA-replicase can be produced by culturing host cells transformed with a clone encoding the desired protein. Using this procedure, inactive RNA-replicase is produced in host cells and inhibits proliferation of phage Q-fl and phase SP-RNA. The amino acid-substituted enzyme may be used to protect host cells from viral infection. This procedure may also be applied to other RNA viruses, particularly plant and animal RNA viruses. The procedure can be used in plant and animal immunization. 064-89
New immunoreactive protein of human B-lymphotropic virus recombinant vaccine production and disease diagnosis Baylor Coll. Med., US Dept. Health Human Serv World 8809 814; 15 December 1988 An isolated immunoreactive human B-lymphotropic virus protein encoded by a portion of a 9 kb HindIII DNA fragment of the virus genome is new. Also new are: (1) a recombinant expression vector, preferably a plasmid, containing the DNA sequence; (2) bacteria and cells transformed with the expression vector; (3) a method for producing recombinant human B-lymphotropic virus immunoreactive protein which comprises transforming a host with a vector and isolating the protein from the host; (4) a method for diagnosing human B-lymphotropic virus diseases in a biological sample using an immunoadsorbent; (5) an immunoadsorbent; (6) a kit for immunoassay containing the immunoadsorbent; and (7) an immunogen comprising an immunogenic amount of recombinant protein and a physiologically acceptable carrier. The bacterial host is preferably Escherichia coli. The immunogens are useful in recombinant vaccine production. 060-89
Recombinant vaccinia virus expressing gB glycoprotein from herpes-simplex virus-1 useful for recombinant vaccine production City of Hope Eur. 297 924; 4 January 1989 A recombinant vaccinia virus which expresses gB glycoprotein identical to that expressed by herpes simplex virus-1 (HSV1) is new. Also new is the recombinant gB glycoprotein. The gB glycoprotein is an important inducer of protective immunity to HSV1 infection and plays an important role in cell penetration. It may be used in recombinant vaccine production. The gB gene is modified at the 5' end by in vitro oligonucleotide directed mutagenesis to conver the gB translation initiation codon (ATG) to an EcoRI site so that ligation to EcoRI-cleaved plasmid pKB results in an in-frame fusion to a vaccinia virus derived ATG codon. The modified gB sequence is inserted into the vaccinia virus genome under the control of the vaccinia virus promoter. The mature gB glycoprotein is glycosylated, expressed at the cell surface and is indistinguishable from authentic HSV1 gB. 061-89
370 Vaccine, Vol. 7, August 1989
New non-infectious HIV virus particles produced in mammal cell culture used as vaccine against AIDS Immune Response World 8809 670; 15 December 1988 An immunogen comprising non-infectious retroviral, specifically HIV virus, particles devoid of outer envelope proteins, specifically gp160 and gpl20, is new. Also new is a monoclonal antibody reactive with one or more retroviral proteins but not reactive with the outer envelope proteins, and procedures for stimulating the immune system of humans with retro virus infection, and for preventing HIV virus infection. The new procedures and reagents can be used in the prevention and therapy of retroviral, e.g. HIV virus, infections. In an example, cells infected with HIV virus were cultured in RPMI 1640 medium containing 10% fetal bovine serum, 25mM HEPES, 50 /~g m l - I gentamicin and 100 gg ml-1 streptomycin. The cultures were expanded and supernatant from 5-7-day-old suspensions of the HIV virus-infected cells were filtered. A fl propiolactone solution was added to the supernatant and incubated at 37°C and pH 7.2-7.4 for 5 h. The supernatant was frozen and exposed to 4.5 mR Co60 gamma-irradiation. The supernatant was then thawed and the new immunogen was isolated free of outer envelope proteins. 062-89
Immunogenic, lymphoproliferative fraction with interleukin stimulation activity isolation from Leishmania infantum useful in vaccine production and diagnosis lnst Pasteur; Uni. Compiegne Fr. 2615 103; 18 November 1988 Fractions extracted from Leishmania spp., e.g. Leishmania infantum, are new. The seven fractions are antigenic and can be used to produce vaccines and for diagnosis of Leishmaniosis. Some of the fractions also have lymphoproliferative activity and induce the release of interleukin-1 from human monocytes. Fractions F1 and F2 contain mannose, glucose, L-fucose and fl-galactose while F3 and F4 contain mannose, glucose, and L-fucose. The fractions are prepared without using toxic reagents. Leishmania spp. promastigotes are cultured at 26°C and pH 6-8 with a partial pressure of 0 2 of 10-80% and agitation. The culture is continued at a partial pressure of 0 2 of 50-100%. The culture is centrifuged to recover the parasites, which are suspended in buffer. The extract is centrifuged and sonicated repeatedly and subject to molecular sieve chromatography. The fractions are eluted with ammonium acetate. 063-89
Production of modified RNA-replicase for protection against viral infection of plant and animal: enzyme engineering by amino acid substitution Yahult Honsha Jpn 3287 483; 24 November 1988 A new process for producing modified RNA-replicase involves substituting amino acid residues for amino acid X in a common amino acid sequence, Tyr-X-Asp-Asp-, which exists in the RNA-replicase protein of RNA virus. More specifically, the RNA-replicase of Q-fl phase is modified by replacing Gly, in the sequence Tyr-Gly-Asp-Asp (amino acid 356-359) of RNA-replicase fl-subunit, with Ala, Ser, Pro, Met or Val. The modified RNA-replicase can be produced by culturing host cells transformed with a clone encoding the desired protein. Using this procedure, inactive RNA-replicase is produced in host cells and inhibits proliferation of phage Q-fl and phase SP-RNA. The amino acid-substituted enzyme may be used to protect host cells from viral infection. This procedure may also be applied to other RNA viruses, particularly plant and animal RNA viruses. The procedure can be used in plant and animal immunization. 064-89