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Recombinant vaccinia viruses for use as vaccines against virus, bacterium and protozoon-mediated disease
Patent Reports
for the prevention and treatment of coccidiosis caused by E. tenella. The antigens are useful as vaccines. Eimera spp. inflict heavy economic losses on the fowl industry. By fusing specific antibody-producing cells (spleen cells) with cells of a myeloma, it is possible to produce hybridoma cells which secrete monoclonal antibodies directed speeilieally against the original sensitizing antigen. An example describes the immunization i.p. of 18-wk-old female BALB/c mice with merozoites from primary cultures of fowl kidney cells infected with E. tenella sporozoites. The animals are boosted i.p. with the meroizoites in medium 199 at 2-5 wk intervals after the initial immunization. Spleen cells are then fused with cells of myeloma P3-X63.Ag8.653 using 30% PEG 1000. Hybridomas are cultured and selected in HAT medium, cloning is effected in soft agarose. 093-87
New envelope protein of Visna virus, its fragments and DNA coding sequences, useful in vaccine preparation and diagnosis Inst. Pasteur Fr. 2586 427; 27 February 1987 New polypeptides of the envelope protein of the Visna virus (which causes inflammation in the central nervous system of sheep) are described along with its fragments. The polypeptides are of use for the production of conjugates for making anti-Visna vaccines and in diagnosis for detecting Visna infection in body fluids. The polypeptides are obtained by recombinant DNA techniques. DNA coding sequences are incorporated into suitable vectors under the control of a promoter and followed by appropriate stop signals and polyadenylation signals. They are especially used to make infectious vaccinia virus hybrids by insertion into the TK gene of this virus. The vectors are used to transform suitable hosts for production of the polypeptide. 094-87
Hepatitis B virus vaccine production involves treating hepatitis B virus genetically engineered surface antigen with formaldehyde Green-Cross Jpn 2022 728; 30 January 1987 A hepatitis B virus vaccine contains hepatitis B virus surface antigen (HBsAg) obtained by genetic engineering as the principal component. The HBsAg-containing aq. solution is treated with formaldehyde at a final concentration of the latter of 0.0(0-4).02 w/v%. The protein concentration of HBsAg is adjusted to 0.02-0.05 w/v%, preferably 10-30 w/ v% sucrose containing 10-50 mM phosphate buffer. The fomaldehyde treatment preferably proceeds at 35--40°C for 94-98 h, and the formaldehyde is then removed. Dialysis is then effected using 10-50 mM phosphate buffer (pH 6-8) for over 30 h and the HBsAg obtained is sterilized by filtration and used for vaccine production. The vaccine material is inactivated without lowering the HBsAg solubility. 095--87
Recombinant vaccinia viruses for use as vaccines against virus, bacterium and protozoon-mediated disease Chibaken: Nippon-Zeon Jpn 2044 178; 26 February 1987 Recombinant vaccinia viruses with weak side effects after vaccination are new. The viruses may be used as vaccines for e.g. herpes virus, vesicular stomatitis virus, hepatitis A virus, hepatitis B virus, non-A non-B hepatitis type virus, foot-andmouth-disease virus, polio virus, rabies virus, encephalitis virus, Vibrio cholerae, Clostridium tetani, Plasmodium spp., Schistosoma spp., etc. The recombinant vaccine viruses comprise a vaccine in which exogenous DNA is inserted into an unnecessary DNA region of weak toxic, Lister, temp.-sensitive variant vaceinia virus strains. In the recombinant strains, propagation is impossible in rabbit kidney cells at 41°C. In addition, pox size on hatching egg chorioallantoic membrane 320
Vaccine, Vol. 5, December 1987
is small (below 3 ram). Examples of recombinant strains for use as vaccines are discussed. 096-87
A process for the purification of recombinant vaccinia virus for use as vaccine Chibaken: Nippon-Zeon Jpn 2044 179; 26 February 1987 A simple operation for the extraction and the purification of recombinant vaceinia virus vaccines is new. Recombinant viruses are prepared by the integration of exogenous DNA into the virus region encoding thymidine kinase (EC 2.7.1.21) of weak toxic, Lister, temp.-sensitive, variant vaccinia viruses (unable to grow above 40.5"C, and with small pox size on hatching egg chorioallantoic membrane). The viruses are infected into thymidine-kinase-negative animal cells and cultured in medium containing 3-23 I~g 5-bromodeoxyuridine for 5 days. The recombinant virus cells can be selected as plaque-forming cells from the medium. In an example, experiments were carried out with a hepatitis B virus surface antigen encoding vaccinia virus. Optimum bromodeoxyuridine concentration was determined. 097-87
Synthetic peptide-based anti-rabies composition prepared by recombinant DNA techniques and useful as vaccine and for construction of therapeutic and diagnostic hybridoma producing monodonal antibody. Salk Inst. US 4652 629; 24 March 1987 A sequence in the coat glycoprotein of rabies virus is identified as the molecular basis for an essential step in the pathogenesis of the virus, the binding of virus to the acetylcholine receptor at neurovascular junctions prior to virus uptake into peripheral nerves. Based on this discovery, peptide-based, anti-rabies vaccines can be prepared. The peptides may be obtained by chemical synthesis or by recombinant DNA techniques. Hybridomas which secrete anti-rabies antibodies, which bind to specific epitopes on or near the acetylcholine receptor binding segment on the rabies virus coat glycoprotein, are prepared with B cells immunized in vitro with the synthetic peptide. The antisera and monoclonal antibodies are useful therapeutically to induce passive anti-rabies immunity in a mammal that has been exposed to rabies. They are also useful for diagnosing whether a mammal is rabid and whether a mammal has been exposed to live rabies virus. The vaccine avoids risks associated with currently available antirabies vaccines. 098--87
Synthetic polypetides and antibodies related to Epstein-Barr virus nuclear antigen useful as vaccine; also monoclonal antibody receptor preparation Scripps US 4654 419; 31 March 1987 Synthetic polypeptides have defined amino acid sequence and their salts and antigenically related variants are new. The sequence include: H-Arg-Ala-Arg-Gly- Arg-Gly-Arg-GlyArg-Gly-Glu-Lys- Arg-Pro-Met-OH and H-Ile-Met-Ser-AspGlu-Gly-Pro-Gyl Thr-Gly-Asn-Gyl- Leu-Gly-Glu-OH. The polypeptides, containing at least 25 reel % Gly residues can be linked to a carder and introduced into a mammalian host to induce production of antibodies which react with EpsteinBarr virus nuclear antigen. Suitable monoclonal receptors, typically whole antibodies, may be raised in mammalian hosts such as mice, rabbits, horses, etc. Typically a mouse of the 129 G1X+ strain is preferred. Suitable mouse myelomas include P3X63-Ag8.653 and Sp2/0-Ag14. Splenocytes are fused with myeloma cells using polyethylene glycol (PEG 1500). Fused hybrids are selected by sensitivity to HAT. Hybridoma cells producing receptor molecules are identified by ELISA. 099-87
for the prevention and treatment of coccidiosis caused by E. tenella. The antigens are useful as vaccines. Eimera spp. inflict heavy economic losses on the fowl industry. By fusing specific antibody-producing cells (spleen cells) with cells of a myeloma, it is possible to produce hybridoma cells which secrete monoclonal antibodies directed speeilieally against the original sensitizing antigen. An example describes the immunization i.p. of 18-wk-old female BALB/c mice with merozoites from primary cultures of fowl kidney cells infected with E. tenella sporozoites. The animals are boosted i.p. with the meroizoites in medium 199 at 2-5 wk intervals after the initial immunization. Spleen cells are then fused with cells of myeloma P3-X63.Ag8.653 using 30% PEG 1000. Hybridomas are cultured and selected in HAT medium, cloning is effected in soft agarose. 093-87
New envelope protein of Visna virus, its fragments and DNA coding sequences, useful in vaccine preparation and diagnosis Inst. Pasteur Fr. 2586 427; 27 February 1987 New polypeptides of the envelope protein of the Visna virus (which causes inflammation in the central nervous system of sheep) are described along with its fragments. The polypeptides are of use for the production of conjugates for making anti-Visna vaccines and in diagnosis for detecting Visna infection in body fluids. The polypeptides are obtained by recombinant DNA techniques. DNA coding sequences are incorporated into suitable vectors under the control of a promoter and followed by appropriate stop signals and polyadenylation signals. They are especially used to make infectious vaccinia virus hybrids by insertion into the TK gene of this virus. The vectors are used to transform suitable hosts for production of the polypeptide. 094-87
Hepatitis B virus vaccine production involves treating hepatitis B virus genetically engineered surface antigen with formaldehyde Green-Cross Jpn 2022 728; 30 January 1987 A hepatitis B virus vaccine contains hepatitis B virus surface antigen (HBsAg) obtained by genetic engineering as the principal component. The HBsAg-containing aq. solution is treated with formaldehyde at a final concentration of the latter of 0.0(0-4).02 w/v%. The protein concentration of HBsAg is adjusted to 0.02-0.05 w/v%, preferably 10-30 w/ v% sucrose containing 10-50 mM phosphate buffer. The fomaldehyde treatment preferably proceeds at 35--40°C for 94-98 h, and the formaldehyde is then removed. Dialysis is then effected using 10-50 mM phosphate buffer (pH 6-8) for over 30 h and the HBsAg obtained is sterilized by filtration and used for vaccine production. The vaccine material is inactivated without lowering the HBsAg solubility. 095--87
Recombinant vaccinia viruses for use as vaccines against virus, bacterium and protozoon-mediated disease Chibaken: Nippon-Zeon Jpn 2044 178; 26 February 1987 Recombinant vaccinia viruses with weak side effects after vaccination are new. The viruses may be used as vaccines for e.g. herpes virus, vesicular stomatitis virus, hepatitis A virus, hepatitis B virus, non-A non-B hepatitis type virus, foot-andmouth-disease virus, polio virus, rabies virus, encephalitis virus, Vibrio cholerae, Clostridium tetani, Plasmodium spp., Schistosoma spp., etc. The recombinant vaccine viruses comprise a vaccine in which exogenous DNA is inserted into an unnecessary DNA region of weak toxic, Lister, temp.-sensitive variant vaceinia virus strains. In the recombinant strains, propagation is impossible in rabbit kidney cells at 41°C. In addition, pox size on hatching egg chorioallantoic membrane 320
Vaccine, Vol. 5, December 1987
is small (below 3 ram). Examples of recombinant strains for use as vaccines are discussed. 096-87
A process for the purification of recombinant vaccinia virus for use as vaccine Chibaken: Nippon-Zeon Jpn 2044 179; 26 February 1987 A simple operation for the extraction and the purification of recombinant vaceinia virus vaccines is new. Recombinant viruses are prepared by the integration of exogenous DNA into the virus region encoding thymidine kinase (EC 2.7.1.21) of weak toxic, Lister, temp.-sensitive, variant vaccinia viruses (unable to grow above 40.5"C, and with small pox size on hatching egg chorioallantoic membrane). The viruses are infected into thymidine-kinase-negative animal cells and cultured in medium containing 3-23 I~g 5-bromodeoxyuridine for 5 days. The recombinant virus cells can be selected as plaque-forming cells from the medium. In an example, experiments were carried out with a hepatitis B virus surface antigen encoding vaccinia virus. Optimum bromodeoxyuridine concentration was determined. 097-87
Synthetic peptide-based anti-rabies composition prepared by recombinant DNA techniques and useful as vaccine and for construction of therapeutic and diagnostic hybridoma producing monodonal antibody. Salk Inst. US 4652 629; 24 March 1987 A sequence in the coat glycoprotein of rabies virus is identified as the molecular basis for an essential step in the pathogenesis of the virus, the binding of virus to the acetylcholine receptor at neurovascular junctions prior to virus uptake into peripheral nerves. Based on this discovery, peptide-based, anti-rabies vaccines can be prepared. The peptides may be obtained by chemical synthesis or by recombinant DNA techniques. Hybridomas which secrete anti-rabies antibodies, which bind to specific epitopes on or near the acetylcholine receptor binding segment on the rabies virus coat glycoprotein, are prepared with B cells immunized in vitro with the synthetic peptide. The antisera and monoclonal antibodies are useful therapeutically to induce passive anti-rabies immunity in a mammal that has been exposed to rabies. They are also useful for diagnosing whether a mammal is rabid and whether a mammal has been exposed to live rabies virus. The vaccine avoids risks associated with currently available antirabies vaccines. 098--87
Synthetic polypetides and antibodies related to Epstein-Barr virus nuclear antigen useful as vaccine; also monoclonal antibody receptor preparation Scripps US 4654 419; 31 March 1987 Synthetic polypeptides have defined amino acid sequence and their salts and antigenically related variants are new. The sequence include: H-Arg-Ala-Arg-Gly- Arg-Gly-Arg-GlyArg-Gly-Glu-Lys- Arg-Pro-Met-OH and H-Ile-Met-Ser-AspGlu-Gly-Pro-Gyl Thr-Gly-Asn-Gyl- Leu-Gly-Glu-OH. The polypeptides, containing at least 25 reel % Gly residues can be linked to a carder and introduced into a mammalian host to induce production of antibodies which react with EpsteinBarr virus nuclear antigen. Suitable monoclonal receptors, typically whole antibodies, may be raised in mammalian hosts such as mice, rabbits, horses, etc. Typically a mouse of the 129 G1X+ strain is preferred. Suitable mouse myelomas include P3X63-Ag8.653 and Sp2/0-Ag14. Splenocytes are fused with myeloma cells using polyethylene glycol (PEG 1500). Fused hybrids are selected by sensitivity to HAT. Hybridoma cells producing receptor molecules are identified by ELISA. 099-87