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Attenuation of Pasteurella multocida by culturing in iron-deficient culture medium; vaccine development
Patent Reports antibodies against a pathogen is new. The (I) is functionally associated with a membrane of a recombinant stable, continuous cell line capable of its production. The vaccine may comprise a membrane-free (I) dissolved free from the membrane after its production. A vaccine comprising a truncated membrane-free derivative of a membrane-bound (I) is also described. The derivative is devoid of membrane-binding domain and the derivative (I) is free from the membrane and has exposed antigenic determinants capable of raising neutralizing antibodies against a pathogen. Membrane-bound (I) and membrane-free (I) are useful as vaccines to give protection against herpes simplex virus 1 and/or 2. The recombinant host cell is a stable eukaryotic cell line or a mammalian cell line, and (I) is especially a glycoprotein of herpes simplex virus type 1 or 2. 098-85
Herpes simplex virus vaccine active against type I and type II containing glycoprotein purified by affinity chromatography Chem. Sero. Ther. Res. Inst. Eur 135-841; 3 April 1985 A herpes simplex virus (HSV) subunit vaccine is described active against HSV type I and type II. The vaccine contains glycoprotein gB (I) as effective component. (I) May be obtained from HSVI or HSVII particles or cells infected with the virus by affinity chromatography using monoclonal antibody against (I). (I) Exhibits immunological activity sufficient to prevent infection by HSVI and HSVII, is the largest component of the HSV envelope and can be easily isolated and purified. The infected ceils or culture supernatant may be first treated ultrasonically or in a homogenizer and then centrifuged to give a dispersion of HSV particles. The subunit glycoprotein may be dissolved with the aid of anionic or preferably nonionic surfactant. For isolation of (I), a (I) solution is passed through an affinity gel column equilibrated with neutral buffer containing surfactant. Elution is performed with, e.g. 3 M KSCN, aq. 5 M MgCI2, aq. 6 M urea etc., containing surfactant. The eluate is then dialysed against neutral buffer containing surfactant. Vaccine formulation is also described. 099-85
Attenuation of Pasteurella multocida by culturing in irondeficient culture medium; vaccine development Akad. Landwirtschaftwiss E. German 216-954; 2 January 1985 A process is described for the attenuation of Pasteurella multocida bacteria and comprises repeatedly culturing the bacteria in an Fe-deficient culture medium, especially for 150 passages. The attenuated strain can be used to prepare vaccines against P. mulocida infections in animals, especially enzootic pneumonia in calves and pigs. In an example, P. multocida strain 383 was cultured in a medium containing 32.3 g/1 NaHPO4.12H20, 1.36 g/l KH2PO4, 1.19 g/l Nacl, 0.25 g/1 MgSO4.7H20, 6 g/1 glucose, 0.2 g/l L-arginine.HCl, 1.6 g/1 Laspartic acid, 0.12 g/l L-Cystine, 0.15 g/1 L-glutamic acid, 0.2 g/l DL-serine, 0.065 g/1 L-isoleucine, 0.065 g/1 L-leucine, 0.085 g/1 Lphenylalanine, 0.09 g/1 L-tyrosine, 0.002 g/l Ca pantothenate, 0.005 g/l nicotinamide, 0.001 g/l thiamine.HC1 and 0.015 g/1 orotic acid. Each passage involved transfer of 0.5 ml of fresh medium and incubating at 37°C for 18-24 h. The virulence (LDs0 dropped from 1 cell/mouse to more than 5 × 10'5 cells/mouse after 150 passages. 100-85 Trypanosoma cruzi surface antigen preparation useful in Chagas disease vaccine and diagnosis produced using a recombinant expression vector Rockefeller-Univ Eur 138-101; 24 April 1985 A recombinant plasmid vector is described containing DNA coding for intact insect stage surface glycoprotein (I) of Trypanosoma cruzi, especially the 75 000 surface glycoprotein (Ia). These vectors are used to transform suitable microorganism hosts, where the (I) gene is expressed. (I) Are used as vaccines
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Vaccine, Vol. 3, December 1985
against Chagas disease. They may also be used in the diagnosis of this disease. A detailed description of vector preparation is given. Briefly, poly A ÷ RNA, isolated from 3 wk old epirnastigote cultures,is purified, translated in vitro and the product immunoprecipitated. The isolated mRNA is then converted to cDNA and the double-stranded product inserted into the PstI site of plasmid pUCI3 by G-C tailing. The recombinant vectors formed are then used to transform Escherichia coli JM83. Transformed colonies are selected by their resistance to ampicillin and inability to produce/~-galactosidase (EC 3.2.1.23). 101-85
New heptatitis A virus surface protein preparation suitable for use in subunit vaccine: hybridoma generation and monoclonal antibody production Merck-USA Eur 138-704; 24 April 1985 A surface structural protein (VP-1) is described, isolated from hepatitis A virus (grown in tissue cultures) by solubilization of the virus in anionic surfactant and reducing agent and separation of the 33 mol.wt, protein by electrophoresis. This protein is suitable for use in the preparation of an HAV polypeptide subunit vaccine. A process is also described for the production of hyhridoma cells producing monoclonal antibodies against HAV or VP-I. These hybridomas are produced by i.p. injecting BALB/c mice with purified whole HAV in Freund's complete adjuvant, priming the mice with a tall vein injection of the HAV in aq. solution, fusing spleen cells from the immunized mice with cells from the SP2/0 mouse myeloma line using PEG, selecting the hybridomas produced in HAT culture medium, cloning monoclonal antibody secreting hybridomas by limiting dilution and i.p. injecting these cloned cells into Pristane-primed mice for the production of ascites fluid. These antibodies are useful in the neutralization, detection and diagnosis of HAV in humans and other susceptible hosts. 102-85
Human cytomegaiovirus antigen preparation by inoculating a human embryo fibroblast cell culture with human cytomegaiic culture collecting centrifuging and further treating the sediment Inst. Virusol. Bucharest Romanian 85-246; 30 October 1984 A process is described for the production of human cytomegalovirus antigen and comprises inoculating a human embryo fibroblast cell culture with a stem of human cytomegalovirus, collecting the culture when the cytopathic effect is at its greatest and centrifuging for 20 rain at 4°C. An improved titre is obtained by resuspending the sediment in a vol. of 0.1 Mglycine buffer at pH 9.5, followed by extraction at RT and ultrasonic treatment. The supernatant liquid is retained after centrifuging. The sediment is then subjected to this treatment for an additional 2 times. The combined supernatant liquids constitute the amount of antigen obtained. 103-85
Vaccine against Bordetella bronchiseptica production by cultivation of several strains of bacteria and incorporation of killed calls into aluminium carrier Staatliehes-Inst. Immunopraep. Naehrmedien E. German 217-144; 9 January 1985 Production of vaccine against Bordetella bronchiseptica infections comprises: (1) cultivation 3 or 4 different strains of the bacterium by a deep-tank method; (2) killing the cells by a conventional method; (3) incorporating them into an aluminium carrier; and (4) preserving with thiomersal. The cells are preferably grown in a culture medium containing casein hydrolysate as N-source; nicotinamide, nicotinic acid and yeast dialysate as growth factors; soluble starch; Mg, Fe and Cu salts; KH2PO4 and cysteine. Preferably, a 80-150 1 glass column fermenter filled to 2/3 capacity is used. The culture medium is aerated and stirred. The bacterial strains selected are in phase I
Herpes simplex virus vaccine active against type I and type II containing glycoprotein purified by affinity chromatography Chem. Sero. Ther. Res. Inst. Eur 135-841; 3 April 1985 A herpes simplex virus (HSV) subunit vaccine is described active against HSV type I and type II. The vaccine contains glycoprotein gB (I) as effective component. (I) May be obtained from HSVI or HSVII particles or cells infected with the virus by affinity chromatography using monoclonal antibody against (I). (I) Exhibits immunological activity sufficient to prevent infection by HSVI and HSVII, is the largest component of the HSV envelope and can be easily isolated and purified. The infected ceils or culture supernatant may be first treated ultrasonically or in a homogenizer and then centrifuged to give a dispersion of HSV particles. The subunit glycoprotein may be dissolved with the aid of anionic or preferably nonionic surfactant. For isolation of (I), a (I) solution is passed through an affinity gel column equilibrated with neutral buffer containing surfactant. Elution is performed with, e.g. 3 M KSCN, aq. 5 M MgCI2, aq. 6 M urea etc., containing surfactant. The eluate is then dialysed against neutral buffer containing surfactant. Vaccine formulation is also described. 099-85
Attenuation of Pasteurella multocida by culturing in irondeficient culture medium; vaccine development Akad. Landwirtschaftwiss E. German 216-954; 2 January 1985 A process is described for the attenuation of Pasteurella multocida bacteria and comprises repeatedly culturing the bacteria in an Fe-deficient culture medium, especially for 150 passages. The attenuated strain can be used to prepare vaccines against P. mulocida infections in animals, especially enzootic pneumonia in calves and pigs. In an example, P. multocida strain 383 was cultured in a medium containing 32.3 g/1 NaHPO4.12H20, 1.36 g/l KH2PO4, 1.19 g/l Nacl, 0.25 g/1 MgSO4.7H20, 6 g/1 glucose, 0.2 g/l L-arginine.HCl, 1.6 g/1 Laspartic acid, 0.12 g/l L-Cystine, 0.15 g/1 L-glutamic acid, 0.2 g/l DL-serine, 0.065 g/1 L-isoleucine, 0.065 g/1 L-leucine, 0.085 g/1 Lphenylalanine, 0.09 g/1 L-tyrosine, 0.002 g/l Ca pantothenate, 0.005 g/l nicotinamide, 0.001 g/l thiamine.HC1 and 0.015 g/1 orotic acid. Each passage involved transfer of 0.5 ml of fresh medium and incubating at 37°C for 18-24 h. The virulence (LDs0 dropped from 1 cell/mouse to more than 5 × 10'5 cells/mouse after 150 passages. 100-85 Trypanosoma cruzi surface antigen preparation useful in Chagas disease vaccine and diagnosis produced using a recombinant expression vector Rockefeller-Univ Eur 138-101; 24 April 1985 A recombinant plasmid vector is described containing DNA coding for intact insect stage surface glycoprotein (I) of Trypanosoma cruzi, especially the 75 000 surface glycoprotein (Ia). These vectors are used to transform suitable microorganism hosts, where the (I) gene is expressed. (I) Are used as vaccines
416
Vaccine, Vol. 3, December 1985
against Chagas disease. They may also be used in the diagnosis of this disease. A detailed description of vector preparation is given. Briefly, poly A ÷ RNA, isolated from 3 wk old epirnastigote cultures,is purified, translated in vitro and the product immunoprecipitated. The isolated mRNA is then converted to cDNA and the double-stranded product inserted into the PstI site of plasmid pUCI3 by G-C tailing. The recombinant vectors formed are then used to transform Escherichia coli JM83. Transformed colonies are selected by their resistance to ampicillin and inability to produce/~-galactosidase (EC 3.2.1.23). 101-85
New heptatitis A virus surface protein preparation suitable for use in subunit vaccine: hybridoma generation and monoclonal antibody production Merck-USA Eur 138-704; 24 April 1985 A surface structural protein (VP-1) is described, isolated from hepatitis A virus (grown in tissue cultures) by solubilization of the virus in anionic surfactant and reducing agent and separation of the 33 mol.wt, protein by electrophoresis. This protein is suitable for use in the preparation of an HAV polypeptide subunit vaccine. A process is also described for the production of hyhridoma cells producing monoclonal antibodies against HAV or VP-I. These hybridomas are produced by i.p. injecting BALB/c mice with purified whole HAV in Freund's complete adjuvant, priming the mice with a tall vein injection of the HAV in aq. solution, fusing spleen cells from the immunized mice with cells from the SP2/0 mouse myeloma line using PEG, selecting the hybridomas produced in HAT culture medium, cloning monoclonal antibody secreting hybridomas by limiting dilution and i.p. injecting these cloned cells into Pristane-primed mice for the production of ascites fluid. These antibodies are useful in the neutralization, detection and diagnosis of HAV in humans and other susceptible hosts. 102-85
Human cytomegaiovirus antigen preparation by inoculating a human embryo fibroblast cell culture with human cytomegaiic culture collecting centrifuging and further treating the sediment Inst. Virusol. Bucharest Romanian 85-246; 30 October 1984 A process is described for the production of human cytomegalovirus antigen and comprises inoculating a human embryo fibroblast cell culture with a stem of human cytomegalovirus, collecting the culture when the cytopathic effect is at its greatest and centrifuging for 20 rain at 4°C. An improved titre is obtained by resuspending the sediment in a vol. of 0.1 Mglycine buffer at pH 9.5, followed by extraction at RT and ultrasonic treatment. The supernatant liquid is retained after centrifuging. The sediment is then subjected to this treatment for an additional 2 times. The combined supernatant liquids constitute the amount of antigen obtained. 103-85
Vaccine against Bordetella bronchiseptica production by cultivation of several strains of bacteria and incorporation of killed calls into aluminium carrier Staatliehes-Inst. Immunopraep. Naehrmedien E. German 217-144; 9 January 1985 Production of vaccine against Bordetella bronchiseptica infections comprises: (1) cultivation 3 or 4 different strains of the bacterium by a deep-tank method; (2) killing the cells by a conventional method; (3) incorporating them into an aluminium carrier; and (4) preserving with thiomersal. The cells are preferably grown in a culture medium containing casein hydrolysate as N-source; nicotinamide, nicotinic acid and yeast dialysate as growth factors; soluble starch; Mg, Fe and Cu salts; KH2PO4 and cysteine. Preferably, a 80-150 1 glass column fermenter filled to 2/3 capacity is used. The culture medium is aerated and stirred. The bacterial strains selected are in phase I