CA2081638A1 - Preparation of antigens of and of vaccines for the virus of mystery disease, antigens and vaccines obtained for the prevention of this disease - Google Patents
Preparation of antigens of and of vaccines for the virus of mystery disease, antigens and vaccines obtained for the prevention of this diseaseInfo
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- CA2081638A1 CA2081638A1 CA002081638A CA2081638A CA2081638A1 CA 2081638 A1 CA2081638 A1 CA 2081638A1 CA 002081638 A CA002081638 A CA 002081638A CA 2081638 A CA2081638 A CA 2081638A CA 2081638 A1 CA2081638 A1 CA 2081638A1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
- A61K39/145—Orthomyxoviridae, e.g. influenza virus
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/525—Virus
- A61K2039/5252—Virus inactivated (killed)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/525—Virus
- A61K2039/5254—Virus avirulent or attenuated
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55505—Inorganic adjuvants
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/10011—Arteriviridae
- C12N2770/10021—Viruses as such, e.g. new isolates, mutants or their genomic sequences
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/10011—Arteriviridae
- C12N2770/10034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
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Abstract
ABSTRACT
Preparation of antigens of and of vaccines for the virus of Mystery Disease, antigens and vaccines obtained for the prevention of this disease.
The process for isolating the virus of Mystery Disease comprises taking samples from organs of sick or infected pigs, milling the samples and passing the milling supernatant through sensitive heterologous or homologous cells and then harvesting the supernatant. For the industrial production of the virus of Mystery Disease, the virus is cultured in sensitive heterologous or homologous cells.
The preparations of purified viral antigen of Mystery Disease comprise living viral particles, optionally atrtenuated, or inactivated particles or subunit antigens or antigens expressed by genetic recombination from genes of the isolated virus. vaccines containing a vaccinating quantity of such an antigen.
Preparation of antigens of and of vaccines for the virus of Mystery Disease, antigens and vaccines obtained for the prevention of this disease.
The process for isolating the virus of Mystery Disease comprises taking samples from organs of sick or infected pigs, milling the samples and passing the milling supernatant through sensitive heterologous or homologous cells and then harvesting the supernatant. For the industrial production of the virus of Mystery Disease, the virus is cultured in sensitive heterologous or homologous cells.
The preparations of purified viral antigen of Mystery Disease comprise living viral particles, optionally atrtenuated, or inactivated particles or subunit antigens or antigens expressed by genetic recombination from genes of the isolated virus. vaccines containing a vaccinating quantity of such an antigen.
Description
2~8~6~
Preparation of antigens of and of vaccines for the virus of Mystery Disease, antigens and vaccines obtained for the prevention of this disease.
The present invention relateæ to the preparation of antigens of and of vaccines for the virus of Mystery Disea~e, as well as to the antigens and to the vaccines obtained.
The disease called Mystery Disea~e (M.D.) or also Porcine Reproductive Re~piratory Syndrome (P.R.R.S.) began to acquire a identity of its own in pigs, in 1986 in the United StateR and in 1990 in Europe. This disease manifests itself essentially in pig~ by ~ign~ of exhaus-tion, anorexia and hyperthermia of the order of 40C, which a~e conventionally observed in ~OW8 in pig farms affected by the disease. The~e signs are accompanied or followed by reproductive disorders (premature or late farrowing and bi.rth of stillborn, mummified or sickly piglets, and return of the 50W~ to heat). A respiratory syndrome can be observed in piglet with interstitial pneumonia lesions. Older pigs can al~o be affected by respiratory di~orders. All this symptomatology can be accompani~d by diseases caused by chance infections conventionally observed in pigs.
The inventors have succeeded Ln isolating and in identifying a new virus, responsible for this disease.
Thi~ virus is of Myxovirus type, according to the analy-sis carried out in the electron microscope, and has the characteristic of not being neutralised by porcine anti-influenza sera HlNl and H3N2.
A strain of this virus identified under the name Pl29-Z9~ was deposited in the Collection Nationale de Cultures de Micro-organismes held at the Institut Pasteur under No. I-1153.
Moreover, it then turns out that this virus has a certain number of features of the virus of Newcastle disease, while being different from the known strains of this virus.
The subject of the pres nt invention is a process for isolating the virus and its use for the preparation of antigens.
Such a process comprises taking samples from S organs of sick or infected animals, milling the samples, and passing the supernatant through sensitive heterologous or homologous cells such as, in particular, cells of the Vero, MDC~ (in the presence of trypsin), ST, or BHK lines or also through chick embryos.
Th~ samples are preferably taken from the lung of the infected animal or else from a pool of organs com-prising, for example, heart, spleen, liver, kidney and lymphoid tissues.
The harvests are not neutralised by porcine anti-influenza sera HlNl and H3N2. The haemagglutination testsare positive from the an~igen produced in chick embryos.
Another subject of the invention is a process for the industrial production of this virus in which the virUc is cultured in sensitive heterologous or homologous cells such as, especially, cells of the Vero, MDCK tin the presence of trypsin), ST or BHR lines or also in chick embryos.
The harvested virus can be used for the prepar-ation of antigens. To thi~ end, it is, preferably, suitably purified accordiny to conventional procedures for the Myxoviruses, for example ultra-centrifugation or chromatography. It can also be concentrated by conven-tional techniques.
According to the use envisaged, the antigen preparations according to the invention can consist of living viral particles, optionally attenuated, of inac-tivated particles, of sub-unit antigens or even of antigens obtained by genetic recombination from genes of the isola~ed virus, which are inserted in order to be expres~ed in the genome of recombining procaryote or eucaryote hosts.
Another sub~ect of the invention is the vaccines made from the abovementioned antigens, containing an 2 Q ~ 3 ~
effective vaccinating quantity of antigens in suitable carriers.
These vaccines can be provided to be administered to pigs but also to other animals which could turn out to be sensitive to this virus.
An attenuated vaccine can be prepared by pas~ing the virus through a cell culture.
The quantity of viru~ per vaccinal dose is preferably of between 103 and lOa TCID50 pex dose.
The living attenuated vaccine can be pre~ented in the liquid or lyophilised form in the presence of stabili~ers of very varied formulations, which can include sugars, proteins and buffer~. Inorganic or organic adjuvants can be added to the vaccine.
The living vaccine can be administered to anLmals to protect them from the beginning of the fattening period or before artificial insemination or covering, or during gestation, in one and preferably two injections at intervals of three or four weeks.
An inactivated vaccine csn be prepar d from viral suspensions obtained by passing through homologous (porcine) or heterologous cell systems and then inac-tivation by conventional chemical inactivating agents such as beta-propiolactone, enzymes or organic solvents or detergent~O Inactivation can also be obtained by physical action such as ultraviolet radiation or gamma or X-ray irradiations. The inactivating agent can be neutrali~ed if neces~ary.
The inactivated vaccine contains preferably at least the equivalent of 105 TCIDso per vaccinal dose, a concentration determined before inactivation.
The inactivated vaccine can be administered to animals to protect them from the beginning of the fattening period, or before artificial insemination or covering, or during gestation, in one and preferably two injections at intervals of three or four weeks. I~ is pre~erable that the vaccine contains an adjuvant of inorganic or organic origin.
20~1~3~
A recombinant vaccine can be obtained, for example, by insertion of the sequence coding for th~
desired antigen of the ~irus in ~he genome of the host.
The host can be a virus, especially for the preparation of a living vaccine.
This host can also be a bacterial system, a yeast system or a system of other eucaryote cells. In this caser the host is preferably used for the production of antigens which are then purified and conditioned in order to make the vaccine.
The vaccine can be presented in the monovalent form or combined with other viral or bacterial agents responsible for diseases in pigs.
Another subject of the invention, as an equi-valent vaccine against Mystery Disease in pig8, iS avaccine characterised in that it comprises a quantity which is therapeutically effective in pigR, of a virulent strain of Newcastle virus, suitably inactivated, ox of vaccinating antigens of such a strain, with, preferably, a conventional ad~uvant.
Other advantages and characteristics of the invention will become apparent on reading the following description, given by way of non-limiting example.
Example 1 : Isol tion of the viral Rtrain Sampleq of organs were taken from a sow origin-ating from ~ pig farm in Germany and identified under No.
29~.
The samples came from the heart, spleen, liver, kidney, lymphoid tissue and lung, and were used without freezing.
a) Method:
Milled preparations of organs: each sample is individually milled in ~EM medium + antibiotics.
Dilution: organ weight per volume of approxi-mately 1 to 10.
Clarifying centrifugation for recovery of thesupernatant liquid (SL).
Filtration at 0.22 ~.
20~1638 b) Tests on cell cultures.
First passages From:
1) SL of milled lung preparation 2) SL of pool of organs: heart, spleen, liver, kidney and lymphoid tissue cell8 u~ed: Vero, IRO.3, ST, MDCK (in the presence of trypsin), BHK, and primary pig kidney cell lines . inoculation in cultures in established layer (25 cm2 Falcons) . ob~ervation of the cells and freezing at -70C.
Second passages From the first unfrozen passages.
On the ~ame type of cells as the first passages;
the cells are used in simultaneous inoculation.
Observation and freezing.
c) Tests on chick embryos . Inoculation in SPF ch}ck embryo~ of 9 days . Inoculum: SL of milled lung prepsration from the sow 294. 0.1 ml of undiluted inoculum or inoculum diluted to 1/10 in PBS + antibiotics is inoculated intra-allantoidally.
After incubating for 3 day~, the eggs are emptied and the allantoic liquid is harve~ted (individual harvesting from each egg).
. A haemagglutination test with respect to chicken red cells (concentration 20-106 red cell~ per ml) i8 carried out on all the harvestéd samples.
. The successive passages are carried out under the condition~ described above.
d) Results 1) Results of cell cultures In two cell types, change~ in the cell layer could be observed:
- in Vero cells: the cells of the inoculated Falcons show, during the second passage, an appearance of the cell lawn different from the cell~ of the standard - ~
.. - . , .
, - . . . .
.. .
2 ~ g Falcon. This abnormal appearance is observed at +7 days after inoculation.
- in MDCK cells in the presence of t~ypsin (concentration 20 ~g/ml): the cells inoculated by the pool of organs show an appearance different from the standard cells at +6 days of culture.
2) Results in egg~
First passage: no haemagglutination could be shown.
Second passags: 1 egg showq haemagglutination.
The haemagglutination titre HA is 640.
Third passage:
. HA titre which can reach 1024 presence of HA not neutxalised by porcine anti-influenza sera HlN1 and H3N2.
Fourth passage: haemagglutination confirmed withtitres which can reach 2048.
Additionally, a significant mortality in the eggs could be observed in the second, third and fourth pas-sages, it being poRsible for the mortality to exceed 50%of the inoculated eggs.
A positive haemagglutination test was carried out with respect to guinea pig xed cells. Using the haemag-glutina~ing antigen of the 3rd passage, cells of ST and Vero lines and E~rimary pig kidney cells SPF were inocu-lated, a cytopal:hogenic effect was observed in these 3 cell~, a cytopal:hogenic effect more marked than in the cells inoculated directly with lung SL. The haemagglutin-ating effect had disappeared after passages through cells.
From the viral suspension of ~he 3rd passage through eggs, 2 pigs with a weight of 30 kg were inoculated intravenously with a volume of 1 cc. These 2 pigs showed signs of exhaustion, anorexia and hyper-thermia of the order of 40C for at most 7 days. Thesesymptoms are those commonly observed in sows in pig farms attacked by P.R.R.S. or M. D .
Preparation of antigens of and of vaccines for the virus of Mystery Disease, antigens and vaccines obtained for the prevention of this disease.
The present invention relateæ to the preparation of antigens of and of vaccines for the virus of Mystery Disea~e, as well as to the antigens and to the vaccines obtained.
The disease called Mystery Disea~e (M.D.) or also Porcine Reproductive Re~piratory Syndrome (P.R.R.S.) began to acquire a identity of its own in pigs, in 1986 in the United StateR and in 1990 in Europe. This disease manifests itself essentially in pig~ by ~ign~ of exhaus-tion, anorexia and hyperthermia of the order of 40C, which a~e conventionally observed in ~OW8 in pig farms affected by the disease. The~e signs are accompanied or followed by reproductive disorders (premature or late farrowing and bi.rth of stillborn, mummified or sickly piglets, and return of the 50W~ to heat). A respiratory syndrome can be observed in piglet with interstitial pneumonia lesions. Older pigs can al~o be affected by respiratory di~orders. All this symptomatology can be accompani~d by diseases caused by chance infections conventionally observed in pigs.
The inventors have succeeded Ln isolating and in identifying a new virus, responsible for this disease.
Thi~ virus is of Myxovirus type, according to the analy-sis carried out in the electron microscope, and has the characteristic of not being neutralised by porcine anti-influenza sera HlNl and H3N2.
A strain of this virus identified under the name Pl29-Z9~ was deposited in the Collection Nationale de Cultures de Micro-organismes held at the Institut Pasteur under No. I-1153.
Moreover, it then turns out that this virus has a certain number of features of the virus of Newcastle disease, while being different from the known strains of this virus.
The subject of the pres nt invention is a process for isolating the virus and its use for the preparation of antigens.
Such a process comprises taking samples from S organs of sick or infected animals, milling the samples, and passing the supernatant through sensitive heterologous or homologous cells such as, in particular, cells of the Vero, MDC~ (in the presence of trypsin), ST, or BHK lines or also through chick embryos.
Th~ samples are preferably taken from the lung of the infected animal or else from a pool of organs com-prising, for example, heart, spleen, liver, kidney and lymphoid tissues.
The harvests are not neutralised by porcine anti-influenza sera HlNl and H3N2. The haemagglutination testsare positive from the an~igen produced in chick embryos.
Another subject of the invention is a process for the industrial production of this virus in which the virUc is cultured in sensitive heterologous or homologous cells such as, especially, cells of the Vero, MDCK tin the presence of trypsin), ST or BHR lines or also in chick embryos.
The harvested virus can be used for the prepar-ation of antigens. To thi~ end, it is, preferably, suitably purified accordiny to conventional procedures for the Myxoviruses, for example ultra-centrifugation or chromatography. It can also be concentrated by conven-tional techniques.
According to the use envisaged, the antigen preparations according to the invention can consist of living viral particles, optionally attenuated, of inac-tivated particles, of sub-unit antigens or even of antigens obtained by genetic recombination from genes of the isola~ed virus, which are inserted in order to be expres~ed in the genome of recombining procaryote or eucaryote hosts.
Another sub~ect of the invention is the vaccines made from the abovementioned antigens, containing an 2 Q ~ 3 ~
effective vaccinating quantity of antigens in suitable carriers.
These vaccines can be provided to be administered to pigs but also to other animals which could turn out to be sensitive to this virus.
An attenuated vaccine can be prepared by pas~ing the virus through a cell culture.
The quantity of viru~ per vaccinal dose is preferably of between 103 and lOa TCID50 pex dose.
The living attenuated vaccine can be pre~ented in the liquid or lyophilised form in the presence of stabili~ers of very varied formulations, which can include sugars, proteins and buffer~. Inorganic or organic adjuvants can be added to the vaccine.
The living vaccine can be administered to anLmals to protect them from the beginning of the fattening period or before artificial insemination or covering, or during gestation, in one and preferably two injections at intervals of three or four weeks.
An inactivated vaccine csn be prepar d from viral suspensions obtained by passing through homologous (porcine) or heterologous cell systems and then inac-tivation by conventional chemical inactivating agents such as beta-propiolactone, enzymes or organic solvents or detergent~O Inactivation can also be obtained by physical action such as ultraviolet radiation or gamma or X-ray irradiations. The inactivating agent can be neutrali~ed if neces~ary.
The inactivated vaccine contains preferably at least the equivalent of 105 TCIDso per vaccinal dose, a concentration determined before inactivation.
The inactivated vaccine can be administered to animals to protect them from the beginning of the fattening period, or before artificial insemination or covering, or during gestation, in one and preferably two injections at intervals of three or four weeks. I~ is pre~erable that the vaccine contains an adjuvant of inorganic or organic origin.
20~1~3~
A recombinant vaccine can be obtained, for example, by insertion of the sequence coding for th~
desired antigen of the ~irus in ~he genome of the host.
The host can be a virus, especially for the preparation of a living vaccine.
This host can also be a bacterial system, a yeast system or a system of other eucaryote cells. In this caser the host is preferably used for the production of antigens which are then purified and conditioned in order to make the vaccine.
The vaccine can be presented in the monovalent form or combined with other viral or bacterial agents responsible for diseases in pigs.
Another subject of the invention, as an equi-valent vaccine against Mystery Disease in pig8, iS avaccine characterised in that it comprises a quantity which is therapeutically effective in pigR, of a virulent strain of Newcastle virus, suitably inactivated, ox of vaccinating antigens of such a strain, with, preferably, a conventional ad~uvant.
Other advantages and characteristics of the invention will become apparent on reading the following description, given by way of non-limiting example.
Example 1 : Isol tion of the viral Rtrain Sampleq of organs were taken from a sow origin-ating from ~ pig farm in Germany and identified under No.
29~.
The samples came from the heart, spleen, liver, kidney, lymphoid tissue and lung, and were used without freezing.
a) Method:
Milled preparations of organs: each sample is individually milled in ~EM medium + antibiotics.
Dilution: organ weight per volume of approxi-mately 1 to 10.
Clarifying centrifugation for recovery of thesupernatant liquid (SL).
Filtration at 0.22 ~.
20~1638 b) Tests on cell cultures.
First passages From:
1) SL of milled lung preparation 2) SL of pool of organs: heart, spleen, liver, kidney and lymphoid tissue cell8 u~ed: Vero, IRO.3, ST, MDCK (in the presence of trypsin), BHK, and primary pig kidney cell lines . inoculation in cultures in established layer (25 cm2 Falcons) . ob~ervation of the cells and freezing at -70C.
Second passages From the first unfrozen passages.
On the ~ame type of cells as the first passages;
the cells are used in simultaneous inoculation.
Observation and freezing.
c) Tests on chick embryos . Inoculation in SPF ch}ck embryo~ of 9 days . Inoculum: SL of milled lung prepsration from the sow 294. 0.1 ml of undiluted inoculum or inoculum diluted to 1/10 in PBS + antibiotics is inoculated intra-allantoidally.
After incubating for 3 day~, the eggs are emptied and the allantoic liquid is harve~ted (individual harvesting from each egg).
. A haemagglutination test with respect to chicken red cells (concentration 20-106 red cell~ per ml) i8 carried out on all the harvestéd samples.
. The successive passages are carried out under the condition~ described above.
d) Results 1) Results of cell cultures In two cell types, change~ in the cell layer could be observed:
- in Vero cells: the cells of the inoculated Falcons show, during the second passage, an appearance of the cell lawn different from the cell~ of the standard - ~
.. - . , .
, - . . . .
.. .
2 ~ g Falcon. This abnormal appearance is observed at +7 days after inoculation.
- in MDCK cells in the presence of t~ypsin (concentration 20 ~g/ml): the cells inoculated by the pool of organs show an appearance different from the standard cells at +6 days of culture.
2) Results in egg~
First passage: no haemagglutination could be shown.
Second passags: 1 egg showq haemagglutination.
The haemagglutination titre HA is 640.
Third passage:
. HA titre which can reach 1024 presence of HA not neutxalised by porcine anti-influenza sera HlN1 and H3N2.
Fourth passage: haemagglutination confirmed withtitres which can reach 2048.
Additionally, a significant mortality in the eggs could be observed in the second, third and fourth pas-sages, it being poRsible for the mortality to exceed 50%of the inoculated eggs.
A positive haemagglutination test was carried out with respect to guinea pig xed cells. Using the haemag-glutina~ing antigen of the 3rd passage, cells of ST and Vero lines and E~rimary pig kidney cells SPF were inocu-lated, a cytopal:hogenic effect was observed in these 3 cell~, a cytopal:hogenic effect more marked than in the cells inoculated directly with lung SL. The haemagglutin-ating effect had disappeared after passages through cells.
From the viral suspension of ~he 3rd passage through eggs, 2 pigs with a weight of 30 kg were inoculated intravenously with a volume of 1 cc. These 2 pigs showed signs of exhaustion, anorexia and hyper-thermia of the order of 40C for at most 7 days. Thesesymptoms are those commonly observed in sows in pig farms attacked by P.R.R.S. or M. D .
3 ~
The isolated agent thus has the following charac-teristics:
- it causes haemagglutination of g~inea pig red cells from antigen multiplied in chick embryos, - it is not inhibited by porcine anti-influenza sera, - it produces a cytopathogenic effect in various cells, - its properties, made visible in the electron iO microsrope, ally it to the Myxovirus group.
These properties are also those of the strain P129~294 deposited in the CNCM.
Example_2 : Preparation of a livinq vaccine The living vaccine i~ prep~red from the strain P129-294. The strain i~ multiplied by passages through Vero cells. The harve~ing is carried out afterward~. The harvested and processed supernatan~ is lyophilised with a stabiliser SPGA.
The vaccine is packaged in doses of 10, 50 and 100.
Example 3 : Prepara~ion of an inactivated vaccine The virus of the strain P129-294 is multiplied in ST cells. Following the fifth passage, the supernatant containing the virus is harvested, concentrated and purified. It is inactivated by beta-propiolactone and then aluminium hydroxide is added thereto.
The vaccine is packaged in doses of 10 and 50.
The isolated agent thus has the following charac-teristics:
- it causes haemagglutination of g~inea pig red cells from antigen multiplied in chick embryos, - it is not inhibited by porcine anti-influenza sera, - it produces a cytopathogenic effect in various cells, - its properties, made visible in the electron iO microsrope, ally it to the Myxovirus group.
These properties are also those of the strain P129~294 deposited in the CNCM.
Example_2 : Preparation of a livinq vaccine The living vaccine i~ prep~red from the strain P129-294. The strain i~ multiplied by passages through Vero cells. The harve~ing is carried out afterward~. The harvested and processed supernatan~ is lyophilised with a stabiliser SPGA.
The vaccine is packaged in doses of 10, 50 and 100.
Example 3 : Prepara~ion of an inactivated vaccine The virus of the strain P129-294 is multiplied in ST cells. Following the fifth passage, the supernatant containing the virus is harvested, concentrated and purified. It is inactivated by beta-propiolactone and then aluminium hydroxide is added thereto.
The vaccine is packaged in doses of 10 and 50.
Claims (13)
1. Process for isolating the virus of Mystery Disease comprising taking samples from organs of sick or infected pigs, milling the samples and passing the milling supernatant through sensitive heterologous or homologous cells and then harvesting the supernatant.
2. Process according to Claim 1, characterised in that the passages are carried out through cells of the Vero, MDCK, ST, or BHK lines or through chick embryos.
3. Process for the industrial production of the virus of Mystery Disease in which the virus is cultured in sensitive heterologous or homologous cells.
4. Process according to Claim 3, characterised in that the virus is cultured in cells of the Vero, MDCK, ST or BHK lines and the last supernatant is harvested.
5. Process according to Claim 3, characterised in that the virus is cultured in chick embryos and the allantoic liquid is recovered.
6. Preparation of purified viral antigen of Mystery Disease comprising living viral particles, optionally attenuated, or inactivated particles or subunit antigens or antigens expressed by genetic recombination from genes of the isolated virus.
7. Preparation of antigen of the viral strain P129-294 deposited in the CNCM under No. I-1153.
8. Vaccine against Mystery Disease, characterised in that it contains, in a suitable carrier, a vaccinating quantity of an antigen according to one of Claims 5 and 6.
9. Attenuated vaccine according to Claim 8, charac-terised in that it is prepared by passages of the virus through a cell culture.
10. Attenuated vaccine according to Claim 9, charac-terised in that the quantity of virus per vaccinal dose is of between 103 and 108 TCID50.
11. Inactivated vaccine according to Claim 8, charac-terised in that the culture supernatant is inactivated by a chemical or physical inactivating agent.
12. Inactivated vaccine according to Claim 11, characterised in that it contains at least the equivalent of 105 TCID50 per vaccinal dose.
13. Vaccine against Mystery Disease, characterised in that it contains a therapeutically effective dose of a virulent strain of Newcastle virus or of vaccinating antigens of such a strain.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR9113338 | 1991-10-29 | ||
| FR9113338A FR2682966B1 (en) | 1991-10-29 | 1991-10-29 | PREPARATION OF MYSTERY DISEASE VIRUS ANTIGENS AND VACCINES, ANTIGENS AND VACCINES OBTAINED FOR THE PREVENTION OF THIS DISEASE. |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2081638A1 true CA2081638A1 (en) | 1993-04-30 |
Family
ID=9418419
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002081638A Abandoned CA2081638A1 (en) | 1991-10-29 | 1992-10-28 | Preparation of antigens of and of vaccines for the virus of mystery disease, antigens and vaccines obtained for the prevention of this disease |
Country Status (4)
| Country | Link |
|---|---|
| EP (1) | EP0541418A1 (en) |
| CA (1) | CA2081638A1 (en) |
| FR (1) | FR2682966B1 (en) |
| MX (1) | MX9206216A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5510258A (en) * | 1993-02-08 | 1996-04-23 | Bayer Corporation | Porcine reproductive and respiratory syndrome virus antigen and processes for the preparation and use of said antigen in vaccines and diagnostics |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2686097B1 (en) * | 1992-01-14 | 1994-12-30 | Rhone Merieux | PREPARATION OF ANTIGENS AND MYSTERY DISEASE VIRUS VACCINES, ANTIGENS AND VACCINES OBTAINED FOR THE PREVENTION OF THIS DISEASE. |
| GB2289279B (en) * | 1994-05-13 | 1998-09-16 | Iberica Cyanamid | Diagnostic kits and vaccines containing recombinant PRRSV proteins |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5052224A (en) * | 1973-09-10 | 1975-05-09 | ||
| DD145705A1 (en) * | 1979-08-31 | 1981-01-07 | Peter Solisch | METHOD OF GENERATING ORGAN SPECIFIC RETROVIRUS IMMUNE PREVENTION AGENTS FOR PROPHYLAXIS, THERAPY AND DIAGNOSTICS |
| JPS5879929A (en) * | 1981-11-05 | 1983-05-13 | Biseibutsu Kagaku Kenkyusho:Kk | Mixed vaccine of japanese encephalitis and swine parvo |
| JPS5939831A (en) * | 1982-08-27 | 1984-03-05 | Biseibutsu Kagaku Kenkyusho:Kk | Inactivated vaccine of influenzal virus for pig |
| IL70704A0 (en) * | 1983-01-19 | 1984-04-30 | Amgen | Methods and materials for development of parvovirus vaccines |
| JPS62198626A (en) * | 1986-02-26 | 1987-09-02 | Biseibutsu Kagaku Kenkyusho:Kk | Vaccine for preventing infectious disease of virus of hemagglutinating encephalomyelitis |
| GB2220941B (en) * | 1988-06-24 | 1991-09-11 | Nat Res Dev | Fowlpox virus non-essential regions |
| EP0351908B1 (en) * | 1988-07-18 | 1994-10-05 | Duphar International Research B.V | Live newcastle disease virus vacccines |
-
1991
- 1991-10-29 FR FR9113338A patent/FR2682966B1/en not_active Expired - Fee Related
-
1992
- 1992-10-22 EP EP92402883A patent/EP0541418A1/en not_active Withdrawn
- 1992-10-28 MX MX9206216A patent/MX9206216A/en unknown
- 1992-10-28 CA CA002081638A patent/CA2081638A1/en not_active Abandoned
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5510258A (en) * | 1993-02-08 | 1996-04-23 | Bayer Corporation | Porcine reproductive and respiratory syndrome virus antigen and processes for the preparation and use of said antigen in vaccines and diagnostics |
| US5587164A (en) * | 1993-02-08 | 1996-12-24 | Bayer Corporation | Porcine reproductive and respiratory syndrome virus antigen and processes for the preparation and use of said antigen in vaccines and diagnostics |
Also Published As
| Publication number | Publication date |
|---|---|
| FR2682966A1 (en) | 1993-04-30 |
| EP0541418A1 (en) | 1993-05-12 |
| FR2682966B1 (en) | 1994-12-02 |
| MX9206216A (en) | 1993-12-01 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| FZDE | Dead |