EP0098867A1 - Vaccin contre le virus de la maladie bleue et son procede de production - Google Patents

Vaccin contre le virus de la maladie bleue et son procede de production

Info

Publication number
EP0098867A1
EP0098867A1 EP83900518A EP83900518A EP0098867A1 EP 0098867 A1 EP0098867 A1 EP 0098867A1 EP 83900518 A EP83900518 A EP 83900518A EP 83900518 A EP83900518 A EP 83900518A EP 0098867 A1 EP0098867 A1 EP 0098867A1
Authority
EP
European Patent Office
Prior art keywords
virus
vaccine
sheep
btv
bluetongue
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
EP83900518A
Other languages
German (de)
English (en)
Inventor
Stewart Texas A & M University McCONNELL
Georgia A. Texas A & M University CUMMINGS
Charles W. Jr. Texas A & M University LIVINGSTON
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Texas A&M University System
Original Assignee
Texas A&M University System
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Texas A&M University System filed Critical Texas A&M University System
Publication of EP0098867A1 publication Critical patent/EP0098867A1/fr
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • A61K39/15—Reoviridae, e.g. calf diarrhea virus
    • 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
    • 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
    • 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
    • A61K2039/70—Multivalent vaccine
    • 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
    • C12N2720/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsRNA viruses
    • C12N2720/00011—Details
    • C12N2720/12011—Reoviridae
    • C12N2720/12111—Orbivirus, e.g. bluetongue virus
    • C12N2720/12134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • This invention relates to a vaccine for immuniz ⁇ ing ruminants against International serotypes of bluetongu virus (BTV) and a method pf m'a ing said vaccine, and a me ⁇ thod of immunizing ruminants with said vaccine.
  • BTV bluetongu virus
  • Blue ⁇ tongue lesions in affected animals resemble infectious bo- vine virus diarrhea, vesicular stomatitis, malignant catar rhal fever, mycotic stomatitis, rinderpest, photosensitiza- tion, and foot and mouth disease.
  • Bluetongue virus has been incriminated as a cause of hydranencephaly in cat ⁇ tle and of infertility, abortion, and birth of defective young in cattle and sheep. Twenty serotypes are reported in the literature as causing problems ranging from inap- parent infection to acute fulminating infection. Chronic, persistent virus shedding cattle have also been recognized.
  • the virus infects the endothelial cells of the blood vas ⁇ cular system.and the periendothelial cells and the peri- cytes of capillaries, pericapillary arteriodes, and venules Stair, et al., 5 Pathologia Vet 164 (1968).
  • the virus is associated with the reticulo-endothelial system and may occur within the spermatozoa of infected bulls. Luede et al, 3 Proc. 20th World Vet Congress 2039 (1976) .
  • BTV-infected sheep wool growt may be impaired by the development of wool breaks which produce a defective or low yielding fleece.
  • the marked debility following BTV infections may result in a lowering .of resistence to secondary bacterial or chlamydial infec ⁇ tions and other predatory factors.
  • the reproductive effi ⁇ ciency of infected animals is also adversely affected. Abortions and defective offspring are observed in infected animals, and some animals may be barren for one or more breeding seasons.
  • the most significant damage inflicted by bluetongue infections is economic loss resul ' ting from embargoes and stringent testing requirements imposed on • producers who export cattle, cattle semen, and sheep from bluetongue endemic areas.
  • the etiologic agent of bluetongue belongs to the family Reoviridae, genus Orbivirus. This name designation was changed from "reo-like" virus to Orbivirus by the In ⁇ ternational Committee on viral nomenclature in 1971. The viral etiology of bluetonge was established by Theiler in 1906. Erasmus, 51 Aust Vet J 165 (1975).
  • the object of the present invention is to provide a vaccine having the aforesaid advantages and in particular to provide a effective vaccine for protecting cattle, sheep, and goats against BTV.
  • bluetongue viruses can be adapted to and propagated in cell cultures derived from tissues that do not originate from cattle, sheep, goats, or wild un- gulates of the family Bovidae and the virulence so modi ⁇ fied and reduced that no symptoms of the bluetongue virus disease are observed upon parenteral inoculation.
  • the present invention provides for a vaccine for immunizing ruminants against International serotypes of bluetongue virus IT 10, IT 11, IT 13, and
  • IT 17 comprising at least about 10 tissue culture infec ⁇ tious doses of one or more of bluetongue virus serotype IT 10 , IT 11, IT 13, or IT 17 per ml, wherein the virus serotype is attenuated so that upon parenteral administra- tion to said ruminants the vaccine is capable of stimulat ⁇ ing production of a host defense mechanism comparable to that produced by natural infections without producing the pathological responses of disease due to the bluetongue virus serotypes.
  • the present invention also provides for a method of preparing a vaccine for parenteral innoculation of ru ⁇ minants against a bluetongue virus, which comprises culti ⁇ vating and attenuating the bluetongue virus by 30 or more serial passages through Vero cell cultures in a nutrient fluid.
  • the present invention also provides for a method of immunizing cattle, sheep, or goats against bluetongue virus which comprises parenterally administering to said cattle, sheep, or goats at least 2 ml of a vaccine compris- ing at least one or more of bluetongue International virus serotype IT 10, IT 11, IT 13, or IT 17, wherein said virus is attenuated by 30 or more serial passages through Vero cell cultures in a nutrient fluid at an incubation tempera ⁇ ture of from about 30°C to about 36°C, each passage lasting from about 1 day to about 10 days.
  • the present invention also provides for a method of immunizing cattle, sheep or goats against bluetongue virus which comprises parenterally administering to said
  • a vaccine compri- sing at least about 10 tissue culture infectious doses of bluetongue virus International serotype 13 per ml, wherein said virus is attenuated by at least 3 blind pas- sages in pathogen-free embryonating hen eggs prior to 'said 30 or more serial passages through Vero cell cultures.
  • the vaccine is safe in that it will not cause overt disease in ruminants that receive it by the paren ⁇ teral route. Further safety is provided in that the viruses will not pass from vaccinated ruminants to other ruminants in contact with those vaccinated, thereby re ⁇ ducing the possibility of reversion to virulence by animal passage in these species. This constitutes a significant advance in the control of BTV.
  • the basic method for producing the present vaccine comprises cultivating and attentuating the blue- tongue virus by at least thirty serial passages through "Vero" cell cultures in a nutrient fluid.
  • the Vero cell line is derived from the African Green Monkey as described in more detail below.
  • the serial passages are conducted at incubation temperatures of from about 30 C to about 36 C, and each passage lasts from about 1 day to about 10 days.
  • the vaccine may be produced in liquid form suit- able for parenteral inoculation into ruminants.
  • Such liquid vaccine contains at least about 10 4 tissue culture infectious doses of each BTV serotype being protected against per ml.
  • a suitable dosage for mmunizing cattle, sheep, or goats is about 2 ml of such liquid vaccine.
  • Live virulent bluetongue viruses can be ob ⁇ tained from cattle and sheep infected with the viruses according to the methods of isolation and identification described in the literature. For example, see Livingston and Moore, 23 AJVR (1962); Goldsmit and Barzilai, 22 efuah Vet 279 (1965); Luedke et al, 30 AJVR 511 (1969).
  • the serotypes of bluetongue virus for which this invention is most useful are International serotypes 10, 11, 13, and 17 which occur in the United States. It is understood that these specific viruses are by way of ill ⁇ stratio only and that the present invention can be adapted to other BTV serotypes. These four viruses are serological- ly related as determined by the conventional agar gel precipitin test (e.g., each virus contains the same group antigen) but do not cross-protect as determined by sheep inoculation tests or by the conventional serum neutrali ⁇ zation tests. These four serotypes are included in a listing of several bluetongue virus serotypes presented by DeV Amsterdam, 3 Intervirology 47 (1974).
  • the BTV IT, 10, IT 11, IT 13 and IT 17 serotypes have been deposited in The American Type Culture Collection (ATCC) Rockville, Maryland, U.S.A., and catologued in the Catalogue of Strains as VR-187, VR-872, VR-873, and VR-875, respect ⁇ ively.
  • BTV IT 13 may be first adapted to em- bryonating hen eggs with three blind passages
  • a susceptible permanent cell line such as "Vero”
  • Vero a continuous cell line derived from the-African Green Monkey (Cercopithecus aethiops) subculture levels 128 through 150, at low temperatures, preferably 30-36 C, and most preferably 32 C.
  • the virus is puri ⁇ fied by filtration through a membrane with an average pore diameter of 220nm, by chloroform treatment prior to use * in master pools, and by standard terminal dilution techniques.
  • whole blood collected from infected sheep such as that supplied by the USDA, Arthropodborne Viral Diseases Laboratory, Denver, Colorado, may be used.
  • the lowest available sheep passage levels of BTV strains are utilized.
  • the serotypes BTV IT 10, 11, and 17, are utilized.
  • the passage time intervals should be such as to allow for maximum virus replication between passages, typically
  • the optimum passage time can be determined by ⁇ ta ⁇ 3ard cell culture procedures, for example, by cyto ⁇ pathic observations wherein the virus is allowed to 5 replicate during each passage to the point where gross cytopathic effect (CPE) can be observed and 75-100% of t cell monolayer is destroyed.
  • CPE gross cytopathic effect
  • a meth 0 for attenuating virulent strains of BTV, whi when used alone or in optimal proportions and parenteral inoculated, preferably intramuscularly, into sheep, goat or cattle, will immunize against homologous BTV.
  • the method comprises introducing an inoculum of virulent BTV 5 into a nutrient cell culture medium at a temperature of from about 30'C to about 36'C, preferably 32 * C, for a period of from.one to ten days, and thereafter serially passing the viruses through additional such Vero cell cultures for a total of at least 30 consecutive passages 0
  • the viral preparations produced by this invention m be diluted to adjust potency and may have added to them stabilizers, such -as lactose, dextrose, or other non-tox substances.
  • stabilizers such -as lactose, dextrose, or other non-tox substances.
  • the viral preparations may also be desiccat -** * e.g., by freeze-dryin , for storage purposes or for sub ⁇ sequent formulation into liquid vaccines.
  • Stabilizers useful in freeze drying of viruses are described in Rightsel et al, 3 Cryobiology 423 (1967) and Greiff et a Advances in Freeze Drying, 103-122 (1966).
  • a 3.0 cc sample of blood was centrifuged at 1000 rpm for 10 minutes. The supernatant fluid was discarded, and the packed red blood cells were resuspended in 1.0 cc of a balanced salt solution fphos- . phate buffered saline ( PBS)J, The cells were disrupted in - a Ten Broeck tissue grinder, and the disrupted cells were ' brought to a final volume of 3.0 cc with PBS.
  • a quantity of 0.5 cc lysed red blood cell suspension was added to a decanted monolayer of Vero cells and allowed to adsorb for 1 hour at 30-34 * C in standard tubes (16 x 125 mm) prepared as described below.
  • the Vero cell line used is the cell line initiated by Yasamura and Kawakita at the Chiba * University in Chiba, Japan. 21 Nippon Rinsho 1201 (1963). This cell line was brought from the Chiba University to the Laboratory of Tropical Virology, National Institute of Allergy and Infectious Diseases, National Institutes of Health,' in the 93rd passage. Subsequently, the cell line was deposited with the American Type Culture Collection, at passage level 113 and distributed by the ATCC as CCL81 Vero passage No. 122.
  • EMEM Eagle's Minimum Essential Medium in Earle's balanced salt solution
  • NEAA non-essential amino acids
  • fetal bovine serum 10% fetal bovine serum 10%
  • sodium pyruvate 1% sodium pyruvate 1%
  • Kanamycin 200 mcg/ml was seeded with 1 cc of suspended Vero cells (ca. 250,000 cells
  • the growth medium was poured off, the cell monolayer washed with prewar ed Hanks balanced salt solution (HBSS), and 0.5 cc of virus-infected disrupted red blood cells added. Adsorption was allowed to proceed at 30 * -34'C for one hour. Two cc of a maintenance medium consisting of Basal Medium Eagle (BME) in KBSS, supplemented with 3% horse serum, 2% tryptose phosphate broth, 100 units of penicillin, and 100 meg streptomycin was added. The tubes were then incubated at 30-34 * C in a roller drum incubator. About 3-5 such tubes were utilized per viral passage. In each passage the CPE was allowed to develop until 75% or more of the cell monolayer was destroyed, and the samples were then rapidly frozen at -85 * C.
  • BME Basal Medium Eagle
  • the contents of the tubes were harvested and sub ⁇ jected to identical serial passages for 30 to 50 addi ⁇ tional passages.
  • the 29th, 39th, and 49th passages were subjected to an additional passage for the purpose of in ⁇ creasing volume.
  • the virus suspension Prior to passage for increasing volume, • the virus suspension was treated with chloroform according to the procedure of Feldman and Wang, 106 Proc. Soc. Exptl. Biol. Med. 736.(1961), and inoculated onto con- fluent monolayers of Vero cells grown in an antibiotic free medium.
  • the maintenance medium in this case was BME(H) supplemented with 10% lamb serum and 10% tryp ⁇ tose broth.
  • the 30th, 40th, and 50th passage pool were harvested, identified, and titrated by conventional methods.
  • the pool thus prepared constituted a bulk vaccine which could he diluted according to the titer or could have added thereto stabilizers or other nontoxic sub- stances.
  • it is combinable with the other serotypes of BTV prepared in a similar manner and desiccated, or it may be prepared in liquid form as a ultivalent vaccine.
  • any nontoxic nutrient fluid tissue culture medium could have been utilized-.
  • any nontoxic nutrient fluid tissue culture medium could also have been used.
  • Example IA The work of Example IA was repeated, substituting a sample of * live virulent bluetongue virus BTV BT262, an IT 7 serotype, (62-45S, Sheep No. 2790, day after infection 7, whole blood in OPG) for BTV Texas Station Strain. This sample, BTV BT262, was processed in the identical manner specified for the BTV Texas Station Strain in " Example IA.
  • Example IA The work of Example IA was repeated, substituting a sample of live virulent bluetongue virus BTV BT8, an IT 10 serotype, (Sheep No. 2617, day after infection 7, whole blood in OPG) for BTV Texas Station Strain. This sample, BTV BT8 (2617), was processed in the identical manner specified for BTV Texas Station Strain in Example IA.
  • Example IA The work of Example IA was repeated, substituting a sample of virulent bluetongue virus BTV 6741-B, an IT 13
  • BTV Texas Station Strain BTV 6741-B
  • This sample was processed in the identical manner specified for BTV Texas Station Strain in Example IA except that, just prior to inoculation onto Vero cell 0 cultures, this sample was adapted to the chicken embryo by inoculating intravenously into twelve-day-old embryos 0.1 ml of a disrupted suspension of washed sheep red cells. These embryos were collected at selected " intervals after the initial inoculation, and heart tissues from the 5 ' . collected embryos were serially passed three times as 20% suspension in chicken embryos using a similar inoculum size, each such passage lasting from three to seven days. After this adaptation to the chicken embryo, this sample was adapted to Vero cells in the same manner described in o Example IA.
  • the antibody titer of the 5 inoculated yearling sheep was negative by the agar gel precipitin (AGP) test, and less than 1:5 by the serum neutralization (SN) test.
  • AGP agar gel precipitin
  • SN serum neutralization
  • BTV IT 11 Live virulent BTV, cultured and isolated by standard tissue culture techniques and designated as BTV IT 11 were serially passed 10 consecutive times (subsequent to plaque- purification) in continuous Vero cell onolayers at 3-7 day intervals at about 32"C. The 10th passage was tested
  • BTV IT 10 Live virulent BTV, cultured and isolated by standard tissue culture techniques and designated as BTV IT 10. The same serial passages were tested in sheep in the same manner and with the same results as tor BTV ⁇ T 11 in Example IIIA.
  • BTV IT 13 Live virulent BTV, cultured, and isolated by standard tissue culture techniques and designated as BTV IT 13 was serially passed in the same manner as described in Example IIIA for BTV IT 11. The same serial passages were tested in sheep in the same manner and with the same results as for BTV IT 11 in Example IIIA.
  • the 20th passage virus material from Example IIIA was serially passed in tubes containing a confluent monolayer of Vero cells at 30-34 * C for a variable number of passages, At the 29th and 39th passage, the viral progeny were further expanded to make vaccine pools of the 30th and 40th passage level materials. Each pool was produced in antibiotic free medium, titrated to contain about 10 4 - *0 TCIDso/ml (determined by CPE) and frozen at - ' 85'C. Prior to inoculation of Vero cell monolayers, the virus was treated with chloroform to inactivate contaminants and enhance viral infectivity.
  • the 30th and 40th passage pools were used as vaccines by the procedure described in Example II. Briefly, five susceptible yearling sheep were vaccinated with 2.0 ml of the 30th passage material administered parenterally. Five other yearling sheep were.maintained as unvaccinated controls. All 10 sheep are previously determined to be sero-negative to BTV. Each sheep was monitored daily for pyrexia and for evidence of clinical disease. After 28 days, all 10 sheep are challenged parenterally with virulent BTV by administering a total dose of 2.0 ml containing approximately 500-1000 SID/ml. The sheep were observed for 4 weeks for evidence of clinical disease.
  • the 20th passage virus material from example IIIB was serially passed in the same manner as in Example IVA to produce 30th and 40th passage pools of BTV IT 10. These 30th and 40th passage pools were tested in the same manne and with the same results as for BTV IT 11 as in Example IVA.
  • Example IVA The 20th passage virus material from example IIIC wa serially passed in the same manner as in Example IVA to 0 produce 30th and 40th passage .pools of BTV IT 10. These 30th and 40th passage" pools were tested in the same manne and with the same results as for BTV IT 11 as in Example IVA.
  • the 20th passage virus material from example II was serially passed in the same manner as in Example IVA to produce 30th and 40th passage pools of BTV IT 10. These o 30th and 40th passage pools were tested in the same manne and with the same results as for BTV IT 11 as in Example IVA.
  • a virus material containing a combination of two serotypes, IT 10 and IT 11, IT 11 and IT 17, and IT 10 and IT 17 was prepared according to the procedure detailed in Example IA-IC.
  • a 20-ml amount of this material containing each of the listed combinations of serotypes obtained at 0 the fiftieth passage level was dispensed into standard vaccine vials and frozen at -90 * C to -80 * C for inoculation purposes.
  • the frozen vials were thawed rapidly, and each of six sheep (2 per dual vaccine) were given 2.0 ml parentally with a titer of about 10 4*5 TCID 5Q /ml.
  • the 5 sheep were maintained in separate isolation facilities a monitored daily for disease. Blood was collected in OPG on the seventh and the fourteenth day after vaccination for virus isolation studies. All sheep were bled for pirevaccination antibody titer.
  • the vaccinated sheep were given a virulent virus challenge (one of each of the pair with a different single virus) dose consisting of 500-100 SID/m of the serotypes represented.
  • a virulent virus challenge one of each of the pair with a different single virus
  • the sheep were observed daily with no clinica signs of disease observed.
  • unvaccinated sheep ' exhibited typical- manifestations of BTV disease.
  • Vaccines of fiftieth passage level material were pre pared for each of the four serotypes as in Examples IA-ID. Each vaccine was undiluted and was dispensed into standar vaccine vials and frozen at -90"C to -80 * C for inoculatio purposes. The frozen vaccines were thawed rapidly, and each of .20 sheep were given ' 2.0 ml parenterally. All sheep were bled for prevaccination antibody titer and was monitored daily for evidence of pyrexia or other untoward response to the vaccine.
  • IT 10 One serotype, failed to protect against virulent virus challenge. The other three strains were protective. All serum samples assayed for antibody were negative. A retrospective titration of the vaccine residue show the vaccine titer of IT 10 to contain less
  • a vaccine was prepared as in Example IA but with the fiftieth passage level material.
  • the vaccine was diluted to contain either 10 4 TCID 50 /ml, 10 3 TCID 50 /ml, or 10 2 TCIDc Q /ml and was dispensed into standard vaccine vials and frozen at -90'C to -80 * C for inoculation purposes.
  • the frozen vials were thawed rapidly, and each of 72 sheep
  • OMP 4 were given 2.0 ml parenterally, 24 with a titer of 10
  • Example VIIA The work of Example VIIA was repeated, substituting sample of fiftieth passage level BTV IT 10 for BTV IT 11.
  • This sample (IT 10) was processed in. a similar manner except that dilutions of 10 TCID-g/ml, 10 2 TCID 5Q /ml, an
  • Example VIIA The work of Example VIIA was repeated substituting a sample of fiftieth passage level BTV IT 17 for BTV IT 11. This sample (IT 17) was processed in a similar manner except that dilutions of 10 5 TCIDso/ml, 10 4 TCID 50 /ml, and 10 3 TCID5o/ml were used. Similar results to those found in Example VIIA were obtained.
  • a virus material containing the four serotypes, BTV IT 10, IT 11, IT 13, and IT 17, is prepared according to the procedure detailed in Example IA-ID.
  • a 2000-ml amoun of this virus material containing all four serotypes ob ⁇ tained at the fiftieth passage level was dispensed into standard vaccine vials and frozen at -90 * C to -80 * C for inoculation purposes.
  • the froze.n vials were thawed rapidl and each of 1000 sheep were given 2.0 ml parenterally, with a titer of about lO 5 - ⁇ ciD5o/ l.
  • the sheep were ' maintained in separate flocks," 500 in “A” flock, 250 in “B” flock, and the -rest in flocks "C” and “D”.
  • a random selection of 50 sheep from flocks A and B were bled for prevaccination antibody titer and marked for future study.
  • Ten percent of flocks C and D were also pre-bled and similarly marked.
  • the tagged vaccinated sheep were given a virulent virus challenge dose consisting of 500-1000 SID/ml of each of the four serotypes. For four weeks after the challenge, the sheep were observed daily with no clinical signs of disease observed. In contrast, unvaccinated sheep exhibited typical manifestations of BTV disease. Antibody determinations before and after vac- cination showed the sheep to be sero-negative by the agar
  • OMPI gel precipitin test and the serum neutralization test (serum dilution 1:5). However, the vaccinated sheep were fully protected from disease as compared to the unvacci ⁇ nated controls.
  • a vaccine was prepared as in Example VIII but with fortieth passage level material.
  • a 2-ml sample of such - vaccine with a virus titer of about 1 ⁇ 5 « 5 TCID5o/ml was administered parenterally to susceptible cattle (5 each) and goats (10 each) which had been previously determined to be sero-negative to BTV. Other cattle and goats are maintained as unvaccinated controls ' .
  • the antibody status of all animals prior to vaccination was less than 1:5 for goats and 1:10 for cattle by the serum neutralization test and negative by the agar gel pre- • cipitin test.
  • the antibody titer varied from negative to positive by the agar gel precipitin test and from 1:10 to 1:40 on serum neutralization assay. All aminmals were challenged parenterally with virulent BTV of all four serotypes, total dose of 2.0 ml containing 500-1000 SID/ml of each of the four serotypes of BTV. The cattle and goats were observed for 30 to 90 days for evidence of clinical disease. All the vaccinated animals remained normal with no clinical disease or symptoms noted.
  • Example IVA-IVD A 200-ml sample of each of the fortieth passage viru materials obtained as described in Example IVA-IVD was used for inoculation purposes. Prior to vaccination, all experimental yearling sheep were found to have an antibod titer of less than 1:5 by the serum neutralization test and to be agar gel precipitin test negative. A 2-ml amount of a polyvalent vaccine containing equal titer equivalents of BTV IT 10, 11, 13 and 17 prepared by diluting mother pools of stock virus of each serotype to contain about 1 ⁇ 5-0 TCIDso l was administered paren ⁇ terally to each of 20 susceptible yearling sheep with other unvaccinated sheep maintained as unvaccinated controls.
  • Each sheep was monitored daily for evidence of pyrexia or other untoward response to the vaccine.
  • the 20 vaccinated yearling sheep were divide into four groups of five each, and each group was chal ⁇ lenged with 500-1000 SID/ml of virulent BTV, i.e., each group of five sheep received BTV IT 10, 11, 13 and 17, respectively. Appropriate unvaccinated control sheep were included for each group.
  • Example X •A 200-ml sample of vaccine prepared as in Example X was used for inoculation purposes. Prior to vaccination, 80% of the experimental cattle were found to be agar gel -c precipitin test negative. A 2-ml dose of a polyvalent ⁇ - vaccine containing equal titer equivalents of BTV IT 10, 11, 13 and 17 prepared by diluting mother pools of stock virus of each serotype to contain about 10 4 TCIDC. Q /I ⁇ 1 was administered parenterally to each of 69 pregnant (termina 0 aspect of gestation period) cows with other cows main ⁇ tained as unvaccinated contact controls. Fifty days later, the cows were bled for serum precipitin antibody assay and for testing for presence of virus in the blood.
  • a 500-ml sample of vaccine prepared as in * Example X was used for inoculation purposes. Prior to vaccination, all cattle were pregnancy palpated, and 58% cows tested were found to be pregnant. A 2-ml dos.e of a polyvalent vaccine containing equal titer equivalents of BTV IT 10, 11, 13 and 17 prepared by diluting mother pools of stock virus of each serotype to contain about 10 4 TCIDso/ml was administered to each of 180 cows in various stages of pregnancy from 45 days to 8 months and to the 4 herd sires. The cattle were monitored daily for evidence of clinical disease and abortion. Sixty days later, the cow were pregnancy palpated, and 72% of the cows were found t be pregnant. This study shows that the vaccine is non- abortogenic and does not cause transient or long term infertility.
  • a 50-ml sample of vaccine prepared as in Example X was used for inoculation purposes. Prior to vaccination.
  • each fawn was challenged with 250-500 SID/ml of virulent BTV i.e., each -deer fawn received BTV IT 10, 11, 13 and 17, respectively. The remaining deer fawn served as contact controls. Eight and 15 days after virulent virus chal ⁇ lenge, each fawn was bled and tested for evidence of virulent virus.
  • Blood samples from three of the white-tail deer faw on the ninth day after vaccination contained low levels of BTV IT 10. No other serotypes of vaccine virus were isolated.
  • blood samples collected from the two challenged deer on the eighth day after challenge contained virulent virus, one BTV IT 10 and one BTV IT 1 None of the deer showed any signs of clinical illness either after vaccination or after challenge. This study shows that attenuated bluetongue virus contained in the vaccine is fully protective but maintains its ability to replicate in very young susceptible reminants.
  • OMP will be sufficiently attenuated for use as a vaccine if the following criteria are met:

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Virology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Chemical & Material Sciences (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

Les ruminants, en particulier le bétail à corne, les moutons et les chèvres peuvent être protégés contre l'infection du virus "de la langue bleue" (BTV), en particulier les Sérotypes Internationaux IT 10, IT 11, IT 13, et IT 17, par inoculation parentérale avec le vaccin de la présente invention. Le vaccin est préparé par atténuation des sérotypes BTV individuels dans au moins trente passages sériels au travers d'une ligne cellulaire continue ne provenant pas des ruminants, en particulier la culture de cellules "Véro" dérivée du Singe Vert Africain.
EP83900518A 1982-01-19 1983-01-14 Vaccin contre le virus de la maladie bleue et son procede de production Ceased EP0098867A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US34071682A 1982-01-19 1982-01-19
US340716 1982-01-19

Publications (1)

Publication Number Publication Date
EP0098867A1 true EP0098867A1 (fr) 1984-01-25

Family

ID=23334637

Family Applications (1)

Application Number Title Priority Date Filing Date
EP83900518A Ceased EP0098867A1 (fr) 1982-01-19 1983-01-14 Vaccin contre le virus de la maladie bleue et son procede de production

Country Status (9)

Country Link
EP (1) EP0098867A1 (fr)
JP (1) JPS58502210A (fr)
AU (1) AU1155583A (fr)
CA (1) CA1226544A (fr)
DK (1) DK422783D0 (fr)
IL (1) IL67633A0 (fr)
NZ (1) NZ203030A (fr)
WO (1) WO1983002394A1 (fr)
ZA (1) ZA83114B (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110117579A (zh) * 2019-05-29 2019-08-13 军事科学院军事医学研究院军事兽医研究所 表达16型蓝舌病病毒vp2基因的重组病毒及其构建方法和应用

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2062594A1 (fr) * 2007-11-21 2009-05-27 Wyeth Farma, S.A. Vaccin de la fièvre catarrhale du mouton et compositions immunogènes, procédé d'utilisation et son procédé de production
CN106636011B (zh) * 2016-11-14 2018-07-03 陕西诺威利华生物科技有限公司 一种猪流行性腹泻病毒及其应用
CN120249227B (zh) * 2025-06-05 2025-12-05 中国农业科学院哈尔滨兽医研究所（中国动物卫生与流行病学中心哈尔滨分中心） 一种牛流行热病毒的分离方法以及获得的牛流行热病毒及其应用

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO8302394A1 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110117579A (zh) * 2019-05-29 2019-08-13 军事科学院军事医学研究院军事兽医研究所 表达16型蓝舌病病毒vp2基因的重组病毒及其构建方法和应用

Also Published As

Publication number Publication date
JPS58502210A (ja) 1983-12-22
DK422783A (da) 1983-09-16
DK422783D0 (da) 1983-09-16
AU1155583A (en) 1983-07-28
ZA83114B (en) 1983-12-28
IL67633A0 (en) 1983-05-15
CA1226544A (fr) 1987-09-08
NZ203030A (en) 1986-12-05
WO1983002394A1 (fr) 1983-07-21

Similar Documents

Publication Publication Date Title
McClurkin et al. Isolation of cytopathic and noncytopathic bovine viral diarrhea virus from the spleen of cattle acutely and chronically affected with bovine viral diarrhea
Huber et al. Cellular immune mechanisms in Coxsackievirus group B, type 3 induced myocarditis in Balb/C mice
Haesebrouck et al. Effect of intratracheal challenge of fattening pigs previously immunised with an inactivated influenza H1N1 vaccine
US3674861A (en) Live attenuated marek{40 s disease virus vaccine for poultry
NO159782B (no) Anordning ved fjaersystem, foerst og fremst paa kjoeretoeyer.
KR100531491B1 (ko) 백신으로서 소 호흡 코로나바이러스
GB1596653A (en) Vaccine
Wood et al. Observations on the ability of avian reovirus vaccination of hens to protect their progeny against the effects of challenge with homologous and heterologous strains
US3479430A (en) Indirect passive immunization against transmissible gastroenteritis virus in nursing piglets at birth by active immunization of sows prior to farrowing with transmissible gastroenteritis vaccine and method of producing the same
Cursiefen et al. Evaluation of a vaccine against infectious bursal disease in field trials
Awad et al. Preliminary studies of a newly developed subunit vaccine for channel catfish virus disease
WO1983002394A1 (fr) Vaccin contre le virus de la maladie bleue, son procede de production, et procede d'immunisation des ruminants avec ce vaccin
Woolcock Duck hepatitis virus type I: studies with inactivated vaccines in breeder ducks
Gupta et al. Role of thymus-dependent immune system in HVT protection against Marek's disease
US4335105A (en) Specific antigen vaccine for TGE in swine
JPH0710775A (ja) ネコ腸内コロナウイルス由来のイヌコロナウイルスワクチン
Crick The vaccination of man and other animals against rabies
US3585108A (en) Transmissible gastroenteritis vaccines and methods of producing the same
Ariyoshi et al. Vaccination against fowlpox virus via drinking water
Lin et al. Isolation of serotype 2 Marek's disease virus from birds belonging to genus Gallus in Japan
Lee et al. Humoral immune responses to inactivated oil-emulsified Marek's disease vaccine
ES2281094T3 (es) Preparaciones de virus del herpes equino de proteccion cruzada y procedimiento de fabricacion y uso de las mismas.
Karunakaran et al. Seroprevalence of infectious bursal disease (IBD) in parts of Tamil Nadu, India
HU214739B (hu) Eljárás kutya coronavírus vakcina előállítására
Biggs Vaccines and vaccination—past, present and future

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

AK Designated contracting states

Designated state(s): AT BE CH DE FR GB LI LU NL SE

17P Request for examination filed

Effective date: 19840120

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION HAS BEEN REFUSED

18R Application refused

Effective date: 19860808

RIN1 Information on inventor provided before grant (corrected)

Inventor name: CUMMINGS, GEORGIA A.TEXAS A & M UNIVERSITY

Inventor name: MCCONNELL, STEWARTTEXAS A & M UNIVERSITY

Inventor name: LIVINGSTON, CHARLES W., JR.TEXAS A & M UNIVERSITY