A novel influenza viral vector based brucella abortus vaccine at the stage of implementation into practice

CONSTRUCTION OF INFLUENZA VIRAL VECTORS
Schematic representation of the influenza virus NS1 gene (A) and recombinant chimeric NS1 genes of recombinant influenza A viral vectors of the subtypes H5N1 and H1N1 containing the genetic sequences of the Brucella proteins L7/L12 or Omp16 (B)
Four influenza viral vectors of the subtypes Н5N1 or H1N1 expressing the Brucella proteins L7/L12 or Omp16 were obtained by a reverse genetics method: Flu-NS1-124-L7/L12-H5N1, Flu-NS1-124-Omp16-H5N1, Flu-NS1-124-L7/L12-H1N1 and Flu-NS1-124-Omp16-H1N1
* B. abortus proteins L7/L12 (GenBank: AAA19863.1) or Omp16 (GenBank: AAA59360.1)

TECHNICAL CHARACTERISTICS OF THE VACCINE
The vaccine is a mixture of recombinant strains of the influenza virus expressing brucellosis antigens (Omp16 and L7/L12) accumulated in 9-11 day old chicken embryos (CE) and lyophilized with stabilizing medium
The vaccine is packaged in 1-10 ml into ampoules or vials
As a solvent we use 10-20% adjuvant Montanide Gel 01 (Seppic, France), packaged in 5-500 ml in vials
Depending on the subtype of influenza viral vectors included in the formulation, the vaccine is marked as "Vaccine 1" (a mixture of influenza viral vectors subtype H5N1) and "Vaccine 2" (a mixture of influenza viral vectors subtype H1N1)

CONSUMER CHARACTERISTICS OF THE VACCINE
The vector vaccine is used for double immunization of cattle (young animals aged 6 months or more, including bulls, heifers or cows, including pregnant) first by "Vaccine 1" (prime vaccination), and then through 21-28 days "Vaccine 2" (booster vaccination)
The vaccine can be used in the Brucella-free, Brucella-infected or in farms (in conjunction with other antiepizootic measures) at the stage of recovery from brucellosis.
Method and dose of administration of vaccine for cattle in Brucella-free farms: subcutaneously in the neck region in a volume of 1.0 ml. In Brucella-infected farms: subcutaneously in the neck region in a volume of 1.0 ml with simultaneous conjunctival administration of the vaccine in a volume of 0.5 ml (0.25 ml per eye)

THE MAIN PROPERTIES OF THE DEVELOPED VACCINE
The vaccine is a live vaccine based on influenza viral vectors, and induces a strong Th-1 immune response in cattle;
Vaccinated cattle do not form Brucella agglutinogen antibodies, making it easy to differentiate vaccinated animals from infected animals;
As the truncated NS1 protein (interferon antagonist) influenza viral vectors has limited replicative capacity (influenza viral vectors subtype H5 was further attenuated by exchanging its polybasic cleavage site with a trypsin-dependent sequence); this attenuated vaccine cannot cause disease in cattle or humans;
The influenza viral vectors are not shed by vaccinated animals into the environment and cannot be transmitted to other animals or humans;

THE MAIN PROPERTIES OF THE DEVELOPED VACCINE (continuation)
The vaccine is genetically stable, as it retains all of its basic biological properties including attenuation markers and does not lose the Brucella protein inserts after repeated passage in its culture system, chicken embryos;
The vaccine in vaccinated cattle provides formation of long-term protective immune response which lasting at least 12 months after booster vaccination;
The vaccine is able to provide cross-protection against Brucella melitensis infection in pregnant heifers;
The vaccine can be used in all sex and age groups of cattle, regardless of the status of pregnancy in animals, both in Brucella-free and Brucella-infected farms.
In the presence of a production site for the production of dry preparations based on the use of chicken embryos, the vaccine can be easily and on a large scale produced

PUBLICATIONS IN INTERNATIONAL PEER-REVIEWED JOURNALS
Mailybayeva A, et al. Improved influenza viral vector based Brucella abortus vaccine induces robust B and T-cell responses and protection against Brucella melitensis infection in pregnant sheep and goats. PLoS One. 2017;12(10):e0186484. (Impact factor-2.8)
Tabynov K, et al. First evaluation of an influenza viral vector based Brucella abortus vaccine in sheep and goats: Assessment of safety, immunogenicity and protective efficacy against Brucella melitensis infection. Vet Microbiol. 2016;197:15-20. (Impact factor-2.5)
Tabynov K., et al. Simultaneous subcutaneous and conjunctival administration of the influenza viral vector based Brucella abortus vaccine to pregnant heifers provides better protection against B. abortus 544 infection than the commercial B. abortus S19 vaccine. Vaccine. 2016;34(42):5049-52. (Impact factor-3.6)
Tabynov K. Influenza viral vector based Brucella abortus vaccine: a novel vaccine candidate for veterinary practice. Expert Rev Vaccines. 2016;15(10):1237-9. (Impact factor-4.2)
Tabynov K, et al. Prime-booster vaccination of cattle with an influenza viral vector Brucella abortus vaccine induces a long-term protective immune response against Brucella abortus infection. Vaccine. 2016. 34:438-444. (Impact factor-3.6)
Tabynov K, et al. Safety of the novel influenza viral vector Brucella abortus vaccine in pregnant heifers. Ciência Rural. 2016. 46(1):114-118. (Impact factor-0.4)
Tabynov K, et al. An influenza viral vector Brucella abortus vaccine induces good cross-protection against Brucella melitensis infection in pregnant heifers. Vaccine. 2015. 33(31):3619-23. (Impact factor-3.6)
Tabynov K, et al. Novel vector vaccine against Brucella abortus based on influenza A viruses expressing Brucella L7/L12 or Omp16 proteins: Evaluation of protection in pregnant heifers. Vaccine. 2014. 32(45):5889-92. (Impact factor-3.6)
Tabynov K, et al. Novel influenza virus vectors expressing Brucella L7/L12 or Omp16 proteins in cattle induce a strong T-cell immune response, as well as high protectiveness against B. abortus infection. Vaccine. 2014. 32(18):2034-41. (Impact factor-3.6)
Tabynov K, et al. Influenza viral vectors expressing the Brucella OMP16 or L7/L12 proteins as vaccines against B. abortus infection. Virology Journal. 2014. 11: 69. (Impact factor-2.18)

COMPETITIVE ADVANTAGES

FIELD AND REGISTRATION TRIALS
According to Order of the Chairman of the Committee of Veterinary Control and Supervision of the Ministry of Agriculture of the Republic of Kazakhstan No. 76 dated 05.24.2018, between May 17 and November 23, 2018, it was conducted field and registration trials of our vaccine.
It has been established that the vaccine in its physical and immunobiological properties fully meets the requirements of the regulatory document.
The vaccine is harmless and protective in heifers after a double immunization
In late December 2018, it is expected to receive a registration certificate for the vaccine, and its inclusion in the State Register of Veterinary Medicines of the Republic of Kazakhstan

CONCLUSION
Thus, since the introduction of the latest commercial B. abortus RB51 vaccine in the United States 20 years ago, we have for the first time developed and offer to commercialize a completely new vaccine against bovine brucellosis, which has better properties (by safety profile) in comparison with commercial preparations