Optimization Of New Recombinant Influenza Virus Expressing M . Tuberculosis Esat 6 2 a Ag 85 a Proteins Cultivation In Vero Cells

This paper presents the results of the development of the optimal conditions for new TB-FLU Esat6 2A Ag85A recombinant strain cultivation in Vero cell culture. The studies revealed that the maximum accumulation of the virus in cell culture can be achieved with 0,001 TCD50/cell infecting dose, 34,0 ± 0,5 °C incubation temperature and 72 hours of incubation time. The highest reproductive activity of the virus was obtained using trypsin (Sigma # T4049, Lot10D169, # Lot020M7022T1426-1g) at a concentration 2 μg/ml. The TB-FLU Esat6 2A Ag85A recombinant strain expressing mycobacterial proteins Esat6 and Ag85A with open reading frame of NS1 protein of the influenza virus is genetically stable and capable to maintain enthetic inserts during 10 consecutive passages in Vero cell culture. Subject to the established cultivation parameters the recombinant virus accumulation level reached up to 7,75 ± 0,08 lg TCD50/cm 3 with 7,0 log2hemagglutinating titer, which is quite suitable for the development of a TB vaccine for human health care. [Kondybaeva Zh.B, Yershebulov Z.D, Sultankulova K.S, Nakhanov A.K, Taranov D.S, Bulatov E.A, Khairullin B.M, Sandybaev N.T, Sansyzbay A.R, Koshemetov Zh.К, Burabaev A.A, Yessirkepov M.M. Optimization of new recombinant influenza virus expressing M. tuberculosis ESAT6 2A AG85A proteins cultivation in vero cells. Life Sci J 2014;11(8):280-287]. (ISSN:1097-8135). http://www.lifesciencesite.com. 38