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Staphylococcal toxic shock syndrome toxin is cytotoxic for the bovine mammary epithelial cells

DOI 10.18127/j20700997-201905-05

Keywords:

M.P. Shchannikova –  Junior Research Scientist, Laboratory of Immunochemistry in Branch, Institute of Bioorganic Chemistry RAS (Pushchino, Moscow Region)
E-mail: mshannikova@mail.ru
E.K. Tomgorova – Ph.D. (Biol.), Senior Research Scientist, Laboratory of Cell Engineering, Ernst All-Russian Research Institute of Animal Husbandry (Dubrovitsy, Moscow Region)
E-mail: tomgorova@rambler.ru
I.V. Shulcheva – Junior Research Scientist, Laboratory of Immunochemistry in Branch of Institute of Bioorganic Chemistry RAS (Pushchino, Moscow Region)
E-mail: loskutova-i@mail.ru
B.S. Melnik – Dr.Sc. (Phys.-Math.), Head of the Group of Spectroscopy of Protein, Institute of Protein Research RAS (Pushchino, Moscow Region)
E-mail: bmelnik@phys.protres.ru
K.K. Fursova – Ph.D. (Biol.), Senior Research Scientist, Laboratory of Immunochemistry in Branch, Institute of Bioorganic Chemistry RAS (Pushchino, Moscow Region)
E-mail: phursova_k@rambler.ru
S.G. Semushina – Junior Research Scientist, Laboratory of Biological Testing, Branch of Institute of Bioorganic Chemistry RAS (Pushchino, Moscow Region)
E-mail: emushina@bibch.ru
A.N. Noskov – Dr.Sc. (Biol.), Leading Research Scientist, Head of Laboratory of Clostridioses in Gamaleya Federal Research Center of Epidemiology and Microbiology (Moscow)
E-mail: a_n_noskov@mail.ru
V.A. Kazakov – Dr.Sc. (Med.), Senior Research Scientist, Laboratory of Biological Testing, Branch of Institute of Bioorganic Chemistry RAS (Pushchino, Moscow Region)
E-mail: vitalij.tomsk@list.ru
Y.V. Kharitonova – Master Student Pushchino State Institute of Natural Science
E-mail: kharitonova-yv@mail.ru
N.A. Volkova – Dr.Sc. (Biol.), Leading Research Scientist, Head of Laboratory of cell Engineering, Ernst All-Russian Research Institute of Animal Husbandry (Dubrovitsy, Moscow Region)
E-mail: natavolkova@inbox.ru
O.A. Artyemieva – Ph.D. (Biol.), Leading Research Scientist, Head of Laboratory of Microbiology, Ernst All-Russian Research Institute of Animal Husbandry (Dubrovitsy, Moscow Region)
E-mail: vijmikrob@mail.ru
D.A. Nikanova – Research Scientist, Laboratory of Microbiology, Ernst All-Russian Research Institute of Animal Husbandry (Dubrovitsy, Moscow Region)
E-mail: dap2189@gmail.com
N.A. Zinovieva – Academician of RAS, Dr.Sc. (Biol.), Director, Ernst All-Russian Research Institute of Animal Husbandry (Dubrovitsy, Moscow Region)
E-mail: n_zinovieva@mail.ru
F.A. Brovko – Dr.Sc. (Biol.), Head of Laboratory of Immunochemistry, Branch of Institute of Bioorganic Chemistry RAS (Pushchino, Moscow Region)
E-mail: brovko@bibch.ru


Formulation of the problem. Protein expression in animal milk is often associated with mastitis An important role in the development of the disease belongs to Staphylococcus spp. and toxins secreted by them, which have a negative effect on the immune system of both the cow itself and the human milk consumer. Infected segments of the mammary gland become unproductive, which leads to tissue degeneration and to a decrease in the yield of recombinant proteins.
Aim of the research. Investigate the effect of recombinant TSST-1 on breast epithelial cells.
Results. Staphylococcus aureus with the tsst1 gene were isolated previously from animals with mastitis. The effect of TSST-1 on mammary epithelial cells was investigated. The tsst1 gene was cloned and expressed in E. coli. TSST-1 was isolated by Ni-NTA affinity chromatography and gel filtration; endotoxin was removed by chromatography on polymyxin B. The preparation had a purity about 95% according to SDS-PAGE. The secondary structure was confirmed by circular dichroism. Cytotoxic effect of TSST-1 on bMECs was determined by the viability in trypan blue staining and by the MTT assay and compared with previously described staphylococcal enterotoxin H. For TSST-1 the cytotoxic effect was more significant.

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