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1

Evaluation of peripheral blood leukocyte subpopulations by flow cytometry in calves treated with Mannheimia haemolytica leukotoxin

100%
Bednarek D., Urban-Chmiel R., Dudek K., Szymanska-Czerwinska M.
Bulletin of the Veterinary Institute in Puławy
|
2009
|
tom 53
|
nr 2
199-203
The aim of the study was to evaluate the effect of Mannheimia haemolytica leukotoxin (Lkt) on cellular immune response in clinically-healthy calves given intravenously 25 µg per animal of M. haemolytica Al Lkt. The alternations of peripheral blood leukocytes were examined with a flow cytometry. The Lkt treated calves were compared with the non-treated controls before (0) and 1, 2, 3, 4, 5, 6, and 24 h after the treatment. The following parameters were assayed: white blood cell (WBC) count, percentage of polymorphonuclear leukocytes (PMNL), mid-size leukocytes, total percentage of lymphocytes and their subsets: CD2⁺ (T lymphocytes), CD4⁺ (T helper lymphocytes), and CD8⁺ (T suppressor/cytotoxic lymphocytes) with CD4⁺: CD8⁺ratio, and also WC4⁺ cells (B lymphocytes). The obtained results showed that in the treated calves, the WBC count, the percentages of PMNL, mid-size leukocytes, and some subpopulations of peripheral lymphocytes (CD2⁺, CD4⁺, CD8⁺) were significantly lower compared with the controls during the first of three hours of the experiment, and then the parameters increased and returned to the physiological level.
2

Evaluation of Mannheimia haemolytica leukotoxin prepared in nonsupplemented and BSA or FBS supplemented RPMI 1640 medium

100%
Urban-Chmiel R., Wernicki A., Puchalski A., Mikucki P.
Polish Journal of Veterinary Sciences
|
2004
|
tom 07
|
nr 1
Mannheimia haemolytica leukotoxin (Lkt) is the major factor that contributes to lung injury in bovine pneumonic pasteurellosis. Supernatant preparations containing Lkt produced by M. haemolytica serotype 1, grown in RPMI 1640 medium supplemented with BSA or FBS and without supplements were evaluated during this study. Analysis of obtained Lkt showed presence of 105kDa antigen (SDS-PAGE electrophoresis). The obtained bacterial protein fraction estimated as Lkt was detected by Western blotting with mouse monoclonal (Mab 605 and Mab 601) anti-Lkt antibodies. No significant differences were found in obtained leukotoxin between wildtype and reference M. haemolytica strains. Our studies showed that growth in media supplemented with BSA or FBS had no significant influence on leukotoxin production. When BSA or FBS supplements were used, additional protein fractions in electrophoregrams SDS-PAGE were observed. These protein bands did not react with Mab 605 and/or Mab 601 in Western blotting analysis. Lkt immunogenicity was detected by immunoblotting with sera from Lkt immunized rabbits and calves.
3

Influence of Staphylococcus aureus exosecretions isolated from bovine mastitis on leukocyte morphology in vitro

84%
Fijalkowski K., Czernomysy-Furowicz D., Nawrotek P.
Advances in Agricultural Sciences
|
2011
|
tom 14
|
nr 1-2
4

Effect of leukotoxin of Mannheimia haemolytica and LPS of E. coli on secretory response of bovine neutrophils in vitro

67%
Wessely-Szponder J., Urban-Chmiel R., Wernicki A., Bobowiec R.
Polish Journal of Veterinary Sciences
|
2005
|
tom 08
|
nr 2
5

Comparison of immunogenic properties of native and inactivated Mannheimia haemolytica Lkt in calves

67%
Wernicki A., Mikucki P., Urban-Chmiel R., Puchalski A.
Polish Journal of Veterinary Sciences
|
2008
|
tom 11
|
nr 2
83-88
The aim of this study was to compare the immunostimulatory properties of Lkt of M. haemolytica inactivated by formaldehyde and glutaraldehyde and to evaluate the neutralizing properties of anti-Lkt antibodies. The experiment was conducted on 20 Black-and-White Lowland calves of 100 kg body weight, assigned to 4 experimental groups. The animals were given subcutaneous vaccine injections with native Lkt, Lkt inactivated by formaldehyde or Lkt inactivated by glutaraldehyde. The anti-Lkt antibody titres were measured using an enzyme-linked immunosorbent assay (ELISA), based on absorb- ance of the sera obtained from the animals immunized with the different forms of Lkt. The protective effects of the antibodies present in the sera isolated from the vaccinated animals were estimated using an MIT assay. Analysis of the ELISA absorbance values in the sera from calves in the vaccinated groups did not show any significant differences between the groups. The highest increase in absorbance of sera was observed in calves from the group that received formaldehyde-inactivated Lkt. In the case of calves immunized with native Lkt, the absorbance values were lower than in the group immunized with Lkt inactivated by formaldehyde. The lowest absorbance values were observed in sera obtained from calves vaccinated with Lkt inactivated by glutaraldehyde. Analysis of the MTT assay results revealed the greatest Lkt-neutralizing properties of antibodies in the sera of calves immunized with two doses of a vaccine containing native Lkt and Lkt inactivated with formaldehyde.
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