Spontaneous and induced cytolysis of leukocytes from bovine mammary gland in the course of cultivation in vitro - the correlation with neutrophil granulocytes apoptosis
2006
Rysanek, D.(Vyzkumny Ustav Veterinarniho Lekarstvi, Brno (Czech Republic)) E-mail:[email protected] | Sladek, Z.(Mendelova Zemedelska a Lesnicka Univ., Brno (Czech Republic)) | Babak, V.(Vyzkumny Ustav Veterinarniho Lekarstvi, Brno (Czech Republic)) | Vasickova, D.(Mendelova Zemedelska a Lesnicka Univ., Brno (Czech Republic)) | Hubackova, M.(Vyzkumny Ustav Veterinarniho Lekarstvi, Brno (Czech Republic))
The process of leukocyte cytolysis and the manifestations of apoptosis and secondary necrosis of neutrophil granulocytes were studied in four virgin heifers after the induction of leukocyte influx into the mammary gland and after their lavage in in vitro conditions. Cytolysis and apoptosis were induced with heat stress, ultraviolet irradiation and spontaneous aging during 24 hours. The fraction of lysed leukocytes rose during the incubation and reached 73-79% after 24 hours. Phosphate buffered saline resulted in the lowest incidence of cytolysis, the lipopolysaccharide in the highest one. Ultraviolet radiation induced neutrophil apoptosis in a dominant way. Spontaneous leukocyte aging during the in vitro incubation resulted in an increasing share of apoptotic neutrophils depending on the duration of incubation. An increase in the share of necrotic neutrophils was only significant after influx induction with the lipopolysaccharide, but not after induction with buffered saline. Highly significant correlation between the percentage representation of apoptotic neutrophils and the percentage proportion of lysed leukocytes was shown. Significant correlation was demonstrated between the proportion of necrotic neutrophils and the share of lysed leukocytes only after influx induction with the lipopolysaccharide (r=0.579; 0.765). After the influx induction with phosphate buffered saline and staining with Annexin V and propidium iodide, significant negative correlation between the percentage share of necrotic neutrophils and the percentage of lysed leukocytes (r=-0.653) was demonstrated.
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