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Flow cytometry was used to evaluate CD2, CD4, and CDS lymphocyte subpopulations in the peripheral blood of gnotobiotic piglets after administration of lactobacilli and followed E. coli K88+. Fourteen 1-day-old white Slovak crossbreed piglets were divided into four groups. Two experimental groups were given per os from days 2 to 4 Lactobacillus-the first (n = 4) L. salivarius and second (n=3) L. casei. On day 5 the same animals were infected by enteropathogenic nonhaemolytic E. coli. The third group was given only E. coli (n = 4) served as positive control and fourth group included bacteria free animals (n = 3) serve as negative control. Two blood samplings were withdrawn from the retroorbital venous sinus of the piglets' eye on days 3 and 6 after administration of E. coli. Indirect immunofluorescence method for flow cytometry was used. Isolated lymphocytes obtained by density gradient were incubated with primary monoclonal antibodies - mouse anti-pig CD2, CD4 and CD8 and followed FITC-conjugated sheep anti-mouse secondary antibody. Ten thousand lymphocytes were analysed by FACScan (Becton Dickinson) and relative percentage of positive cells was recalculated to the absolute numbers (G(.)l(-1)) by the differential counts of leukocytes. Total numbers of leukocytes and absolute number of lymphocytes were significantly increased (P < 0.05) in both groups treated with lactobacilli, but at the different times, in sampling 1 after application oft. casei and in sampling 2 after L. salivarius. Significant increase in CD2, CD4 and CD8 positive T-cells (P < 0.05) were seen only in the group L. salivarius and E. coli compared to the control group. In our experiment L. salivarius was revealed to be more effective in enhancing the immune response against the subsequent administration of E. coli.

The Influence of Colonization by Lactobacillus Sp. and E. Coli K 88+ on T-Lymphocyte Subpopulations in the Peripheral Blood of Gnotobiotic Piglets