In vitro immunomodulatory effect of nisin on porcine leucocytes.

Abstract

Nisin, a lantibiotic bacteriocin, has been used for years as a natural food preservative. In addition to its antimicrobial activity, nisin also shows immunomodulatory properties, and the nisin-producing Lactococcus lactis strain has been successfully tested as a probiotic in weaned piglets. However, the impact of nisin on porcine immune cells has not yet been explored. The objective of the present study was to examine the in vitro immunomodulatory effect of nisin on porcine peripheral blood leucocytes. The whole heparinized blood samples or freshly isolated peripheral blood mononuclear cells (PBMCs) were incubated with different nisin concentrations (0, 1.56, 3.125, 6.25, 12.5, 25 or 50\xa0µg/ml) for 1, 24, 48 or 72\xa0hr. Escherichia coli bacteria were used to stimulate blood phagocytes, while concanavalin A and lipopolysaccharide from E.\xa0coli were used as mitogens. Control cells remained unstimulated. MTT colorimetric assay was used to evaluate PBMCs viability and mitogenic response. Phagocyte activity and T-cell proliferation were measured by flow cytometry. Flow cytometer was also used for immunophenotyping of T cells. Cytokine levels in the culture media were determined using commercial immunoassay (ELISA) kits. The highest concentration of nisin exhibited proliferative activity (p\xa0˂\xa00.05), stimulated interleukin-1 beta (IL-1β) and interleukin-6 (IL-6) production (both at p\xa0˂\xa00.001), and increased the percentage of CD4+ CD8+ T cells (p\xa0˂\xa00.001) among unstimulated leucocytes. After cell stimulation, however, the highest nisin concentration showed antiproliferative activity (p\xa0˂\xa00.05), decreased phagocytic functions (p\xa0˂\xa00.05) and inhibited the synthesis of IL-6 (time- and concentration-dependent effect). As a typical bacterial product, nisin had a stronger impact on innate immune cells, and its effect on T cells was likely a consequence of the modulation of the activity of antigen-presenting cells. Nisin may be a good candidate as an immunomodulator in pig breeding.