Comparison of Cytokine and Nitric Oxide Induction in Murine Macrophages between Whole Cell and Enzymatically Digested Bifidobacterium sp. Obtained from Monogastric Animals

다국어 초록 (Multilingual Abstract)

The principal objective of this study was to compare the effects of whole and hydrolyzed cells(bifidobacteria) treated with gastrointestinal digestive enzymes on the activation of cloned macrophages. Seven different strains of Bifidobacterium obtained from swine, chickens, and rats, were digested with pepsin followed by pancreatin and the precipitate (insoluble fraction) and supernatant (soluble fraction) obtained via centrifugation. The RAW 264.7 murine macrophages were incubated with either whole cells, the precipitate, or supernatant at various concentrations. Pronounced increases in the levels of nitric oxide (NO), interleukin (IL)-1β, IL-6, IL-12, and tumor necrosis factor (TNF)-α were observed in the whole cells and precipitates, but these effects were less profound in the supernatants. The precipitates also evidenced a slight, but significant, inductive activity for NO and all tested cytokines, with the exception of TNF-α in the macrophage model as compared with the whole cells. By way of contrast, TNF-α production when cultured with whole cells (100 ng/ml) resulted in marked increases as compared with what was observed with the precipitates. The results of this study indicated, for the first time, that digested Bifidobacterium sp. can induce the production of NO and several cytokines in RAW 264.7 murine macrophage cells. In the current study, it was demonstrated that Bifidobacterium strains treated with digestive enzymes, as compared with whole cells, are capable of stimulating the induction of macrophage mediators, which reflects that they may be able to modulate the gastrointestinal immune functions of the host.

참고문헌 (Reference)

1 Rafter, "The role of lactic acid bacteria in colon cancer prevention" 497-502,

2 Salminen, "The effect of orally administered viable probiotic and dairy lactobacilli on mouse lymphocyte proliferation" fems immu (fems immu): 131-135,

3 I, "Survival of Lactobacillus acidophilus and Bifidobacterium sp in the small intestine folowing ingestion in fermented milk A rational basis for the use of probiotics in man" 25-28, 1992

4 Clark, "Selection of bifidobacteria for use as dietary adjuncts in cultured dairy foods III-Tolerance to simulated bile concentrations of human small intestines" 199418-21

5 SAS, "SAS/STAT users guide" SAS Institute Inc., Cary, N.C., USA 2000

6 Ding, "Release of reactive nitrogen intermediates and reactive oxygen intermediates from mouse peritoneal macrophages" immu (immu): 2407-2412, 1988

7 Sakai, T, "Protective effects of digested bacterial cell powder on diarrhea in suckling piglets" 17 : 23-27, 1996

8 Tejada-Simon, M.V, "Proinflammatory cytokine and nitric oxide induction in murine macrophages by cell wall and cytoplasmic extracts of lactic acid bacteria" 62 : 1435-1444, 1999

9 Tejada-Simon, M.V, "Proinflammatory cytokine and nitric oxide induction in murine macrophages by cell wall and cytoplasmic extracts of lactic acid bacteria" 62 : 1435-1444, 1999

10 Miettinen, M, "Production of human tumor necrosis factor alpha, interleukin-6, and interleukin 10 is induced by lactic acid bacteria" 64 : 5403-5405, 1996

11 Fuller, "Probiotics in human medicine" 439-442, 1991

12 Nomoto, K, "Prevention of infections by probiotics" 100 : 583-592, 2005

13 Boisen, S, "Prediction of the apparent ileal digestibility of protein and amino acids in feedstuffs and feed mixtures for pigs by in vitro analyses" 51 : 29-43, 1995

14 Park, S.Y, "Potentiation of hydrogen peroxide, nitric oxide, and cytokine production in RAW 264.7 macrophage cell exposed to human and commercial isolates of Bifidobacter" 46 : 231-241, 1999

15 Cros, M, "Patterns of cytokine induction by Gram-positive and Gram-negative probiotic bacteria" 42 : 173-180, 2004

16 Kimoto, H, "New Lactococcus strain with immunomodulatory activity: enhancement of Th1-type imune response" 48 : 75-82, 2004

17 Adams, "Molecular transductional mechanisms by which IFN-γ and other signals regulate macrophage development Effects of bifidobacterial cytoplasm cell wall and exopolysaccharide on mouse lymphocyte proliferation and cytokine production" immu (immu): 5 70-27 80, 1987

18 Rochat, "Modulation of a specific humoral immune response and changes in intestinal flora mediated through fermented milk intake" fems immu (fems immu): 5-64,

19 Kang, "Immunostimulation effect of cell wall components isolated from Lactobacillus plantarum" 1994195-199

20 Takahashi, "Immune response of mice to orally administered lactic acid bacteria Proinflammatory cytokine and nitric oxide induction in murine macrophages by cell wall and cytoplasmic extracts of lactic acid bacteria" 1557 1435-1560 1444, 1993

21 Takahashi, "Immune response of mice to orally administered lactic acid bacteria" 1557-1560, 1993

22 Gilliland, "Health and nutritional benefits from lactic acid bacteria" 175-188,

23 Uemura, J, "Functional alteration of murine macrophages stimulated with extracellular polysaccharides from Lactobacillus delbrueckii ssp. bulgaricus OLL1073R-1" 20 : 267-273, 2003

24 Uemura, J, "Functional alteration of murine macrophages stimulated with extracellular polysaccharides from Lactobacillus delbrueckii ssp" 20 : 267-273, 2003

25 Sasaki, "Enhanced resistance of mice to Escherichia coli infection induced by administration of peptidoglycan derived from Bifidobacterium thermophilum" 443-437, 1994

26 Dong, "Elevated gene expression and production of interleukins 2 and 6 during exposure to vomitoxin and cycloheximide in the EL-4 thymoma" deoxynivale (deoxynivale): 282-290, 1994

27 Takahashi, T, "Effects of orally ingested Bifidobacterium longum on the mucosal IgA response of mice to dietary antigens" 62 : 10-15, 1998

28 Takahashi, T, "Effects of orally ingested Bifidobacterium longum on the mucosal IgA response of mice to dietary antigens" 62 : 10-15, 1998

29 Amrouche, T, "Effects of bifidobacterial cytoplasm, cell wall and exopolysaccharide on mouse lymphocyte proliferation and cytokine production" 16 : 70-80, 2006

30 Fukushima, Y, "Effect of bifidobacteria feeding on fecal flora and production of immunoglobulins in lactating mouse" 46 : 193-197, 1999

31 Hashimoto, "Comparison of the TNF-α level induced by human-derived Bifidobacterium longum and rat-derived Bifidobacterium animalis in mouse peritoneal cells" 79-89, 1994

32 Hatcher, "Augmentation of macrophage phagocytic activity by cell-free extracts of selected lactic acid-producing bacteria" 2485-2492, 1993

33 Sekine, K, "Analysis of antitumor properties of effector cells stimulated with a cell wall preparation" 18 : 148-153, 1995

34 Ouwehand, A.C, "Adhesion of probiotic microrganisms to intestinal mucus" 9 : 623-630, 1999