Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
권호 표지

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

  • Kim, Dong-Woon (Animal Nutrition and Physiology Division, National Institute of Animal Science, RDA) ;
  • Cho, Sung-Back (Animal Nutrition and Physiology Division, National Institute of Animal Science, RDA) ;
  • Lee, Hyun-Jeong (Animal Nutrition and Physiology Division, National Institute of Animal Science, RDA) ;
  • Chung, Wan-Tae (Animal Nutrition and Physiology Division, National Institute of Animal Science, RDA) ;
  • Kim, Kyoung-Hoon (Animal Nutrition and Physiology Division, National Institute of Animal Science, RDA) ;
  • HwangBo, Jong (Animal Nutrition and Physiology Division, National Institute of Animal Science, RDA) ;
  • Nam, In-Sik (Animal Nutrition and Physiology Division, National Institute of Animal Science, RDA) ;
  • Cho, Yong-Il (Animal Nutrition and Physiology Division, National Institute of Animal Science, RDA) ;
  • Yang, Mhan-Pyo (Department of Veterinary Medicine, College of Veterinary Medicine and Research Institute of Veterinary Medicine, Chungbuk National University) ;
  • Chung, Il-Byung (Animal Nutrition and Physiology Division, National Institute of Animal Science, RDA)
  • Published : 2007.08.30
Download PDF
⟨ Previous Next ⟩

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 $(IL)-1{\beta}$, IL-6, IL-12, and tumor necrosis factor $(TNF)-{\alpha}$ 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 $(TNF)-{\alpha}$ in the macrophage model as compared with the whole cells. By way of contrast, $(TNF)-{\alpha}$ 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.

Keywords

References

  1. Adams, D.O. and T.A. Hamilton. 1987. Molecular transductional mechanisms by which IFN-${\gamma}$ and other signals regulate macrophage development. Immunol. Rev. 97, 5-27 https://doi.org/10.1111/j.1600-065X.1987.tb00514.x
  2. Amrouche, T., Y. Boutin, G. Prioult, and I. Fliss. 2006. Effects of bifidobacterial cytoplasm, cell wall and exopolysaccharide on mouse lymphocyte proliferation and cytokine production. Int. Dairy J. 16, 70-80 https://doi.org/10.1016/j.idairyj.2005.01.008
  3. Boisen, S. and J.A. Fernandez. 1995. Prediction of the apparent ileal digestibility of protein and amino acids in feedstuffs and feed mixtures for pigs by in vitro analyses. Anim. Feed Sci. Techno. 51, 29-43 https://doi.org/10.1016/0377-8401(94)00686-4
  4. Clark, P.A. and J.H. Martin. 1994. Selection of bifidobacteria for use as dietary adjuncts in cultured dairy foods: III-Tolerance to simulated bile concentrations of human small intestines. Cult. Dairy Prod. J. 29, 18-21
  5. Cross, M., A. Ganner, D. Teilab, and L. Fray. 2004. Patterns of cytokine induction by Gram-positive and Gram-negative probiotic bacteria. FEMS Immunol. Med. Microbiol. 42, 173-180 https://doi.org/10.1016/j.femsim.2004.04.001
  6. Ding, A.H., C.F. Nathan, and D.J. Stuehr. 1988. Release of reactive nitrogen intermediates and reactive oxygen intermediates from mouse peritoneal macrophages. J. Immunol. 141, 2407-2412
  7. Dong, W., J.L. Azcona-Olivera, K.H. Brooks, J.E. Linz, and J.J. Pestka. 1994. Elevated gene expression and production of interleukins 2, 4, 5, and 6 during exposure to vomitoxin (deoxynivalenol) and cycloheximide in the EL-4 thymoma. Toxicol. Appl. Pharmacol. 127, 282-290 https://doi.org/10.1006/taap.1994.1163
  8. Fukushima, Y., Y. Kawata, K. Mizumachi, J. Kurisaki, and T. Mitsuoka. 1999. Effect of bifidobacteria feeding on fecal flora and production of immunoglobulins in lactating mouse. Int. J. Food Microbiol. 46, 193-197 https://doi.org/10.1016/S0168-1605(98)00183-4
  9. Fuller, R. 1991. Probiotics in human medicine. Gut. 32, 439-442 https://doi.org/10.1136/gut.32.4.439
  10. Gilliland, S.E. 1990. Health and nutritional benefits from lactic acid bacteria. Fed. Eur. Microbial. Rev. 87, 175-188
  11. Hatcher, G.E. and R.S. Lambrecht. 1993. Augmentation of macrophage phagocytic activity by cell-free extracts of selected lactic acid-producing bacteria. J. Dairy Sci. 76, 2485-2492 https://doi.org/10.3168/jds.S0022-0302(93)77583-9
  12. Kang, K.Y., S.H. Park, and T.B. Choe. 1994. Immunostimulation effect of cell wall components isolated from Lactobacillus plantarum. J. Microbiol. Biotechnol. 4, 195-199
  13. Kimoto, H., K. Mizumachi, T. Okamoto, and J. Kurisaki. 2004. New Lactococcus strain with immunomodulatory activity: enhancement of Th1-type immune response. Microbiol. Immunol. 48, 75-82 https://doi.org/10.1111/j.1348-0421.2004.tb03490.x
  14. Kirjavainen, P.V., H.S. El-Nezami, S.J. Salminen, J.T. Ahokas, and P.F. Wright. 1999. The effect of orally administered viable probiotic and dairy lactobacilli on mouse lymphocyte proliferation. FEMS Immunol. Med. Microbiol. 26, 131-135 https://doi.org/10.1111/j.1574-695X.1999.tb01380.x
  15. Link-Amster, H., F. Rochat, K.Y. Saudan, O. Mignot, and J.M. Aeschlimann. 1994. Modulation of a specific humoral immune response and changes in intestinal flora mediated through fermented milk intake. FEMS Immunol. Med. Microbiol. 10, 55-64 https://doi.org/10.1111/j.1574-695X.1994.tb00011.x
  16. Marteau, P., P. Pochart, Y. Bouhnik, S. Zidi, I. Goderel, and J.C. Rambaud. 1992. Survival of Lactobacillus acidophilus and Bifidobacterium sp. in the small intestine following ingestion in fermented milk. A rational basis for the use of probiotics in man. Gastroenterol. Clin. Biol. 16, 25-28
  17. Miettinen, M., J. Voupio-Varkila, and K. Varkila. 1996. Production of human tumor necrosis factor alpha, interleukin-6, and interleukin 10 is induced by lactic acid bacteria. Infec. Immun. 64, 5403-5405
  18. Nomoto, K. 2005. Prevention of infections by probiotics. J. Biosci. Bioeng. 100, 583-592 https://doi.org/10.1263/jbb.100.583
  19. Ouwehand, A.C., P.V. Kirjavainen, M.M. Gronlund, and S.J. Salminen. 1999. Adhesion of probiotic microorganisms to intestinal mucus. Int. Dairy J. 9, 623-630 https://doi.org/10.1016/S0958-6946(99)00132-6
  20. Park, S.Y., G.E. Ji, Y.T. Ko, H.K. Hoo, Z. Ustunol, and J.J. Pestka. 1999. Potentiation of hydrogen peroxide, nitric oxide, and cytokine production in RAW 264.7 macrophage cell exposed to human and commercial isolates of Bifidobacterium. Int. J. Food Microbiol. 46, 231-241 https://doi.org/10.1016/S0168-1605(98)00197-4
  21. Rafter, J.J. 1995. The role of lactic acid bacteria in colon cancer prevention. Scand. J. Gastroenterol. 30, 497-502 https://doi.org/10.3109/00365529509089779
  22. Sakai, T., M. Hamakawa, and K. Shirai. 1996. Protective effects of digested bacterial cell powder on diarrhea in suckling piglets. Agri-Practice. 17, 23-27
  23. SAS. 2000. SAS/STAT user's guide. Version 8, SAS Institute Inc., Cary, N.C., USA
  24. Sasaki, T., S. Fukami, and S. Namioka. 1994. Enhanced resistance of mice to Escherichia coli infection induced by administration of peptidoglycan derived from Bifidobacterium thermophilum. J. Vet. Med. Sci. 53, 443-437
  25. Sekine, K., T. Kasashima, and Y. Hashimoto. 1994. Comparison of the TNF-$\alpha$ level induced by human-derived Bifidobacterium longum and rat-derived Bifidobacterium animalis in mouse peritoneal cells. Bifidobact. Microfl. 13, 79-89 https://doi.org/10.12938/bifidus1982.13.2_79
  26. Sekine, K., J. Ohta, M. Onishi, T. Tatsuki, Y. Shimokawa, T. Toida, T. Kawashima, and Y. Hashimoto. 1995. Analysis of antitumor properties of effector cells stimulated with a cell wall preparation (WPG) of Bifidobacterium infantis. Biol. Pharm. Bull. 18, 148-153 https://doi.org/10.1248/bpb.18.148
  27. Takahashi, T., E. Nakagawa, and T. Nara. 1998. Effects of orally ingested Bifidobacterium longum on the mucosal IgA response of mice to dietary antigens. Biosci Biotech. Biochem. 62, 10-15 https://doi.org/10.1271/bbb.62.10
  28. Takahashi, T., T. Oka, H. Iwana, T. Kuwata, and Y. Yamamoto. 1993. Immune response of mice to orally administered lactic acid bacteria. Biosci. Biotech. Biochem. 57, 1557-1560 https://doi.org/10.1271/bbb.57.1557
  29. Tejada-Simon, M.V. and J.J. Pestka. 1999. Proinflammatory cytokine and nitric oxide induction in murine macrophages by cell wall and cytoplasmic extracts of lactic acid bacteria. J. Food Prot. 62, 1435-1444 https://doi.org/10.4315/0362-028X-62.12.1435
  30. Uemura, J., H. Kitazawa, Y. Kawai, T. Itoh, M. Oda, and T. Saito. 2003. Functional alteration of murine macrophages stimulated with extracellular polysaccharides from Lactobacillus delbrueckii ssp. bulgaricus OLL1073R-1. Food Microbiol. 20, 267-273 https://doi.org/10.1016/S0740-0020(02)00177-6