Cytoskeleton Reorganization and Cytokine Production of Macrophages by Bifidobacterial Cells and Cell-Free Extracts

  • Lee, Myung-Ja (Department of Food and Nutrition, Seoul National University) ;
  • Zang, Zhen-Ling (Institute of Labor Hygiene, Shandong Academy of Medical Sciences) ;
  • Choi, Eui-Yul (Department of Genetic Engineering, Hallym University) ;
  • Shin, Hyun-Kyung (Department of Food Science and Nutrition, Hallym University) ;
  • Ji, Geun-Eog (Department of Food and Nutrition, Seoul National University)
  • Published : 2002.06.01

Abstract

Bifidobacteria have been previously shown to stimulate the immune functions and cytokine production in macrophages and T-lymphocytes. Accordingly, the RAW 264.7 murine macrophage cell line was used to assess the effects of Bifidobacterium on the proliferation and cytoskeleton reorganization of the cells. Cytokine production after exposure to Bifidobacterium was also monitored in both whole cells and cell-free extracts. When RAW 264.7 cells were cultured for 24 h in the presence of heat-killed Bifidobacterium bifidum BGN4, the proliferation of macrophages was slowed down in a dose-dependent manner and cell differentiation was observed by staining with the actin-specific fluorescent dye, rhodamin-conjugated phalloidin. Although EL-4 cells, a T-cell line, stimulated RAW 264.7 cells to produce TNF-${\alpha}$ and IL-6, the stimulatory activity of B. bifidum BGN4 decreased as the EL-4 cell number increased. When disrupted and fractionated BGN4 was used, the whole cell fraction was more effective than the other fractions for the TNF-${\alpha}$ production. In contrast, the cell-free extract exhibited the highest IL-6 production level among the fractions, which was evident even at a $1{\mu}g/ml$ concentration. The current results demonstrate that Bifidobacterium induced differentiation of the macrophages from the fast proliferative stage and that the cytokine production was differentially induced by the whole cells and cell-free extracts. The in vitro approaches employed herein are expected to be useful in further characterization of the effects of bifidobacteria with regards to gastrointestinal and systemic immunity.

References

  1. FEMS Microbiol. Lett. v.56 Exocellular products from Bifidobacterium adolescentis as immunomodifiers in the lymphoproliferative rexponsis of mouse splenocytes Gomez, E.;M. M. Melgar, G. P. Silva;A. Portoles;I. Gil.
  2. J. Dairy Sci. v.76 Augmentation of macrophage phagocytic activity by cell-free dxtracts of selected lactic acid-producing bacteria Hatcher, G. E.;R. S. Lambrecht https://doi.org/10.3168/jds.S0022-0302(93)77583-9
  3. Scand. J. Gastroenterol. v.30 The role of lactic acid bacteria in colon cancer prevention Sarih, M.;V. Souvannavong;A. Adam https://doi.org/10.3109/00365529509089779
  4. Bifidobact. Microfl. v.13 Comparison of the TNF-α levels induced by human-derived Bifidobacterium longrm and rat-derived Bifidobacterium animalis in mouse peritoneal cells Sekine, K.;T. Kasashima;Y. Hashimoto https://doi.org/10.12938/bifidus1982.13.2_79
  5. Current Protocols in Immunolgy Trypan blue exclusion test for cell viability Strober, W.;Coligan, J. E.(ed.);Strober, W.(ed.);Marguiles, D. H.(ed.);Shevach, E. M.(ed.);Strober, W.(ed.)
  6. J. Dairy Sci. v.74 Enhancement of immune response in Peyer's patch cells cultured with Bifidobacterium breve Yasui, H.;M. Ohwaki https://doi.org/10.3168/jds.S0022-0302(91)78272-6
  7. Microbial Ecol. Health Dis. v.5 Detection of Bifidobacterium strains that induce large quantities of IgA Yasui, H. N. Nagaoka;A. Mike;K. Hayakawa;M. Ohwaki https://doi.org/10.3109/08910609209141310
  8. Nutr. Res. v.13 Induction of human cytokines by bacteria used in dairy foods Solis Pereyra, B.;D. Lemonnier https://doi.org/10.1016/S0271-5317(05)80737-7
  9. Clin. Immunol. Immunopathol. v.78 The induction of cardioangitis by Lactobacillus casei cell wall in mice. Ⅰ. The cytokine production from murine macrophages by Lactobacillus casei cell wall extract Okitsu-Negishi S.;I. Nakano;K. Suzuki;S. Hashira;T. Abe;K. Yoshino https://doi.org/10.1006/clin.1996.0005
  10. J. Vet. Med. Sci. v.53 Enhanced resistance of mice to Escherichia coli infection induced by administration of peptidoglycan derived from Bifidobacterium thermophilum Sasaki, T.;S. Fukami;S. Namioka
  11. J. Microbiol. Biotechnol. v.9 Comparison of nitric oxkde, hbdrogen peroxide and cytokine production on RAW 264.7 cells gy Bifidobacterium and other intestinal bacteria Om, A. S.;S. Y. Park;G. E. Ji
  12. J. Food Prot. v.53 Anticarcinogenic and immunological proterties of dietary lactobacilli Fernandes, C. F.;K. M. Shagani
  13. Immunotoxicol. v.16 Abjuvant activity of the cell wall of Bifidobacterium infantis for in vivo immune responses in mice Sekine, K.;E. Uatanabe-Sekine;T. Toida;T. Kasashima;T. Kataoka;Y. Hashimoto
  14. Bifidobact. Microfl. v.4 Immunoresponsiveness of newborn piglets and peptidoglycan derived from Bifidobacterium Namioka, S. https://doi.org/10.12938/bifidus1982.4.1_3
  15. Milchwissenshaft v.46 Effects of bifidobacteria cells on mitogenic response of splenocytes and several functions of phagocytes Kado-Oka;Y. S. Fujiwara;U. Hirota
  16. Biol. Pharm. Bull. v.18 Analysis of antitumor properties of effector cells stimulated with a cell wall preparation (WPO) of Bifidobacterium infantis Sekine, K.;J. Ohta;M. Onishi;T. Tatsuki;Y. Shimokawa;T. Toida;T. Kawashima;Y. Hashimoto https://doi.org/10.1248/bpb.18.148
  17. Mycopathologia v.113 Cytotoxic and immunotoxic effects of Fusarium mycotoxins using a rapid colorimetuic gioassay Visconti, A.;F. Mivervini;L. G. Lucivero;V. Gambatesa https://doi.org/10.1007/BF00436128
  18. Int. J. Food Microbiol. v.46 Potentiation of hydrogen peroxide, nitric oxide, and cytokine production in RAW 264.7 macrophage cells exposed to human and commercial isolates of Bifidobacterium Park, S. Y.;G. E. Ji;Y. T. Ko;H. K. Jung;Z. Ustunol;J. J. Pestka https://doi.org/10.1016/S0168-1605(98)00197-4
  19. J. Dairy Sci. v.80 Differential cytokine production in clonal macrophage and T-cell lines cultured with bifidobacteria Marin, M. L.;J. H. Lee;J. Murtha;Z. Stunol;J. J. Pestka https://doi.org/10.3168/jds.S0022-0302(97)76232-5
  20. Biosci. Biotech. Biochem v.57 Screening for the immunopotentiating activity of food microorganisms and engancement of the immune response by Bifidobacterium adolescentis M101-4 Lee, J.;A. Ametani;A. Enomoto;Y. Sato;H. Motoshima;F. Ike;S. Kaminogawa https://doi.org/10.1271/bbb.57.2127
  21. Exp. Cell. Res. v.190 Tubular lysosomes and their drug reactivity on cultured resident macrophages and in cell-free medium Young, M. R.;A. H. Gordon;P.H. Hart https://doi.org/10.1016/0014-4827(90)90198-J
  22. Pediatr. Res. v.35 Effeciveness of Bifidobacterium bifidum in mediating the climical course of murine rotavirus diarrhea Duffy, L. C.;M. A. Zielizny;M. Riepenhoff-Talty;D. Dryja;S. Sayagtagery-Altaie;E. Griffiths;D. Ruffin;GH. Barreyy;J. Rossman;P. L. Ogra https://doi.org/10.1203/00006450-199406000-00014
  23. Biophys. Res. Commun. v.191 Nitric oxide synthase induces macrophage death by apoptosis Sarih, M.;V. Souvannavong;A. Adam https://doi.org/10.1006/bbrc.1993.1246
  24. J. Clin. Invest. v.95 Nitric oxidc mediates cytotoxicity and basic fibroblast frowth factor release in cultured vascular smooth muscle cells. A possible mechanism of neo vascularization in atherosclerotic plaques Fukuo, K.;T. Inoue;S. Morimoto;T. Nakahashi;O. Yasuda;S. Kitano;R. Sasada;T. Ogihara https://doi.org/10.1172/JCI117712
  25. Nature v.372 Signal transduction and segulation in smooth muscle Somlyo, A.;A. Somlyo https://doi.org/10.1038/372231a0
  26. Annu. Rev. Pharmacal. Toxicol. v.35 Macrophages and inflmmatory mediators in tissue injury Laskin, D. L.;J. Pendino https://doi.org/10.1146/annurev.pa.35.040195.003255
  27. J. Inf. Dis. v.172 Passive protection against rotavirus-1 induced diarrhea of mouse pups born to and nursed by dams fed Bifidobacterium breve YIT 4064 Yasui, H.;J. Kiyoshima;H. Ushijima https://doi.org/10.1093/infdis/172.2.403
  28. Cell v.78 No at work Schmidt, H.H.H.W.;W. Walter https://doi.org/10.1016/0092-8674(94)90267-4
  29. Toxicol. Appl. Pharmacol. v.27 Elevated gene expression and production of interleukins 2,4,5, and 6 during exposurd to vomitoxin(deoxynivalenol) and cycloheximide in the EL-4 thymoma Dong, W.;J. I. Azcona-Olivera;K. H. Brooks;J. E. Linz;J. J. Pestka
  30. J. Biol. Chem. v.268 Exprossion of the nitric oxide synthase gene in mouse macrophages activated for tumor cell killing. Molecular basis for the synergy between interferon-gamma and lipopolysaccharide Lorsbach, R. B.;W. J. Murphy;C. J. Lowenstein;S. H. Snyder;S. W. Russell
  31. Cytokine v.3 Peyer's patch B cells with memory cell characteristics undergo terminal differentiation within 24 hours in response to interleukin-6 Beagley, K. W.;J. H. Eldridge;W. K. Aicher;J. Mestecky;S. Di Fabio;H. Kiyono;J. R. McGhee https://doi.org/10.1016/1043-4666(91)90030-H
  32. Infect. Immun v.64 Production of human necrosis factor alpha, interleukin-6 and interleukin-10 is induced by lactic acid bacteria Miettinen, M.;J. Vuopio-Varkila;K. Varkila
  33. FEMS Immunol. Med. Microbiol. v.10 Modulation of a specific humoral immune response and changes in intestinal flora metiated through fermented milk intake Link-Amster, H.;F. Rochat;K. Y. Saudan;O. Mignot;J. M. Aeschlimann https://doi.org/10.1111/j.1574-695X.1994.tb00011.x
  34. Biosci. Biotech. Biochem. v.57 Immune response of mice to orally administered lactic acid bacteria Takahashi, T.;T. Oka;H. Iwana;T. Kuwata;Y. Yamamoto https://doi.org/10.1271/bbb.57.1557
  35. Biosci-Biotech. Biochem. v.61 Characterization of a water-soluble polysaccharide fraction with immunopotentiating activity from Bifidobacterium adolescentis Hosono, A.;J. Lee;A. Ametani;M. Natsume;M. Hirayama;T. Adachi;S. Kaminogawa https://doi.org/10.1271/bbb.61.312
  36. Sci. Amer. v.266 Biological roles of nitric oxide Snyder, S. H.;D. S. Bredt
  37. Bifidobact. Microfl. v.13 Induction and activation of tumoricidal cells in vivo and in vitro by the bacterial cell wall of Bifidobacterium infantis Sekine, K.;E. Watanabe-Sekine; J. Ohta;T. Toida;T. Tatsuki;T. Kawashima;Y. Hashimoto https://doi.org/10.12938/bifidus1982.13.2_65
  38. Lactic Acid Bacteria Microbiology and Functional Aspects Bifidobacteria and probiotic action Ballongue, J.;Salminen S.(ed.);von Wright A.(ed.)