Advances in Clinical and Experimental Medicine
2014, vol. 23, nr 6, November-December, p. 871–876
Publication type: original article
Bovine Lactoferrin Enhances Proliferation of Human Peripheral Blood Lymphocytes and Induces Cytokine Production in Whole Blood Cultures
1 Laboratory of Immunobiology, Institute of Immunology and Experimental Therapy, Polish Academy of Science, Wrocław, Poland
Background. . Lactoferrin belongs to the immunoregulatory milk proteins involved in iron metabolism as well in providing innate immunity to newborns. The protein has been the subject of numerous clinical studies.
Objectives. The aim of this investigation was to evaluate the effects of bovine lactoferrins (bLF), differing in source and iron content, on spontaneous proliferation of human peripheral blood mononuclear cells (PBMC) and cytokine production by human whole blood cultures.
Material and Methods. The following bLF preparations were used: partially iron saturated or devoid of iron bLF from milk and bLF from colostrum. The study was conducted on 12 healthy volunteers (men, 20–24 years old). The effects of bLFs on the proliferation of PBMC in four-day cultures was studied at 50–0.6 µg/mL concentration range and the rate of proliferation was determined using the MTT colorimetric method. TNF α and IL-6 levels, induced by the bLFs in 24 h whole blood cultures, were measured by ELISA.
Results. The lactoferrins stimulated autologous proliferation of human peripheral blood mononuclear cells (PBMC) in a dose-dependent manner, with a comparable efficacy. This stimulation occurred both in the constant presence of bLFs in the cultures and also upon preincubation of PBMC with bLFs with subsequent exhaustive wash of cells. Only bLF from colostrum induced production of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) in cultures of whole blood cells. This phenomenon took place predominantly at concentration of 50 µg/mL.
Conclusion. The results showed potent stimulation of the proliferative response of PBMC by bovine lactoferrin, associated with the induction of proinflammatory cytokines only in the case of colostral bLF. This observation may be of importance when high doses of bLF are used in therapy and by designing diet supplementation with this protein.
IL-6, proliferation, lactoferrin, TNF-α, autologous PBMC culture.
- Vogel HJ: Lactoferrin, a bird’s eye view. Biochem Cell Biol 2012, 90, 233–244.
- Masson PL, Heremans JF, Schonne E: Lactoferrin, an iron-binding protein in neutrophilic leukocytes. J Exp Med 1969, 130, 643–658.
- Johnson EE, Wessling-Resnick M: Iron metabolism and the innate immune response to infection. Microbes Infect 2012, 14, 207–216.
- Zimecki M, Mazurier J, Machnicki M, Wieczorek Z, Montreuil J, Spik G: Immunostimulatory activity of lactotransferrin and maturation of CD4- CD8-murine thymocytes. Immunol Lett 1991, 30, 119–123.
- Zimecki M, Mazurier J, Spik G, Kapp JA: Human lactoferrin induces phenotypic and functional changes in murine splenic B cells. Immunology 1995, 86, 122–127.
- Debbabi H, Dubarry M, Rautureau M, Tome D: Bovine lactoferrin induces both mucosal and systemic immune response in mice. J Dairy Res 1998, 65, 283–293.
- Suzuki YA, Lopez V, Lonnerdal B: Mammalian lactoferrin receptors: structure and function. Cell Mol Life Sci 2005, 62, 2560–2575.
- Zimecki M, Kocieba M, Kruzel M: Immunoregulatory activities of lactoferrin in the delayed type hypersensitivity in mice are mediated by a receptor with affinity to mannose. Immunobiology 2002, 205, 120–131.
- Curran CS, Demick KP, Mansfield JM: Lactoferrin activates macrophages via TLR4-dependent and -independent signaling pathways. Cell Immunol 2006, 242, 23–30.
- Chien YJ, Chen WJ, Hsu WL, Chiou SS: Bovine lactoferrin inhibits Japanese encephalitis virus by binding to heparan sulfate and receptor for low density lipoprotein. Virology 2008, 379, 143–151.
- Baveye S, Elass E, Fernig DG, Blanquart C, Mazurier J, Legrand D: Human lactoferrin interacts with soluble CD14 and inhibits expression of endothelial adhesion molecules, E-selectin and ICAM-1, induced by the CD14- lipopolysaccharide complex. Infect Immun 2000, 68, 6519–6525.
- Legrand D, Vigie K, Said EA, Elass E, Masson M, Slomianny MC, Carpentier M, Briand JP, Mazurier J, Hovanessian AG: Surface nucleolin participates in both the binding and endocytosis of lactoferrin in target cells. Eur J Biochem 2004, 271, 303–317.
- Shin K, Wakabayashi H, Yamauchi K, Yaeshima T, Iwatsuki K: Recombinant human intelectin binds bovine lactoferrin and its peptides. Biol Pharm Bull 2008, 31, 1605–1608.
- Choi BK, Actor JK, Rios S, d’Anjou M, Stadheim TA, Warburton S, Giaccone E, Cukan M, Li H, Kull A, Sharkey N, Gollnick P, Kocieba M, Artym J, Zimecki M, Kruzel ML, Wildt S: Recombinant human lactoferrin expressed in glycoengineered Pichia pastoris: effect of terminal N-acetylneuraminic acid on in vitro secondary humoral immune response. Glycoconj J 2008, 25, 581–593.
- Zimecki M, Artym J, Kocięba M, Duk M, Kruzel ML: The effect of carbohydrate moiety structure on the immunoregulatory activity of lactoferrin in vitro. Cell Mol Biol Lett 2014, 19, 284–296.
- Djeha A, Brock JH: Effect of transferrin, lactotransferrin and chelated iron on human T-lymphocytes. Br J Haematol 1992, 80, 235–241.
- Miyauchi H, Kaino A, Shinoda I, Fukuwatari Y, Hayasawa H: Immunomodulatory effect of bovine lactoferrin pepsin hydrolysate on murine splenocytes and Peyer’s patch cells. J Dairy Sci 1997, 80, 2330–2339.
- Slater K, Fletcher J: Lactoferrin derived from neutrophils inhibits the mixed lymphocyte reaction: Blood 1987, 69, 1328–1333.
- Zimecki M, Właszczyk A, Cheneau P, Brunel AS, Mazurier J, Spik G, Kübler A: Immunoregulatory effects of a nutritional preparation containing bovine lactoferrin taken orally by healthy individuals. Arch Immunol Ther Exp 1998, 46, 231–240.
- Zimecki M, Stępniak D, Szynol A, Kruzel ML: Lactoferrin Regulates Proliferative Response of Human Peripheral Blood Mononuclear Cells to Phytohemagglutinin and Mixed Lymphocyte Reaction. Arch Immunol Ther Exp 2001, 49, 147–154.
- Zimecki M, Właszczyk A, Wojciechowski R, Dawiskiba J, Kruzel M: Lactoferrin regulates the immune responses in post-surgical patients. Arch Immunol Ther Exp 2001, 49, 325–333.
- Zimecki M, Kruzel ML: Milk-derived proteins and peptides of potential therapeutic and nutritive value. J Exp Ther Oncol 2007, 6, 89–106.
- Artym J, Zimecki M: The role of lactoferrin in the proper development of newborns. Postepy Hig Med Dosw (Online) 2005, 59, 421–432.
- Machnicki M, Zimecki M, Zagulski T: Lactoferrin regulates the release of tumour necrosis factor alpha and interleukin 6 in vivo. Int J Exp Pathol 1993, 74, 433–439.
- Adamik B, Zimecki M, Właszczyk A, Berezowicz P, Kübler A: Lactoferrin effects on the in vitro immune response in critically ill patients. Arch Immunol Ther Exp 1998, 46, 169–176.
- Smolen JS, Luger TA, Chused TM, Steinberg AD: Responder cells in the human autologous mixed lymphocyte reaction. J Clin Invest 1981, 68, 1601–1604.
- Zimecki M, Międzybrodzki R, Mazurier J, Spik G: Regulatory Effects of Lactoferrin and Lipopolysaccharide on LFA-1 Expression on Human Peripheral Blood Mononuclear Cells. Arch Immunol Ther Exp 1999, 47, 257–264.
- Frydecka I, Zimecki M, Boćko D, Kosmaczewska A, Teodorowska R, Ciszak L, Kruzel M, Włodarska-Polinsk J, Kuliczkowski K, Kornafel J: Lactoferrin-induced up-regulation of zeta (zeta) chain expression in peripheral blood T lymphocytes from cervical cancer patients. Anticancer Res 2002, 22, 1897–1901.
- Duthille I, Masson M, Spik G, Mazurier J: Lactoferrin stimulates the mitogen-activated protein kinase in the human lymphoblastic T Jurkat cell line. Adv Exp Med Biol 1998, 443, 257–260.
- Hou JM, Chen EY, Wei SC, Lin F, Lin QM, Lan XH, Xue Y, Wu M: Lactoferrin inhibits apoptosis through insulin-like growth factor I in primary rat osteoblasts. Acta Pharmacol Sin 2014, 35, 523–530.
- Ogasawara Y, Imase M, Oda H, Wakabayashi H, Ishii K: Lactoferrin directly scavenges hydroxyl radicals and undergoes oxidative self-degradation: a possible role in protection against oxidative DNA damage. Int J Mol Sci 2014, 15, 1003–1013.
- Bi BY, Leveugle B, Liu JL, Collard A, Coppe P, Roche AC, Nillesse N, Capron M, Spik G, Mazurier J: Immunolocalization of the lactotransferrin receptor on the human T lymphoblastic cell line Jurkat. Eur J Cell Biol 1994, 65, 164–171.
- Yokota S, Geppert TD, Lipsky PE: Enhancement of antigenand mitogen-induced human T lymphocyte proliferation by tumor necrosis factor-alpha. J Immunol 1988, 140, 531–536.
- Takimori S, Shimaoka H, Furukawa J, Yamashita T, Amano M, Fujitani N, Takegawa Y, Hammarström L, Kacskovics I, Shinohara Y, Nishimura S: Alteration of the N-glycome of bovine milk glycoproteins during early lactation. FEBS J 2011, 278, 3769–3781.
- Le Parc A, Dallas DC, Duaut S, Leonil J, Martin P, Barile D: Characterization of goat milk lactoferrin N-glycans and comparison with the N-glycomes of human and bovine milk. Electrophoresis 2014, 35, 1560–1570.