Soluble Branched β-(1,4)Glucans from Acetobacter Species Show Strong Activities to Induce Interleukin-12 in Vitro and Inhibit T-helper 2 Cellular Response with Immunoglobulin E Production in Vivo

Abstract

An Extracellular Polysaccharide, Ac-1, Produced By Acetobacter Polysaccharogenes Is Composed Of β-(1,4)GluCan With Branches Of Glucosyl Residues. We Found That Ac-1 Showed A Strong Activity To Induce Production Of Interleukin-12 P40 And Tumor Necrosis Factor-α By MacroPhage Cell Lines In Vitro. Cellulase Treatment Completely Abolished The Activity Of Ac-1 To Induce Tumor Necrosis Factor-α Production By Macrophages, Whereas Treatment Of Ac-1 With Polymyxin B Or Proteinase Did Not Affect The Activity. Results Of Experiments Using Toll-Like Receptor (Tlr) 4-Deficient Mice And Tlr4-Transfected Human Cell Line Indicated That Tlr4 Is Involved In Pattern RecogniTion Of Ac-1. In Vivo Administration Of Ac-1 Significantly Reduced The Serum Levels Of Ovalbumin (Ova)-Specific Ige And Interleukin-4 Production By T Cells In Response To Ova In Mice Immunized With Ova. Ac-1, A Soluble Branched β-(1,4)Glucan May Be Useful In Prevention And Treatment Of Allergic Disorders With Ige Production. An Extracellular Polysaccharide, Ac-1, Produced By Acetobacter Polysaccharogenes Is Composed Of β-(1,4)GluCan With Branches Of Glucosyl Residues. We Found That Ac-1 Showed A Strong Activity To Induce Production Of Interleukin-12 P40 And Tumor Necrosis Factor-α By MacroPhage Cell Lines In Vitro. Cellulase Treatment Completely Abolished The Activity Of Ac-1 To Induce Tumor Necrosis Factor-α Production By Macrophages, Whereas Treatment Of Ac-1 With Polymyxin B Or Proteinase Did Not Affect The Activity. Results Of Experiments Using Toll-Like Receptor (Tlr) 4-Deficient Mice And Tlr4-Transfected Human Cell Line Indicated That Tlr4 Is Involved In Pattern RecogniTion Of Ac-1. In Vivo Administration Of Ac-1 Significantly Reduced The Serum Levels Of Ovalbumin (Ova)-Specific Ige And Interleukin-4 Production By T Cells In Response To Ova In Mice Immunized With Ova. Ac-1, A Soluble Branched β-(1,4)Glucan May Be Useful In Prevention And Treatment Of Allergic Disorders With Ige Production. Toll was first identified as a protein that controls dorsoventral pattern formation in the early stage of development of Drosophila (1Belvin M.P. Anderson K.V. Annu. Rev. Cell Dev. Biol. 1996; 12: 393-416Crossref PubMed Scopus (685) Google Scholar) and was shown to participate in antimicrobial immune responses (2Lemaitre B. Nicolas E. Michaut L. Reichhart J.M. Hoffmann J.A. Cell. 1996; 86: 973-983Abstract Full Text Full Text PDF PubMed Scopus (2993) Google Scholar). 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The of was by using an to of not in a of peritoneal of In was with 10 of B for 2 for of activity with for and for of was with of in and for was in the with with a from Acetobacter species to using and with in an to for of The was with a 2 containing 10 as an in a with a a of the in and the to The in for by a system with a The system was with a for and in was used as the in the was to To formation in the which to a of the a of was to the and with the Cell cell in in and 2 to mouse macrophage cell and and a human embryonic kidney cell HEK embryonic kidney from the of and Cell and in with from the peritoneal of used as a of mouse peritoneal exudate in containing for 2 been was to the with in the of of the RNA was from cell using a was with to using a 2 The to with The used as TLR2 TLR2 TLR6 TLR6 TLR9 TLR9 was and to and with as a DNA was using a DNA and a with the was by and the used in using an A mouse and a have been T. K. T. Y. 2000; 95: PubMed Google Scholar). A TLRs and mouse by the and the by in 10 10 and 10 The in and to The for in Tween with in for with TBST, and for with in in TBST, the was using an system and using a in a of HEK with of containing the binding of as an and of using the HEK with for The and in of and the activity was using a with a to the activity was with the and the of and by using an enzyme-linked immunosorbent of from and and for with lipoprotein A in the of IFN-γ, and the was and in the by for and using from and with of in of the the with of in of The with a of of and and and of Production of a in for The cell from the T as by with T and in cell with of of the and the of and in the by used to the of and to the of and of and by of an with and and with in saline for for for and and The with saline with Tween and with IgE for for with of with and the was of the was by A of was as of the of Acetobacter species that produce a of polysaccharide and the of components was using with a A from the of A. is of and in the of The of the components in is shown in I. that the in (30Minakami H. 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The structural of polysaccharide that of polysaccharide for the in the Thus, is a of β-(1,4)glucan with of of components of in a Cellulose from Acetobacter to and derived from Acetobacter species by the of in the mouse macrophage cell and the various and all of which are derived from A. to produce derived from Acetobacter a of the of in mouse macrophage cell the B and not produce in to not was used in the an was to of and by of macrophages from shown in and of and by peritoneal from To the from with and to of the of activity to as that of 3, A and Thus, the protein of is not for the activity of Acetobacter species are Gram-negative bacteria containing LPS, the of of with which the of by shown in B the activity of but not the activity of Thus, the that of the activity of is to by is of β-(1,4)Glucan Is for shown in has a branched containing a of of the to of To the of of in of was with which β-(1,4)glucan shown in the of to of by macrophages was by the of to of by macrophages was not by the We the of a which binding of to receptors R.B. Williams D.L. J. Exp. Google Scholar), the activity of with in the of was not by of that β-(1,4)glucan is for the activity of Is Involved in Production by Cells with have been identified as ancient receptors that confer specificity to the host innate immune the recognition of pathogen-associated molecular patterns. We the of in the recognition of by using The peritoneal from with with and in the by The that the produced in to in with in a and We by the peritoneal of with to is in by shown in the from to a for but not to A to produce a of TNF-α, that is in by the We the of in the of the peritoneal from with shown in the from not to lipoprotein to produce TNF-α, from produced in to as as although to a with from that TLR2 is not in cellular of in a Human Cell the of in cellular the of a mammalian with a The was in a HEK cell The of the was by of Cells with of the with and and the activity was shown in of the not the of that the HEK to was the TLRs including 2 not to of TLR2 with TLR6 not with but with zymosan Polymyxin also activation in HEK that of is not in the activities of Administration of Response with IgE Production to the of in the of immune in the of in with with and Mice from to The of with was The IgE and in the the of was in in with We T from the of with shown in T from with produced in to OVA, the of was in with The of in to was in the of T from with Thus, responses but responses by of in We have identified an derived from A. that activity to of and by is a of β-(1,4)glucan with of and has a molecular of that the β-(1,4)glucan of is important for the activation of The of by peritoneal from in to was and the of HEK to was by with that is in the cellular to containing β-(1,4)glucan with of produced by Acetobacter Toll was first identified as a protein that controls dorsoventral pattern formation in the early stage of development of Drosophila (1Belvin M.P. Anderson K.V. Annu. Rev. Cell Dev. Biol. 1996; 12: 393-416Crossref PubMed Scopus (685) Google Scholar) and was shown to participate in antimicrobial immune responses (2Lemaitre B. Nicolas E. Michaut L. Reichhart J.M. Hoffmann J.A. Cell. 1996; 86: 973-983Abstract Full Text Full Text PDF PubMed Scopus (2993) Google Scholar). mammalian Toll have been identified and have been shown to play important roles in the recognition of various components (3Rock F.L. Hardiman G. Timans J.C. Kastelein R.A. Bazan J.F. Proc. Natl. Acad. Sci. U. S. A. 1998; 95: 588-593Crossref PubMed Scopus (1451) Google Scholar). has been shown to recognize derived from Gram-negative bacteria, TLR2 recognizes peptidoglycan and derived from bacteria and zymosan containing produced by yeast (4Takeuchi O. Hoshino K. Kawai T. Sanjo H. Takada H. Ogawa T. Takeda K. Akira S. Immunity. 1999; 11: 443-451Abstract Full Text Full Text PDF PubMed Scopus (2785) Google Scholar, 5Poltorak A. He X. Smirnova I. Liu M. Huffel C.V. Du X. Birdwell D. Alejos E. Silva M. Galanos C. Freudenberg M. Castagnoli P.R. Layton B. Beutler B. Science. 1998; 282: 2085-2088Crossref PubMed Scopus (6444) Google Scholar, 6Poltorak A. Ricciardi-Castagnoli P. Citterio S. Beutler B. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 2163-2167Crossref PubMed Scopus (396) Google Scholar, 7Hoshino K. Takeuchi O. Kawai T. Ogawa T. Takeda K. Akira S. J. Immunol. 1999; 162: 3749-3752Crossref PubMed Google Scholar, H.D. Libraty D.H. Krutzik S.R. Yang R.B. Belisle J.T. Bleharski J.R. Maitland M. Norgard M.V. Plevy S.E. Smale S.T. Brennan P.J. Bloom B.R. Godowski P.J. Modlin R.L. Science. 1999; 285: 732-736Crossref PubMed Scopus (1406) Google Scholar, 9Michel T. Reichhart J.M. Hoffmann J.A. Royet J. Nature. 2001; 414: 756-759Crossref PubMed Scopus (613) Google Scholar, 10Lien E. Sellati T.J. Yoshimura A. Flo T.H. Rawadi G. Finberg R.W. Carroll J.D. Espevik T. Ingalls R.R. Radolf J.D. Golenbock D.T. J. Biol. Chem. 1999; 274: 33419-33425Abstract Full Text Full Text PDF PubMed Scopus (787) Google Y. Faure E. Thomas L. Equils O. Arditi M. J. Immunol. 2001; 167: 987-994Crossref PubMed Scopus (346) Google Scholar). TLR9 has recently been shown to play a in the recognition of CpG DNA (17Hemmi H. Takeuchi O. Kawai T. Kaisho T. Sato S. Sanjo H. Matsumoto M. Hoshino K. Wagner H. Takeda K. Akira S. Nature. 2000; 408: 740-745Crossref PubMed Scopus (5378) Google Scholar). the of TLRs are not has been shown that TLR6 a with TLR2 A.M. Ozinsky A. Underhill D.M. Aderem A. Wilson C.B. J. Immunol. 2001; 166: PubMed Scopus Google Scholar). The of the that of in to in from HEK with TLR2 TLR2 with TLR6 not to The activity of was not by with the of by the protein in the cellular to is not to B not the activity of but the activity of in a is a receptor for pattern recognition of are major structural components of fungal cell walls that modulate innate immunity in part by macrophage activation in mammalians. There have many reports in which of fungal cell walls that contain has been (21Chihara G. Maeda Y. Hamuro J. Sasaki T. Fukuoka F. Nature. 1969; 222: 687-688Crossref PubMed Scopus (625) Google Scholar, D.C. Parsegian V.A. Science. 1990; 249: 1278-1281Crossref PubMed Scopus (120) Google Scholar, Y. Nio Y. Yoshida O. J. Urol. 1992; 148: 1571-1576Crossref PubMed Scopus (13) Google Scholar). Zymosan, which consists of β-(1,3)glucans S.H. Ye J. Frazer D.G. Shi X. Castranova V. J. Biol. Chem. 2001; 276: 20781-20787Abstract Full Text Full Text PDF PubMed Scopus (130) Google Scholar, 26Sorenson W.G. Shahan T. Simpson J. Ann. Agric. Environ. Med. 1998; 5: 65-71PubMed Google Scholar, has been reported to bind to membrane components such as complement receptor 3, a scavenger receptor, lactosylceramide, and dectin-1 (27Taylor P.R. Brown G.D. Reid D.M. Willment J.A. Martinez P.L. Gordon S. Wong S.Y. J. Immunol. 2002; 169: 3876-3882Crossref PubMed Scopus (529) Google Scholar, 28Brown G.D. Taylor P.R. Reid D.M. Willment J.A. Williams D.L. Martinez P.L. Wong S.Y. Gordon S. J. Exp. Med. 2002; 196: 407-412Crossref PubMed Scopus (814) Google Scholar). It has been suggested that zymosan signals through heterodimers of TLR2 and TLR6 (15Ozinsky A. Underhill D.M. Fontenot J.D. Hajjar A.M. Smith K.D. Wilson C.B. Schroeder L. Aderem A. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 13766-13771Crossref PubMed Scopus (1682) Google Scholar, 16Bulut Y. Faure E. Thomas L. Equils O. Arditi M. J. Immunol. 2001; 167: 987-994Crossref PubMed Scopus (346) Google Scholar), although zymosan is thought to contain multiple stimulators for macrophages in addition to β-(1,3)glucans. In the HEK with TLR2 with TLR6 not to It has recently been reported that a linear β-(1,3)glucan, the binding of macrophage to pattern-recognition receptors using MyD88 for its signal and (29Kataoka K. Muta T. Yamazaki S. Takeshige K. J. Biol. Chem. 2002; 277: 36825-36831Abstract Full Text Full Text PDF PubMed Scopus (186) Google Scholar). We that a that the binding of to not that pattern recognition receptors for are not in the activity of is a of β-(1,4)glucan with of and and the of that is for recognition by is was with a which β-(1,4)glucan the of to of by macrophages was the not the of to that the β-(1,4)glucan is for the activity of It is that immune responses not in but also in In the that of to the of IgE and The of IgE and is to of response, the of is of T in the of to that produce IFN-γ, which are not for of immunity and tumor but are also for of responses J. J. Immunol. Google Scholar, R. P. M.P. E. G. S. J. Exp. Med. PubMed Scopus Google Scholar, T.R. R.L. Annu. Rev. Immunol. Scopus Google Scholar). in are thought to of by a major by which to derived from Acetobacter species have and not for with responses but also for tumor development and of which In have that branched from A. strong activities to the of and in and with IgE in is for cellular responses to for the of We S. Akira for for and for for and for A