Summary
While tumor cell-derived factors have been demonstrated to hamper the in vitro differentiation and maturation of dendritic cells (DCs) from hematopoietic stem cells, their effects on DC differentiation from CD14+ plastic-adherent monocytic precursors have been controversial. To address this issue, we examined the effects of the culture supernatants from six tumor cell lines on in vitro DC differentiation and maturation from monocytes. Two tumor cell supernatants, MDA468 and 293T, were found to be able to affect the in vitro differentiation of DCs from monocytic precursors, leading to the generation of a distinct type of DC with markedly reduced expression of DC-SIGN, downregulation of CD11c, HLA-DR and CD1a, and upregulation of CD123, HLA-ABC, CD80, CD40, CD86, CD54, CD83, CD25 and CCR7. Functionally, these DCs exhibited reduced phagocytosis and enhanced allostimulatory capacity. Further investigation demonstrated that the changes in DC phenotype and functions were due to the presence of mycoplasmas in these two cell lines; eradication of mycoplasmas completely abolished the observed effects, and importantly, pure mycoplasmas in the absence of tumor cell supernatants were able to produce the same effects. Since mycoplasmas are common contamination agents in routine tissue culture, our results caution that many reported effects of DCs in culture warrant re-evaluation. The distinct effects of mycoplasmas on DC differentiation described in this report could potentially benefit future development of DC-based vaccination and therapeutic applications.
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References
R.M. Steinman (1991) ArticleTitleThe dendritic cell system and its role in immunogenicity Annu. Rev. Immunol. 9 271–296
P. Allavena L. Piemonti D. Longoni S. Bernasconi A. Stoppacciaro L. Ruco A. Mantovani (1998) ArticleTitleIL-10 prevents the differentiation of monocytes to dendritic cells but promotes their maturation to macrophages Eur. J. Immunol. 28 359–369
C. Buelens V. Verhasselt D. De Groote K. Thielemans M. Goldman F. Willems (1997) ArticleTitleHuman dendritic cell responses to lipopolysaccharide and CD40 ligation are differentially regulated by interleukin-10 Eur. J. Immunol. 27 1848–1852
T. De Smedt M. Van Mechelen G. De Becker J. Urbain O. Leo M. Moser (1997) ArticleTitleEffect of interleukin-10 on dendritic cell maturation and function Eur. J. Immunol. 27 1229–1235
D.I. Gabrilovich H.L. Chen K.R. Girgis H.T. Cunningham G.M. Meny S. Nadaf D. Kavanaugh D.P. Carbone (1996) ArticleTitleProduction of vascular endothelial growth factor by human tumors inhibits the functional maturation of dendritic cells Nat. Med. 2 1096–103
C. Menetrier-Caux G. Montmain M.C. Dieu C. Bain M.C. Favrot C. Caux J.Y. Blay (1998) ArticleTitleInhibition of the differentiation of dendritic cells from CD34(+) progenitors by tumor cells: role of interleukin-6 and macrophage colony-stimulating factor Blood 92 4778–4791
T. Oyama S. Ran T. Ishida S. Nadaf L. Kerr D.P. Carbone D.I. Gabrilovich (1998) ArticleTitleVascular endothelial growth factor affects dendritic cell maturation through the inhibition of nuclear factor-kappa B activation in hemopoietic progenitor cells J. Immunol. 160 1224–1232
J. Peguet-Navarro I. Sportouch Popa O. Berthier D. Schmitt J. Portoukalian (2003) ArticleTitleGangliosides from human melanoma tumors impair dendritic cell differentiation from monocytes and induce their apoptosis J. Immunol. 170 3488–3494
S.M. Kiertscher J. Luo S.M. Dubinett M.D. Roth (2000) ArticleTitleTumors promote altered maturation and early apoptosis of monocyte-derived dendritic cells J. Immunol. 164 1269–1276
F. Sallusto M. Cella C. Danieli A. Lanzavecchia (1995) ArticleTitleDendritic cells use macropinocytosis and the mannose receptor to concentrate macromolecules in the major histocompatibility complex class II compartment: downregulation by cytokines and bacterial products J. Exp. Med. 182 389–400
C. Ardavin G. Martinezdel Hoyo P. Martin F. Anjuere C.F. Arias A.R. Marin S. Ruiz V. Parrillas H. Hernandez (2001) ArticleTitleOrigin and differentiation of dendritic cells Trends Immunol. 22 691–700
K.P. MacDonald D.J. Munster G.J. Clark A. Dzionek J. Schmitz D.N. Hart (2002) ArticleTitleCharacterization of human blood dendritic cell subsets Blood 100 4512–4520
S.P. Robinson S. Patterson N. English D. Davies S.C. Knight C.D. Reid (1999) ArticleTitleHuman peripheral blood contains two distinct lineages of dendritic cells Eur. J. Immunol. 29 2769–2778
M.F. Barile S. Rottem (1993) Mycoplasmas in Cell Culture Plenum Press New York
H.G. Drexler C.C. Uphoff (2000) The Encyclopedia of Cell Technology J. Wiley New York
M. Salio V. Cerundolo A. Lanzavecchia (2000) ArticleTitleDendritic cell maturation is induced by mycoplasma infection but not by necrotic cells Eur. J. Immunol. 30 705–708
H. Weigt P.F. Muhlradt A. Emmendorffer N. Krug A. Braun (2003) ArticleTitleSynthetic mycoplasma-derived lipopeptide MALP-2 induces maturation and function of dendritic cells Immunobiology 207 223–233
S. Rottem (2003) ArticleTitleInteraction of mycoplasmas with host cells Physiol. Rev. 83 417–432
O. Takeuchi A. Kaufmann K. Grote T. Kawai K. Hoshino M. Morr P.F. Muhlradt S. Akira (2000) ArticleTitleCutting edge: preferentially the R-stereoisomer of the mycoplasmal lipopeptide macrophage-activating lipopeptide-2 activates immune cells through a toll-like receptor 2- and MyD88-dependent signaling pathway J. Immunol. 164 554–557
C. Link R. Gavioli T. Ebensen A. Canella E. Reinhard C.A. Guzman (2004) ArticleTitleThe Toll-like receptor ligand MALP-2 stimulates dendritic cell maturation and modulates proteasome composition and activity Eur. J. Immunol. 34 899–907
J. Xie J. Qian S. Wang M.E. Freeman SuffixIII J. Epstein Q. Yi (2003) ArticleTitleNovel and detrimental effects of lipopolysaccharide on in vitro generation of immature dendritic cells: involvement of mitogen-activated protein kinase p38 J. Immunol. 171 4792–4800
A. Dzionek A. Fuchs P. Schmidt S. Cremer M. Zysk S. Miltenyi D.W. Buck J. Schmitz (2000) ArticleTitleBDCA-2, BDCA-3, and BDCA-4: three markers for distinct subsets of dendritic cells in human peripheral blood J. Immunol. 165 6037–6046
S. Grabbe E. Kampgen G. Schuler (2000) ArticleTitleDendritic cells: multi-lineal and multi-functional Immunol. Today 21 431–433
M.J. Kwakkenbos G.W. Chang H.H. Lin W. Pouwels E.C. Jong Particlede R.A. Lier Particlevan S. Gordon J. Hamann (2002) ArticleTitleThe human EGF-TM7 family member EMR2 is a heterodimeric receptor expressed on myeloid cells J. Leukoc. Biol. 71 854–862
M.F. Lipscomb B.J. Masten (2002) ArticleTitleDendritic cells: immune regulators in health and disease Physiol. Rev. 82 97–130
T.B. Geijtenbeek R. Torensma S.J. Vliet Particlevan G.C. Duijnhoven Particlevan G.J. Adema Y. Kooyk Particlevan C.G. Figdor (2000) ArticleTitleIdentification of DC-SIGN, a novel dendritic cell-specific ICAM-3 receptor that supports primary immune responses Cell 100 575–585
Steinman R.M, DC-SIGN: a guide to some mysteries of dendritic cells. Cell 100: 491–494, 2000.
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Received 21 April 2004; accepted in revised form 1 August 2004 © 2005 National Science Council, Taipei
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Chen, X., Chang, LJ. Mycoplasma-mediated alterations of in vitro generation and functions of human dendritic cells. J Biomed Sci 12, 31–46 (2005). https://doi.org/10.1007/s11373-004-8181-9
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DOI: https://doi.org/10.1007/s11373-004-8181-9