Cell Biology and Toxicology

, Volume 22, Issue 4, pp 243–255 | Cite as

Improvement of human dendritic cell culture for immunotoxicological investigations



A toxic injury such as a decrease in the number of immature dendritic cells caused by a cytotoxic effect or a disturbance in their maturation process can be responsible for immunodepression. There is a need to improve in vitro assays on human dendritic cells used to detect and evaluate adverse effects of xenobiotics. Two aspects were explored in this work: cytotoxic effects of xenobiotics on immature dendritic cells, and the interference of xenobiotics with dendritic cell maturation. Dendritic cells of two different origins were tested. Dendritic cells obtained either from umbilical cord blood CD34+ cells or, for the first time, from umbilical cord blood monocytes. The cytotoxicity assay on immature dendritic cells has been improved. For the study of the potential adverse effects of xenobiotics on the maturation process of dendritic cells, several parameters were selected such as expression of markers (CD86, CD83, HLA-DR), secretion of interleukins 10 and 12, and proliferation of autologous lymphocytes. The relevance and the efficiency of the protocol applied were tested using two mycotoxins, T-2 toxin and deoxynivalence, DON, which are known to be immunosuppressive, and one phycotoxin, domoic acid, which is known not to have any immunotoxic effect. Assays using umbilical cord monocyte dendritic cell cultures with the protocol defined in this work, which involves a cytotoxicity study followed by evaluation of several markers of adverse effects on the dendritic cell maturation process, revealed their usefulness for investigating xenobiotic immunotoxicity toward immune primary reactions.


cytotoxicity dendritic cell maturation trichothecenes 



dendritic cell




fetal bovine serum




granulocyte-macrophage colony-stimulating factor


monocyte-derived dendritic cells


phosphate-buffered saline




Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Caux C, Saeland S, Favre C, Duvert V, Mannomi P, Banchereau J. TNF-uα strongly potentiates interleukin-3 and GM-CSF induced proliferation of human CD34 hematopoietic progenitor cell. Blood. 1990;75:2292–8.PubMedGoogle Scholar
  2. Caux C, Dezutter-Dambuyant C, Schmitt D, Banchereau J. GM-CSF and TNF-uα cooperate in the generation of dendritic Langerhans cells. Nature. 1992;360:258–61.PubMedCrossRefGoogle Scholar
  3. Clark GJ, Angel N, Kato M, et al. The role of dendritic cells in the innate immune system. Microbes Infect. 2000;2:257–72.PubMedCrossRefGoogle Scholar
  4. Coutant KD, De Brugerolle De Fraissinette A, Cordier A, Ulrich P. Modulation of the activity of human monocyte-derived dendritic cells by chemical haptens, a metal allergen and a staphylococcal superantigene. Toxicol Sci. 1998;52:189–98.CrossRefGoogle Scholar
  5. Froquet R, Sibiril Y, Parent-Massin D. Improvement of megakaryocytic progenitor culture for toxicological investigations.Toxicol In Vitro. 2001;15:691–9.PubMedCrossRefGoogle Scholar
  6. Froquet R, Arnold F, Batina P, Parent-Massin D. Do trichothecenes reduce viability of circulating blood cells and modify haemostasis parameters? Mycopathologia. 2003;156:349–56.PubMedCrossRefGoogle Scholar
  7. Laupeze B, Amiot L, Sparfel L, Le Ferrec E, Fauchet R, Fardel O. Polycyclic aromatic hydrocarbons affect functional differentiation and maturation of human monocyte-derived dendritic cells. J Immunol. 2002;168:2652–8.PubMedGoogle Scholar
  8. Lautraite S, Parent-Massin D, Rio B, Hoellinger H. Comparison of toxicity induced by T-2 toxin on human and rat granulo-monocytic progenitors with an in vitro model. Hum Exp Toxicol. 1995;14: 672–8.PubMedGoogle Scholar
  9. Lautraite S, Parent-Massin D, Rio B, Hoellinger H. In vitro toxicity induced by deoxynivalenol (DON) on human and rat granulo-monocytic progenitors. Cell Biol Toxicol. 1997;13: 175–83.PubMedCrossRefGoogle Scholar
  10. Parent-Massin D. Haematotoxicity of trichothecenes. Toxicol Lett. 2004;153:75–81.PubMedCrossRefGoogle Scholar
  11. Pestka J, Zhou HR, Moon Y, Chung YJ. Cellular and molecular mechanisms for immune modulation by deoxynivalenol and other trichothecenes: unraveling a paradox. Toxicol lett. 2004;153:61–73.PubMedCrossRefGoogle Scholar
  12. Rio B, Lautraite S, Parent-Massin D. In vitro toxicity of trichothecenes on human erythroblastic progenitors. Hum Exp Toxicol. 1997;16:673–9.PubMedCrossRefGoogle Scholar
  13. Rio B, Froquet R, Sibiril Y, Parent-Massin D. Relevance of erythroblast progenitor clonogenic assays in toxicological studies. ATLA. 1999;27:363. [Abstract].Google Scholar
  14. Romani N, Gruner S, Brang D, et al. Proliferating dendritic cell progenitors in human blood. J Exp Med. 1994;180:83–93.PubMedCrossRefGoogle Scholar
  15. Romani N, Reider D, Heuer M, et al. Generation of mature dendritic cells from human blood. An improved method with special regard to clinical applicability. J Immunol Methods. 1996;196:137–51.Google Scholar
  16. Sallusto F, Lanzavecchia A. Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor alpha. J Exp Med. 1994;179:1109–18.PubMedCrossRefGoogle Scholar
  17. Steinman RM. The dendritic cell system and its role in immunogenecity. Annu Rev Immunol. 1991;9: 271–96.PubMedCrossRefGoogle Scholar
  18. Verhasselt V, Buelens C, Willems F, De Groote D, Haeffner-Cavaillon N, Goldman M. Bacterial lipopolysaccharide stimulates the production of cytokines and the expression of costimulatory molecules by human peripheral blood dendritic cells: evidence for a CD14-dependent pathway. J Immunol. 1997;158:2919–25.PubMedGoogle Scholar
  19. Whittle K, Gallacher S. Marine toxins. Br Med Bul. 2000;56:236–53.CrossRefGoogle Scholar
  20. Zhou LJ, Tedder, T.F. Human blood dendritic cells selectively express CD83, a member of the immunoglobulin superfamily. J Immunol. 1995;154:3821–35.PubMedGoogle Scholar

Copyright information

© Springer Science + Business Media, Inc. 2006

Authors and Affiliations

  1. 1.Laboratoire de Toxicologie Alimentaire, Technopôle Brest-IroiseUniversité de Bretagne OccidentalePlouzanéFrance

Personalised recommendations