Discrimination of the heterogeneity of bone marrow‑derived dendritic cells

Wang,Jing; Dai,Xiaomin; Hsu,Chiaching; Ming,Changsheng; He,Ying; Zhang,Ji; Wei,Lai; Zhou,Ping; Wang,Cong‑Yi; Yang,Jun; Gong,Nianqiao

doi:10.3892/mmr.2017.7448

Discrimination of the heterogeneity of bone marrow‑derived dendritic cells

Authors:
- Jing Wang
- Xiaomin Dai
- Chiaching Hsu
- Changsheng Ming
- Ying He
- Ji Zhang
- Lai Wei
- Ping Zhou
- Cong‑Yi Wang
- Jun Yang
- Nianqiao Gong
View Affiliations

Affiliations: Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Key Laboratory of The Ministry of Health, Key Laboratory of The Ministry of Education, Wuhan, Hubei 430030, P.R. China, Pediatric Department, Kuang Tien General Hospital, Taichung, Taiwan 433, R.O.C.
Published online on: September 8, 2017 https://doi.org/10.3892/mmr.2017.7448
Pages: 6787-6793
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The present study aimed to discriminate different subsets of cultured dendritic cells (DCs) to evaluate their immunological characteristics. DCs offer an important foundation for immunological studies, and mouse bone marrow (BM) cells cultured with granulocyte‑macrophage colony‑stimulating factor (GM‑CSF) have been used extensively to generate CD11c+/major histocompatibility complex II+ BM‑derived DCs (BMDCs). Immature DCs are considered to have strong migration and phagocytic antigen‑capturing abilities, whereas mature DCs are thought to activate naive T cells and express high levels of costimulatory cytokines and adhesion molecules. In most culture systems, non‑adherent cells are collected as mature and qualified DCs, and the remaining adherent cells are discarded. The output from GM‑CSF cultures comprises mostly adherent cells, and only a small portion of them is non‑adherent. This situation has resulted in ambiguities in the attempts to understand results from the use of cultured DCs. In the present study, DCs were divided into three subsets: i) Non‑adherent cells; ii) adherent cells and iii) mixed cells. The heterogeneous features of cultured DCs were identified by evaluating the maturation status, cytokine secretion and the ability to activate allogeneic T cells according to different subsets. Results from the study demonstrated that BMDC culture systems were a heterogeneous group of cells comprising non‑adherent cells, adherent cells, mixed cells and firmly adherent cells. Non‑adherent cells may be used in future studies that require relatively mature DCs such as anticancer immunity. Adherent cells may be used to induce tolerance DCs, whereas mixed cells may potentiate either tolerogenicity or pro‑tumorigenic responses. Firmly adherent cells were considered to have macrophage‑like properties. The findings may aid in immunological studies that use cultured DCs and may lead to more precise DC research.

View Figures

View References

1	Banchereau J and Steinman RM: Dendritic cells and the control of immunity. Nature. 392:245–252. 1998. View Article : Google Scholar : PubMed/NCBI
2	Wang J, Zhao LB, Chang S, Ming CS, Yang J and Gong NQ: Dynamic changes of phenotypes and secretory functions during the differentiation of pre-DCs to mature DCs. J Huazhong Univ Sci Technolog Med Sci. 37:191–196. 2017.PubMed/NCBI
3	Satpathy AT, Wu X, Albring JC and Murphy KM: Re(de)fining the dendritic cell lineage. Nat Immunol. 13:1145–1154. 2012. View Article : Google Scholar : PubMed/NCBI
4	Cortez-Retamozo V, Etzrodt M and Pittet MJ: Regulation of macrophage and dendritic cell responses by their lineage precursors. J Innate Immun. 4:411–423. 2012. View Article : Google Scholar : PubMed/NCBI
5	Merad M, Sathe P, Helft J, Miller J and Mortha A: The dendritic cell lineage: Ontogeny and function of dendritic cells and their subsets in the steady state and the inflamed setting. Annu Rev Immunol. 31:563–604. 2013. View Article : Google Scholar : PubMed/NCBI
6	Shurin MR, Pandharipande PP, Zorina TD, Haluszczak C, Subbotin VM, Hunter O, Brumfield A, Storkus WJ, Maraskovsky E and Lotze MT: FLT3 ligand induces the generation of functionally active dendritic cells in mice. Cell Immunol. 179:174–184. 1997. View Article : Google Scholar : PubMed/NCBI
7	Naik SH, Proietto AI, Wilson NS, Dakic A, Schnorrer P, Fuchsberger M, Lahoud MH, O'Keeffe M, Shao QX, Chen WF, et al: Cutting edge: Generation of splenic CD8⁺ and CD8⁺ dendritic cell equivalents in Fms-like tyrosine kinase 3 ligand bone marrow cultures. J Immunol. 174:6592–6597. 2005. View Article : Google Scholar : PubMed/NCBI
8	Yi HJ and Lu GX: Adherent and non-adherent dendritic cells are equivalently qualified in GM-CSF, IL-4 and TNF-α culture system. Cell Immunol. 277:44–48. 2012. View Article : Google Scholar : PubMed/NCBI
9	Kalantari T, Kamali-Sarvestani E, Zhang GX, Safavi F, Lauretti E, Khedmati ME and Rostami A: Generation of large numbers of highly purified dendritic cells from bone marrow progenitor cells after co-culture with syngeneic murine splenocytes. Exp Mol Pathol. 94:336–342. 2013. View Article : Google Scholar : PubMed/NCBI
10	Delirezh N and Shojaeefar E: Phenotypic and functional comparison between flask adherent and magnetic activated cell sorted monocytes derived dendritic cells. Iran J Immunol. 9:98–108. 2012.PubMed/NCBI
11	Abdi K, Singh NJ and Matzinger P: Lipopolysaccharide-activated dendritic cells: ‘Exhausted’ or alert and waiting? J Immunol. 188:5981–5989. 2012. View Article : Google Scholar : PubMed/NCBI
12	Kushwah R, Wu J, Oliver JR, Jiang G, Zhang J, Siminovitch KA and Hu J: Uptake of apoptotic DC converts immature DC into tolerogenic DC that induce differentiation of Foxp3⁺ Treg. Eur J Immunol. 40:1022–1035. 2010. View Article : Google Scholar : PubMed/NCBI
13	Pantel A, Teixeira A, Haddad E, Wood EG, Steinman RM and Longhi MP: Direct type I IFN but not MDA5/TLR3 activation of dendritic cells is required for maturation and metabolic shift to glycolysis after poly IC stimulation. PLoS Biol. 12:e10017592014. View Article : Google Scholar : PubMed/NCBI
14	Wang Y, Cao Y, Meng Y, You Z, Liu X and Liu Z: The novel role of thymopentin in induction of maturation of bone marrow dendritic cells (BMDCs). Int Immunopharmacol. 21:255–260. 2014. View Article : Google Scholar : PubMed/NCBI
15	Li R, Zheng X, Popov I, Zhang X, Wang H, Suzuki M, Necochea-Campion RD, French PW, Chen D, Siu L, et al: Gene silencing of IL-12 in dendritic cells inhibits autoimmune arthritis. J Transl Med. 10:192012. View Article : Google Scholar : PubMed/NCBI
16	Song Q, Meng Y, Wang Y, Li M, Zhang J, Xin S, Wang L and Shan F: Maturation inside and outside bone marrow dendritic cells (BMDCs) modulated by interferon-α (IFN-α). Int Immunopharmacol. 17:843–849. 2013. View Article : Google Scholar : PubMed/NCBI
17	Inaba K, Inaba M, Romani N, Aya H, Deguchi M, Ikehara S, Muramatsu S and Steinman RM: Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor. J Exp Med. 176:1693–1702. 1992. View Article : Google Scholar : PubMed/NCBI
18	Schmid MA, Kingston D, Boddupalli S and Manz MG: Instructive cytokine signals in dendritic cell lineage commitment. Immunol Rev. 234:32–44. 2010. View Article : Google Scholar : PubMed/NCBI
19	Kunieda Y, Tamura Y, Sasaki H, Kurokawa Y, Miyagishima T, Ohae Y, Hige S, Gotohda Y, Tanaka J, Higuchi A, et al: Carcinoid of the papilla of Vater-somatostatinoma-a case report. Gan No Rinsho. 32:831–836. 1986.(In Japanese). PubMed/NCBI
20	Sela U, Olds P, Park A, Schlesinger SJ and Steinman RM: Dendritic cells induce antigen-specific regulatory T cells that prevent graft versus host disease and persist in mice. J Exp Med. 208:2489–2496. 2011. View Article : Google Scholar : PubMed/NCBI
21	Lyons AB and Parish CR: Determination of lymphocyte division by flow cytometry. J Immunol Methods. 171:131–137. 1994. View Article : Google Scholar : PubMed/NCBI
22	Pinto LA, Galvão Castro B, Soares MB and Grassi MF: An evaluation of the spontaneous proliferation of peripheral blood mononuclear cells in HTLV-1-infected individuals using flow cytometry. ISRN Oncol. 2011:3267192011.PubMed/NCBI
23	Haniffa M, Collin M and Ginhoux F: Ontogeny and functional specialization of dendritic cells in human and mouse. Adv Immunol. 120:1–49. 2013. View Article : Google Scholar : PubMed/NCBI
24	Steinman RM and Cohn ZA: Identification of a novel cell type in peripheral lymphoid organs of mice. I. Morphology, quantitation, tissue distribution. J Exp Med. 137:1142–1162. 1973. View Article : Google Scholar : PubMed/NCBI
25	Lutz MB, Kukutsch N, Ogilvie AL, Rössner S, Koch F, Romani N and Schuler G: An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow. J Immunol Methods. 223:77–92. 1999. View Article : Google Scholar : PubMed/NCBI
26	Merad M and Manz MG: Dendritic cell homeostasis. Blood. 113:3418–3427. 2009. View Article : Google Scholar : PubMed/NCBI
27	Weigel BJ, Nath N, Taylor PA, Panoskaltsis-Mortari A, Chen W, Krieg AM, Brasel K and Blazar BR: Comparative analysis of murine marrow-derived dendritic cells generated by Flt3L or GM-CSF/IL-4 and matured with immune stimulatory agents on the in vivo induction of antileukemia responses. Blood. 100:4169–4176. 2002. View Article : Google Scholar : PubMed/NCBI
28	Dudek AM, Martin S, Garg AD and Agostinis P: Immature, Semi-Mature, and fully mature dendritic cells: Toward a DC-cancer cells interface that augments anticancer immunity. Front Immunol. 4:4382013. View Article : Google Scholar : PubMed/NCBI
29	Strome SE, Voss S, Wilcox R, Wakefield TL, Tamada K, Flies D, Chapoval A, Lu J, Kasperbauer JL, Padley D, et al: Strategies for antigen loading of dendritic cells to enhance the antitumor immune response. Cancer Res. 62:1884–1889. 2002.PubMed/NCBI
30	Beriou G, Moreau A and Cuturi MC: Tolerogenic dendritic cells: Applications for solid organ transplantation. Curr Opin Organ Transplant. 17:42–47. 2012. View Article : Google Scholar : PubMed/NCBI
31	Kim SJ and Diamond B: Modulation of tolerogenic dendritic cells and autoimmunity. Semin Cell Dev Biol. 41:49–58. 2015. View Article : Google Scholar : PubMed/NCBI