Skip to main content
SpringerLink
Log in

A Basis for Distinguishing Cultured Dendritic Cells and Macrophages in Cytospins and Fixed Sections

  • Published:
Pediatric and Developmental Pathology

Abstract

There is a burgeoning literature on the contrasting role of intratumoral dendritic cells (DCs) and tumor-associated macrophages, making reliable identification of both cell types in clinical and experimental tissue sections important. However, because these cell types are closely related and share several differentiation antigens, their absolute distinction in tissue sections is difficult. We differentiated DCs and macrophages from monocytes in vitro, prepared cytospins and paraffin-embedded sections of the various cell populations, and tested a variety of antibodies that purportedly recognize monocytes and DCs for their capacity to react and distinguish cells after conventional formalin fixation. Cultured DCs but not macrophages were detected by fascin, DC-LAMP, and CD83 with a predictable increase in staining that paralleled their maturation. Staining by CD1a was found on immature DCs but was weak and absent on mature DCs and macrophages, respectively. CD14 and CD163 were characteristic for macrophages and absent on DCs. CD68, HLA-DR, and S100 did not discriminate between DCs and macrophages. We conclude that antigens such as HLA-DR and S100 are not in themselves sufficient for identification of DCs in formalin-fixed tissue sections, but that additional macrophage-specific (CD14, CD163) and DC-specific (CD1a, CD83, fascin, DC-LAMP) antigens should be used to distinguish cell types from each other and to provide information on their state of maturation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3

Similar content being viewed by others

References

  1. MG Manz D Traver T Miyamoto IL Weissman K Akashi (2001) ArticleTitleDendritic cell potentials of early lymphoid, and myeloid progenitors Blood 97 3333–3341 Occurrence Handle10.1182/blood.V97.11.3333 Occurrence Handle1:CAS:528:DC%2BD3MXktFOkt7w%3D Occurrence Handle11369621

    Article  CAS  PubMed  Google Scholar 

  2. C Robert RC Fuhlbrigge JD Kieffer et al. (1999) ArticleTitleInteraction of dendritic cells with skin endothelium: a new perspective on immunosurveillance J Exp Med 189 627–636

    Google Scholar 

  3. RM Steinman (2003) ArticleTitleSome interfaces of dendritic cell biology APMIS 111 675–697

    Google Scholar 

  4. Y Delneste P Charbonnier N Herbault et al. (2003) ArticleTitleInterferon-gamma switches monocyte differentiation from dendritic cells to macrophages Blood 101 143–150

    Google Scholar 

  5. G Hausser B Ludewig HR Gelderblom Y Tsunetsugu-Yokota K Akagawa A Meyerhans (1997) ArticleTitleMonocyte-derived dendritic cells represent a transient stage of differentiation in the myeloid lineage Immunobiology 197 534–542

    Google Scholar 

  6. K Steinbrink M Wolfl H Jonuleit J Knop AH Enk (1997) ArticleTitleInduction of tolerance by IL-10–treated dendritic cells J Immunol 159 4772–4780 Occurrence Handle1:CAS:528:DyaK2sXntlKnt7s%3D Occurrence Handle9366401

    CAS  PubMed  Google Scholar 

  7. D Fortsch M Rollinghoff S Stenger (2000) ArticleTitleIL-10 converts human dendritic cells into macrophage-like cells with increased antibacterial activity against virulent Mycobacterium tuberculosis J Immunol 165 978–987

    Google Scholar 

  8. Banchereau J. Converging and diverging properties of human interleukin-4 and interleukin-10. Behring Inst Mitt 1995;58–77

  9. Z Qin G Noffz M Mohaupt T Blankenstein (1997) ArticleTitleInterleukin-10 prevents dendritic cell accumulation and vaccination with granulocyte-macrophage colony-stimulating factor gene-modified tumor cells J Immunol 159 770–776

    Google Scholar 

  10. M Cella D Jarrossay F Facchetti et al. (1999) ArticleTitlePlasmacytoid monocytes migrate to inflamed lymph nodes and produce large amounts of type I interferon Nat Med 5 919–923

    Google Scholar 

  11. FP Siegal N Kadowaki M Shodell et al. (1999) ArticleTitleThe nature of the principal type 1 interferon-producing cells in human blood Science 284 1835–1837 Occurrence Handle1:CAS:528:DyaK1MXjvFSqsbg%3D Occurrence Handle10364556

    CAS  PubMed  Google Scholar 

  12. M Gilliet YJ Liu (2002) ArticleTitleHuman plasmacytoid-derived dendritic cells and the induction of T-regulatory cells Hum Immunol 63 1149–1155

    Google Scholar 

  13. A Wakkach N Fournier V Brun JP Breittmayer F Cottrez H Groux (2003) ArticleTitleCharacterization of dendritic cells that induce tolerance and T regulatory 1 cell differentiation in vivo Immunity 18 605–617

    Google Scholar 

  14. GV Mazariegos AF Zahorchak J Reyes et al. (2003) ArticleTitleDendritic cell subset ratio in peripheral blood correlates with successful withdrawal of immunosuppression in liver transplant patients Am J Transplant 3 689–696

    Google Scholar 

  15. J Vakkila AW Thomson K Vettenranta H Sariola UM Saarinen-Pihkala (2004) ArticleTitleDendritic cell subsets in childhood and in children with cancer: relation to age and disease prognosis Clin Exp Immunol 135 455–461

    Google Scholar 

  16. M Bauer V Redecke JW Ellwart et al. (2001) ArticleTitleBacterial CpG-DNA triggers activation and maturation of human CD11c−, CD123+ dendritic cells J Immunol 166 5000–5007

    Google Scholar 

  17. A Boonstra C Asselin-Paturel M Gilliet et al. (2003) ArticleTitleFlexibility of mouse classical and plasmacytoid-derived dendritic cells in directing T helper type 1 and 2 cell development: dependency on antigen dose and differential toll-like receptor ligation J Exp Med 197 101–109

    Google Scholar 

  18. K Inoue M Furihata Y Ohtsuki Y Fujita (1993) ArticleTitleDistribution of S-100 protein–positive dendritic cells and expression of HLA-DR antigen in transitional cell carcinoma of the urinary bladder in relation to tumour progression and prognosis Virchows Arch A Pathol Anat Histopathol 422 351–355

    Google Scholar 

  19. Y Kakeji Y Maehara D Korenaga et al. (1993) ArticleTitlePrognostic significance of tumor-host interaction in clinical gastric cancer: relationship between DNA ploidy and dendritic cell infiltration J Surg Oncol 52 207–212

    Google Scholar 

  20. M Ikeguchi M Ikeda S Tatebe M Maeta N Kaibara (1998) ArticleTitleClinical significance of dendritic cell infiltration in esophageal squamous cell carcinoma Oncol Rep 5 1185–1189

    Google Scholar 

  21. L Lespagnard D Gancberg G Rouas et al. (1999) ArticleTitleTumor-infiltrating dendritic cells in adenocarcinomas of the breast: a study of 143 neoplasms with a correlation to usual prognostic factors and to clinical outcome Int J Cancer 84 309–314

    Google Scholar 

  22. TE Reichert C Scheuer R Day W Wagner TL Whiteside (2001) ArticleTitleThe number of intratumoral dendritic cells and zeta-chain expression in T cells as prognostic and survival biomarkers in patients with oral carcinoma Cancer 91 2136–2147

    Google Scholar 

  23. S Ishigami S Natsugoe K Tokuda et al. (2000) ArticleTitleClinical impact of intratumoral natural killer cell and dendritic cell infiltration in gastric cancer Cancer Lett 159 103–108 Occurrence Handle10.1016/S0304-3835(00)00542-5 Occurrence Handle1:CAS:528:DC%2BD3cXmtF2qu7w%3D Occurrence Handle10974412

    Article  CAS  PubMed  Google Scholar 

  24. A Mantovani S Sozzani M Locati P Allavena A Sica (2002) ArticleTitleMacrophage polarization: tumor-associated macrophages as a paradigm for polarized M2 mononuclear phagocytes Trends Immunol 23 549–555 Occurrence Handle10.1016/S1471-4906(02)02302-5 Occurrence Handle1:CAS:528:DC%2BD38XotVKktrc%3D Occurrence Handle12401408

    Article  CAS  PubMed  Google Scholar 

  25. TR Jerrells JH Dean G Richardson GB Cannon RB Herberman (1979) ArticleTitleIncreased monocyte-mediated cytostasis of lymphoid cell lines in breast and lung cancer patients Int J Cancer 23 768–776

    Google Scholar 

  26. A Mantovani A Tagliabue JH Dean TR Jerrells RB Herberman (1979) ArticleTitleCytolytic activity of circulating human monocytes on transformed and untransformed human fibroblasts Int J Cancer 23 28–31

    Google Scholar 

  27. A Mantovani TR Jerrells JH Dean RB Herberman (1979) ArticleTitleCytolytic and cytostatic activity on tumor cells of circulating human monocytes Int J Cancer 23 18–27

    Google Scholar 

  28. N Vanderheyde E Aksoy Z Amraoui P Vandenabeele M Goldman F Willems (2001) ArticleTitleTumoricidal activity of monocyte-derived dendritic cells: evidence for a caspase-8–dependent, Fas-associated death domain-independent mechanism J Immunol 167 3565–3569

    Google Scholar 

  29. SM Santini T Di Pucchio C Lapenta S Parlato M Logozzi F Belardelli (2003) ArticleTitleA new type I IFN-mediated pathway for the rapid differentiation of monocytes into highly active dendritic cells Stem Cells 21 357–362

    Google Scholar 

  30. JL Yu JW Rak (2003) ArticleTitleHost microenvironment in breast cancer development: inflammatory and immune cells in tumour angiogenesis and arteriogenesis Breast Cancer Res 5 83–88

    Google Scholar 

  31. P Kalinski P Vieira JH Schuitemaker Q Cai M Kapsenberg (2003) ArticleTitleGeneration of human type 1- and type 2-polarized dendritic cells from peripheral blood Methods Mol Biol 215 427–436

    Google Scholar 

  32. F Sallusto A Lanzavecchia (1994) ArticleTitleEfficient 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 179 1109–1118 Occurrence Handle10.1084/jem.179.4.1109 Occurrence Handle1:CAS:528:DyaK2cXhs1Wjurg%3D Occurrence Handle8145033

    Article  CAS  PubMed  Google Scholar 

  33. W Zou J Borvak F Marches S Wei T Isaeva TJ Curiel (2001) Isolation, culture and propagation of dendritic cells MT Lotze AW Thomson (Eds) Dendritic Cells EditionNumber2nd Academic Press San Diego 82

    Google Scholar 

  34. PC Rodriguez AH Zea J DeSalvo et al. (2003) ArticleTitleL-arginine consumption by macrophages modulates the expression of CD3 zeta chain in T lymphocytes J Immunol 171 1232–1239

    Google Scholar 

  35. DH Munn E Shafizadeh JT Attwood I Bondarev A Pashine AL Mellor (1999) ArticleTitleInhibition of T cell proliferation by macrophage tryptophan catabolism J Exp Med 189 1363–1372 Occurrence Handle10.1084/jem.189.9.1363 Occurrence Handle1:CAS:528:DyaK1MXivFGku7w%3D Occurrence Handle10224276

    Article  CAS  PubMed  Google Scholar 

  36. U O’Doherty M Peng S Gezelter et al. (1994) ArticleTitleHuman blood contains two subsets of dendritic cells, one immunologically mature and the other immature Immunology 82 487–493

    Google Scholar 

Download references

Acknowledgment

Jukka Vakkila received support from the Nona and Kullervo Vare Foundation and the Finnish Cultural Foundation. This work was supported by the Marjorie K. Harmer Pediatric Pathology Endowment, Children’s Hospital of Pittsburgh.

Author information

Authors and Affiliations

  1. Hospital for Children and Adolescents, University of Helsinki, Biomedicum, 4th Floor, Suite B431b, P.O. Box 700, FIN-00029, Hus, Finland

    Jukka Vakkila

  2. Molecular Medicine Institute, University of Pittsburgh School of Medicine, Room 411, 300 Technology Drive, Pittsburgh, PA, 5219, USA

    Jukka Vakkila & Michael T. Lotze

  3. Children’s Hospital of Pittsburgh, University of Pittsburgh School of Medicine, 3705 Fifth Avenue, Pittsburgh, PA, 15213, USA

    Connie Riga & Ronald Jaffe

Authors
  1. Jukka Vakkila
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michael T. Lotze
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Connie Riga
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ronald Jaffe
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ronald Jaffe.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vakkila, J., Lotze, M.T., Riga, C. et al. A Basis for Distinguishing Cultured Dendritic Cells and Macrophages in Cytospins and Fixed Sections. Pediatr Dev Pathol 8, 43–51 (2005). https://doi.org/10.1007/s10024-004-5045-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10024-004-5045-2

Keywords

Search

Navigation