Pediatric and Developmental Pathology

, Volume 8, Issue 1, pp 43–51

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

  • Jukka Vakkila
  • Michael T. Lotze
  • Connie Riga
  • Ronald Jaffe
Article

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.

Keywords

dendritic cells immunohistochemistry macrophages 

References

  1. 1.
    Manz, MG, Traver, D, Miyamoto, T, Weissman, IL, Akashi, K 2001Dendritic cell potentials of early lymphoid, and myeloid progenitorsBlood9733333341CrossRefPubMedGoogle Scholar
  2. 2.
    Robert, C, Fuhlbrigge, RC, Kieffer, JD,  et al. 1999Interaction of dendritic cells with skin endothelium: a new perspective on immunosurveillanceJ Exp Med189627636Google Scholar
  3. 3.
    Steinman, RM 2003Some interfaces of dendritic cell biologyAPMIS111675697Google Scholar
  4. 4.
    Delneste, Y, Charbonnier, P, Herbault, N,  et al. 2003Interferon-gamma switches monocyte differentiation from dendritic cells to macrophagesBlood101143150Google Scholar
  5. 5.
    Hausser, G, Ludewig, B, Gelderblom, HR, Tsunetsugu-Yokota, Y, Akagawa, K, Meyerhans, A 1997Monocyte-derived dendritic cells represent a transient stage of differentiation in the myeloid lineageImmunobiology197534542Google Scholar
  6. 6.
    Steinbrink, K, Wolfl, M, Jonuleit, H, Knop, J, Enk, AH 1997Induction of tolerance by IL-10–treated dendritic cellsJ Immunol15947724780PubMedGoogle Scholar
  7. 7.
    Fortsch, D, Rollinghoff, M, Stenger, S 2000IL-10 converts human dendritic cells into macrophage-like cells with increased antibacterial activity against virulent Mycobacterium tuberculosisJ Immunol165978987Google Scholar
  8. 8.
    Banchereau J. Converging and diverging properties of human interleukin-4 and interleukin-10. Behring Inst Mitt 1995;58–77Google Scholar
  9. 9.
    Qin, Z, Noffz, G, Mohaupt, M, Blankenstein, T 1997Interleukin-10 prevents dendritic cell accumulation and vaccination with granulocyte-macrophage colony-stimulating factor gene-modified tumor cellsJ Immunol159770776Google Scholar
  10. 10.
    Cella, M, Jarrossay, D, Facchetti, F,  et al. 1999Plasmacytoid monocytes migrate to inflamed lymph nodes and produce large amounts of type I interferonNat Med5919923Google Scholar
  11. 11.
    Siegal, FP, Kadowaki, N, Shodell, M,  et al. 1999The nature of the principal type 1 interferon-producing cells in human bloodScience28418351837PubMedGoogle Scholar
  12. 12.
    Gilliet, M, Liu, YJ 2002Human plasmacytoid-derived dendritic cells and the induction of T-regulatory cellsHum Immunol6311491155Google Scholar
  13. 13.
    Wakkach, A, Fournier, N, Brun, V, Breittmayer, JP, Cottrez, F, Groux, H 2003Characterization of dendritic cells that induce tolerance and T regulatory 1 cell differentiation in vivoImmunity18605617Google Scholar
  14. 14.
    Mazariegos, GV, Zahorchak, AF, Reyes, J,  et al. 2003Dendritic cell subset ratio in peripheral blood correlates with successful withdrawal of immunosuppression in liver transplant patientsAm J Transplant3689696Google Scholar
  15. 15.
    Vakkila, J, Thomson, AW, Vettenranta, K, Sariola, H, Saarinen-Pihkala, UM 2004Dendritic cell subsets in childhood and in children with cancer: relation to age and disease prognosisClin Exp Immunol135455461Google Scholar
  16. 16.
    Bauer, M, Redecke, V, Ellwart, JW,  et al. 2001Bacterial CpG-DNA triggers activation and maturation of human CD11c−, CD123+ dendritic cellsJ Immunol16650005007Google Scholar
  17. 17.
    Boonstra, A, Asselin-Paturel, C, Gilliet, M,  et al. 2003Flexibility 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 ligationJ Exp Med197101109Google Scholar
  18. 18.
    Inoue, K, Furihata, M, Ohtsuki, Y, Fujita, Y 1993Distribution 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 prognosisVirchows Arch A Pathol Anat Histopathol422351355Google Scholar
  19. 19.
    Kakeji, Y, Maehara, Y, Korenaga, D,  et al. 1993Prognostic significance of tumor-host interaction in clinical gastric cancer: relationship between DNA ploidy and dendritic cell infiltrationJ Surg Oncol52207212Google Scholar
  20. 20.
    Ikeguchi, M, Ikeda, M, Tatebe, S, Maeta, M, Kaibara, N 1998Clinical significance of dendritic cell infiltration in esophageal squamous cell carcinomaOncol Rep511851189Google Scholar
  21. 21.
    Lespagnard, L, Gancberg, D, Rouas, G,  et al. 1999Tumor-infiltrating dendritic cells in adenocarcinomas of the breast: a study of 143 neoplasms with a correlation to usual prognostic factors and to clinical outcomeInt J Cancer84309314Google Scholar
  22. 22.
    Reichert, TE, Scheuer, C, Day, R, Wagner, W, Whiteside, TL 2001The number of intratumoral dendritic cells and zeta-chain expression in T cells as prognostic and survival biomarkers in patients with oral carcinomaCancer9121362147Google Scholar
  23. 23.
    Ishigami, S, Natsugoe, S, Tokuda, K,  et al. 2000Clinical impact of intratumoral natural killer cell and dendritic cell infiltration in gastric cancerCancer Lett159103108CrossRefPubMedGoogle Scholar
  24. 24.
    Mantovani, A, Sozzani, S, Locati, M, Allavena, P, Sica, A 2002Macrophage polarization: tumor-associated macrophages as a paradigm for polarized M2 mononuclear phagocytesTrends Immunol23549555CrossRefPubMedGoogle Scholar
  25. 25.
    Jerrells, TR, Dean, JH, Richardson, G, Cannon, GB, Herberman, RB 1979Increased monocyte-mediated cytostasis of lymphoid cell lines in breast and lung cancer patientsInt J Cancer23768776Google Scholar
  26. 26.
    Mantovani, A, Tagliabue, A, Dean, JH, Jerrells, TR, Herberman, RB 1979Cytolytic activity of circulating human monocytes on transformed and untransformed human fibroblastsInt J Cancer232831Google Scholar
  27. 27.
    Mantovani, A, Jerrells, TR, Dean, JH, Herberman, RB 1979Cytolytic and cytostatic activity on tumor cells of circulating human monocytesInt J Cancer231827Google Scholar
  28. 28.
    Vanderheyde, N, Aksoy, E, Amraoui, Z, Vandenabeele, P, Goldman, M, Willems, F 2001Tumoricidal activity of monocyte-derived dendritic cells: evidence for a caspase-8–dependent, Fas-associated death domain-independent mechanismJ Immunol16735653569Google Scholar
  29. 29.
    Santini, SM, Di Pucchio, T, Lapenta, C, Parlato, S, Logozzi, M, Belardelli, F 2003A new type I IFN-mediated pathway for the rapid differentiation of monocytes into highly active dendritic cellsStem Cells21357362Google Scholar
  30. 30.
    Yu, JL, Rak, JW 2003Host microenvironment in breast cancer development: inflammatory and immune cells in tumour angiogenesis and arteriogenesisBreast Cancer Res58388Google Scholar
  31. 31.
    Kalinski, P, Vieira, P, Schuitemaker, JH, Cai, Q, Kapsenberg, M 2003Generation of human type 1- and type 2-polarized dendritic cells from peripheral bloodMethods Mol Biol215427436Google Scholar
  32. 32.
    Sallusto, F, Lanzavecchia, A 1994Efficient 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 alphaJ Exp Med17911091118CrossRefPubMedGoogle Scholar
  33. 33.
    Zou, W, Borvak, J, Marches, F, Wei, S, Isaeva, T, Curiel, TJ 2001

    Isolation, culture and propagation of dendritic cells

    Lotze, MTThomson, AW eds. Dendritic Cells2ndAcademic PressSan Diego82
    Google Scholar
  34. 34.
    Rodriguez, PC, Zea, AH, DeSalvo, J,  et al. 2003L-arginine consumption by macrophages modulates the expression of CD3 zeta chain in T lymphocytesJ Immunol17112321239Google Scholar
  35. 35.
    Munn, DH, Shafizadeh, E, Attwood, JT, Bondarev, I, Pashine, A, Mellor, AL 1999Inhibition of T cell proliferation by macrophage tryptophan catabolismJ Exp Med18913631372CrossRefPubMedGoogle Scholar
  36. 36.
    O’Doherty, U, Peng, M, Gezelter, S,  et al. 1994Human blood contains two subsets of dendritic cells, one immunologically mature and the other immatureImmunology82487493Google Scholar

Copyright information

© Society for Pediatric Pathology 2005

Authors and Affiliations

  • Jukka Vakkila
    • 1
    • 2
  • Michael T. Lotze
    • 2
  • Connie Riga
    • 3
  • Ronald Jaffe
    • 3
  1. 1.Hospital for Children and AdolescentsUniversity of HelsinkiBiomedicumFinland
  2. 2.Molecular Medicine Institute, University of Pittsburgh School of MedicinePittsburghUSA
  3. 3.Children’s Hospital of Pittsburgh, University of Pittsburgh School of MedicinePittsburghUSA

Personalised recommendations