Flt3-ligand, IL-4, GM-CSF, and adherence-mediated isolation of murine lung dendritic cells: Assessment of isolation technique on phenotype and function

Kena A. Swanson, Yan Zheng, Kathleen M. Heidler, Zhen Du Zhang, Tonya J. Webb, David S. Wilkes

Research output: Contribution to journalArticle

34 Scopus citations

Abstract

Lung dendritic cells (DCs) are difficult to study due to their limited quantities and the complexities required for isolation. Although many procedures have been used to overcome this challenge, the effects of isolation techniques on lung DCs have not been reported. The current study shows that freshly isolated DCs (CD11c+) have limited ability to induce proliferation in allogeneic T cells, and are immature as indicated by low cell surface expression of costimulatory molecules compared with liver or splenic DCs. DCs isolated after overnight culture or from mice treated with Flt3L are phenotypically mature and potent stimulators of allogeneic T cells. DCs could not be propagated from lung mononuclear cells in response to IL-4 and GM-CSF. Contrary to data reported for nonpulmonary DCs, expression of CCR6 was decreased on mature lung DCs, and only a subset of mature DCs expressed higher levels of CCR7. Absence of CD8α expression indicates that freshly isolated DCs are myeloid-type, whereas mature DCs induced by overnight culture are both "lymphoid" (CD8α+) and "myeloid" (CD8α-). DCs from mice genetically deficient in CD8α expression were strong simulators of allogeneic T cells which was consistent with data showing that CD8α- DCs from CD8α-sufficient mice are better APCs compared with CD8α+ DCs from the same mice. These data show that freshly isolated lung DCs are phenotypically and functionally distinct, and that the isolation technique alters the biology of these cells. Therefore, lung DC phenotype and function must be interpreted relative to the technique used for isolation.

Original languageEnglish (US)
Pages (from-to)4875-4881
Number of pages7
JournalJournal of Immunology
Volume173
Issue number8
DOIs
StatePublished - Oct 15 2004

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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