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Blood, 1 February 2005, Vol. 105, No. 3, pp. 1319-1328. Prepublished online as a Blood First Edition Paper on September 21, 2004; DOI 10.1182/blood-2004-05-1749. This Article Full Text (PDF) Supplemental Materials and Methods All Versions of this Article: 2004-05-1749v1 105/3/1319    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Schleicher, U. Articles by Bogdan, C. Search for Related Content PubMed PubMed Citation Articles by Schleicher, U. Articles by Bogdan, C. Social Bookmarking                   What's this?  Previous Article  |  Next Article  Submitted May 6, 2004 Accepted September 15, 2004 Minute numbers of contaminant CD8+ T cells or CD11b+CD11c+ NK cells are the source of IFN- in IL-12/IL-18-stimulated mouse macrophage populations Ulrike Schleicher, Andrea Hesse, and Christian Bogdan* Institute of Clinical Microbiology, Immunology and Hygiene, University of Erlangen, Erlangen, Germany; Institute of Medical Microbiology and Hygiene, University of Freiburg, Freiburg, Germany * Corresponding author; email: christian.bogdan{at}uniklinik-freiburg.de. Macrophages were reported to be strong producers of IFN after stimulation by IL-12 plus IL-18, which gave rise to a novel concept of autocrine macrophage activation. Here, we show that peritoneal exudate and bone marrow-derived mouse macrophages generated by conventional techniques contain small quantities of CD11b+CD11c+CD31+DX5+NK1.1+ NK cells or CD3+CD8+TCR++ T cells, respectively. Intracellular cytokine staining, purification of macrophages by sorting, and the analysis of macrophages from alymphoid RAG2-/--chain-/- mice revealed that the high amount of IFN protein in the supernatants of unseparated IL-12/IL-18-stimulated macrophage populations originates exclusively from the contaminating lymphoid cells. Notably, IL-12/IL-18 still induced IFN mRNA in highly purified macrophages from wild-type mice and in macrophages from RAG2-/--chain-/- mice, whereas nuclear translocation of STAT4 and production of IFN protein were no longer detectable. These results question the concept of autocrine macrophage activation by secreted IFN, suggest differences in the expression of IFN mRNA and protein between macrophages and lymphoid cells, and illustrate that the limited purity of most myeloid cell populations (98%) might lead to false conclusions. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? 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