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© by The Society for Leukocyte Biology
Journal of Leukocyte Biology, doi:10.1189/jlb0302124

Received for publication March 12, 2002.
Revised May 19, 2003.
Accepted for publication June 4, 2003.

Article

Immature macrophages derived from mouse bone marrow produce large amounts of IL-12p40 after LPS stimulation

M. A. P. Oliveira *{ddagger}, G. M. A. C. Lima *, M. T. Shio *, P. J. M. Leenen {dagger}, and I. A. Abrahamsohn *@

*Departamento de Imunologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, SP, Brasil; {dagger}Department of Immunology, Erasmus MC, Rotterdam, The Netherlands; {ddagger}Present address: Departamento de Microbiologia, Imunologia, Parasitologia e Patologia, Instituto de Patologia Tropical e Saúde Pública, Universidade Federal de Goiás, Goiânia, GO, Brasil

@ To whom correspondence should be addressed. E-mail: iabraham{at}usp.br.


  Abstract

Production of IL-12 is an important indicator of the macrophage's ability to regulate immune responses. In this study, we investigated the IL-12 production by macrophages in different developmental stages. To this end, macrophages were generated in vitro from precursors stimulated with M-CSF, GM-CSF or IL-3. Density separation yielded populations enriched in different maturation stages. Invariably, only cells banding at the 40-50% Percoll interface produced large amounts of IL-12p40 when stimulated with LPS, whereas only low levels of IL-12p70 were produced. These cells represented immature macrophages, as indicated by the absence of precursor markers CD31/ER-MP12, Ly-6C/ER-MP20 and ER-MP58, and by the low level of expression of mature-cell markers like ER-HR3, scavenger receptor and CD11b/Mac-1. Upon further maturation, the macrophages' ability to produce IL-12p40 decreased, coinciding with increased nitric oxide production upon LPS stimulation. These results show that immature macrophages produce high levels of IL-12p40 and thus may either contribute to IL-12p70 production or regulate it.

Key Words: monocyte/macrophage differentiation • IL-12 • nitric oxide • M-CSF • GM-CSF • IL-3




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