Journal of Leukocyte Biology BioLegend: Treg, Th17, Stem Cell
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Originally published online as doi:10.1189/jlb0302124 on August 1, 2003

Published online before print August 1, 2003
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(Journal of Leukocyte Biology. 2003;74:857-867.)
© 2003 by Society for Leukocyte Biology

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*,1

* 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

1 Correspondence: Dept. Imunologia, ICB/USP Av. Prof. Lineu Prestes 1730, CEP 05508-900, SP, Brasil. Phone: 55-11-3091-7383; Fax: 55-11-3091-7224; E-mail: iabraham{at}usp.br

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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