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Published online before print August 11, 2006

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© by The Society for Leukocyte Biology
Journal of Leukocyte Biology, doi:10.1189/jlb.0406249

Received for publication April 5, 2006.
Revised June 2, 2006.
Accepted for publication June 27, 2006.

Article

Biochemical and functional characterization of three activated macrophage populations

Department of Cell Biology and Molecular Genetics, University of Maryland, College Park

^@ To whom correspondence should be addressed. E-mail: dmosser{at}umd.edu.

	Abstract

We generated three populations of macrophages (M) in vitro and characterized each. Classically activated M (Ca-M) were primed with IFN- and stimulated with LPS. Type II-activated M (M-II) were also similarly primed but stimulated with LPS plus immune complexes. Alternatively activated M (AA-M) were primed overnight with IL-4. Here, we present a side-by-side comparison of the three cell types but focus primarily on differences between M-II and AA-M, as both have been classified as M2 M, distinct from Ca-M. We show that M-II are more similar to Ca-M than they are to AA-M. M-II and Ca-M, but not AA-M, produce high levels of NO and have low arginase activity. AA-M express FIZZ1, whereas neither M-II nor Ca-M does. M-II and Ca-M express relatively high levels of CD86, whereas AA-M are virtually devoid of this costimulatory molecule. Ca-M and M-II are efficient APC, whereas AA-M fail to stimulate efficient T cell proliferation. The differences between Ca-M and M-II are more subtle. Ca-M produce IL-12 and give rise to Th1 cells, whereas M-II produce high levels of IL-10 and thus, give rise to Th2 cells secreting IL-4 and IL-10. M-II express two markers that may be used to identify them in tissue. These are sphingosine kinase-1 and homologous to lymphotoxins, shows inducible expression and competes with herpes simplex virus glycoprotein D for herpes virus entry mediator/TNF-related 2 (LIGHT; TNF superfamily 14), a member of the TNF superfamily. Thus, Ca-M, M-II, and AA-M represent three populations of cells with different biological functions.

Key Words: IL-10 • IL-12 • sphingosine kinase • LIGHT

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