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Interleukin-33 contributes to both M1 and M2 chemokine marker expression in human macrophages.

Joshi AD, Oak SR, Hartigan AJ, Finn WG, Kunkel SL, Duffy KE, Das A, Hogaboam CM - BMC Immunol. (2010)

Bottom Line: Human macrophages constitutively expressed the membrane-associated (i.e. ST2L) and the soluble (i.e. sST2) ST2 receptors.M2 (IL-4 + IL-13) skewing stimuli markedly increased the expression of ST2L, but neither polarizing cytokine treatment promoted the release of sST2 from these cells.Together, these data suggest that primary responses to IL-33 in bone marrow derived human macrophages favors M1 chemokine generation while its addition to polarized human macrophages promotes or amplifies M2 chemokine expression.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, University of Michigan Medical School, Ann Arbor, MI, USA.

ABSTRACT

Background: Interleukin-33 is a member of the IL-1 cytokine family whose functions are mediated and modulated by the ST2 receptor. IL-33-ST2 expression and interactions have been explored in mouse macrophages but little is known about the effect of IL-33 on human macrophages. The expression of ST2 transcript and protein levels, and IL-33-mediated effects on M1 (i.e. classical activation) and M2 (i.e. alternative activation) chemokine marker expression in human bone marrow-derived macrophages were examined.

Results: Human macrophages constitutively expressed the membrane-associated (i.e. ST2L) and the soluble (i.e. sST2) ST2 receptors. M2 (IL-4 + IL-13) skewing stimuli markedly increased the expression of ST2L, but neither polarizing cytokine treatment promoted the release of sST2 from these cells. When added to naïve macrophages alone, IL-33 directly enhanced the expression of CCL3. In combination with LPS, IL-33 blocked the expression of the M2 chemokine marker CCL18, but did not alter CCL3 expression in these naive cells. The addition of IL-33 to M1 macrophages markedly increased the expression of CCL18 above that detected in untreated M1 macrophages. Similarly, alternatively activated human macrophages treated with IL-33 exhibited enhanced expression of CCL18 and the M2 marker mannose receptor above that detected in M2 macrophages alone.

Conclusions: Together, these data suggest that primary responses to IL-33 in bone marrow derived human macrophages favors M1 chemokine generation while its addition to polarized human macrophages promotes or amplifies M2 chemokine expression.

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Effect of IL-33 on human macrophages previously exposed to either M1 or M2 conditions. M1- (i.e. CCR7 and CCL3) and M2- (i.e. CCL18 and mannose receptor) associated markers in human macrophages previously exposed to M1 (A) or M2 (B) conditions for 24 h, which were secondarily challenged with IL-33 alone (10 ng/ml), IL-33 + M1 or M2 polarizing mediators, or M1 or M2 polarizing mediators for an additional 24 h. Results are expressed as fold increase in transcript levels compared with levels of these transcripts in human macrophages exposed to media alone (i.e. control condition). Data shown are mean ± SEM, and these data are representative of three separate experiments with 3 donor macrophage lines. ** P ≤ 0.01 compared with transcript levels in appropriate control macrophages.
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Figure 6: Effect of IL-33 on human macrophages previously exposed to either M1 or M2 conditions. M1- (i.e. CCR7 and CCL3) and M2- (i.e. CCL18 and mannose receptor) associated markers in human macrophages previously exposed to M1 (A) or M2 (B) conditions for 24 h, which were secondarily challenged with IL-33 alone (10 ng/ml), IL-33 + M1 or M2 polarizing mediators, or M1 or M2 polarizing mediators for an additional 24 h. Results are expressed as fold increase in transcript levels compared with levels of these transcripts in human macrophages exposed to media alone (i.e. control condition). Data shown are mean ± SEM, and these data are representative of three separate experiments with 3 donor macrophage lines. ** P ≤ 0.01 compared with transcript levels in appropriate control macrophages.

Mentions: To investigate the effect of IL-33 on human macrophages following their exposure for 24 h to either M1 or M2 polarizing conditions, IL-33 alone, IL-33 with M1 or M2 polarizing mediators, or M1 or M2 polarizing mediators alone were added for an additional 24 h to primary human macrophage cultures. As shown in Figure 6A, the addition of IL-33 alone macrophages exposed to M1 mediators did not induce the expression of CCR7 to the same magnitude as that observed in M1 polarized cultures exposed to M1 mediators with IL-33 or M1 mediators. Similar results were observed with CCL3 transcript expression although IL-33 alone did not induce CCL3 transcript expression in M1 polarized human macrophages (Figure 6A). However, the presence of IL-33 with M1 polarizing mediators significantly increased the expression of the M2 marker CCL18 in macrophages compared with M1 macrophages alone (Figure 6A). In cultures of M2 polarized macrophages, transcript levels of CCL18 were markedly induced after the secondary addition of IL-33 alone or IL-33 with M2 mediators for an additional 24 h (Figure 6B). Interestingly, the secondary addition of M2 mediators did not further enhance CCL18 expression in M2 polarized macrophages (Figure 6B). The effect of IL-33 on mannose receptor transcript expression was also examined and similar results were observed; the secondary addition of the combination of IL-33 and M2 mediators further enhanced mannose receptor expression on M2 polarized macrophages. Overall, these data suggest that IL-33 appears to promote M2 chemokine expression in polarized macrophages since it promoted CCL18 in M1 human macrophages and appears to further enhance the expression of M2 markers in M2-polarized macrophages.


Interleukin-33 contributes to both M1 and M2 chemokine marker expression in human macrophages.

Joshi AD, Oak SR, Hartigan AJ, Finn WG, Kunkel SL, Duffy KE, Das A, Hogaboam CM - BMC Immunol. (2010)

Effect of IL-33 on human macrophages previously exposed to either M1 or M2 conditions. M1- (i.e. CCR7 and CCL3) and M2- (i.e. CCL18 and mannose receptor) associated markers in human macrophages previously exposed to M1 (A) or M2 (B) conditions for 24 h, which were secondarily challenged with IL-33 alone (10 ng/ml), IL-33 + M1 or M2 polarizing mediators, or M1 or M2 polarizing mediators for an additional 24 h. Results are expressed as fold increase in transcript levels compared with levels of these transcripts in human macrophages exposed to media alone (i.e. control condition). Data shown are mean ± SEM, and these data are representative of three separate experiments with 3 donor macrophage lines. ** P ≤ 0.01 compared with transcript levels in appropriate control macrophages.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2967528&req=5

Figure 6: Effect of IL-33 on human macrophages previously exposed to either M1 or M2 conditions. M1- (i.e. CCR7 and CCL3) and M2- (i.e. CCL18 and mannose receptor) associated markers in human macrophages previously exposed to M1 (A) or M2 (B) conditions for 24 h, which were secondarily challenged with IL-33 alone (10 ng/ml), IL-33 + M1 or M2 polarizing mediators, or M1 or M2 polarizing mediators for an additional 24 h. Results are expressed as fold increase in transcript levels compared with levels of these transcripts in human macrophages exposed to media alone (i.e. control condition). Data shown are mean ± SEM, and these data are representative of three separate experiments with 3 donor macrophage lines. ** P ≤ 0.01 compared with transcript levels in appropriate control macrophages.
Mentions: To investigate the effect of IL-33 on human macrophages following their exposure for 24 h to either M1 or M2 polarizing conditions, IL-33 alone, IL-33 with M1 or M2 polarizing mediators, or M1 or M2 polarizing mediators alone were added for an additional 24 h to primary human macrophage cultures. As shown in Figure 6A, the addition of IL-33 alone macrophages exposed to M1 mediators did not induce the expression of CCR7 to the same magnitude as that observed in M1 polarized cultures exposed to M1 mediators with IL-33 or M1 mediators. Similar results were observed with CCL3 transcript expression although IL-33 alone did not induce CCL3 transcript expression in M1 polarized human macrophages (Figure 6A). However, the presence of IL-33 with M1 polarizing mediators significantly increased the expression of the M2 marker CCL18 in macrophages compared with M1 macrophages alone (Figure 6A). In cultures of M2 polarized macrophages, transcript levels of CCL18 were markedly induced after the secondary addition of IL-33 alone or IL-33 with M2 mediators for an additional 24 h (Figure 6B). Interestingly, the secondary addition of M2 mediators did not further enhance CCL18 expression in M2 polarized macrophages (Figure 6B). The effect of IL-33 on mannose receptor transcript expression was also examined and similar results were observed; the secondary addition of the combination of IL-33 and M2 mediators further enhanced mannose receptor expression on M2 polarized macrophages. Overall, these data suggest that IL-33 appears to promote M2 chemokine expression in polarized macrophages since it promoted CCL18 in M1 human macrophages and appears to further enhance the expression of M2 markers in M2-polarized macrophages.

Bottom Line: Human macrophages constitutively expressed the membrane-associated (i.e. ST2L) and the soluble (i.e. sST2) ST2 receptors.M2 (IL-4 + IL-13) skewing stimuli markedly increased the expression of ST2L, but neither polarizing cytokine treatment promoted the release of sST2 from these cells.Together, these data suggest that primary responses to IL-33 in bone marrow derived human macrophages favors M1 chemokine generation while its addition to polarized human macrophages promotes or amplifies M2 chemokine expression.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, University of Michigan Medical School, Ann Arbor, MI, USA.

ABSTRACT

Background: Interleukin-33 is a member of the IL-1 cytokine family whose functions are mediated and modulated by the ST2 receptor. IL-33-ST2 expression and interactions have been explored in mouse macrophages but little is known about the effect of IL-33 on human macrophages. The expression of ST2 transcript and protein levels, and IL-33-mediated effects on M1 (i.e. classical activation) and M2 (i.e. alternative activation) chemokine marker expression in human bone marrow-derived macrophages were examined.

Results: Human macrophages constitutively expressed the membrane-associated (i.e. ST2L) and the soluble (i.e. sST2) ST2 receptors. M2 (IL-4 + IL-13) skewing stimuli markedly increased the expression of ST2L, but neither polarizing cytokine treatment promoted the release of sST2 from these cells. When added to naïve macrophages alone, IL-33 directly enhanced the expression of CCL3. In combination with LPS, IL-33 blocked the expression of the M2 chemokine marker CCL18, but did not alter CCL3 expression in these naive cells. The addition of IL-33 to M1 macrophages markedly increased the expression of CCL18 above that detected in untreated M1 macrophages. Similarly, alternatively activated human macrophages treated with IL-33 exhibited enhanced expression of CCL18 and the M2 marker mannose receptor above that detected in M2 macrophages alone.

Conclusions: Together, these data suggest that primary responses to IL-33 in bone marrow derived human macrophages favors M1 chemokine generation while its addition to polarized human macrophages promotes or amplifies M2 chemokine expression.

Show MeSH
Related in: MedlinePlus