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Characterization and partial purification of Candida albicans Secretory IL-12 Inhibitory Factor.

Wang M, Mukherjee PK, Chandra J, Lattif AA, McCormick TS, Ghannoum MA - BMC Microbiol. (2008)

Bottom Line: The minimal inhibitory dose of CA-SIIF was found to be 200 mug/ml.CA-SIIF-GP produced a higher inhibitory effect on IL-12 production compared to CA-SIIF-NGP and CA-SIIF crude (P < 0.01), proving that CA-SIIF is a glycoprotein in nature.These results suggest important role for CA-SIIF in interactions of C. albicans with the host immune system.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center for Medical Mycology, Department of Dermatology, University Hospitals of Cleveland and Case Western Reserve University, Cleveland, Ohio, USA. mingyuewang@gmail.com

ABSTRACT

Background: We have previously shown that supernatant from Candida albicans (CA) culture contains a Secretory Interleukin (IL)-12 Inhibitory Factor (CA-SIIF), which inhibits IL-12 production by human monocytes. However, the effect of CA-SIIF on secretion of other cytokines by monocytes is unknown, and detailed characterization of this factor has not been performed.

Results: In this study, we demonstrate that the IL-12 inhibitory activity of CA-SIIF was serum-independent, based on the reduction of IL-12 levels in monocytes stimulated under serum-independent conditions. The minimal inhibitory dose of CA-SIIF was found to be 200 mug/ml. Investigation of CA-SIIF's effect on macrophages IL-12 production in vitro and in vivo also showed that CA-SIIF inhibited IL-12 production by murine macrophages both in vitro (from 571 +/- 24 pg/ml to 387 +/- 87 pg/ml; P = 0.05) and in vivo (from 262 +/- 6 pg/ml to 144 +/- 30 pg/ml; P < 0.05). In addition to IL-12, cytokine array analysis revealed that CA-SIIF induced differential production of other cytokines also. In this regard, reduction in levels were observed for IL-8, IL-10, IL-13, monocyte chemoattractant protein (MCP)-1, MCP-2, macrophage inflammatory protein (MIP)-1, RANTES, etc. In contrast, levels of other chemokines e.g. MCP-4, MIF and MIP-3alpha (P < 0.05) were increased. We also found that CA-SIIF suppressed the maturation of human monocytes to dendritic cells (CD1a expression = 13 +/- 3% vs 36 +/- 2% of the control; P < 0.01). Next, to identify the biochemical nature of CA-SIIF, we separated this factor into a Concanavalin A (ConA)-binding glycoprotein fraction (CA-SIIF-GP) and a non-ConA-binding protein fraction (CA-SIIF-NGP) using ConA affinity chromatography. Both fractions were then tested for this inhibitory effect on human monocyte IL-12 production. CA-SIIF-GP produced a higher inhibitory effect on IL-12 production compared to CA-SIIF-NGP and CA-SIIF crude (P < 0.01), proving that CA-SIIF is a glycoprotein in nature.

Conclusion: CA-SIIF is a glycoprotein which exhibits serum-independent inhibition of IL-12 production from monocytes in vitro and in vivo, and also modulates differentiation of monocytes into dendritic cells. These results suggest important role for CA-SIIF in interactions of C. albicans with the host immune system.

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CA-SIIF dramatically reduces CD1a expression on human monocyte-derived dendritic cells. (A) Impact of CA-SIIF on GM-CSF/IL-4 induced monocyte-derived dendritic cells. (a) CD14 expression on fresh monocytes, (b) CD1a expression on monocytes after 5 day culture with GM-CSF/IL-4 in absence of CA-SIIF, (c) CD1a expression on monocytes after 5 day culture with GM-CSF/IL-4 in presence of CA-SIIF. Flow histograms are representative of 3 independent experiments. (B) Cumulative dramatic reduction of CD1a expression on human monocytes-derived dendritic cells cultured with CA-SIIF. Cell density: 5 × 105/ml. n = 3.
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Figure 5: CA-SIIF dramatically reduces CD1a expression on human monocyte-derived dendritic cells. (A) Impact of CA-SIIF on GM-CSF/IL-4 induced monocyte-derived dendritic cells. (a) CD14 expression on fresh monocytes, (b) CD1a expression on monocytes after 5 day culture with GM-CSF/IL-4 in absence of CA-SIIF, (c) CD1a expression on monocytes after 5 day culture with GM-CSF/IL-4 in presence of CA-SIIF. Flow histograms are representative of 3 independent experiments. (B) Cumulative dramatic reduction of CD1a expression on human monocytes-derived dendritic cells cultured with CA-SIIF. Cell density: 5 × 105/ml. n = 3.

Mentions: Our previous data suggested CA-SIIF IL-12 inhibition involves the Extracellular signal-regulated Kinase (ERK) Mitogen-Activated Protein Kinase (MAPK) signaling pathway [8]. Since ERK and p38 MAPK signaling pathways are known to differentially regulate the maturation of monocyte-derived human dendritic cells [13], we hypothesized that CA-SIIF inhibits differentiation of human monocytes into dendritic cells. As shown in Figure 5, flow cytometry analysis revealed significantly reduced expression of CD1a (a surface marker specific for dendritic cells) in monocytes incubated with CA-SIIF compared to control media (13 ± 3% vs 36 ± 2%; P < 0.01). These studies demonstrated that CA-SIIF inhibits differentiation of human monocytes to dendritic cells.


Characterization and partial purification of Candida albicans Secretory IL-12 Inhibitory Factor.

Wang M, Mukherjee PK, Chandra J, Lattif AA, McCormick TS, Ghannoum MA - BMC Microbiol. (2008)

CA-SIIF dramatically reduces CD1a expression on human monocyte-derived dendritic cells. (A) Impact of CA-SIIF on GM-CSF/IL-4 induced monocyte-derived dendritic cells. (a) CD14 expression on fresh monocytes, (b) CD1a expression on monocytes after 5 day culture with GM-CSF/IL-4 in absence of CA-SIIF, (c) CD1a expression on monocytes after 5 day culture with GM-CSF/IL-4 in presence of CA-SIIF. Flow histograms are representative of 3 independent experiments. (B) Cumulative dramatic reduction of CD1a expression on human monocytes-derived dendritic cells cultured with CA-SIIF. Cell density: 5 × 105/ml. n = 3.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2289826&req=5

Figure 5: CA-SIIF dramatically reduces CD1a expression on human monocyte-derived dendritic cells. (A) Impact of CA-SIIF on GM-CSF/IL-4 induced monocyte-derived dendritic cells. (a) CD14 expression on fresh monocytes, (b) CD1a expression on monocytes after 5 day culture with GM-CSF/IL-4 in absence of CA-SIIF, (c) CD1a expression on monocytes after 5 day culture with GM-CSF/IL-4 in presence of CA-SIIF. Flow histograms are representative of 3 independent experiments. (B) Cumulative dramatic reduction of CD1a expression on human monocytes-derived dendritic cells cultured with CA-SIIF. Cell density: 5 × 105/ml. n = 3.
Mentions: Our previous data suggested CA-SIIF IL-12 inhibition involves the Extracellular signal-regulated Kinase (ERK) Mitogen-Activated Protein Kinase (MAPK) signaling pathway [8]. Since ERK and p38 MAPK signaling pathways are known to differentially regulate the maturation of monocyte-derived human dendritic cells [13], we hypothesized that CA-SIIF inhibits differentiation of human monocytes into dendritic cells. As shown in Figure 5, flow cytometry analysis revealed significantly reduced expression of CD1a (a surface marker specific for dendritic cells) in monocytes incubated with CA-SIIF compared to control media (13 ± 3% vs 36 ± 2%; P < 0.01). These studies demonstrated that CA-SIIF inhibits differentiation of human monocytes to dendritic cells.

Bottom Line: The minimal inhibitory dose of CA-SIIF was found to be 200 mug/ml.CA-SIIF-GP produced a higher inhibitory effect on IL-12 production compared to CA-SIIF-NGP and CA-SIIF crude (P < 0.01), proving that CA-SIIF is a glycoprotein in nature.These results suggest important role for CA-SIIF in interactions of C. albicans with the host immune system.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center for Medical Mycology, Department of Dermatology, University Hospitals of Cleveland and Case Western Reserve University, Cleveland, Ohio, USA. mingyuewang@gmail.com

ABSTRACT

Background: We have previously shown that supernatant from Candida albicans (CA) culture contains a Secretory Interleukin (IL)-12 Inhibitory Factor (CA-SIIF), which inhibits IL-12 production by human monocytes. However, the effect of CA-SIIF on secretion of other cytokines by monocytes is unknown, and detailed characterization of this factor has not been performed.

Results: In this study, we demonstrate that the IL-12 inhibitory activity of CA-SIIF was serum-independent, based on the reduction of IL-12 levels in monocytes stimulated under serum-independent conditions. The minimal inhibitory dose of CA-SIIF was found to be 200 mug/ml. Investigation of CA-SIIF's effect on macrophages IL-12 production in vitro and in vivo also showed that CA-SIIF inhibited IL-12 production by murine macrophages both in vitro (from 571 +/- 24 pg/ml to 387 +/- 87 pg/ml; P = 0.05) and in vivo (from 262 +/- 6 pg/ml to 144 +/- 30 pg/ml; P < 0.05). In addition to IL-12, cytokine array analysis revealed that CA-SIIF induced differential production of other cytokines also. In this regard, reduction in levels were observed for IL-8, IL-10, IL-13, monocyte chemoattractant protein (MCP)-1, MCP-2, macrophage inflammatory protein (MIP)-1, RANTES, etc. In contrast, levels of other chemokines e.g. MCP-4, MIF and MIP-3alpha (P < 0.05) were increased. We also found that CA-SIIF suppressed the maturation of human monocytes to dendritic cells (CD1a expression = 13 +/- 3% vs 36 +/- 2% of the control; P < 0.01). Next, to identify the biochemical nature of CA-SIIF, we separated this factor into a Concanavalin A (ConA)-binding glycoprotein fraction (CA-SIIF-GP) and a non-ConA-binding protein fraction (CA-SIIF-NGP) using ConA affinity chromatography. Both fractions were then tested for this inhibitory effect on human monocyte IL-12 production. CA-SIIF-GP produced a higher inhibitory effect on IL-12 production compared to CA-SIIF-NGP and CA-SIIF crude (P < 0.01), proving that CA-SIIF is a glycoprotein in nature.

Conclusion: CA-SIIF is a glycoprotein which exhibits serum-independent inhibition of IL-12 production from monocytes in vitro and in vivo, and also modulates differentiation of monocytes into dendritic cells. These results suggest important role for CA-SIIF in interactions of C. albicans with the host immune system.

Show MeSH
Related in: MedlinePlus