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Culture of mouse peritoneal macrophages with mouse serum induces lipid bodies that associate with the parasitophorous vacuole and decrease their microbicidal capacity against Toxoplasma gondii.

Mota LA, Roberto Neto J, Monteiro VG, Lobato CS, Oliveira MA, Cunha Md, D'Ávila H, Seabra SH, Bozza PT, DaMatta RA - Mem. Inst. Oswaldo Cruz (2014)

Bottom Line: LBs were found in association with the parasitophorous vacuole, suggesting that T. gondii may benefit from this lipid source.Moreover, increased numbers of macrophage LBs correlated with high prostaglandin E2 (PGE2) production and decreased nitric oxide (NO) synthesis.Collectively, these results suggest that culture with MS caused a decrease in microbicidal activity of macrophages against T. gondii by increasing PGE2 while lowering NO production.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Biologia Celular e Tecidual, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ, Brasil.

ABSTRACT
Lipid bodies [lipid droplets (LBs)] are lipid-rich organelles involved in lipid metabolism, signalling and inflammation. Recent findings suggest a role for LBs in host response to infection; however, the potential functions of this organelle in Toxoplasma gondii infection and how it alters macrophage microbicidal capacity during infection are not well understood. Here, we investigated the role of host LBs in T. gondii infection in mouse peritoneal macrophages in vitro. Macrophages cultured with mouse serum (MS) had higher numbers of LBs than those cultured in foetal bovine serum and can function as a model to study the role of LBs during intracellular pathogen infection. LBs were found in association with the parasitophorous vacuole, suggesting that T. gondii may benefit from this lipid source. Moreover, increased numbers of macrophage LBs correlated with high prostaglandin E2 (PGE2) production and decreased nitric oxide (NO) synthesis. Accordingly, LB-enriched macrophages cultured with MS were less efficient at controlling T. gondii growth. Treatment of macrophages cultured with MS with indomethacin, an inhibitor of PGE2 production, increased the microbicidal capacity against T. gondii. Collectively, these results suggest that culture with MS caused a decrease in microbicidal activity of macrophages against T. gondii by increasing PGE2 while lowering NO production.

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: nitric oxide (NO) (μM) (A) and prostaglandin E2 (PGE2) (ng/mL) (B)production of activated macrophages cultured with foetal bovine serum (blackbars) or mouse serum (MS) (white bars). After 1, 24 or 48 h of culture,macrophages were activated with interferon-γ and lipopolysaccharide andmediators evaluated in the supernatant after 24 h of culture. Asterisks meansignificantly different from respective values for macrophages cultured with MSas calculated by the Student t test (p > 0.05). Onerepresentative experiment out of three.
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f07: : nitric oxide (NO) (μM) (A) and prostaglandin E2 (PGE2) (ng/mL) (B)production of activated macrophages cultured with foetal bovine serum (blackbars) or mouse serum (MS) (white bars). After 1, 24 or 48 h of culture,macrophages were activated with interferon-γ and lipopolysaccharide andmediators evaluated in the supernatant after 24 h of culture. Asterisks meansignificantly different from respective values for macrophages cultured with MSas calculated by the Student t test (p > 0.05). Onerepresentative experiment out of three.

Mentions: Macrophages cultured in MS showed decreased microbicidal against T.gondii - Next, the effect of culturing macrophages in different sera on theproduction of NO and PGE2 and their microbicidal capacity against T.gondii was determined. Because LBs were present in low numbers inapproximately 50% of the recently obtained macrophages, these cells were cultured for 1,24 and 48 h with both sera, classically activated and after 24 h, and NO andPGE2 production in the culture supernatants were measured. At all threetime points, MoMS produced significantly less NO than MoFBS (Fig. 7A). Furthermore, NO production decreased over timeindependently of the serum used (Fig. 7A). Incontrast, PGE2 production in MoMS increased significantly, reaching aproduction peak when macrophages were cultured for 24 h (Fig. 7B).


Culture of mouse peritoneal macrophages with mouse serum induces lipid bodies that associate with the parasitophorous vacuole and decrease their microbicidal capacity against Toxoplasma gondii.

Mota LA, Roberto Neto J, Monteiro VG, Lobato CS, Oliveira MA, Cunha Md, D'Ávila H, Seabra SH, Bozza PT, DaMatta RA - Mem. Inst. Oswaldo Cruz (2014)

: nitric oxide (NO) (μM) (A) and prostaglandin E2 (PGE2) (ng/mL) (B)production of activated macrophages cultured with foetal bovine serum (blackbars) or mouse serum (MS) (white bars). After 1, 24 or 48 h of culture,macrophages were activated with interferon-γ and lipopolysaccharide andmediators evaluated in the supernatant after 24 h of culture. Asterisks meansignificantly different from respective values for macrophages cultured with MSas calculated by the Student t test (p > 0.05). Onerepresentative experiment out of three.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4238769&req=5

f07: : nitric oxide (NO) (μM) (A) and prostaglandin E2 (PGE2) (ng/mL) (B)production of activated macrophages cultured with foetal bovine serum (blackbars) or mouse serum (MS) (white bars). After 1, 24 or 48 h of culture,macrophages were activated with interferon-γ and lipopolysaccharide andmediators evaluated in the supernatant after 24 h of culture. Asterisks meansignificantly different from respective values for macrophages cultured with MSas calculated by the Student t test (p > 0.05). Onerepresentative experiment out of three.
Mentions: Macrophages cultured in MS showed decreased microbicidal against T.gondii - Next, the effect of culturing macrophages in different sera on theproduction of NO and PGE2 and their microbicidal capacity against T.gondii was determined. Because LBs were present in low numbers inapproximately 50% of the recently obtained macrophages, these cells were cultured for 1,24 and 48 h with both sera, classically activated and after 24 h, and NO andPGE2 production in the culture supernatants were measured. At all threetime points, MoMS produced significantly less NO than MoFBS (Fig. 7A). Furthermore, NO production decreased over timeindependently of the serum used (Fig. 7A). Incontrast, PGE2 production in MoMS increased significantly, reaching aproduction peak when macrophages were cultured for 24 h (Fig. 7B).

Bottom Line: LBs were found in association with the parasitophorous vacuole, suggesting that T. gondii may benefit from this lipid source.Moreover, increased numbers of macrophage LBs correlated with high prostaglandin E2 (PGE2) production and decreased nitric oxide (NO) synthesis.Collectively, these results suggest that culture with MS caused a decrease in microbicidal activity of macrophages against T. gondii by increasing PGE2 while lowering NO production.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Biologia Celular e Tecidual, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ, Brasil.

ABSTRACT
Lipid bodies [lipid droplets (LBs)] are lipid-rich organelles involved in lipid metabolism, signalling and inflammation. Recent findings suggest a role for LBs in host response to infection; however, the potential functions of this organelle in Toxoplasma gondii infection and how it alters macrophage microbicidal capacity during infection are not well understood. Here, we investigated the role of host LBs in T. gondii infection in mouse peritoneal macrophages in vitro. Macrophages cultured with mouse serum (MS) had higher numbers of LBs than those cultured in foetal bovine serum and can function as a model to study the role of LBs during intracellular pathogen infection. LBs were found in association with the parasitophorous vacuole, suggesting that T. gondii may benefit from this lipid source. Moreover, increased numbers of macrophage LBs correlated with high prostaglandin E2 (PGE2) production and decreased nitric oxide (NO) synthesis. Accordingly, LB-enriched macrophages cultured with MS were less efficient at controlling T. gondii growth. Treatment of macrophages cultured with MS with indomethacin, an inhibitor of PGE2 production, increased the microbicidal capacity against T. gondii. Collectively, these results suggest that culture with MS caused a decrease in microbicidal activity of macrophages against T. gondii by increasing PGE2 while lowering NO production.

Show MeSH
Related in: MedlinePlus