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Neisseria gonorrhoeae Modulates Immunity by Polarizing Human Macrophages to a M2 Profile.

Ortiz MC, Lefimil C, Rodas PI, Vernal R, Lopez M, Acuña-Castillo C, Imarai M, Escobar A - PLoS ONE (2015)

Bottom Line: Until now there are no reports of the gonococcus effects on human MФ polarization.Moreover, functional results showed that N. gonorrhoeae-treated MФ are unable to induce proliferation of human T-cells, suggesting a more likely regulatory phenotype.This study has important implications for understanding the mechanisms of clearance versus long-term persistence of N. gonorroheae infection and might be applicable for the development of new therapeutic strategies.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Investigación en Ciencias Odontológicas, Facultad de Odontología. Universidad de Chile, Santiago, Chile.

ABSTRACT
Current data suggest that Neisseria gonorrhoeae is able to suppress the protective immune response at different levels, such as B and T lymphocytes and antigen-presenting cells. The present report is focused on gonococcus evasion mechanism on macrophages (MФ) and its impact in the subsequent immune response. In response to various signals MФ may undergo classical-M1 (M1-MФ) or alternative-M2 (M2-MФ) activation. Until now there are no reports of the gonococcus effects on human MФ polarization. We assessed the phagocytic ability of monocyte-derived MФ (MDM) upon gonococcal infection by immunofluorescence and gentamicin protection experiments. Then, we evaluated cytokine profile and M1/M2 specific-surface markers on MФ challenged with N. gonorrhoeae and their proliferative effect on T cells. Our findings lead us to suggest N. gonorrhoeae stimulates a M2-MФ phenotype in which some of the M2b and none of the M1-MФ-associated markers are induced. Interestingly, N. gonorrhoeae exposure leads to upregulation of a Programmed Death Ligand 1 (PD-L1), widely known as an immunosuppressive molecule. Moreover, functional results showed that N. gonorrhoeae-treated MФ are unable to induce proliferation of human T-cells, suggesting a more likely regulatory phenotype. Taken together, our data show that N. gonorroheae interferes with MФ polarization. This study has important implications for understanding the mechanisms of clearance versus long-term persistence of N. gonorroheae infection and might be applicable for the development of new therapeutic strategies.

No MeSH data available.


Related in: MedlinePlus

Hyporesponsive alloantigen T-cell responses induced by MФ infected with N. gonorrhoeae.CFSE-labeled CD4+ cells proliferation after non-adherent cells were co-cultured with human MФ treated for 24 hours with LPS-IFN-γ (M1-MΦ), IL-4 (M2-MΦ) or N. gonorrhoeae (MOI = 10, 100, 1000) for 7 days at the ratio of 2:1. As a positive control of proliferation we used PBL stimulated with 150 U/mL of IL-2 and 20 μg/mL of anti-CD3 (OKT-3). (A) Representative T CD4+ cell proliferation dot plots from one of the donors are shown. (B) CD4 + cells proliferation average under different conditions. ** p < 0.01 indicates that only M1-MФ profile is able to significantly induce proliferation of CD4+ cells in a mixed lymphocyte reaction.
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pone.0130713.g005: Hyporesponsive alloantigen T-cell responses induced by MФ infected with N. gonorrhoeae.CFSE-labeled CD4+ cells proliferation after non-adherent cells were co-cultured with human MФ treated for 24 hours with LPS-IFN-γ (M1-MΦ), IL-4 (M2-MΦ) or N. gonorrhoeae (MOI = 10, 100, 1000) for 7 days at the ratio of 2:1. As a positive control of proliferation we used PBL stimulated with 150 U/mL of IL-2 and 20 μg/mL of anti-CD3 (OKT-3). (A) Representative T CD4+ cell proliferation dot plots from one of the donors are shown. (B) CD4 + cells proliferation average under different conditions. ** p < 0.01 indicates that only M1-MФ profile is able to significantly induce proliferation of CD4+ cells in a mixed lymphocyte reaction.

Mentions: Since the surface markers and cytokines profile induced by N. gonorrhoeae on infected MΦ are well-matched with M2 profile, we addressed to study the capacity of GC-treated MФ to stimulate T cells. PBL from a single donor were co-cultured with GC-treated MФ (or M1/M2-MФ for the controls) from another donor in a mixed lymphocyte reaction (MLR) assay. After 7 days of co-culture with N. gonorrhoeae-treated MФ, CD4+ cells exhibited no significant proliferation, evaluated trough CFSE dilution, as compared to M1-MΦ-exposed cells (Fig 5). Although T cell proliferation was evaluated at three different MOIs (10, 100 and 1000), we did not observe significant differences between them.


Neisseria gonorrhoeae Modulates Immunity by Polarizing Human Macrophages to a M2 Profile.

Ortiz MC, Lefimil C, Rodas PI, Vernal R, Lopez M, Acuña-Castillo C, Imarai M, Escobar A - PLoS ONE (2015)

Hyporesponsive alloantigen T-cell responses induced by MФ infected with N. gonorrhoeae.CFSE-labeled CD4+ cells proliferation after non-adherent cells were co-cultured with human MФ treated for 24 hours with LPS-IFN-γ (M1-MΦ), IL-4 (M2-MΦ) or N. gonorrhoeae (MOI = 10, 100, 1000) for 7 days at the ratio of 2:1. As a positive control of proliferation we used PBL stimulated with 150 U/mL of IL-2 and 20 μg/mL of anti-CD3 (OKT-3). (A) Representative T CD4+ cell proliferation dot plots from one of the donors are shown. (B) CD4 + cells proliferation average under different conditions. ** p < 0.01 indicates that only M1-MФ profile is able to significantly induce proliferation of CD4+ cells in a mixed lymphocyte reaction.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4488386&req=5

pone.0130713.g005: Hyporesponsive alloantigen T-cell responses induced by MФ infected with N. gonorrhoeae.CFSE-labeled CD4+ cells proliferation after non-adherent cells were co-cultured with human MФ treated for 24 hours with LPS-IFN-γ (M1-MΦ), IL-4 (M2-MΦ) or N. gonorrhoeae (MOI = 10, 100, 1000) for 7 days at the ratio of 2:1. As a positive control of proliferation we used PBL stimulated with 150 U/mL of IL-2 and 20 μg/mL of anti-CD3 (OKT-3). (A) Representative T CD4+ cell proliferation dot plots from one of the donors are shown. (B) CD4 + cells proliferation average under different conditions. ** p < 0.01 indicates that only M1-MФ profile is able to significantly induce proliferation of CD4+ cells in a mixed lymphocyte reaction.
Mentions: Since the surface markers and cytokines profile induced by N. gonorrhoeae on infected MΦ are well-matched with M2 profile, we addressed to study the capacity of GC-treated MФ to stimulate T cells. PBL from a single donor were co-cultured with GC-treated MФ (or M1/M2-MФ for the controls) from another donor in a mixed lymphocyte reaction (MLR) assay. After 7 days of co-culture with N. gonorrhoeae-treated MФ, CD4+ cells exhibited no significant proliferation, evaluated trough CFSE dilution, as compared to M1-MΦ-exposed cells (Fig 5). Although T cell proliferation was evaluated at three different MOIs (10, 100 and 1000), we did not observe significant differences between them.

Bottom Line: Until now there are no reports of the gonococcus effects on human MФ polarization.Moreover, functional results showed that N. gonorrhoeae-treated MФ are unable to induce proliferation of human T-cells, suggesting a more likely regulatory phenotype.This study has important implications for understanding the mechanisms of clearance versus long-term persistence of N. gonorroheae infection and might be applicable for the development of new therapeutic strategies.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Investigación en Ciencias Odontológicas, Facultad de Odontología. Universidad de Chile, Santiago, Chile.

ABSTRACT
Current data suggest that Neisseria gonorrhoeae is able to suppress the protective immune response at different levels, such as B and T lymphocytes and antigen-presenting cells. The present report is focused on gonococcus evasion mechanism on macrophages (MФ) and its impact in the subsequent immune response. In response to various signals MФ may undergo classical-M1 (M1-MФ) or alternative-M2 (M2-MФ) activation. Until now there are no reports of the gonococcus effects on human MФ polarization. We assessed the phagocytic ability of monocyte-derived MФ (MDM) upon gonococcal infection by immunofluorescence and gentamicin protection experiments. Then, we evaluated cytokine profile and M1/M2 specific-surface markers on MФ challenged with N. gonorrhoeae and their proliferative effect on T cells. Our findings lead us to suggest N. gonorrhoeae stimulates a M2-MФ phenotype in which some of the M2b and none of the M1-MФ-associated markers are induced. Interestingly, N. gonorrhoeae exposure leads to upregulation of a Programmed Death Ligand 1 (PD-L1), widely known as an immunosuppressive molecule. Moreover, functional results showed that N. gonorrhoeae-treated MФ are unable to induce proliferation of human T-cells, suggesting a more likely regulatory phenotype. Taken together, our data show that N. gonorroheae interferes with MФ polarization. This study has important implications for understanding the mechanisms of clearance versus long-term persistence of N. gonorroheae infection and might be applicable for the development of new therapeutic strategies.

No MeSH data available.


Related in: MedlinePlus