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Naturally-acquired cellular immune response against Plasmodium vivax merozoite surface protein-1 paralog antigen.

Changrob S, Leepiyasakulchai C, Tsuboi T, Cheng Y, Lim CS, Chootong P, Han ET - Malar. J. (2015)

Bottom Line: Recombinant PvMSP1P antigen results in production of a significant Th1 cytokine response in immunized mice.PvMSP1P-19 strongly induces a specific cellular immune response for protection against P. vivax compared with PvDBPII as the antigen induces activation of IFN-γ-producing effector cells following natural P. vivax exposure.Upon stimulation, PvMSP1P-19 has the potential to activate the recall response of Th1 effector memory cells that play a role in killing the parasite.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Microbiology and Applied Technology, Faculty of Medical Technology, Mahidol University, Bangkok, Thailand. siriruk.ch@gmail.com.

ABSTRACT

Background: Plasmodium vivax merozoite surface protein-1 paralog (PvMSP1P) is a glycosylphosphatidylinositol-anchored protein expressed on the merozoite surface. This molecule is a target of natural immunity, as high anti-MSP1P-19 antibody levels were detected during P. vivax infection and the antibody inhibited PvMSP1P-erythrocyte binding. Recombinant PvMSP1P antigen results in production of a significant Th1 cytokine response in immunized mice. The present study was performed to characterize natural cellular immunity against PvMSP1P-19 and PvDBP region II in acute and recovery P. vivax infection.

Methods: Peripheral blood mononuclear cells (PBMCs) from acute and recovery P. vivax infection were obtained for lymphocyte proliferation assay upon PvMSP1P-19 and PvDBP region II antigen stimulation. The culture supernatant was examined for the presence of the cytokines IL-2, TNF, IFN-γ and IL-10 by enzyme-linked immunosorbent assay (ELISA). To determine whether Th1 or Th2 have a memory response against PvMSP1P-19 and PvDBPII protein antigen, PBMCs from subjects who had recovered from P. vivax infection 8-10 weeks prior to the study were obtained for lymphocyte proliferation assay. Cytokine-producing cells were analysed by flow cytometry.

Results: IL-2 was detected at high levels in lymphocyte cultures from acutely infected P. vivax patients upon PvMSP1P-19 stimulation. Analysis of the Th1 or Th2 memory response in PBMC cultures from subjects who had recovered from P. vivax infection showed significantly elevated levels of PvMSP1P-19 and PvDBPII-specific IFN-γ-producing cells (P  <  0.05). Interestingly, the response of IFN-γ-producing cells in PvMSP1P stimulation was fourfold greater in recovered subjects than that in acute-infection patients. CD4(+) T cells were the major cell phenotype involved in the response to PvMSP1P-19 and PvDBPII antigen.

Conclusions: PvMSP1P-19 strongly induces a specific cellular immune response for protection against P. vivax compared with PvDBPII as the antigen induces activation of IFN-γ-producing effector cells following natural P. vivax exposure. Upon stimulation, PvMSP1P-19 has the potential to activate the recall response of Th1 effector memory cells that play a role in killing the parasite.

No MeSH data available.


Related in: MedlinePlus

T-cell responses to PvMSP1P-19 antigen using multiparameter flow cytometry. Intracellular cytokine assay demonstrating the T-cell response of P. vivax-recovered subjects to PvMSP1P-19 or PvDBPII with negative (media) and positive controls (PMA/Ionomycin). PBMCs from individuals who had recovered from P. vivax infection 8–10 weeks prior to the study were removed for lymphocyte proliferation assay and intracellular cytokine detection by flow cytometric analysis. The data are shown as the average levels of cytokine-producing cells in individual subjects (n  =  6). This shows the gating strategy to identify IL-2/IFN-γ/IL-10-producing cells.
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Fig3: T-cell responses to PvMSP1P-19 antigen using multiparameter flow cytometry. Intracellular cytokine assay demonstrating the T-cell response of P. vivax-recovered subjects to PvMSP1P-19 or PvDBPII with negative (media) and positive controls (PMA/Ionomycin). PBMCs from individuals who had recovered from P. vivax infection 8–10 weeks prior to the study were removed for lymphocyte proliferation assay and intracellular cytokine detection by flow cytometric analysis. The data are shown as the average levels of cytokine-producing cells in individual subjects (n  =  6). This shows the gating strategy to identify IL-2/IFN-γ/IL-10-producing cells.

Mentions: Many different IFN-γ-producing cell subsets, including to αβ T cells, γδ T cells and NK cells, have been shown to be capable of responding to Plasmodium parasites. Here, the phenotype of cells involved in the IFN-γ and IL-10 response upon PvMSP1P-19 stimulation was evaluated by flow cytometric analysis (Figure 3). Upon PvMSP1P-19 stimulation, CD4+ T cells were the major source of IL-2 and IFN-γ production. PvMSP1P-19 stimulation showed significantly higher IL-2 and IFN-γ levels than the medium control (IL-2, PvMSP1P-19  =  0.070%  ±  0.041%, unstimulated  =  0.041%  ±  0.023%, P  <  0.05; IFN-γ, PvMSP1P-19  =  0.112%  ±  0.053%, unstimulated  =  0.037%  ±  0.022%, P  <  0.05, Figures 4a-b). The effector CD4+ IFN-γ+ response to PvMSP1P-19 antigen was double that of CD8+ T cells (Figure 4a). Similarly, PvDBP region II antigen also had a high potential to induce CD4+ IFN+ T cell response (PvDBPII  =  0.094%  ±  0.046%, unstimulated  =  0.037%  ±  0.022%, P  <  0.05, Figure 4b). Phenotyping of IL-10-producing cells upon PvMSP1P-19 stimulation showed that CD4+ and CD8+ T cells were not major sources of IL-10 (CD4+, PvMSP1P-19  =  0.008%  ±  0.012%, unstimulated  =  0.002%  ±  0.002%, CD8+, PvMSP1P-19  =  0.008%  ±  0.009%, unstimulated  =  0.005%  ±  0.007%, P  >  0.05, Figure 5). These data suggest that CD4+ T cells dominate over CD8+ T cells in the responses against PvMSP1P-19 and PvDBP region II antigens.Figure 3


Naturally-acquired cellular immune response against Plasmodium vivax merozoite surface protein-1 paralog antigen.

Changrob S, Leepiyasakulchai C, Tsuboi T, Cheng Y, Lim CS, Chootong P, Han ET - Malar. J. (2015)

T-cell responses to PvMSP1P-19 antigen using multiparameter flow cytometry. Intracellular cytokine assay demonstrating the T-cell response of P. vivax-recovered subjects to PvMSP1P-19 or PvDBPII with negative (media) and positive controls (PMA/Ionomycin). PBMCs from individuals who had recovered from P. vivax infection 8–10 weeks prior to the study were removed for lymphocyte proliferation assay and intracellular cytokine detection by flow cytometric analysis. The data are shown as the average levels of cytokine-producing cells in individual subjects (n  =  6). This shows the gating strategy to identify IL-2/IFN-γ/IL-10-producing cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4403936&req=5

Fig3: T-cell responses to PvMSP1P-19 antigen using multiparameter flow cytometry. Intracellular cytokine assay demonstrating the T-cell response of P. vivax-recovered subjects to PvMSP1P-19 or PvDBPII with negative (media) and positive controls (PMA/Ionomycin). PBMCs from individuals who had recovered from P. vivax infection 8–10 weeks prior to the study were removed for lymphocyte proliferation assay and intracellular cytokine detection by flow cytometric analysis. The data are shown as the average levels of cytokine-producing cells in individual subjects (n  =  6). This shows the gating strategy to identify IL-2/IFN-γ/IL-10-producing cells.
Mentions: Many different IFN-γ-producing cell subsets, including to αβ T cells, γδ T cells and NK cells, have been shown to be capable of responding to Plasmodium parasites. Here, the phenotype of cells involved in the IFN-γ and IL-10 response upon PvMSP1P-19 stimulation was evaluated by flow cytometric analysis (Figure 3). Upon PvMSP1P-19 stimulation, CD4+ T cells were the major source of IL-2 and IFN-γ production. PvMSP1P-19 stimulation showed significantly higher IL-2 and IFN-γ levels than the medium control (IL-2, PvMSP1P-19  =  0.070%  ±  0.041%, unstimulated  =  0.041%  ±  0.023%, P  <  0.05; IFN-γ, PvMSP1P-19  =  0.112%  ±  0.053%, unstimulated  =  0.037%  ±  0.022%, P  <  0.05, Figures 4a-b). The effector CD4+ IFN-γ+ response to PvMSP1P-19 antigen was double that of CD8+ T cells (Figure 4a). Similarly, PvDBP region II antigen also had a high potential to induce CD4+ IFN+ T cell response (PvDBPII  =  0.094%  ±  0.046%, unstimulated  =  0.037%  ±  0.022%, P  <  0.05, Figure 4b). Phenotyping of IL-10-producing cells upon PvMSP1P-19 stimulation showed that CD4+ and CD8+ T cells were not major sources of IL-10 (CD4+, PvMSP1P-19  =  0.008%  ±  0.012%, unstimulated  =  0.002%  ±  0.002%, CD8+, PvMSP1P-19  =  0.008%  ±  0.009%, unstimulated  =  0.005%  ±  0.007%, P  >  0.05, Figure 5). These data suggest that CD4+ T cells dominate over CD8+ T cells in the responses against PvMSP1P-19 and PvDBP region II antigens.Figure 3

Bottom Line: Recombinant PvMSP1P antigen results in production of a significant Th1 cytokine response in immunized mice.PvMSP1P-19 strongly induces a specific cellular immune response for protection against P. vivax compared with PvDBPII as the antigen induces activation of IFN-γ-producing effector cells following natural P. vivax exposure.Upon stimulation, PvMSP1P-19 has the potential to activate the recall response of Th1 effector memory cells that play a role in killing the parasite.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Microbiology and Applied Technology, Faculty of Medical Technology, Mahidol University, Bangkok, Thailand. siriruk.ch@gmail.com.

ABSTRACT

Background: Plasmodium vivax merozoite surface protein-1 paralog (PvMSP1P) is a glycosylphosphatidylinositol-anchored protein expressed on the merozoite surface. This molecule is a target of natural immunity, as high anti-MSP1P-19 antibody levels were detected during P. vivax infection and the antibody inhibited PvMSP1P-erythrocyte binding. Recombinant PvMSP1P antigen results in production of a significant Th1 cytokine response in immunized mice. The present study was performed to characterize natural cellular immunity against PvMSP1P-19 and PvDBP region II in acute and recovery P. vivax infection.

Methods: Peripheral blood mononuclear cells (PBMCs) from acute and recovery P. vivax infection were obtained for lymphocyte proliferation assay upon PvMSP1P-19 and PvDBP region II antigen stimulation. The culture supernatant was examined for the presence of the cytokines IL-2, TNF, IFN-γ and IL-10 by enzyme-linked immunosorbent assay (ELISA). To determine whether Th1 or Th2 have a memory response against PvMSP1P-19 and PvDBPII protein antigen, PBMCs from subjects who had recovered from P. vivax infection 8-10 weeks prior to the study were obtained for lymphocyte proliferation assay. Cytokine-producing cells were analysed by flow cytometry.

Results: IL-2 was detected at high levels in lymphocyte cultures from acutely infected P. vivax patients upon PvMSP1P-19 stimulation. Analysis of the Th1 or Th2 memory response in PBMC cultures from subjects who had recovered from P. vivax infection showed significantly elevated levels of PvMSP1P-19 and PvDBPII-specific IFN-γ-producing cells (P  <  0.05). Interestingly, the response of IFN-γ-producing cells in PvMSP1P stimulation was fourfold greater in recovered subjects than that in acute-infection patients. CD4(+) T cells were the major cell phenotype involved in the response to PvMSP1P-19 and PvDBPII antigen.

Conclusions: PvMSP1P-19 strongly induces a specific cellular immune response for protection against P. vivax compared with PvDBPII as the antigen induces activation of IFN-γ-producing effector cells following natural P. vivax exposure. Upon stimulation, PvMSP1P-19 has the potential to activate the recall response of Th1 effector memory cells that play a role in killing the parasite.

No MeSH data available.


Related in: MedlinePlus