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Immunologic activation of human syncytiotrophoblast by Plasmodium falciparum.

Lucchi NW, Peterson DS, Moore JM - Malar. J. (2008)

Bottom Line: How ST responds to this interaction remains poorly understood.Following iRBC/ST interaction, ST C-Jun N-terminal kinase 1 (JNK1) was activated and modest increases in the mRNA expression of TGF-beta and IL-8/CXCL8 were observed.In addition, this interaction increased secretion of MIF and MIP-1alpha/CCL3 by ST and induced migration of PBMC towards iRBC-stimulated ST.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Infectious Diseases and Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA 30602, USA. frd9@cdc.gov

ABSTRACT

Background: Malaria during pregnancy is characterized by the sequestration of malaria-infected red blood cells (iRBC) in the intervillous spaces of the placenta, often accompanied by the infiltration of maternal mononuclear cells, causing substantial maternal and foetal/infant morbidity. The iRBC bind to receptors expressed by the syncytiotrophoblast (ST). How ST responds to this interaction remains poorly understood. Because it is known that ST is immunoactive and can respond to infectious agents, the consequences of this ST-iRBC interaction should be investigated.

Methods: An in vitro system was used to assess the biochemical and immunological changes induced in ST by ST-adherent iRBCs. Changes in ST mitogen-activated protein kinase (MAPK) activation were assessed by immunoblotting and mRNA expression levels of selected cytokine and chemokines in primary ST bound by iRBC were determined using real-time, reverse transcription PCR. In addition, secreted cytokine and chemokine proteins were assayed by standard ELISA, and chemotaxis of PBMC was assessed using a two-chamber assay system.

Results: Following iRBC/ST interaction, ST C-Jun N-terminal kinase 1 (JNK1) was activated and modest increases in the mRNA expression of TGF-beta and IL-8/CXCL8 were observed. In addition, this interaction increased secretion of MIF and MIP-1alpha/CCL3 by ST and induced migration of PBMC towards iRBC-stimulated ST.

Conclusion: Results from this study provide the first evidence that ST participates in shaping the local immunological milieu and in the recruitment of maternal immune cells to the maternal blood space during placental malaria infection.

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Chemokine Secretion by primary ST cells upon the binding of iRBCST. Primary ST was stimulated with iRBCST (iRBC), uRBCs or left unstimulated (MED) over given time courses as indicated. (A) While stimulation with uRBC did not lead to any increase in MIF secretion more than that observed for unstimulated ST, stimulation with CS2-iRBCST led to an increased time-dependent secretion. This difference in MIF production among the different stimulant was statistically significant (p < 0.0032). (B) Two out of five placentas tested showed a time dependent increase in MIP-1α/CCL3 secretion upon interaction with iRBCST but not with uRBC or unstimulated this difference being statistically significant (p < 0.0040). (C) Secretion of IL-8/CXCL8 increased over time even in unstimulated cells, with no significant additional increase with iRBCST stimulation. Independent results from two placentas are shown.
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Figure 3: Chemokine Secretion by primary ST cells upon the binding of iRBCST. Primary ST was stimulated with iRBCST (iRBC), uRBCs or left unstimulated (MED) over given time courses as indicated. (A) While stimulation with uRBC did not lead to any increase in MIF secretion more than that observed for unstimulated ST, stimulation with CS2-iRBCST led to an increased time-dependent secretion. This difference in MIF production among the different stimulant was statistically significant (p < 0.0032). (B) Two out of five placentas tested showed a time dependent increase in MIP-1α/CCL3 secretion upon interaction with iRBCST but not with uRBC or unstimulated this difference being statistically significant (p < 0.0040). (C) Secretion of IL-8/CXCL8 increased over time even in unstimulated cells, with no significant additional increase with iRBCST stimulation. Independent results from two placentas are shown.

Mentions: In addition to gene expression analysis, functional ST activation was assessed via the measurement of cytokine and chemokine secretion. Supernatants from stimulated primary ST were used. Similar to BeWoST [35], substantial time-dependent secretion of MIF was observed upon stimulation of primary ST with iRBCST but not with uRBC or unstimulated cells (Figure 3A). The differences in MIF secretion were significantly different among the stimuli (p < 0.0032). MIP-1α/CCL3 was not consistently detected, but in 2 of 5 ST cultures, secretion was upregulated in response to iRBCST (Figure 3B) (p < 0.0040). The production of IL-8/CXCL8 did not increase with iRBCST binding, although a time-dependent increase, clearly representing constitutive expression, was observed (Figure 3C). No secretion of TNF-α, MIP-1β/CCL4, or IL-10 was detected.


Immunologic activation of human syncytiotrophoblast by Plasmodium falciparum.

Lucchi NW, Peterson DS, Moore JM - Malar. J. (2008)

Chemokine Secretion by primary ST cells upon the binding of iRBCST. Primary ST was stimulated with iRBCST (iRBC), uRBCs or left unstimulated (MED) over given time courses as indicated. (A) While stimulation with uRBC did not lead to any increase in MIF secretion more than that observed for unstimulated ST, stimulation with CS2-iRBCST led to an increased time-dependent secretion. This difference in MIF production among the different stimulant was statistically significant (p < 0.0032). (B) Two out of five placentas tested showed a time dependent increase in MIP-1α/CCL3 secretion upon interaction with iRBCST but not with uRBC or unstimulated this difference being statistically significant (p < 0.0040). (C) Secretion of IL-8/CXCL8 increased over time even in unstimulated cells, with no significant additional increase with iRBCST stimulation. Independent results from two placentas are shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2268702&req=5

Figure 3: Chemokine Secretion by primary ST cells upon the binding of iRBCST. Primary ST was stimulated with iRBCST (iRBC), uRBCs or left unstimulated (MED) over given time courses as indicated. (A) While stimulation with uRBC did not lead to any increase in MIF secretion more than that observed for unstimulated ST, stimulation with CS2-iRBCST led to an increased time-dependent secretion. This difference in MIF production among the different stimulant was statistically significant (p < 0.0032). (B) Two out of five placentas tested showed a time dependent increase in MIP-1α/CCL3 secretion upon interaction with iRBCST but not with uRBC or unstimulated this difference being statistically significant (p < 0.0040). (C) Secretion of IL-8/CXCL8 increased over time even in unstimulated cells, with no significant additional increase with iRBCST stimulation. Independent results from two placentas are shown.
Mentions: In addition to gene expression analysis, functional ST activation was assessed via the measurement of cytokine and chemokine secretion. Supernatants from stimulated primary ST were used. Similar to BeWoST [35], substantial time-dependent secretion of MIF was observed upon stimulation of primary ST with iRBCST but not with uRBC or unstimulated cells (Figure 3A). The differences in MIF secretion were significantly different among the stimuli (p < 0.0032). MIP-1α/CCL3 was not consistently detected, but in 2 of 5 ST cultures, secretion was upregulated in response to iRBCST (Figure 3B) (p < 0.0040). The production of IL-8/CXCL8 did not increase with iRBCST binding, although a time-dependent increase, clearly representing constitutive expression, was observed (Figure 3C). No secretion of TNF-α, MIP-1β/CCL4, or IL-10 was detected.

Bottom Line: How ST responds to this interaction remains poorly understood.Following iRBC/ST interaction, ST C-Jun N-terminal kinase 1 (JNK1) was activated and modest increases in the mRNA expression of TGF-beta and IL-8/CXCL8 were observed.In addition, this interaction increased secretion of MIF and MIP-1alpha/CCL3 by ST and induced migration of PBMC towards iRBC-stimulated ST.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Infectious Diseases and Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA 30602, USA. frd9@cdc.gov

ABSTRACT

Background: Malaria during pregnancy is characterized by the sequestration of malaria-infected red blood cells (iRBC) in the intervillous spaces of the placenta, often accompanied by the infiltration of maternal mononuclear cells, causing substantial maternal and foetal/infant morbidity. The iRBC bind to receptors expressed by the syncytiotrophoblast (ST). How ST responds to this interaction remains poorly understood. Because it is known that ST is immunoactive and can respond to infectious agents, the consequences of this ST-iRBC interaction should be investigated.

Methods: An in vitro system was used to assess the biochemical and immunological changes induced in ST by ST-adherent iRBCs. Changes in ST mitogen-activated protein kinase (MAPK) activation were assessed by immunoblotting and mRNA expression levels of selected cytokine and chemokines in primary ST bound by iRBC were determined using real-time, reverse transcription PCR. In addition, secreted cytokine and chemokine proteins were assayed by standard ELISA, and chemotaxis of PBMC was assessed using a two-chamber assay system.

Results: Following iRBC/ST interaction, ST C-Jun N-terminal kinase 1 (JNK1) was activated and modest increases in the mRNA expression of TGF-beta and IL-8/CXCL8 were observed. In addition, this interaction increased secretion of MIF and MIP-1alpha/CCL3 by ST and induced migration of PBMC towards iRBC-stimulated ST.

Conclusion: Results from this study provide the first evidence that ST participates in shaping the local immunological milieu and in the recruitment of maternal immune cells to the maternal blood space during placental malaria infection.

Show MeSH
Related in: MedlinePlus