Apoptosis Activity And Mitogen Activated Protein Kinase Expression Of The Mouse Macrophage Cell Line J774a.1 Infected With A Recombinant Bcg Expressing The C-Terminus Of Merozoite Surface Protein-1 Of Plasmodium falciparum

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Date
2016-12
Authors
Zulkipli, Anis Fadhilah
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Publisher
Universiti Sains Malaysia
Abstract
Macrophage apoptosis exerts an efficient mechanism in controlling intracellular infection during innate immune response against various pathogens including malaria parasites. This study was carried out to determine the apoptosis activity and mitogen activated protein kinase (MAPK) expression in mouse macrophage cell line J774A.1 infected with a Mycobacterium bovis bacille Calmette-Guerin (BCG) clone and a recombinant BCG (rBCG) clone expressing the C-terminus of merozoite surface protein-1 (MSP-1C) of Plasmodium falciparum for 48 hours. The nuclear staining with Hoechst 33342 showed that the rBCG clone was capable of increasing the nuclear condensation and morphological stages of apoptosis in the infected cells compared to the BCG-infected cells and the LPS-stimulated cells. The flow cytometric analysis using Annexin-V and PI staining confirmed that the rBCG clone significantly increased the percentage of early apoptotic activity in the infected macrophage higher than the one stimulated by the parent BCG clone and LPS. This apoptotic response corresponded with the reduction of the anti-apoptotic Bcl-2 protein expression and higher p53 expression. The colorimetric assay demonstrated that the BCG clone is capable of stimulating higher production of caspase-1, -3, -8 and -9 while the rBCG clone only stimulated the expression of caspase-1 and -9 in the infected macrophages, suggesting the involvement of mitochondrial-mediated (intrinsic) pathway of apoptosis. In addition, an infection with BCG and rBCG clones stimulated higher extracellular signal-regulated kinase 2 (ERK2) protein expression in the infected cells but significantly reduced the expression of p38 and cjun N-terminal kinase (JNK) proteins, suggesting the involvement of certain MAPK in the apoptosis activity of the infected macrophages. In conclusion, both the BCG and rBCG clones are capable of inducing macrophage apoptosis activity in the mouse macrophage cell line J774A.1. This mechanism is important for the elimination of pathogens such as malaria parasite during the phagocytosis activity of macrophage. However, the rBCG clone showed higher apoptosis activity than those produced by the parent BCG clone.
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Macrophage apoptosis exerts an efficient mechanism , against various pathogens including malaria parasites.
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