Immunomodulatory effects of recombinant BCG expressing MSP-1C of<i>Plasmodium falciparum</i>on LPS- or LPS+IFN-γ-stimulated J774A.1 cells

Mohamad, Dhaniah; Suppian, Rapeah; Nor, Norazmi Mohd

doi:10.4161/hv.28695

Cited by 8 publications

(9 citation statements)

References 85 publications

(90 reference statements)

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“…Moreover, serum MAF had a 2.9-fold higher macrophage activation rate than IFN-γ. The results of this evaluation using our novel assay are very consistent with previous findings of increased phagocytic activity in macrophages when exposed to LPS, IFN-γ, and serum MAF [20] .…”

Section: Resultssupporting

confidence: 90%

A high-throughput quantitative assay system for macrophage phagocytic activity

et al. 2018

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Innate immunity is at the frontline of defense against pathogens. Macrophages are a group of phagocytes that secrete cytokines to mediate the transmission of information on pathogen infectiousness to the acquired immune system. Our previous study established an assay for quantifying phagocytic activity in two human monocytic cell lines, U937 and THP-1. Overall, this assay system was quick, easy, and robust, except for the time-consuming quantification step using image analysis. Thus, in this study, we improved our previous assay system. For evaluating phagocytic activity, fluorescently labeled magnetic beads were phagocytosed and detected with a fluorescent-multi well plate reader. This system facilitates high-throughput screening of macrophage activating factors.

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Section: Resultssupporting

confidence: 90%

A high-throughput quantitative assay system for macrophage phagocytic activity

et al. 2018

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“…The results showed a robust production of the proinflammatory mediator, NO and the enzyme iNOS which were highest in the rBCG immunised group. This was similar to their production in J774.1A macrophage cell lines infected with rBCG [9,10]. LPS stimulation of macrophages [14] cardiomyoblasts [15], cardiac myocytes [16] also led to a significant expression of iNOS and NO production.…”

Section: Discussionsupporting

confidence: 72%

“…Along this line, our group had earlier developed a recombinant BCG (rBCG) malaria vaccine candidate expressing the MSP-1 of P. falciparum which elicited increased nitric oxide (NO) production and significant inducible nitric oxide synthase (iNOS) expression alongside increased inflammatory cytokine release [9] with a possible role of TLR-4 [10]. The current study explored the role of the TLR-4 attachment to MSP-1 in eliciting the production of the pro-inflammatory mediator, NO and its inducing enzyme, iNOS via TAK-242-based inhibition.…”

Section: Introductionmentioning

confidence: 99%

The role of TLR-4 in the immunomodulatory effects of recombinant BCG expressing MSP-1C of Plasmodium falciparum

Abbas

Suppian

2019

J Infect Dev Ctries

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Introduction: An earlier constructed recombinant BCG expressing the MSP-1C of Plasmodium falciparum, induced inflammatory responses leading to significant production of nitric oxide (NO) alongside higher expression of the enzyme inducible nitric oxide synthase (iNOS) and significant production of the regulatory cytokine, IL-10, indicating significant immunomodulatory effects of the construct. The mechanism of these responses had not been established but is thought to involve toll-like receptor 4 (TLR-4). Methodology: The present study was carried out to determine the role of TLR-4 on eliciting the immunomodulatory effects of recombinant BCG expressing MSP-1C of Plasmodium falciparum leading to the production of NO and IL-10, as well as the expression of iNOS. Six groups of mice (n = 6 per group) were immunised thrice, three weeks apart with intraperitoneal phosphate buffered saline T80 (PBS-T80), BCG or rBCG in the presence or absence of a TLR-4 inhibitor; TAK-242, given one hour prior to each immunisation. Peritoneal macrophages were harvested from the mice and cultured for the determination of NO, iNOS and IL-10 via Griess assay, ELISA and Western blot respectively. Results: The results showed significant inhibition of the production of NO and IL-10 and the expression of iNOS in all groups of mice in the presence of TAK-242. Conclusions: These results presented evidence of the role of TLR-4/rBCG attachment mechanism in modulating the production of NO and IL-10 and the expression of iNOS in response to our rBCG-based malaria vaccine candidate expressing MSP-1C of P. falciparum.

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“…The BCG-MSP1C containing the MSP-1C of Plasmodium falciparum was constructed by Nurul (2007) using assembly Polymerase chain reaction (PCR) in the previous study. The stability of the MSP-1C gene of Plasmodium falciparum in BCG-MSP1C cells was determined using PCR and the protein expression was determined by immunocytochemistry (ICC) analysis using specific antibody against MSP-1C ( Dhaniah et al . 2014 ).…”

Section: Methodsmentioning

confidence: 99%

Apoptosis Activity of the Mouse Macrophage Cell Line J774A.1 Infected with a Recombinant BCG consisting the C-Terminus of Merozoite Surface Protein-1 of Plasmodium falciparum

Anis

Zakaria

Abdikarim

et al. 2018

tlsr

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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 in mouse macrophage cell line J774A.1 infected with a Mycobacterium bovis bacille Calmette-Guerin (BCG) clone and a recombinant BCG clone expressing the C-terminus of merozoite surface protein-1 (BCG-MSP1C) of Plasmodium falciparum for 48 h. In this study, a parent BCG cells was used as a control. The nuclear staining with Hoechst 33342 showed that the BCG-MSP1C cells was capable of increasing the nuclear condensation and morphological stages of apoptosis in the infected cells compared to the BCG-infected cells and the lipopolysaccharide (LPS)-stimulated cells. The flow cytometric analysis using Annexin-V and Propidium iodide (PI) staining confirmed that the BCG-MSP1C cells significantly increased the percentage of early apoptotic activity in the infected macrophage higher than the one stimulated by the parent BCG cells 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 cells capable of stimulating higher production of caspase-1, –3, –8 and –9 while the BCG-MSP1C cells stimulated the expression of caspase-1 and -9 in the infected macrophages, suggesting the involvement of mitochondrial-mediated (intrinsic) pathway of apoptosis. In conclusion, both the BCG and BCG-MSP1C cells 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 BCG-MSP1C cells showed higher apoptosis activity than those produced by the parent BCG cells.

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Immunomodulatory effects of recombinant BCG expressing MSP-1C ofPlasmodium falciparumon LPS- or LPS+IFN-γ-stimulated J774A.1 cells

Cited by 8 publications

References 85 publications

A high-throughput quantitative assay system for macrophage phagocytic activity

A high-throughput quantitative assay system for macrophage phagocytic activity

The role of TLR-4 in the immunomodulatory effects of recombinant BCG expressing MSP-1C of Plasmodium falciparum

Apoptosis Activity of the Mouse Macrophage Cell Line J774A.1 Infected with a Recombinant BCG consisting the C-Terminus of Merozoite Surface Protein-1 of Plasmodium falciparum

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