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Quantification of Intracellular Growth Inside Macrophages is a Fast and Reliable Method for Assessing the Virulence of Leishmania Parasites
Published on: March 16, 2018
Monarch-1 Activation in Murine Macrophage Cell Line (J774 A.1) Infected with Iranian Strain of Leishmania major
A Fata1, Mr Mahmoudian, A Varasteh
1Research Center for Skin Diseases and Cutaneous Leishmaniasis, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iarn.
Background:
Leishmania major is an intracellular parasite transmitted through the bite of the female phlebotomine sand flies. Leishmania major is able to escape the host immune defense and survive within macrophages. Modulation of the NF-κB (Nuclear Factor-Kappa B) activation and suppression of the pro-inflammatory cytokines by L. major are the main evasion mechanisms that remain to be explored. This study aims to examine the expression level of the Monarch-1 in L. major-infected macrophages, as a negative regulator of the NF-κB activation.
Methods:
Murine macrophage cell line (J774 A.1) was infected by metacyclic form of Leishmania promastigotes at macrophage/parasite ratio of 1:10. After harvesting infected cells at different times, total RNA was extracted and converted to cDNA. Semi-quantitative RT-PCR was performed for Monarch-1 by specific primers. Hypoxanthine Phospho-Ribosyl Transferase (HPRT) was used as an internal control to adjust the amount of mRNA in each sample.
Results:
Semiquantitive analysis of Monarch-1 mRNA expression level showed a significant expression increase within 6 to 30 hours after L. major infection of macrophages when compared to the control macrophages.
Conclusion:
Monarch-1 expression level reveals a significant increase in the early phase of macrophage infection with L. major, which in turn may suppress IL-12 production in Leishmania infected macrophages and deeply influence the relationship between host and parasite.
Insights
Leishmania major infection significantly increases Monarch-1 expression in macrophages early on. This may suppress crucial immune responses like IL-12 production, impacting host-parasite interactions.
Area of Science:
- Immunology
- Parasitology
- Molecular Biology
Background:
- Leishmania major is an intracellular parasite evading host immunity within macrophages.
- Understanding Leishmania evasion mechanisms, including NF-κB modulation, is crucial.
- Monarch-1, a negative regulator of NF-κB, is investigated as a key player.
Purpose of the Study:
- To investigate the expression level of Monarch-1 in macrophages infected with Leishmania major.
- To explore Monarch-1's role in the host-parasite interaction during early infection phases.
Main Methods:
- Murine macrophage cell line (J774 A.1) infected with Leishmania promastigotes.
- Semi-quantitative RT-PCR used to analyze Monarch-1 mRNA expression.
- Hypoxanthine Phospho-Ribosyl Transferase (HPRT) served as an internal control.
Main Results:
- Monarch-1 mRNA expression significantly increased 6 to 30 hours post-infection.
- A notable upregulation of Monarch-1 was observed in Leishmania-infected macrophages compared to controls.
Conclusions:
- Monarch-1 expression is significantly upregulated during the early stages of Leishmania major macrophage infection.
- Increased Monarch-1 may suppress Interleukin-12 (IL-12) production, influencing host-parasite dynamics.
