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Updated: Jul 15, 2026
Lentiviral Vector Preparation for Efficient Gene and MicroRNA Modulation of Peritoneal Cavity Tissue-Resident Macrophages In Vivo in Mice
Published on: February 16, 2024
[Effect of transfected Rv0901 gene on the activity of mice macrophages]
Qi Zhong1, Lang Bao, Zheng-Ling Shang
1Department of Infection and Immunity, West China school of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Objective:
To test the effect of Rv0901 gene of Mycobacterium tuberculosis on the activity of mice macrophages.
Methods:
Peritoneal macrophages of mice were isolated and transfected with pcDNA3. 1 or pcDNA3. 1-Rv0901 plasmid DNA, along with the GFP DNA. The effectiveness of the transfection was detected by RT-PCR. The expression of the GFP and the apoptosis ratio of the macrophages were detected by FCM 72 hours after the transfection. The level of nitric oxide and IFN-gamma in cultural supernatant were also measured 72 hours after transfection.
Results:
All of the macrophages being transfected had the expression of GFP. After being transfected with pcDNA3. 1-Rv0901, the 528 bp gene of Rv0901 was amplified by RT-PCR. The macrophages transfected with pcDNA3. 1-Rv0901 had higher apoptosis ratio [(56.4 +/- 2.0)% vs (19.9 +/- 1.5)%] and released more nitric oxide [(40.4 +/- 3.0) micromol/L vs (27.5 +/- 3.2) micromol/L] and IFN-gamma [(2.11 +/- 0.031) ng/mL vs (0.62 +/- 0.025) ng/mL] in the cultural supernatants than those transfected with pcDNA3. 1 (P < 0.05).
Conclusion:
Transient transfection of pcDNA3. 1-Rv0901 increases the apoptosis ratio of the macrophages, which could increase the release of nitric oxide and IFN-gamma.
Insights
The Mycobacterium tuberculosis Rv0901 gene increases macrophage apoptosis, leading to higher nitric oxide and IFN-gamma release. This study investigates the impact of Rv0901 on immune cell activity.
Area of Science:
- Immunology
- Microbiology
- Molecular Biology
Context:
- Tuberculosis remains a significant global health challenge, driven by Mycobacterium tuberculosis.
- Understanding the molecular mechanisms of host-pathogen interaction is crucial for developing new therapeutic strategies.
- Macrophages play a central role in the innate and adaptive immune response to M. tuberculosis infection.
Purpose:
- To investigate the functional role of the Rv0901 gene from Mycobacterium tuberculosis in modulating murine macrophage activity.
- To determine the effect of Rv0901 expression on macrophage apoptosis, nitric oxide production, and interferon-gamma release.
Summary:
- Murine peritoneal macrophages were transfected with a plasmid encoding Rv0901 or a control plasmid.
- RT-PCR confirmed Rv0901 gene expression, and flow cytometry assessed apoptosis and GFP expression.
- Macrophages expressing Rv0901 exhibited significantly increased apoptosis ratios and elevated levels of nitric oxide and IFN-gamma in culture supernatants compared to controls.
Impact:
- The findings suggest that Rv0901 contributes to host immune responses by inducing macrophage apoptosis.
- Increased apoptosis may enhance the release of key immune mediators like nitric oxide and IFN-gamma, potentially influencing pathogen control.
- This research provides insights into the immunomodulatory functions of M. tuberculosis virulence factors.
