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Isolating Human Peripheral Blood Mononuclear Cells and CD4+ T cells from Sézary Syndrome Patients for Transcriptomic Profiling
Published on: October 14, 2021
MicroRNA expression profiling of peripheral blood mononuclear cells associated with syphilis
Tao Huang1,2, Jun Zhang1, Wujian Ke1
1Dermatology Hospital, Southern Medical University, Guangzhou, China.
Background:
Treponema pallidum (T. pallidum) infection evokes significant immune responses, resulting in tissue damage. The immune mechanism underlying T. pallidum infection is still unclear, although microRNAs (miRNAs) have been shown to influence immune cell function and, consequently, the generation of antibody responses during other microbe infections. However, these mechanisms are unknown for T. pallidum.
Methods:
In this study, we performed a comprehensive analysis of differentially expressed miRNAs in healthy individuals, untreated patients with syphilis, patients in the serofast state, and serologically cured patients. miRNAs were profiled from the peripheral blood of patients obtained at the time of serological diagnosis. Then, both the target sequence analysis of these different miRNAs and pathway analysis were performed to identify important immune and cell signaling pathways. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was performed for microRNA analysis.
Results:
A total of 74 differentially regulated miRNAs were identified. Following RT-qPCR confirmation, three miRNAs (hsa-miR-195-5p, hsa-miR-223-3p, hsa-miR-589-3p) showed significant differences in the serofast and serologically cured states (P < 0.05). One miRNA (hsa-miR-195-5p) showed significant differences between untreated patients and healthy individuals.
Conclusions:
This is the first study of miRNA expression differences in peripheral blood mononuclear cells (PBMCs) in different stages of T. pallium infection. Our study suggests that the combination of three miRNAs has great potential to serve as a non-invasive biomarker of T. pallium infections, which will facilitate better diagnosis and treatment of T. pallium infections.
Insights
This study identifies specific microRNAs (miRNAs) in syphilis patients, revealing potential non-invasive biomarkers for Treponema pallidum infection. These findings could improve diagnosis and treatment strategies for syphilis.
Area of Science:
- Immunology
- Molecular Biology
- Genetics
Background:
- Treponema pallidum (T. pallidum) infection triggers immune responses and tissue damage, but underlying mechanisms remain unclear.
- MicroRNAs (miRNAs) are known to modulate immune cell function in other infections, but their role in T. pallidum infection is uninvestigated.
Purpose of the Study:
- To investigate microRNA (miRNA) expression profiles in different stages of T. pallidum infection.
- To identify potential non-invasive biomarkers for syphilis diagnosis and management.
Main Methods:
- Comprehensive analysis of differentially expressed miRNAs in healthy individuals and syphilis patients (untreated, serofast, cured).
- miRNA profiling from peripheral blood using quantitative reverse transcription-polymerase chain reaction (RT-PCR).
- Target sequence and pathway analyses to identify relevant immune and cell signaling pathways.
Main Results:
- Identified 74 differentially regulated miRNAs.
- Confirmed significant differences in hsa-miR-195-5p, hsa-miR-223-3p, and hsa-miR-589-3p between serofast and cured states.
- Found hsa-miR-195-5p significantly differed between untreated patients and healthy individuals.
Conclusions:
- This is the first study to analyze miRNA expression differences in peripheral blood mononuclear cells (PBMCs) across T. pallidum infection stages.
- A combination of three specific miRNAs shows potential as a non-invasive biomarker for T. pallidum infections.
- These miRNA biomarkers could aid in the diagnosis and treatment of syphilis.
