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Gene Expression Profiling of Infecting Microbes Using a Digital Bar-coding Platform
Published on: January 13, 2016
Host gene expression analysis in Sri Lankan melioidosis patients
Shivankari Krishnananthasivam1, Nimanthi Jayathilaka2, Harindra Darshana Sathkumara1
1Genetech Research Institute, Colombo, Sri Lanka.
Background:
Melioidosis is a life threatening infectious disease caused by the gram-negative bacillus Burkholderia pseudomallei predominantly found in southeast Asia and northern Australia. Studying the host transcription profiles in response to infection is crucial for understanding disease pathogenesis and correlates of disease severity, which may help improve therapeutic intervention and survival. The aim of this study was to analyze gene expression levels of human host factors in melioidosis patients and establish useful correlation with disease biomarkers, compared to healthy individuals and patients with sepsis caused by other pathogens.
Methods:
The study population consisted of 30 melioidosis cases, 10 healthy controls and 10 sepsis cases caused by other pathogens. Total RNA was extracted from peripheral blood mononuclear cells (PBMC's) of study subjects. Gene expression profiles of 25 gene targets including 19 immune response genes and 6 epigenetic factors were analyzed by real time quantitative polymerase chain reaction (RT-qPCR).
Principal Findings:
Inflammatory response genes; TLR4, late onset inflammatory mediator HMGB1, genes associated with antigen presentation; MICB, PSMB2, PSMB8, PSME2, epigenetic regulators; DNMT3B, HDAC1, HDAC2 were significantly down regulated, whereas the anti-inflammatory gene; IL4 was up regulated in melioidosis patients compared to sepsis cases caused by other pathogens. Septicaemic melioidosis cases showed significant down regulation of IL8 compared to sepsis cases caused by other pathogens. HMGB1, MICB, PSMB8, PSMB2, PSME2, HDAC1, HDAC2 and DNMT3B showed consistent down regulation of gene expression in melioidosis patients compared to other sepsis infection, irrespective of comorbidities such as diabetes, duration of clinical symptoms and antibiotic treatment.
Significance:
Specific immune response genes and epigenetic regulators are differentially expressed among melioidosis patients and patients with sepsis caused by other pathogens. Therefore, these genes may serve as biomarkers for disease diagnosis to distinguish melioidosis from cases of sepsis due to other infections and therapeutic intervention for melioidosis.
Insights
Gene expression analysis reveals distinct immune and epigenetic profiles in melioidosis patients, differentiating them from other sepsis cases. These findings highlight potential biomarkers for diagnosing melioidosis and guiding therapeutic interventions.
Area of Science:
- Infectious Diseases
- Genomics
- Immunology
Background:
- Melioidosis, a severe infectious disease caused by Burkholderia pseudomallei, is prevalent in Southeast Asia and Northern Australia.
- Understanding host gene expression in melioidosis is vital for elucidating disease mechanisms and severity.
- This study aimed to compare host gene expression in melioidosis patients with healthy individuals and other sepsis cases.
Purpose of the Study:
- To analyze gene expression profiles of human host factors in melioidosis patients.
- To identify correlations between gene expression and disease biomarkers.
- To differentiate melioidosis from other sepsis infections based on host transcription profiles.
Main Methods:
- The study included 30 melioidosis patients, 10 healthy controls, and 10 sepsis patients with other causes.
- Total RNA was extracted from peripheral blood mononuclear cells (PBMCs).
- Gene expression of 25 targets (19 immune, 6 epigenetic) was analyzed using real-time quantitative PCR (RT-qPCR).
Main Results:
- Downregulation of inflammatory genes (TLR4, HMGB1) and epigenetic regulators (DNMT3B, HDAC1, HDAC2) was observed in melioidosis patients compared to other sepsis cases.
- Upregulation of the anti-inflammatory gene IL4 and downregulation of IL8 in septicaemic melioidosis were noted.
- Consistent downregulation of HMGB1, MICB, PSMB8, PSMB2, PSME2, HDAC1, HDAC2, and DNMT3B occurred in melioidosis, independent of comorbidities.
Conclusions:
- Differential expression of immune response genes and epigenetic regulators distinguishes melioidosis from other sepsis types.
- These genes show potential as diagnostic biomarkers for melioidosis.
- Identifying these gene expression patterns may aid in developing targeted therapeutic interventions for melioidosis.
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