Isolation and Functional Characterization of Myeloid-Derived Suppressor Cells in Infections Under High Containment
Leigh A Kotze1, Vinzeigh N Leukes1, Nelita Du Plessis2
1Division of Molecular Biology and Human Genetics, Faculty of Medical and Health Sciences, DST-NRF Centre of Excellence for Biomedical Tuberculosis Research, South African Medical Research Council for Tuberculosis Research, Stellenbosch University, Cape Town, South Africa.
Abstract:
The current absence of markers unique to MDSC, particularly those expanded during human infection, necessitate concurrent demonstration of their suppressive capacity to ensure unequivocal identification. This is further complicated by the array of heterogeneous markers used to characterize MDSC in various conditions and models. Standardization of phenotypic and functional characterization, as well as isolation, from infectious biological samples of patients, are critical for accurately reporting MDSC dynamics, function, organ abundance, and establishment of their therapeutic value in infectious diseases. To illustrate, we report on our established method for MDSC isolation from bronchoalveolar lavage fluid and peripheral blood of pulmonary TB patients, as well as functional impact on T cells by measuring T cell activation, proliferation, and cytokine production.
Insights
Identifying myeloid-derived suppressor cells (MDSC) during infection is challenging due to a lack of unique markers. This study details a standardized method for MDSC isolation and functional assessment in tuberculosis patients.
Area of Science:
- Immunology
- Infectious Diseases
- Cell Biology
Background:
- Myeloid-derived suppressor cells (MDSC) play a crucial role in immune regulation during infection.
- Current methods for identifying MDSC are hampered by the lack of unique markers, especially in human infections.
- Heterogeneous markers and varied isolation techniques complicate the accurate characterization of MDSC.
Purpose of the Study:
- To address the need for standardized methods for identifying and characterizing MDSC in infectious diseases.
- To establish a reliable protocol for isolating MDSC from patient samples.
- To evaluate the functional impact of MDSC on T cell responses in the context of pulmonary tuberculosis.
Main Methods:
- Development and application of a standardized protocol for MDSC isolation from bronchoalveolar lavage fluid and peripheral blood.
- Characterization of MDSC using phenotypic markers.
- Functional assessment of MDSC by measuring their impact on T cell activation, proliferation, and cytokine production.
Main Results:
- Successfully isolated MDSC from pulmonary tuberculosis patients' bronchoalveolar lavage fluid and peripheral blood.
- Demonstrated the functional capacity of isolated MDSC to suppress T cell responses.
- Provided a validated method for assessing MDSC dynamics and function in infectious disease contexts.
Conclusions:
- Standardized isolation and functional characterization are essential for accurate MDSC assessment in infectious diseases.
- The developed method facilitates the study of MDSC in pulmonary tuberculosis.
- This approach is critical for understanding MDSC's role and therapeutic potential in infectious diseases.
