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A Microscopic Phenotypic Assay for the Quantification of Intracellular Mycobacteria Adapted for High-throughput/High-content Screening
Published on: January 17, 2014
Immunophenotypic characterization of peripheral T lymphocytes in Mycobacterium tuberculosis infection and disease
D S S Rodrigues1, E A S Medeiros, L Y Weckx
1Infectious Diseases Discipline and Immunization Center, Federal University of São Paulo, São Paulo, Brazil.
Insights
Cellular immune responses, specifically T-lymphocyte subsets, are crucial in tuberculosis (TB) outcomes. Active TB patients show reduced CD4+ and CD8+ T cells and increased activation markers compared to healthy individuals.
Area of Science:
- Immunology
- Infectious Diseases
- Cell Biology
Background:
- The cellular immune response is critical for managing Mycobacterium tuberculosis infection.
- Understanding T-lymphocyte subset dynamics is key to predicting tuberculosis (TB) clinical outcomes.
Purpose of the Study:
- To investigate T-lymphocyte subset distribution and activation phenotypes in individuals with varying Mycobacterium tuberculosis exposure and disease states.
- To correlate peripheral T-cell profiles with active TB, latent infection, and post-treatment recovery.
Main Methods:
- Multi-parameter flow cytometry was used to analyze T CD4+ and CD8+ lymphocyte numbers and phenotypes (activation, memory/naïve).
- Blood samples from 71 volunteers were analyzed, categorized into PPD-negative healthy, PPD-positive healthcare workers, active TB patients, and treated TB patients.
- Statistical analyses compared T-cell subset distributions and activation markers across the four groups.
Main Results:
- Active TB patients had significantly lower T CD4+ and CD8+ lymphocyte counts compared to healthy controls.
- A higher proportion of naïve T CD4+ cells was observed in the PPD-negative group, while the active TB group had fewer.
- T-cell activation markers (CD38, HLA-DR) were elevated in active TB patients, indicating increased cellular activation.
Conclusions:
- Peripheral T-lymphocyte profiles, including subset numbers and activation states, reflect cellular immune responses during active tuberculosis.
- Treatment appears to restore T-cell numbers, suggesting a dynamic redistribution of naïve, memory, and effector T-cell phenotypes in response to M. tuberculosis infection and disease.
Abstract:
The cellular immune response probably plays a pivotal role in determining the clinical outcome after exposure to Mycobacterium tuberculosis. We used multi-parameter flow-cytometry to evaluate the distribution of T-lymphocyte subsets during infection and disease caused by M. tuberculosis. Samples were obtained from 71 volunteers to identify the T CD4+ and CD8+ lymphocyte numbers, and the activation plus memory/naïve phenotypes, as defined by CD38, HLA-DR, CD45RA and CD27 markers. Subjects were divided into 18 healthy volunteers without detectable reaction to purified protein derivative (PPD-), 18 health care workers with a recent conversion to PPD, 20 patients with active pulmonary tuberculosis (TBC) and 15 patients with treated TBC at 6 months of therapy. By multiple-comparison analyses, the T CD4+ lymphocyte number of the TBC group was lower than the PPD- group (P < 0.05). This difference was apparently lost after treatment. The higher and the lower number of naïve T CD4+ cells was observed in the PPD- and TBC group, respectively. CD8+ T lymphocytes were also statistically different among the four groups (P = 0.0002), lower in the TBC group (P < 0.05). CD8+ T lymphocyte activation was evaluated by the CD38 and HLA-DR surface expression. The percentage distribution of these markers was statistically different between the four groups (P = 0.0055). TBC patients had a higher percentage of CD38+ cells and mean fluorescence index, suggesting an overall increase of cell activation. These results suggest that peripheral T lymphocytes reflect cellular activation during TBC, along with possible redistribution of naïve, memory/effector and late differentiated memory/effector phenotypes in the peripheral blood after infection and disease caused by M. tuberculosis.
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