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Isolation of Tonsillar Mononuclear Cells to Study Ex Vivo Innate Immune Responses in a Human Mucosal Lymphoid Tissue
Published on: June 14, 2020
Quantification of cells expressing markers of proliferation and apoptosis in chronic tonsilitis
V Avramović1, V Petrović1, M Jović1
1Institute of Histology and Embryology, Faculty of Medicine University of Niš, Serbia;
Insights
Chronic tonsillitis disrupts lymphocyte balance, impacting immunity. This study quantifies proliferation and apoptosis markers, revealing distinct patterns in hypertrophic versus recurrent tonsillitis, influencing treatment decisions.
Area of Science:
- Immunology
- Cell Biology
- Pathology
Background:
- Chronic tonsillitis can impair palatine tonsil immunocompetence by disturbing lymphocyte proliferation and apoptosis.
- Understanding these cellular dynamics is crucial for assessing the tonsil's role in systemic immunity.
Purpose of the Study:
- To quantify cells expressing proliferation and apoptosis markers in palatine tonsil follicles.
- To compare these cellular dynamics between hypertrophic and recurrent tonsillitis.
Main Methods:
- Immunohistochemical staining (LSAB/HRP) for Ki-67 (proliferation), Bcl-2, survivin (anti-apoptotic), and Fas/CD95, caspase-3, Bax (pro-apoptotic) markers.
- Quantitative image analysis to determine numerical areal density of immunopositive cells.
- Study included tonsils from patients with hypertrophic (n=6) and recurrent (n=5) tonsillitis.
Main Results:
- Ki-67 positive cells (proliferation) were significantly higher in hypertrophic tonsillitis (14681.4 ± 1460.5) than in recurrent tonsillitis (12491.4 ± 2321.6).
- Survivin (117.4 ± 14.5) and caspase-3 (210 ± 24) positive cells were significantly lower in hypertrophic tonsillitis compared to recurrent tonsillitis (survivin, 406.9 ± 98.4; caspase-3, 350.4 ± 119.4).
- Localization of markers was similar across both tonsillitis types.
Conclusions:
- The rates of lymphocyte proliferation and apoptosis differ significantly between hypertrophic and recurrent tonsillitis.
- These distinct cellular profiles suggest varying immunological potential, impacting clinical management choices between conservative and surgical treatments.
Abstract:
During chronic tonsillitis, the relationship between proliferation and apoptosis of lymphocytes in tonsillar follicles can be disturbed, which gives rise to attenuation of tonsil immunocompetence and diminishing its contribution in systemic immunity. In this study, we have quantified the cells expressing the markers of proliferation and apoptosis in the follicles of the palatine tonsil. Six tonsils from patients aged 10-29 years with hypertrophic tonsillitis and five tonsils from patients aged 18-22 years with recurrent tonsillitis were studied. The sections of paraffin blocks of tonsillar tissue were stained by the immunohistochemical LSAB/HRP method with the utilisation of antibodies for: Ki-67 antigen-cell marker of proliferation; Bcl-2 and survivin anti-apoptotic factors and Fas/CD95, caspase-3 and Bax pro-apoptotic factors. The size of lymphoid follicles, i.e. mean follicle area and number of lymphoid follicle immunopositive cells per mm2 of a slice area, i.e. numerical areal density were determined by the quantitative image analysis. The localisation of Ki-67, Bcl-2, survivin, Fas/CD95, caspase-3 and Bax- immunopositive cells inside the palatine tonsil was similar in both types of tonsillitis. The number of Ki-67 immunopositive cells was significantly (p < 0.01) larger in the tonsils with hypertrophic tonsillitis (14681.4 ± 1460.5) in comparison to those with recurrent tonsillitis (12491.4 ± 2321.6), although the number of survivin and caspase-3 immunopositive cells was significantly (p < 0.05) larger in recurrent tonsillitis (survivin, 406.9 ± 98.4; caspase-3, 350.4 ± 119.4) when compared to those with hypertrophic tonsillitis (survivin, 117.4 ± 14.5; caspase-3, 210 ± 24). Our results show that the rate of the proliferation and apoptosis of follicular lymphocytes is different in various types of tonsillitis. This suggests that the immunological potential of the palatine tonsil varies in patients with hypertrophic and recurrent tonsillitis, which in practice poses a dilemma over the choice of conservative or surgical treatment.
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