Immunohistological and morphometric analysis of intra-epithelial lymphocytes and Langerhans cells in healthy and diseased human gingival tissues

Abstract

Periodontal diseases are histologically characterized by an infiltration of several inflammatory cell populations into the gingival epithelium and connective tissue, associated with degradation of extracellular matrix components. The purpose of this in situ study was to evaluate the inflammatory state of gingival tissues by the number of intra-epithelial lymphocyte (IEL) subsets and the area fraction ( A A %) occupied by collagen fibres in the upper gingival connective tissue, and also to evaluate the number of CD1a+ Langerhans cells (LC) in order to show correlation(s), if any, between these histological findings. The gingival samples were from 10 clinically healthy controls (group C), 8 patients with gingivitis (group G) and 9 with chronic adult periodontitis (group P). A quantitative evaluation of the number of cell populations (CD1a+, CD45RB+, CD3+, CD8+, CD20+, TIA-1+ and GrB+ cells) and the area fraction ( A A %) occupied by collagen fibres in the upper gingival connective tissue was made by morphometric and automated image analysis. The results showed that, compared with group C, all IEL subset numbers were significantly increased ( p<0.05) in G and P groups, CD20+ excepted. In addition, there was a significant increase in the cytotoxic TIA-1+ IEL number ( p<0.05) in group P when compared with group G. The study also showed a significant decrease in the number of CD1a+ LC in groups G and P ( p<0.02 and p<0.001, respectively) when compared with group C. No significant difference was found in CD1a+ LC number between groups G and P. The determination of coefficients of correlation ( r) with data obtained for each patient showed that in group G, CD1a+ LC number was significantly correlated with CD45RB+ ( p<0.05) and CD3+ ( p<0.01) IEL numbers whereas during periodontitis, CD1a+ LC number was significantly and inversely correlated with CD20+ ( p<0.01), cytotoxic TIA-1+ ( p<0.01) and with activated cytotoxic GrB+ ( p<0.01) IEL numbers. Moreover, in group P a significant ( p<0.05) positive correlation was shown between CD1a+ LC number and the A A % occupied by collagen fibres. This work demonstrates a decrease in CD1a+ LC number according to the severity of the periodontal disease estimated by the number of IEL and by the area fraction occupied by collagen fibres in human gingiva. The decrease of such cells could represent a way to avoid immune overstimulation.