engleski

Flow cytometric determination of the leukocyte subsets in paraffinstimulated whole human saliva

Human saliva has an important role in the oral immune system which is a part of an extensive and specialized mucosa-associated lymphoid tissue. Antimicrobial properties of saliva were primarily investigated through its soluble components and little is known about its cellular components. We developed a method to quantify leukocytes in paraffin-stimulated whole human saliva (SWS) using four color flow cytometry and performed reliability study on 8 healthy volunteers (4 males, 4 females, mean age 26.1, SD 1.1). Based on the stains we used (CD45 FITC, CD14 PE, CD20 PerCP, CD3APC)and FITC threshold gating, we could discriminate major leukocyte populations(granulocytes, monocytes and lymphocytes), as well as T and B cells, and measure their percentages. Saliva values were compared to the whole blood values analyzed according to standard laboratory procedure. Predominant leukocyte population in saliva were granulocytes (97.5%, SD 2.1) followed by very low frequencies of monocytes (1.3%, SD 0.7) and lymphocytes (1.2%, SD 1.5). Percentages of T (72.32%, SD 9.23) or B (11.87%, SD 6.33) cells in saliva did not differ significantly from the peripheral blood values. Intra-class coefficients of correlation (ICC) of assessed parameters ranged from 0.6 to 0.9 suggesting generally good reliability of the measurement. Flow cytometry can be used to reliably quantify leukocytes and its subsets in paraffin SWS. The identification and complete characterization of salivary leukocytes could be used to better understand pathogenesis of many oral diseases.

flow cytometry; saliva; leukocyte subsets

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