Simultaneous evaluation of functional and phenotypic characteristics of T lymphocytes in patients with schistosomiasis

Abstract

Flow cytometry enables the detection of surface markers and intracellular cytokines at the single cell level. This technique was used in this study to assess cell surface molecule and intracellular cytokines in patients with schistosomiasis.

Nineteen patients with S. mansoni infection and ten normal subjects were assessed. Patients were divided according to the ultrasonographic examination and rectal snip biopsy into intestinal and hepatosplenic groups. Ten healthy subjects were studied as control.

Peripheral blood mononuclear cells was isolated using Ficoll-Hypaque ahd stimulated with phorbol myristate acetate and ionomycin in presence of Brefeldin A Unstimulated cells were stained for surface markers and intracellular cytokines and stimulated cells were stained for intracellular cytokines. 10,000 cells were acquired and analyzed using FACScan flow cytometer (Lysis II). An electrolyte gate was set on the lymphocytes on the forward scatter and side scatter plot. CD3, CD19, CD4, IFN-y and IL-4 were analyzed by detection of FITC and PE staining as percent of positively stained lymphocytes.

Statistical analysis using ANOVA and "t" test showed non significant difference in the mean percentage of CD3+ cells in intestinal, hepatosplenic and control groups. Similarly there