The critical role of cytokines for Th cells in allergic diseases is widely accepted. However, the cytokine profiles and immune regulatory roles of Tr, Tc and B cells on cytokine secretion in allergic diseases are not well defined. One pertinent problem with the use of peripheral blood mononuclear cells (PBMC) is the low yields and purity of such cells. We examined the use of tonsillar mononuclear cells (MC) for such studies.

Tonsillar MC from 6 allergic and non-allergic rhinitis patients were isolated by gradient centrifuge. Crude mite extracts at 10μg/mL were applied to stimulate the cells in vitro. Subsets of lymphocyte were isolated by MACS. Cytokine FACS assays and real-time PCR were used to analyze the cytokine secretion and gene expression.

Large numbers of tonsillar MC (greater than 1×10⁹) were obtained from each sample. FACS analysis indicated that CD4+ T cells accounted for approximately 36%, CD8+ T cells 10% and B cells 54% of the tonsillar MC. MACS isolated cells were routinely 92–97% pure as assessed by FACS analysis. The viability of T cell was greater than 90% after 24h culture. No cytokine secretion was detected from MC without antigen stimulation from both allergic and non-allergic patients. When the cells were challenged with crude mite extracts, both INF-gamma and IL-4 were detected. The cytokine profiles/ratios were similar to those obtained from PBMCs.

Human tonsillar MC is a suitable source of T and B cells for functional study on antigen specific cytokine secretion in allergic diseases.