T helper cell polarisation as a measure of the maturation of the immune response

BACKGROUND: T helper cell polarisation is important under chronic immune stimulatory conditions and drives the type of the evolving immune response. Mice treated with superantigens in vivo display strong effects on T-h subset differentiation. The aim of the study was to detect the intrinsic capacity of T cells to polarise under various ex vivo conditions. Methods: Purified CD4(+) T cells obtained from super-antigen-treated mice were cultured under Th polarising conditions in vitro. By combining intracellular cytokine staining and subsequent flow cytometric analysis with quantitative cytokine measurements in culture supernatants by enzyme-linked immunosorbent assay ( ELISA), the differential Th polarising capacity of the treatment can be detected in a qualitative and quantitative manner. Results and conclusions: BALB/c mice were shown to be biased to develop strong T(h)2 polarised immune responses using T(h)0 stimulation of purified CD4(+) T cells from phosphate-buffered saline-treated mice. Nevertheless, our analysis methodology convincingly showed that even in these mice, Toxic Shock Syndrome Toxin-1 treatment in vivo resulted in a significantly stronger T(h)1 polarising effect than control treatment. Our results indicate that populations of T-h cells can be assessed individually for their differential T(h)1 or T(h)2 maturation capacity in vivo by analysing robust in vitro polarisation cultures combined with intracellular cytokine staining and ELISA