Open Access Highly Accessed Open Badges Research article

Characterisation of CD154+ T cells following ex vivo birch allergen stimulation defines a close relationship between T cell subsets in healthy volunteers

Karen A Smith1, Nicola J Gray2, Elizabeth Cheek3, Femi Saleh4, Jo Lavender4, Anthony J Frew2, Florian Kern1 and Michael D Tarzi1*

Author Affiliations

1 Brighton and Sussex Medical School, Division of Clinical Medicine, Pathogen-Host-Interactions Group, University of Sussex, Brighton, BN1 9PS, UK

2 Department of Respiratory Medicine, Royal Sussex County Hospital, Brighton, BN2 5BE, UK

3 Department of Computing, Engineering and Mathematics, University of Brighton, Brighton, BN2 4GJ, UK

4 Diagnostic Immunology Laboratory, Blood Sciences Department, Royal Sussex County Hospital, Brighton, BN2 5BE, UK

For all author emails, please log on.

BMC Immunology 2013, 14:14  doi:10.1186/1471-2172-14-14

Published: 22 March 2013



Allergic sensitisation has been ascribed to a dysregulated relationship between allergen-specific Th1, Th2 and regulatory T cells. We hypothesised that the relationship between these T cell subsets could be better defined using a short-term allergen stimulation system followed by direct analysis of CD154-positive T cells. Using peripheral blood samples from birch pollinosis patients and healthy non-atopic controls, we sought to explore the frequencies and phenotype of birch-stimulated CD154-positive T helper cells following ex vivo birch allergen stimulation.


Activated CD154-positive Th1, Th2 and Tr1-like cells, that co-expressed IFNγ, IL-4 and IL-10 respectively, were identified in both birch-allergic and non-allergic participants. We observed a close correlation between Th1, Th2 and Tr1-like cell frequency in non-allergic volunteers, such that the three parameters increased together to maintain a low Th2: Th1 ratio. The relationship between Th1, Th2 and Tr1-like responses was dysregulated in birch-allergic patients, with abrogation of the IL-10 response and a higher Th2: Th1 ratio. A close correlation was observed between Th2 cell frequency and the absolute concentration of birch-specific IgE within the birch-allergic group, and we confirmed previous reports of a more differentiated T cell phenotype in allergic subjects.


The findings demonstrate an important balance between IFNγ, IL-4 and IL-10 T cell responses to birch allergen in health, where Th2 responses to allergens were frequently observed, but apparently balanced by Th1 and regulatory responses. The detection of CD154 positive T cells after short-term antigen stimulation may be a useful method for the detection of T cell responses to allergens when cost, speed and convenience are priorities.

Allergen-specific T cell; Birch pollen allergy; Ex vivo phenotyping; Flow cytometry