[1, 2] Moreover, the allergen-specific CD4+ T cells of non-allergic subjects were mostly either unpolarized or produced low levels of interferon-γ (IFN-γ) and interleukin-10 (IL-10).[1, 2] In the current study, we sought to confirm these findings by examining the CD4+ T-cell response to the major horse allergen Equ c 1, an important lipocalin allergen[8] with the prevalence of IgE reactivity close to 80% among horse

dust-allergic subjects.[9, 10] For this purpose, we analysed the CD4+ T-cell responses of horse dust-exposed Equ c 1-sensitized and healthy subjects focusing on the dominant epitope region of the allergen. This region is strongly recognized by the T cells of almost all Equ c 1-sensitized subjects examined.[11] As with the major allergen LY294002 in vitro of dog, Can f 1[1], and the major allergen of cow, Bos d 2[2], the frequency of Equ c 1-specific CD4+ T cells in the peripheral blood is very low. In allergic subjects, it is mostly higher than in non-allergic ones. Moreover, the function and phenotype of Equ c 1-specific CD4+ T cells differ between these two subject groups. p143–160 (GIVKENIIDLTKIDRCFQ), an 18-mer peptide containing the immunodominant

epitope Poziotinib in vivo region of Equ c 1, was synthesized and purified by GL Biochem (Shanghai, China). Recombinant (r) Equ c 1 was produced in Pichia pastoris, as described previously.[11] Fourteen clinically diagnosed horse-allergic subjects (subjects A–N) with positive (≥ 3 mm) skin prick tests with rEqu c 1 and nine horse dust-exposed non-atopic control Janus kinase (JAK) subjects (subjects O–W) with negative skin prick tests were recruited to the study. The subjects were characterized

at the Pulmonary Clinic of Kuopio University Hospital, as described in detail previously.[11] In brief, the allergic subjects exhibited a positive horse UniCAP result (FEIA; Pharmacia, Uppsala, Sweden; > 0·7 kU/l) and a positive skin prick test (≥ 3 mm) with a commercial horse epithelial extract (ALK Abellò, Hørsholm, Denmark), whereas the control subjects were negative in these tests. The non-atopic control subjects had horse riding as a hobby, and were therefore constantly exposed to horse allergens. Human leucocyte antigen (HLA) class II genotyping for the DQ and DR alleles of the subjects was performed in the Clinical Laboratory of the Finnish Red Cross Blood Service (Helsinki, Finland[12]) or in the Immunogenetics Laboratory of the University of Turku (Turku, Finland[13]) with PCR-based lanthanide-labelled sequence-specific oligonucleotide hybridization (Supplementary material, Table S1). Signed informed consent was provided by all subjects participating in the study and the study was approved by the Ethics Committee of Kuopio University Hospital, permission # 182/99.