Supplementary Material for: Cellular and Humoral Responses to Cte f 2, a Cat Flea Allergen, in Children with Papular Urticaria

<b><i>Background:</i></b> Papular urticaria (PU) is a common insect bite skin hypersensitivity in tropical countries. In order to gain insight into its causal allergens, we aimed to evaluate cellular and humoral immune responses to the recombinant salivary antigen Cte f 2 from the cat flea <i>Ctenocephalides felis</i>. <b><i>Method:</i></b> Sixty patients with PU and 27 healthy controls were included in this study. Specific IgE, IgG, IgG1, and IgG4 against Cte f 2 and <i>C. felis</i> extract were determined by ELISA. The T-cell response was analyzed using a carboxyfluorescein succinimidyl ester (CFSE)-based dilution assay and Th1/Th2/Th17 cytokine measurements. In addition, a proteomic analysis of IgG and IgE reactive spots of <i>C. felis</i> extract was performed. <b><i>Results:</i></b> The frequency of IgE sensitization to Cte f 2 was similar between patients (36.7%) and controls (40.7%). The specific IgE, IgG1, and IgG4 responses to Cte f 2 and <i>C. felis</i> extract were not significantly different between patients and controls. Among the 3 conditions (i.e., Cte f 2, <i>C. felis</i> extract, and only medium) Cte f 2 was the strongest inducer of CD3+CD4+ proliferation in the patients; however, the mean response was not significantly different from those in controls (Cte f 2: 4.5 vs. 2.5%; <i>p</i> = 0.46). No salivary proteins were identified in <i>C. felis</i>, and most of the spots were identified as muscle-skeletal components (tropomyosin, actin, myosin, and ankirin). <b><i>Conclusions:</i></b> Cte f 2 induces IgE and IgG production as well as T-cell proliferation in children living in a geographical area where PU induced by a flea bite is common. The use of <i>C. felis</i> extract is not recommended for the study of bite-induced hypersensitivity disease since salivary antigens are not well represented.