Immune response to Toxocara canis infection in C57BL/6 mice

Experimental Design:Two different experiments were conducted.In the first experiment, eight-week-old female C57BL/6J mice (n = 20) were orally infected with 1,000 infective eggs of T. canis suspended in a 0.9% NaCl solution. Non-infected mice (n = 5) served as controls. Five infected mice were euthanized at 0 (control group), 3, 5, 14, and 28 days post-infection (dpi).In the second experiment, eight-week-old female C57BL/6J mice (n = 18) were treated intranasally with 2 µg of TES in 20 µl of PBS, while control mice (n = 5) received an intranasal treatment with the same volume of PBS. Nasal administration was applied once per day for 3 days. Six TES-treated mice were euthanized at each of the following time points: 3, 14, and 28 days post-first intranasal administration (dpfi). The control mice were euthanized at 3 dpfi.After sacrifice the following procedures were performed:Lung images were taken - folder Lung_imagesLung sections were submitted to hematoxylin-eosin and periodic-acid Schiff staining - folder HEMultiparametric flow cytometric analyses were performed on isolated lung cell suspensions - folder Flow_cytometryTES specific antibodies were measured in sera using ELISA - folder AntibodiesTotal IgE was measured in sera and in lung tissue lysates using ELISA Antibody Pair Kit (StemCell Technologies, Canada) - folder AntibodiesCytokine concentration was measured in splenocyte culture media and lung tissue lysates using commercial DuoSet Elisa kits (R&D Systems, USA) - folders: splenocytes, lung_lysatesCytokine expression in lung tissue lysates was analyzed using the Proteome Profiler™ Mouse XL Cytokine Array (R&D Systems, USA) - folder Proteome_Profiler_Mouse_XL_Cytokine_ArrayGene expression in lungs was assesed using qPCR - folder qPCRExplanation of abbreviations used in attached files:   d0_1-5 – Mice from the control group (not infected, n=5).   d3_1-5 – Mice euthanized 3 days post-infection (n=5).   d5_1-5 – Mice euthanized 5 days post-infection (n=5).   d14_1-5 – Mice euthanized 14 days post-infection (n=5).   d28_1-5 – Mice euthanized 28 days post-infection (n=5).   PBS_d3_1-5 – Control group mice euthanized 3 days after the first PBS intranasal administration (n=5).   TES_d3_1-6 – Mice euthanized 3 days after the first TES intranasal administration (n=6).   TES_d14_1-6 – Mice euthanized 14 days after the first TES intranasal administration (n=6).   TES_d28_1-6 – Mice euthanized 28 days after the first TES intranasal administration (n=6)