C57bl 6 wild type mice

Female BALB/c and C57BL/6 wild-type (wt) mice were purchased from Janvier Laboratories (Le Genest-Saint-Isle, France) and acclimatized for at least 10 days before challenge. BALB/c ST2-KO mice [10 (link)] and C57BL/6 IL-33-KO mice were backcrossed for at least 10 generations. ST2-KO mice were originally obtained from Dr Andrew McKenzie (MRC Laboratory of Molecular Biology, Cambridge, UK) and IL-33-KO mice were kindly provided by Jean-Philippe Girard (Université de Toulouse, Toulouse, France). Mice were bred and housed in our animal facilities. Mice were 7 to 10 weeks-old when challenged with L. donovani. Naïve congenic mice, matched according to age, were used as non-infected controls. The results were obtained in two to three independent experiments, with a total of 5 to 13 mice per time point.

C57BL/6 wildtype (WT) mice were obtained from Janvier Labs (Saint Berthevin) or from Charles River Laboratories (Sulzfeld). All mice were maintained at the Walter Brendel Centre of Experimental Medicine, LMU, Munich, or at the core facility animal models at the Biomedical Center, LMU, Planegg-Martinsried, Germany and at the Zentrale Versuchstierhaltung Innenstadt (ZVH), LMU Munich, Germany. Eight to 25 weeks old male and female mice were used for all experiments. Neonatal WT mice were used for unilateral ureteral obstruction experiments.

C57BL/6 wild-type mice obtained from Janvier Labs were used for all experiments. Mice were housed in a specific pathogen-free facility and were aged of 8 weeks at the beginning of experiments. All animal experiments were approved by the local institutional ethic committee.
Adenosine 5’-tri-phosphate disodium salt (A2383) and β-nicotinamide adenine dinucleotide hydrate (N7004) were purchased from Sigma Aldrich. Red blood cell (RBC) lysis/fixation Solution, True-Nuclear transcription factor buffer set, fluorochrome-conjugated streptavidin, and antibodies to CD45 (clone 30-F11), CD4 (RM4-5), CD8 (53-6.7), CD25 (PC-61), CD19 (1D3/CD19), B220 (RA3-6B2), FoxP3 (MF-14), CD27 (LG.3A10), CD62L (MEL-14), CD69 (H1.2F3) or P2X7R (1F11), and purified CD16/CD32 (TruStain FcX) were obtained from Biolegend or Sony Biotechnology. Rabbit polyclonal antibody K1G, specific to mouse P2X7, was described in our previous studies (12 (link), 13 (link)). K1G was used to stain P2X7 at the surface of blood myeloid cells as illustrated in Supplemental Figure 3, using a secondary donkey anti-rabbit IgG from Jackson ImmunoResearch. Biotinylated polyclonal antibody specific to mouse IgGa was obtained from Jackson ImmunoResearch and monoclonal antibody to ARTC2.2 (Nika102) from Novus Biologicals.

Eight to ten weeks old female C57BL/6 wild-type mice (Janvier, Le Genest St. Isle, France) were separated into two groups of four mice each. Animals were injected intravenously into the tail vein with ∼7 nmol/kg body weight (100 μl, 1.4 μM) of Eu-labeled C[ABD]SL[Eu] or the control CSL[Eu]. Blood was drawn from the tail vein after 0.25, 24, 72, 120, and 144 h. Heparinized blood was centrifuged and plasma was collected. Mice were euthanized at 144 or 216 h, and organs were homogenized using a TissueLyser (Qiagen, Valencia, CA, United States) in PBS (1:1 weight/volume ratio). The Europium content in the blood and organ samples was determined using an Infinite M1000 PRO (Tecan, Durham, NC, United States) time-resolved fluorescence plate reader. Obtained values were correlated to a spiked standard dilution series of respective proteins in mouse serum or a homogenate of respective organs.
Serum concentrations were plotted against time, and the four last values were used to calculate the beta half-life of the protein using a one phase decay model in Prism (GraphPad Software, San Diego, CA, United States). Student’s t-test was used to compare protein concentrations in the serum at the last time point at 144 h. For comparing protein concentrations in organ samples, a two-way ANOVA on log-transformed data followed by Sidak’s multiple comparisons test was performed.

Female BALB/c and C57BL/6 wild-type mice were purchased from Janvier Labs (Saint-Berthevin, France) and maintained under standard specific pathogen-free conditions. All animal experiments were performed in accordance with the National Animal Protection Guidelines and approved by the German Animal Ethics Committee for the Protection of Animals (G0113/15, H0438/17). G. muris cysts were originally purchased from Waterborne, Inc. (USA) and were later maintained by passage in BALB/c mice. Cysts were isolated from fresh fecal samples via a sucrose gradient, as described previously^{55 (link)} and were administered to mice via oral gavage at a dosage of 1,000 cysts per mouse suspended in 200 μl distilled water. After two weeks, G. muris-infected mice and age/sex-matched naïve controls were sacrificed by isoflurane inhalation, followed by cervical dislocation. For faecal cyst counts, 3–4 faecal pellets were collected individually from each mouse 3 times a week, followed by weighing and homogenization in 2 mL distilled water. Cyst numbers were counted in a Neubauer hemocytometer chamber and for weekly cyst shedding over the course of a 6 week infection period as presented in Fig. 1a, the mean value of cyst shedding was calculated across the three collection time points per week.

Experiments were performed with female C57Bl/6 wild type mice (Janvier), ${\text{sPLA}}_2^{\text{tg}}$ mice (23 (link)), Trpa1 KO mice [Jackson; (24 (link))] and male Swiss wild type mice (Federal University of Parana colony). The ${\text{sPLA}}_2^{\text{tg}}$ mouse model contains a transgene for secreted phospholipase A2. The background strain is C57Bl/6. The Trpa1 KO mouse background strain is a mix of C57Bl/6J with FVB 129P2/OlaHsd. All mice were aged 8 weeks to 8 months. The difference in age is considered to have no influence on the nociception and itch responses. Mice were housed conventionally with ad libitum water and food, consisting of regular chow (Teklad 2916, Envigo). Experiments were conducted according to the institutional Animal Care and Use Committee regulations. All methods were carried out in accordance with relevant guidelines and regulations. The study was also carried out in compliance with the ARRIVE guidelines.