Microtainer blood collection tubes with k2edta

For blood isolation, mice (7 weeks of age) were terminally anesthetized with ketamine-xylazine based on the weight of the animal, then the posterior vena cava was severed, and blood collected by heparinized capillary (Fisherbrand) to BD Microtainer blood collection tubes with K₂EDTA (BD) and additional tissues were isolated for RNA extraction. All relevant ethical regulations for work with human participants were complied with and patient blood samples were collected following written informed consent as approved by the University of Florida Institutional Review Board (IRB). Sampling age is included in Supplementary Data 1. Blood was collected into DNA/RNA Shield Blood Collection Tubes (Zymo Research) or BD Vacutainer Tubes, ACD A (BD Biosciences). Peripheral blood mononuclear cells (PBMC) were isolated from the buffy coat of collected whole blood, followed by density gradient separation using Lympholyte-H according to manufacturer’s provided protocol (Cedarlane). Red blood cells were lysed and removed by using the RBC Lysis Buffer (Roche). PBMCs were washed with PBS and used for either genomic DNA isolation (Flexigene kit; Qiagen), total RNA extraction (TRIzol).

Spleen were harvested at sacrifice, resuspended in PBS, and homogenized using gentleMACS™ Octo Dissociator (Miltenyi Biotec, Auburn, CA). Whole blood was collected using BD Microtainer® Blood Collection Tubes with K2EDTA (BD Biosciences). Splenic homogenates and whole blood were then combined and subjected to 1-Step Polymorphs density gradient centrifugation (Accurate Chemical, Westbury, NY). Both PBMCs and PMN buffy coats were harvested and combined. Immune cells were then stained using CellTrace™ CFSE Cell Proliferation Kit according to manufacturer’s specifications (ThermoFisher). Viable cells were counted and ~1 × 10⁷ immune cells were given to each recipient mouse via IP injection.