B6 sjl ptprca pepcb boyj cd45.1

All mice were maintained and bred in-house in an American Association of Laboratory Animal Care-approved facility at the University of California, Berkeley. All procedures were approved by the University of California, Berkeley Animal Use and Care Committee. C57BL/6, C57BL/6-Tg(Ins2-TFRC/OVA)296Wehi/WehiJ (RIPmOVA), B6(Cg)-Rag2tm1.1Cgn/J (Rag2^−/−), B6.C-H2-Kbm1/ByJ (Kbm1), B6.129P2-Il2tm1Hor/J (IL-2^−/−), and B6.SJL-Ptprca Pepcb/BoyJ (CD45.1) were from Jackson Labs. OT-I Rag2^−/− mice were from Taconic Farms. OT-I Rag2^−/− CD45.1 mice were generated by crossing OT-I Rag2^−/− mice to CD45.1 mice. F5 Rag1^−/− mice⁵⁴ , CD11cYFP mice^{55 (link)}, and AireGFP mice^{56 (link)} have been previously described. CD11cYFP RIPmOVA mice and AireGFP RIPmOVA mice were generated by crossing CD11cYFP or AireGFP mice to RIPmOVA mice, respectively. IL15Rα^−/−mice were generated by crossing B6.C-Tg(CMV-cre)1Cgn/J (Cmv-cre) mice with C57BL/6-Il15ratm2.1Ama/J (IL15R flox/flox) mice, both from Jackson Labs. To generate bone marrow chimeras, bone marrow was collected from the tibias and femurs of donor mice, then treated with ammonium chloride–potassium bicarbonate buffer to lyse red blood cells. Host mice were lethally irradiated (900rads) prior to intravenous injection of 5×10⁶ bone marrow cells.

BALB/c, C57BL/6 (B6), B6.129S7-Il1r1^tm1Imx/J (Il1r1^−/−), B6.C3(Cg)-Rora^sg/J (Rora^sg/sg), B6.SJL-Ptprc^aPepc^b/BoyJ (CD45.1), and C.129S7(B6)-Rag1^tm1Mom/J (Rag1^−/−) mice were purchased from Jackson Laboratory (Bar Harbor, ME). Il13^egfp/egfp mice (IL-13-deficient mice) (17 (link), BALB/c background) and Rora^fl/flIl7r-Cre mice (13 (link), B6 background) were maintained in specific pathogen-free conditions at Mayo Clinic (Rochester, MN). ILC2-deficient mice were generated by reconstituting lethally irradiated CD45.1 mice with 2–3 million bone marrow (BM) cells isolated from WT or Rora^sg/sg littermates (18 ). Animals used in this study were female and ranged from 7 to 12 weeks of age. All protocols and procedures for the handling of the mice were reviewed and approved by the Mayo Institutional Animal Care and Use Committee, Mayo Clinic.

Enhanced GFP mice were bred in-house on a C57BL/6 background. C57BL/6J and B6.SJL-Ptprc^a Pepc^b/BoyJ (CD45.1) mice were obtained from Jackson Laboratories. Mice matched for sex, age (+/− 7 days), and body mass (+/− 3 g) were co-housed from the age of 4–5 weeks, with a density no greater than 5 mice per cage. The animals were monitored daily for evidence of fighting and were exposed to gel food diet supplement at least 2 times, 2–6 weeks before performance of parabiotic attachment. Animals were randomly assigned a pair member, with the appropriate transgene or CD45 congenic allotype, within their co-housed cohort. Pairs established for parabiotic attachment were “cage paired,” meaning they were housed only with their future parabiotic pair member, 2 weeks before the parabiosis procedure, and the mice were observed again for aggressive behavior. Aggressive and injured mice were removed from the study. Aggression at this stage was highly uncommon, occurring in only 1 pair out of the 21 included in this study.

Specific pathogen-free 8-12 week old female C57Bl/6J, B6.PL-Thy1a/CyJ (Thy1.1+), C57BL/6-Tg(TcraTcrb)1100Mjb/J (OT-I mice), B6.SJL-Ptprca Pepcb/BoyJ (CD45.1+), and CD4-cre mice were purchased from Jackson Laboratories. p110δ−/− mice and p110δ^fl/fl mice (9 (link), 14 , 29 (link)) were a kind gift from Dr. Martin Turner (Babraham Institute, UK). OT-I mice were backcrossed with p110δ−/− mice to generate p110δ−/−OT-I mice (all on C57BL/6 background). The Drexel University and Drexel University College of Medicine, Institutional Animal Care and Use Committee (IACUC) reviewed and approved the animal care and use protocol for this study. All mice were maintained in AAALAC certified barrier facilities at Drexel University College of Medicine. Mice were anesthetized with 2-2-2-tribromoethanol (avertin; 250 mg/Kg i.p., Acros), and infected intranasally (i.n.) with a sublethal dose (10 TCID₅₀) of A/Puerto Rico/8/34 (PR8 H1N1) viral strain (generous gift of Dr. W. Gerhard, Wistar Institute, Philadelphia, PA) or with a sublethal dose (1.87x10³ TCID₅₀) of the OVA_257-264-expressing influenza A/WSN/33 (WSN-OVA H1N1) virus strain (kind gift from Dr. David Topham, University of Rochester). Some mice were intravenously infected with 10⁴ cfu OVA-expressing Listeria monocytogenes (L.m.-OVA) (kind gift from Dr. Hao Shen, University of Pennsylvania).