March 1, 2019
Plos Pathogens
Product: Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae
Mice:
C57BL/6J (stock #000664), MHCIIfl/fl (stock #013181) [89], and Arg1fl/fl (stock #008817) [90] mice were purchased from the Jackson Laboratory (JAX). CCR2-/- mice (JAX stock #004999) [34] were generously provided by Dr. E Pamer (MSKCC). Rosa26flSTOP-tdRFP mice [36] were generously provided by Dr. J. Sun (MSKCC) [91]. The dblGATA mice [50] on a C57BL/6 background were generously provided by Dr. H. Rosenberg (NIH). Vav1-Cre mice (JAX stock #008610) [92] were generously provided by Dr. F Geissmann (MSKCC). RAG2-/-c-/- mice (stock #4111) [93, 94] were purchased from Taconic. CD45.1+ mice (stock #564) were purchased from Charles River Laboratories. The CCR2-DTR depleter mice and CCR2-GFP reporter mice were generated as previously described [5, 95]. All mouse strains were bred and housed in the Memorial Sloan Kettering Cancer Centers (MSKCC) Research Animal Resource Center under specific pathogen-free conditions. Mice on the RAG-/-c-/- background were maintained on amoxicillin- and Vitamin E-containing chow. To ablate monocytes, CCR2-DTR or CCR2-DTR RAG-/-c-/- mice and their control littermates were injected intraperitoneally with 200 ng (10 ng/g body weight) of diphtheria toxin (List Biological Laboratories) every other day for three doses, starting the day before infection (Fig 1B), unless otherwise noted. All experiments were conducted using male and female mice at age 68 weeks with sex- and age-matched mice in experimental and control groups. Experiments with CCR2-DTR, CCR2-DTR RAG-/-c-/-, CCR2-Cre and Vav1-Cre mice used littermate control mice that were weaned from the same litters and co-housed.