Ccr2 mice

Per1^−/− mice^{45 (link)} were obtained from Dr. CC Lee at Baylor College of Medicine, Houston, TX, USA. Ccr2^−/− mice were obtained from Jackson Lab (Bar Harbor, ME, USA). The Per1^−/−; Ccr2^−/− (DKO) mice were generated at the expected Mendelian ratios and developed normally. All of the animals were backcrossed for at least five generations before the first pilot study to ensure a largely homogenous C57BL/6J background. Male WT C57BL/6J mice and gene knockout mice at 8–10 weeks of age were used in this work. The animals were maintained in cycles of 12 h of light and 12 h of darkness with free access to food and water ad libitum. All animal care and use procedures were in accordance with the guidelines of the Institutional Animal Care and Use Committee at Nanjing University of Science and Technology.

C57BL/6 and CCR2−/− mice were purchased from Jackson Laboratory (Bar Harbor, ME, USA). IFNAR1−/− mice were obtained from Dr. Chien-Kuo Lee (Graduate Institute of Immunology, National Taiwan University, Taipei, Taiwan). MyD88−/− mice were obtained from Hui-Chen Chen (Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan). Mice were maintained under specific pathogen free conditions in Chang Gung University. All animal experiments were performed according to the animal protocol approved by the Institutional Animal Care and User Committee of Chang Gung University and in accordance with the guidelines of Animal Care and Use of Laboratory Animals of the Taiwanese Council of Agriculture.

Myd88^−/− mice were used with permission of Dr. S. Akira (34 (link)), Mincle^−/− mice have been described (59 (link)) and were used with permission by the Consortium for Functional Glycomics. Ccr2^−/− mice were obtained from Jackson Laboratories. C57BL/6, Mincle^−/−, Myd88^−/−, and Ccr2^−/− mice were bred at the animal facility of the Medical Faculty in Erlangen. Mcl-1^flox/flox;LysM-Cre conditional knockout mice were bred and immunized at St. Jude Children’s Research Hospital. For some experiments, C57BL/6 mice were purchased from Charles River. Liposomes and recombinant H1 were provided by the Statens Serum Institut. Adjuvant formulations were prepared as described (10 (link)). Mice were injected with 100 µl i.p. or 2 × 50 µl liposomes or 10 nmol CpG 1826 s.c. into the hind footpads. Two micrograms of H1 was only added if antigen-specific re-stimulation was performed. In some experiments, 7-nitrobenzo-2-oxa-1,3-diazole (NBD)-fluorescently labeled liposomes were injected in order to visualize the adjuvant.

C57BL/6J, B6.129S7-Ifng^tm1Ts/J (Ifng^-/-), and B6.129S1-Il12b^tm1Jm/J (Il12p40^-/-) mice were purchased from The Jackson Laboratory. Ccr2^-/- mice [54 (link)], YET40 mice [58 ], and Il12p40^-/- mice were purchased from The Jackson Laboratory and maintained and bred in a specific pathogen-free facility at the University of Pennsylvania.

Specific pathogen-free AKR, BALB/c, and C57BL/6 mice were purchased from Harlan. CX3CR1^gfp/+ mice were bred at the University of Manchester. CCR2^−/− mice were purchased from The Jackson Laboratory. All strains of mouse were maintained in individually ventilated cages. Only the males were used in experiments when they were 6–12 wk old. The mouse studies were reviewed and approved by the Home Office and performed under the strict legal requirements of the Animal (Scientific Procedures) Act 1986 (as amended).

Ccr2^–/– mice (strain 004999) were obtained from The Jackson Laboratory. To generate KPC-CCR2^–/– mice, Kras^LSL-G12DPdx1-Cre (KC) mice were crossed with p53^LSL-R172H/+ or p53^{LSL-R172H/LSL-R172H} mice to generate KPC mice. We crossed CCR2^–/– mice with KC mice to generate KC-CCR2^–/– mice, and these were then crossed with p53^R172H/+ or p53^{LSL-R172H/LSL-R172H} mice to generate KPC-CCR2^–/– mice. The KPC-CCR2^–/– mice were of C57BL/6 background. KPC mice on the C57BL/6 background were used as the control for the comparative studies. The progeny were born in an expected Mendelian ration, with no obvious functional defects. In vivo drug studies were done using a KPC genetically engineered mouse model (68 (link)) on a mixed background as previously described. At the endpoint (or, for the early studies, at 10–11 weeks), full necropsies were performed on all study animals and included a gross examination of all organs for macroscopic disease as previously described (15 (link), 67 (link), 69 (link)).