Open Access Highly Accessed Research article

Important role of CCR2 in a murine model of coronary vasculitis

Hernan G Martinez1,2, Marlon P Quinones3, Fabio Jimenez1,2, Carlos Estrada1, Kassandra M Clark1, Kazuo Suzuki4, Noriko Miura5, Naohito Ohno5, Sunil K Ahuja2,6 and Seema S Ahuja1*

1 Department of Medicine (MC 7870), University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX, 78229-3900, USA

2 South Texas Veterans Health Care System, Audie L. Murphy Division, San Antonio, TX, 78229-3900, USA

3 Department Psychiatry, University of Texas Health Science Center at San Antonio (UTHSCSA), San Antonio, TX, 78229-3900, USA

4 Inflammation Program, Department of Immunology, Chiba University Graduate School of Medicine, Chiba, 260-8670, Japan

5 Laboratory for Immunopharmacology of Microbial Products, School of Pharmacy, Tokyo University of Pharmacy and Life Science, Hachioji Tokyo, 192-0392, Japan

6 The Veterans Administration Center for Aids and HIV-1 infection, South Texas Veterans Health Care System, San Antonio, TX, USA

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BMC Immunology 2012, 13:56 doi:10.1186/1471-2172-13-56

Published: 17 October 2012

Additional files

Additional file 1:

Figure S1. Protocol for coronary/aortic inflammation in mice using CAWS. A full cycle of CAWS included two rounds of intra-peritoneal (I.P.) injections (1 mg/mouse/day for five consecutive days) administered four weeks apart as previously described47-51. Early experiments were conducted with a dose of 4 mg instead of 1 mg of CAWS per day. Results derived from either dose had identical disease incidence and severity. In some experiments, mice were sacrificed 10 days after the first five days of CAWS injections (cycle one). At this time point we were unable to identify ongoing inflammation in the coronary or aortic walls (data not shown). We identified inflammation clearly and consistently in 100% of mice 30 days after the completion of the first cycle.

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Additional file 2:

Figure S2. Coronary and aortic analysis of macrophages. Coronary and aortic macrophages were immunostained with the ER-HR3 antibody in Ccr2+/+ and Ccr2−/− mice including isotype control.

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Additional file 3:

Figure S3. Serum levels of Myeloperoxidase (MPO). MPO levels were detected by ELISA after full cycle of CAWS in Ccr2+/+ and Ccr2−/− mice.

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Additional file 4:

Figure S4. Antibody against MPO (anti-MPO), IgG1 and IgG2a levels Ccr2+/+ and Ccr2−/− mice after full cycles of CAWS. A. Serum antibodies against MPO detected by ELISA in Ccr2+/+ and Ccr2−/− mice after two cycles of CAWS, including PBS control (values expressed as optical density). B-C. Serum levels of anti-IgG1 and anti-IgG2a, in Ccr2+/+ and Ccr2−/− mice after two cycles of CAWS.

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Additional file 5:

Figure S5. Th1 and Th2 response in spleen of Ccr2+/+ and Ccr2−/− mice. Percentage of IFNγ (Th1) and IL-4 (Th2) in splenocytes after full cycle of CAWS in Ccr2+/+ and Ccr2−/− mice. Each bar represents the mean ± SE from a representative experiment with 6–8 mice per group.

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Additional file 6:

Table S1. Flow cytometric markers used to identify specific cell type.

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