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Subcutaneous Infection with S

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1 Subcutaneous Infection with S
Subcutaneous Infection with S. aureus in Mice Reveals Association of Resistance with Influx of Neutrophils and Th2 Response  Nadine Nippe, Georg Varga, Dirk Holzinger, Bettina Löffler, Eva Medina, Karsten Becker, Johannes Roth, Jan M. Ehrchen, Cord Sunderkötter  Journal of Investigative Dermatology  Volume 131, Issue 1, Pages (January 2011) DOI: /jid Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Susceptibility to Staphylococcus aureus-induced skin infection in inbred mouse strains. Different inbred mouse strains were inoculated subcutaneously with 2 × 107 CFU of S. aureus strain SH1000 or phosphate-buffered saline (PBS). (a) Time course of footpad swelling of S. aureus-infected BALB/c (open squares), C57BL/6 (black squares), and DBA/2 (gray squares) mice is shown. (b–d) Bacterial loads in systemic organs of S. aureus-infected BALB/c (open bars), C57BL/6 (black bars), and DBA/2 (gray bars) mice at different time points after infection. (b) Bacterial loads of S. aureus-infected footpads, (c) draining popliteal lymph nodes, and (d) inguinal lymph nodes of S. aureus-infected mice. (e) Bacterial loads in kidneys of S. aureus-infected mice. Bars represent the mean ± SEM of 10 mice per group of three independent experiments. *P<0.05; **P<0.001. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Acute inflammatory response to skin infection with Staphylococcus aureus. Myeloperoxidase (MPO) activity was determined as a parameter for the amount of neutrophils in footpad tissue homogenates (a) 7 hours and (b) 48 hours after injection of 2 × 107 CFU of S. aureus strain SH1000 or phosphate-buffered saline (PBS) into the footpad of BALB/c (open bars), C57BL/6 (black bars), and DBA/2 (gray bars) mice. Data represent the mean ± SEM of 12 mice per group from three independent experiments. *P<0.05. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Influx of polymorphonuclear leukocytes in Staphylococcus aureus-infected skin. (a) Representative sections of staining for Ly-6G of footpads 48 hours after infection with 2 × 107 CFU of S. aureus strain SH1000 or phosphate-buffered saline (PBS). (b) Numbers of infiltrated Ly-6G+ cells per field. (c) Percentage of Ly-6G+ cells compared with total numbers of cells. BALB/c (open squares), C57BL/6 (black squares), and DBA/2 (gray squares) mice. n=4 per group, *P<0.05. Bar=0.2mm. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Expression of chemokines in Staphylococcus aureus-infected tissue. Time course of (a) CXCL-1 and (b) CXCL-2 mRNA expression in footpad tissue of BALB/c (open squares), C57BL/6 (black squares), and DBA/2 (gray squares) mice subcutaneously injected with 2 × 107 CFU of S. aureus strain SH1000. PCR data were normalized to ribosomal protein L9 expression and relative copies and mean ± SEM were calculated of nine mice per group of three independent experiments. (c) Secreted CXCL-2 concentration in lavage of footpad tissues of BALB/c (open bars), C57BL/6 (black bars), and DBA/2 (gray bars) mice 48 hours after infection with 2 × 107 CFU of S. aureus strain SH1000 or phosphate-buffered saline (PBS). Results represent mean ± SEM of nine mice per group of three independent experiments. *P<0.05. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Cytokine production by CD4+ T cells isolated from Staphylococcus aureus-infected mice. CD4+ T cells were isolated from draining lymph nodes of S. aureus-infected BALB/c (open bars), C57BL/6 (black bars), and DBA/2 (gray bars) mice 16 days after infection and were co-cultured with syngeneic bone marrow-derived dendritic cells (bmDCs) stimulated with heat-killed S. aureus (SH1000 AG; ratio of 40:1 CD4+ T cells: bmDCs) or unstimulated syngeneic bmDCs (w/o AG). After 7 days, the cytokine production was assessed from supernatants of the co-cultures. (a) IFN-γ, (b) IL-4, (c) IL-13, and (d) IL-17. Results represent the mean ± SEM of 10 mice per group of three independent experiments. *P<0.05. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

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