Sustained increases in numbers of pulmonary dendritic cells after respiratory syncytial virus infection Marc Beyer, MD, Holger Bartz, MD, Katharina Hörner,

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Sustained increases in numbers of pulmonary dendritic cells after respiratory syncytial virus infection Marc Beyer, MD, Holger Bartz, MD, Katharina Hörner, MD, Sandra Doths, Cordula Koerner-Rettberg, MD, Jürgen Schwarze, MD Journal of Allergy and Clinical Immunology Volume 113, Issue 1, Pages 127-133 (January 2004) DOI: 10.1016/j.jaci.2003.10.057

FIG 1 Low dose RSV infection results in peribronchial and perivascular inflammation in the lung. Mice were infected with RSV or UV-RSV and killed after 6 and 21 days. Lungs were inflated with formalin-saline, excised, and embedded in paraffin wax. Sections were stained with hematoxylin-eosin, and representative images were taken at ×100 magnification. Journal of Allergy and Clinical Immunology 2004 113, 127-133DOI: (10.1016/j.jaci.2003.10.057)

FIG 2 MHC II and CD86 expression on pulmonary CD11c+ cells after RSV infection. Mice were naïve or infected intranasally with either RSV or UV-RSV. Lung MNCs from naïve mice and on days 4, 6, 10, 14, and 21 after infection were stained with antibodies to CD11c, MHC II, and CD86 and analyzed by FACS. A, Absolute numbers of CD11c+ MHC IIhi cells (DC) per lung after inoculation with either RSV (■) or UV-RSV (□) are expressed as mean ± SD (n = 4, ∗P < .05 vs UV-RSV). B, Expression of CD11c vs MHC II in pulmonary MNCs after RSV infection is presented in dot plots from a representative experiment. Numbers indicate the percentage of cells in the appropriate quadrant. C, Expression of CD86 (gray bars) and MHC IIhi (open bars) on pulmonary CD11c+ cells of naïve mice or after RSV infection are expressed as mean ± SD (n = 4, ∗MHC II, §CD86; P < .05 vs naïve). Journal of Allergy and Clinical Immunology 2004 113, 127-133DOI: (10.1016/j.jaci.2003.10.057)

FIG 3 Flow cytometric analysis of isolated CD11c+ lung cells. CD11c+ lung cells isolated 10 days after mock or RSV infection were stained with anti-CD11c-PE and anti-F4/80-FITC or with anti-CD11c-FITC and anti-CD4-PE or anti-CD8α-PE antibodies (open histogram) or isotype controls (shaded histogram). Histograms illustrate expression of these surface markers on cells gated for expression of CD11c from a representative experiment (n = 4, 4 mice pooled per experiment). Percentages of double positive cells are indicated. Journal of Allergy and Clinical Immunology 2004 113, 127-133DOI: (10.1016/j.jaci.2003.10.057)

FIG 4 Phagocytosis by isolated pulmonary CD11c+ cells. Pulmonary CD11c+ cells were isolated on day 10 after mock (open bars) or RSV infection (grey bars). The remaining negative fraction was designated as CD11c− cells. Cells of each population were incubated with FITC-DX at 37°C or 4°C, and uptake was determined by FACS analysis as MFI. Values of ΔMFI (MFI-37°C – MFI-4°C) are expressed as mean ± SD (n = 3, ∗P < .05 RSV vs mock). Journal of Allergy and Clinical Immunology 2004 113, 127-133DOI: (10.1016/j.jaci.2003.10.057)

FIG 5 T-cell stimulatory capacity of isolated pulmonary CD11c+ cells. CD11c+ cells were isolated from lungs on day 10 after mock (□, ○) or RSV infection (■, ●); the remaining negative fraction was designated as CD11c− cells. Splenocytes of C57Bl/6 mice were enriched for T cells, and 105 cells were incubated alone or with increasing numbers of CD11c+ (□, ■) or CD11c− (○, ●) cells. T-cell proliferation was determined by 3H-thymidine incorporation after 5 days of culture. Cpm from a representative experiment are expressed as mean ± SD (n = 3, P < .05; ∗CD11c+ from RSV/mock vs T cells alone, §CD11c+ from RSV vs CD11c+ from mock and T cells alone). Journal of Allergy and Clinical Immunology 2004 113, 127-133DOI: (10.1016/j.jaci.2003.10.057)