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. Author manuscript; available in PMC: 2011 Apr 26.
Published in final edited form as: Cancer Cell. 2010 Feb 4;17(2):121–134. doi: 10.1016/j.ccr.2009.12.019

Figure 1. B Cells Are Critical Regulators of Premalignant Progression in HPV16 Mice.

Figure 1

(A) Percentage of CD45+ cells in skin single cell suspensions isolated from negative littermates (−LM), HPV16/JH+/−, and HPV16/JH−/− mice at 1, 4, and 6 months of age assessed by flow cytometry.

(B and C) Mast cells (B, blue staining) and Gr1+ myeloid cells (C, brown staining) in skin of HPV16/JH+/− and HPV16/JH−/− mice at 1, 4, and 6 months of age assessed quantitatively after chloroacetate esterase histochemistry or Gr1 immunohistochemistry (IHC), respectively.

(D) Flow cytometric analysis of immune cell lineages expressed as percentages of total CD45+ leukocyte infiltrates in ear tissue of negative littermates (−LM), HPV16, HPV16/JH+/−, HPV16/JH−/−, and HPV16/RAG1−/− mice at 1, 4, and 6 months of age.

(E) Dendritic (CD11c+) and macrophage (F4/80+) lineage cell composition of CD11b+Gr1+ (left) and CD11b+Gr1− (right) myeloid populations evaluated by flow cytometry in skin of negative littermates (−LM), HPV16, and HPV16/JH−/− mice at 4 months of age.

(F) Angiogenic vasculature in skin tissue sections from negative littermates (−LM), HPV16/JH+/−, and HPV16/JH−/− mice at 1, 4, and 6 months of evaluation by CD31/PECAM-1 IHC revealing endothelial cells (brown staining).

(G) Reduced VEGF-A and active MMP-9 protein levels in skin extracts from HPV16/JH−/− versus HPV16/JH+/− mice (4 and 6 months) as assessed by ELISA.

(H) Keratinocyte proliferation in skin of negative littermates (−LM), HPV16/JH+/−, and HPV16/JH−/− mice at 1, 4, and 6 months of age, as evaluated by quantitation of bromodeoxyuridine (BrdU)-positive keratinocytes (red staining).

(I) Percentage of ear skin in HPV16/JH−/− and HPV16/JH+/− mice developing hyperplastic lesions by 1 month of age (Hyp) or dysplasia by 4 and 6 months of age (Dys).

(A–I) Results shown represent mean ± SEM (n = 5–8 mice) and asterisks (*) indicate statistically significant differences (p < 0.05, Mann-Whitney). Representative images of HPV16/JH+/− and HPV16/JH−/− mouse skin at 4 months of age are shown. Values represent average of five high-power fields of view per mouse and five mice per category. FOV, field of view; solid red line, epidermal-dermal interface; e, epidermis; d, dermis. Scale bars represent 50 μm. See also Figure S1.