Transduction of the E6 and E7 Genes of Epidermodysplasia-Verruciformis-Associated Human Papillomaviruses Alters Human Keratinocyte Growth and Differentiation.

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Presentation transcript:

Transduction of the E6 and E7 Genes of Epidermodysplasia-Verruciformis-Associated Human Papillomaviruses Alters Human Keratinocyte Growth and Differentiation in Organotypic Cultures Ingeborg L.A. Boxman, Linda H.C. Mulder, Jan ter Schegget Journal of Investigative Dermatology Volume 117, Issue 6, Pages 1397-1404 (December 2001) DOI: 10.1046/j.0022-202x.2001.01602.x Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Histology of epithelial raft cultures developed from uninfected PHKs and PHKs infected with recombinant retroviruses containing HPV E6 and E7 genes. Uninfected PHKs and PHKs infected with the vector-only virus (pLXSN) or with retroviruses containing E6 and E7 genes of various HPV types were seeded on fibroblast populated collagen lattices and subsequently grown for 9 d at the air–medium interface. Five micron sections were stained with hematoxylin and eosin. The HPV types are indicated at each panel. Two enlargements with details of papilla-like structures are shown (third and fourth panel in bottom row). Scale bars: 100 μm. Journal of Investigative Dermatology 2001 117, 1397-1404DOI: (10.1046/j.0022-202x.2001.01602.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 EV-HPV E6 and E7 transduction delays or disrupts squamous differentiation in epithelial raft cultures. Five micron serial sections as described in Figure 1 were stained with antibodies to K10 (left two columns) and to SPRR2 (right two columns). The recombinant retroviruses used in the infection are indicated; PHK, uninfected keratinocytes; pLXSN, keratinocytes infected with the vector-only virus. Scale bar: 200 μm. Journal of Investigative Dermatology 2001 117, 1397-1404DOI: (10.1046/j.0022-202x.2001.01602.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 EV-HPV E6 and E7 transduction induces PCNA without BrdU incorporation in suprabasal cells in epithelial raft cultures. Five micron serial sections as described in Figure 1 were stained with antibodies to PCNA (left two columns) and to BrdU (right two columns). The recombinant retroviruses used in the infection are indicated; PHK, uninfected keratinocytes; pLXSN, keratinocytes infected with the vector-only virus. Scale bar: 100 μm. Journal of Investigative Dermatology 2001 117, 1397-1404DOI: (10.1046/j.0022-202x.2001.01602.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Transcription of EV-HPV E6 and E7 genes in raft cultures as detected by in situ hybridization. Five micron serial sections as described in Figure 1 were hybridized with 35S-labeled riboprobes and photographed with bright-field illumination. Riboprobes applied to the sections were HPV5 E6E7 antisense (5AS), HPV5 E6E7 sense (5S), HPV17 E6E7 antisense (17AS), and HPV17 E6E7 sense (17S). Cultures shown were developed from PHKs infected with the vector-only virus (pLXSN), with pLXSN5E6E7, or with pLXSN17E6E7 as indicated. Sections were exposed for the number of days indicated. Scale bar: 200 μm. Journal of Investigative Dermatology 2001 117, 1397-1404DOI: (10.1046/j.0022-202x.2001.01602.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions