Journal: Virology

Article Title: Cells of human aminopeptidase N (CD13) transgenic mice are infected by human coronavirus-229E in vitro, but not in vivo

doi: 10.1016/j.virol.2005.02.023

Figure Lengend Snippet: Immunohistochemistry of cryosections shows that the hAPN has the same cellular distribution as mouse APN in transgenic animals. Serial sections from the small intestine or kidney of age matched non-transgenic and hAPN-transgenic mice were tested for reactivity with a rat anti mouse APN (mAPN) MAb-R3-242 or with a mouse anti-hAPN MAb-DW1 and stained using Vector Red and counter stained with hematoxylin. The Vector Red ® substrate is also fluorescent and these images are shown in the panels below the hematoxylin counter-stained images.

Article Snippet: Homozygous, heterozygous, and hemizygous hAPN-transgenic mice all express hAPN with a similar expression pattern and hAPN was expressed on the epithelial surface of the lungs. (A) Serial cryosections of small intestine from age matched non-transgenic females (XX), heterozygous females (X h X), homozygous females (X h X h ), non-transgenic males (XY), and hemizygous males (X h Y) were analyzed for expression of mouse APN (mAPN) using rat MAb-R3-242 to mAPN or with a mouse anti-hAPN MAb-DW1 and binding was determined by a fluorescent substrate (Vector Red ® ). (B) Cryosections of lungs from age matched non-transgenic and homozygous hAPN-transgenic mice were analyzed with a MAb-R3-242 specific to mouse APN (mAPN) or a MAb-DW1 against hAPN.

Techniques: Immunohistochemistry, Transgenic Assay, Staining, Plasmid Preparation

Journal: Virology

Article Title: Cells of human aminopeptidase N (CD13) transgenic mice are infected by human coronavirus-229E in vitro, but not in vivo

doi: 10.1016/j.virol.2005.02.023

Figure Lengend Snippet: Homozygous, heterozygous, and hemizygous hAPN-transgenic mice all express hAPN with a similar expression pattern and hAPN was expressed on the epithelial surface of the lungs. (A) Serial cryosections of small intestine from age matched non-transgenic females (XX), heterozygous females (X h X), homozygous females (X h X h ), non-transgenic males (XY), and hemizygous males (X h Y) were analyzed for expression of mouse APN (mAPN) using rat MAb-R3-242 to mAPN or with a mouse anti-hAPN MAb-DW1 and binding was determined by a fluorescent substrate (Vector Red ® ). (B) Cryosections of lungs from age matched non-transgenic and homozygous hAPN-transgenic mice were analyzed with a MAb-R3-242 specific to mouse APN (mAPN) or a MAb-DW1 against hAPN. Binding of the MAbs was revealed by a fluorescent substrate (Vector Red ® ).

Article Snippet: Homozygous, heterozygous, and hemizygous hAPN-transgenic mice all express hAPN with a similar expression pattern and hAPN was expressed on the epithelial surface of the lungs. (A) Serial cryosections of small intestine from age matched non-transgenic females (XX), heterozygous females (X h X), homozygous females (X h X h ), non-transgenic males (XY), and hemizygous males (X h Y) were analyzed for expression of mouse APN (mAPN) using rat MAb-R3-242 to mAPN or with a mouse anti-hAPN MAb-DW1 and binding was determined by a fluorescent substrate (Vector Red ® ). (B) Cryosections of lungs from age matched non-transgenic and homozygous hAPN-transgenic mice were analyzed with a MAb-R3-242 specific to mouse APN (mAPN) or a MAb-DW1 against hAPN.

Techniques: Transgenic Assay, Expressing, Binding Assay, Plasmid Preparation

Journal: Virology

Article Title: Cells of human aminopeptidase N (CD13) transgenic mice are infected by human coronavirus-229E in vitro, but not in vivo

doi: 10.1016/j.virol.2005.02.023

Figure Lengend Snippet: Some embryonic cells from the hAPN-transgenic mice express hAPN protein and are infected by HCoV-229E. (A) Cells derived from non-transgenic and hAPN-transgenic embryos were inoculated with HCoV-229E at an MOI of 1 and fixed 20 h post-inoculation. Cells were tested for hAPN expression (red) with a mouse anti-hAPN MAb-DW1 and for expression of HCoV-229E proteins (green) with polyclonal goat antisera to HCoV-229E. In addition, the overlay of these two images is shown (merged). (B) Embryonic cells from non-transgenic and hAPN-transgenic mice were inoculated with HCoV-229E at an MOI of 1, unbound virus was removed by washing, and the titer of the virus in the supernatant at 4 h and 24 h post-infection was determined by TCID 50 in MRC5 cells. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Homozygous, heterozygous, and hemizygous hAPN-transgenic mice all express hAPN with a similar expression pattern and hAPN was expressed on the epithelial surface of the lungs. (A) Serial cryosections of small intestine from age matched non-transgenic females (XX), heterozygous females (X h X), homozygous females (X h X h ), non-transgenic males (XY), and hemizygous males (X h Y) were analyzed for expression of mouse APN (mAPN) using rat MAb-R3-242 to mAPN or with a mouse anti-hAPN MAb-DW1 and binding was determined by a fluorescent substrate (Vector Red ® ). (B) Cryosections of lungs from age matched non-transgenic and homozygous hAPN-transgenic mice were analyzed with a MAb-R3-242 specific to mouse APN (mAPN) or a MAb-DW1 against hAPN.

Techniques: Transgenic Assay, Infection, Derivative Assay, Expressing

Journal: Virology

Article Title: Cells of human aminopeptidase N (CD13) transgenic mice are infected by human coronavirus-229E in vitro, but not in vivo

doi: 10.1016/j.virol.2005.02.023

Figure Lengend Snippet: Dendritic cells derived from the bone marrow of hAPN-transgenic mice express hAPN and are infected by HCoV-229E. (A) Dendritic cells were isolated from non-transgenic (a, c, and e) and hAPN-transgenic (b, d, and f) male littermates and analyzed by cytofluorometry. The controls were cells that were not treated with primary antibody (gray line) or treated with antibodies (hamster, rat, or mouse) to an irrelevant antigen (dotted line). The surface expression of CD11c (a and b, solid line with gray shading), mouse APN (c and d, solid line with gray shading), MHC class I (e and f, dashed line), and human APN (e and f, solid line with gray shading) were identified with hamster MAb-HL3, rat MAb-R3-242, mouse MAb-KH117, and MAb-DW1, respectively. Dendritic cells isolated from non-transgenic and hAPN-transgenic mice were mock inoculated (M) or inoculated (I) with HCoV-229E at an MOI of 1.0 at 4 °C. Unbound virus was removed by washing and cells were incubated at 34 °C for 20 h. (B) Immunoblot analysis of protein lysates was performed using polyclonal goat anti-HCoV-229E that reacts strongly with nucleocapsid protein (arrow). (C) IFA of acetone fixed hAPN-transgenic dendritic cells was done using anti-hAPN MAb-DW1 (red) and with polyclonal goat anti-HCoV-229E (green). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Homozygous, heterozygous, and hemizygous hAPN-transgenic mice all express hAPN with a similar expression pattern and hAPN was expressed on the epithelial surface of the lungs. (A) Serial cryosections of small intestine from age matched non-transgenic females (XX), heterozygous females (X h X), homozygous females (X h X h ), non-transgenic males (XY), and hemizygous males (X h Y) were analyzed for expression of mouse APN (mAPN) using rat MAb-R3-242 to mAPN or with a mouse anti-hAPN MAb-DW1 and binding was determined by a fluorescent substrate (Vector Red ® ). (B) Cryosections of lungs from age matched non-transgenic and homozygous hAPN-transgenic mice were analyzed with a MAb-R3-242 specific to mouse APN (mAPN) or a MAb-DW1 against hAPN.

Techniques: Derivative Assay, Transgenic Assay, Infection, Isolation, Expressing, Incubation, Western Blot