Skip to main content
. 2017 May 8;13(5):e1006371. doi: 10.1371/journal.ppat.1006371

Fig 3. Microscopic and macroscopic detection of fluorescence in donor ferrets.

Fig 3

(A) WBC infection percentages of the rCDVs in directly inoculated (D1-6) ferrets were determined by flow cytometry at different time-points. The circular inset shows which reporter virus was administered to a donor ferret via which route. After necropsy, infection percentages of the rCDVs in single cell suspensions of lymphoid tissues were determined by flow cytometry. rCDVs were detected in all lymphoid tissues (man LN: mandibular lymph node; ax LN: axillary LN; ing LN: inguinal LN; TB LN: tracheo-bronchial LN; RP LN: retropharyngeal LN; mes LN: mesenteric LN; GALT: gut-associated lymphoid tissue; PP: Peyer’s patches; BM: bone marrow; ND: not determined). Results obtained from lymph nodes corresponded to the kinetics of the rCDVs in WBC. In 5/6 ferrets the IT delivered virus became dominant (D2 –D6). (B, C) Donor ferrets D1 and D2 were chosen as representative examples. Virus was isolated from WBC, BAL, CSF and throat, nose, eye and rectal swabs by titration on Vero-cCD150 cells. (Left panels) Screening of virus isolations by CLSM allowed determination of the dominance of rCDVs at 8 DPI. (Centre panels) Relative contribution of the various reporter viruses to the total amount of rCDV present in the sample collected 8 DPI or at necropsy (CSF and BAL). (Right panels) Direct CLSM of various tissues at necropsy revealed fluorescence in the epithelium of the trachea (dotted line marks basement membrane, Lu marks lumen), and B-cell follicles in lymphoid tissues (delineated by dotted line). (D) Macroscopic fluorescence produced by rCDVSHVenus(6) and rCDVSHdTom(6) was directly detected in live ferrets and during necropsy at the nose, eyes, mouth and skin. The inset in the second panel clearly shows that green and red foci of infected cells can be discerned (#). During necropsy, macroscopic fluorescence was detected in the tongue (arrows show tonsils), lungs and Peyer’s patches. Again, the inset in the third panel shows separate foci of green and red cells (#).