Reovirus therapy of tumors with activated Ras pathway

Science

Volumn 282, Issue 5392, 1998, Pages 1332-1334

  Coffey, Matthew C ^a   Strong, James E ^a   Forsyth, Peter A ^b   Lee, Patrick W K ^a  

^a Health Sciences Centre Calgary   (Canada)

^b UNIVERSITY OF CALGARY   (Canada)

Author keywords

[No Author keywords available]

Indexed keywords

ANIMAL CELL; ANTINEOPLASTIC ACTIVITY; ARTICLE; CANCER; CANCER THERAPY; GLIOBLASTOMA; HUMAN; HUMAN CELL; NONHUMAN; PRIORITY JOURNAL; REOVIRUS; VIRUS CELL INTERACTION;

EID: 0032515141     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.282.5392.1332     Document Type: Article

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31. 7 PFUs of reovirus or UV-inactivated reovirus in 20 μl of sterile PBS. Tumor size was measured twice weekly for 2 to 4 weeks. All animals were killed when the control mice showed severe morbidity due to excessive tumor burden.
32. Immunofluorescence analysis was carried out on formalin-fixed, paraffin-embedded tumor sections mounted on cover slips. After removal of paraffin by xylene, the sections were rehydrated and exposed to the primary antibody (rabbit polydonal antiserum to reovirus type 3 diluted 1/100 in PBS) for 2 hours at room temperature. After three washes with PBS, the sections were exposed to the secondary antibody (goat antibody to rabbit immunoglobulin G, coupled to fluorescein isothiocyanate conjugate, diluted 1/100 in PBS containing 10% goat serum and 0.005% Evan's Blue counter-stain) for 1 hour at room temperature. Finally, the fixed and treated sections were washed three more times with PBS and then once with water. The sections were dried, mounted on slides in 90% glycerol containing 0.1% phenylenediamine, and viewed with a Zeiss Axiophot microscope.
33. We thank H.-J. Kung for THC-11 cells, W. Yong for U87 cells, D. Edwards for C3 cells, Q. Shi for excellent technical assistance, and D. Demetrick, P. Brasher, and B. Rewcastle for helpful discussions. Supported by the Medical Research Council of Canada (P.W.K.L).