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VR-1605 is recombinant Vaccinia virus strain rVV-hgp100 that expresses human melanoma antigen gp100. It is considered a potent immunogen to activate tumor reactive T cells in both human and mice, and was developed by NCI.

VR-1605 is recombinant Vaccinia virus strain rVV-hgp100 that expresses human melanoma antigen gp100. It is considered a potent immunogen to activate tumor reactive T cells in both human and mice, and was developed by NCI.

Cell enlargement and clustering of rounded cells in cell culture; plaques with rounding and eventual sloughing.

Production Host: Human TK- cells (provided by the depositor (Dr. Zhiya Yu). Virus can be propagated in numerous cell lines (see Alternate Hosts)).

Alternate Hosts: BS-C-1 cells (ATCC CCL-26), Vero cells (ATCC CCL-81), and HeLa cells (ATCC CCL-2)

Plant cells 24-48 hours in advance and infect when cultures are 80-90% confluent. Process virus by freeze/ thawing it three times by vapor phase of liquid nitrogen and 37.0°C water bath (vortex mixing on high speed for 20 seconds between each freeze/ thaw), sonicating with 40% power for 30 seconds, and then a final vortex mixing on high speed for 30 seconds. Remove cell growth medium from the flask, and inoculate with a small volume of virus stock diluted in virus growth medium to provide a MOI of about 1-3. Adsorb 1-2 hours at 37.0°C in a humidified 5% CO2 atmosphere, rocking every 15-20 minutes to redistribute inoculum. End adsorption by adding virus growth medium.

Incubate infected culture for 1-3 days at 37.0°C in a humidified 5% CO2 atmosphere, until CPE are well advanced through 90% of the culture.

rVV-hgp100 was constructed in Dr. Nicolas P. Restifo's laboratory of Surgery Branch/ NCI/ NIH in the early 1990's.