Detection of rKSHV.219 DNA in clonogenic colonies derived from infected CD34+ HPCs
. | rKSHV.219+ colonies/total colonies analyzed . | . | . | ||
|---|---|---|---|---|---|
. | CFU-GM . | BFU-E . | HPP . | ||
| rKSHV.219 | 21% (12 of 56)* | 0% (0 of 9) | 40% (4 of 10)† | ||
| UV-rKSHV.219 | 0% (0 of 33) | 0% (0 of 10) | 0% (0 of 7) | ||
| Heat-treated rKSHV.219 | 0% (0 of 35) | 0% (0 of 7) | 0% (0 of 7) | ||
| Mock | 0% (0 of 36) | 0% (0 of 3) | 0% (0 of 2) | ||
. | rKSHV.219+ colonies/total colonies analyzed . | . | . | ||
|---|---|---|---|---|---|
. | CFU-GM . | BFU-E . | HPP . | ||
| rKSHV.219 | 21% (12 of 56)* | 0% (0 of 9) | 40% (4 of 10)† | ||
| UV-rKSHV.219 | 0% (0 of 33) | 0% (0 of 10) | 0% (0 of 7) | ||
| Heat-treated rKSHV.219 | 0% (0 of 35) | 0% (0 of 7) | 0% (0 of 7) | ||
| Mock | 0% (0 of 36) | 0% (0 of 3) | 0% (0 of 2) | ||
CD34+ HPCs were exposed to rKSHV.219, UV-irradiated rKSHV.219 (10 μW/cm2 for 1 hour), or heat-inactivated rKSHV.219 (65°C for 1 hour), and plated into MethoCult H4433. Colonies were visually scored, randomly isolated, and DNA was analyzed by real-time PCR analysis at 12 to 15 days after plating. Experiments were performed twice. Colonies with less than 1.0 calculated viral copy per cell were scored as negative for KSHV infection. Colonies demonstrating less than 2 human β-globin copies were excluded from analysis.
5.3 ± 1.2 mean rKSHV.219 genomic copies per human cell in positive CFU-GM colonies.
1.7 ± 0.4 mean rKSHV.219 genomic copies per human cell in positive HPP colonies.