Figure 5.
Figure 5. Drug treatment of monkey RQ2282 that received a transplant with cells containing a monocistronic DHFR resistance gene. (A) Changes in peripheral blood counts with 2 treatment courses. The first treatment consisted of 8 days of peg-SCF and G-CSF given concurrently with TMTX 6 mg/kg/d × 5, and NBMRP-P 2 mg/kg/d × 5. The second treatment course also included peg-SCF and G-CSG, as in the first treatment cycle, but with TMTX increased to 8 mg/kg/d × 5 and NBMPR-P 2 mg/kg/d × 5. Note the increased myelosuppression following the second course. (B) PCR analysis for quantitation of proviral DNA copy in samples from Ficoll-separated peripheral blood mononuclear cells (M) and granulocytes (G), at various time points after the second treatment course. The top row shows signals from the vector genome (DHFR primers), and the bottom row shows internal control for actin DNA sequences. The right-hand lanes show varying dilutions of producer cell and PG-13 DNA, with dilution ranging from 0 to 10% producer cell DNA. Samples from a monkey that did not receive a transplant (Untx PB) serve as negative controls. Samples from 4 days before treatment (d-4) and various days after the start of the treatment cycle are shown. Below each column is the actin-corrected, percentage copy number of cell as determined by PhosphorImager analysis.

Drug treatment of monkey RQ2282 that received a transplant with cells containing a monocistronic DHFR resistance gene. (A) Changes in peripheral blood counts with 2 treatment courses. The first treatment consisted of 8 days of peg-SCF and G-CSF given concurrently with TMTX 6 mg/kg/d × 5, and NBMRP-P 2 mg/kg/d × 5. The second treatment course also included peg-SCF and G-CSG, as in the first treatment cycle, but with TMTX increased to 8 mg/kg/d × 5 and NBMPR-P 2 mg/kg/d × 5. Note the increased myelosuppression following the second course. (B) PCR analysis for quantitation of proviral DNA copy in samples from Ficoll-separated peripheral blood mononuclear cells (M) and granulocytes (G), at various time points after the second treatment course. The top row shows signals from the vector genome (DHFR primers), and the bottom row shows internal control for actin DNA sequences. The right-hand lanes show varying dilutions of producer cell and PG-13 DNA, with dilution ranging from 0 to 10% producer cell DNA. Samples from a monkey that did not receive a transplant (Untx PB) serve as negative controls. Samples from 4 days before treatment (d-4) and various days after the start of the treatment cycle are shown. Below each column is the actin-corrected, percentage copy number of cell as determined by PhosphorImager analysis.

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