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Quantitative assessment of retroviral transfer of the human multidrug resistance 1 gene to human mobilized peripheral blood progenitor cells engrafted in nonobese diabetic/severe combined immunodeficient mice.
Blood. 2000 Feb 15; 95(4):1237-48.Blood

Abstract

Mobilized peripheral blood progenitor cells (PBPC) are a potential target for the retrovirus-mediated transfer of cytostatic drug-resistance genes. We analyzed nonobese diabetic/severe combined immunodeficient (NOD/SCID) mouse-repopulating CD34+ PBPC from patients with cancer after retroviral transduction in various cytokine combinations with the hybrid vector SF-MDR, which is based on the Friend mink cell focus-forming/murine embryonic stem-cell virus and carries the human multidrug resistance 1 (MDR1) gene. Five to 13 weeks after transplantation of CD34+ PBPC into NOD/SCID mice (n = 84), a cell dose-dependent multilineage engraftment of human leukocytes up to an average of 33% was observed. The SF-MDR provirus was detected in the bone marrow (BM) and in its granulocyte fractions in 96% and 72%, respectively, of chimeric NOD/SCID mice. SF-MDR provirus integration assessed by quantitative real-time polymerase chain reaction (PCR) was optimal in the presence of Flt-3 ligand/thrombopoietin/stem-cell factor, resulting in a 6-fold (24% +/- 5% [mean +/- SE]) higher average proportion of gene-marked human cells in NOD/SCID mice than that achieved with IL-3 alone (P <.01). A population of clearly rhodamine-123(dull) human myeloid progeny cells could be isolated from BM samples from chimeric NOD/SCID mice. On the basis of PCR and rhodamine-123 efflux data, up to 18% +/- 4% of transduced cells were calculated to express the transgene. Our data suggest that the NOD/SCID model provides a valid assay for estimating the gene-transfer efficiency to repopulating human PBPC that may be achievable in clinical autologous transplantation. P-glycoprotein expression sufficient to prevent marrow aplasia in vivo may be obtained with this SF-MDR vector and an optimized transduction protocol. (Blood. 2000;95:1237-1248)

Authors+Show Affiliations

German Cancer Research Center, Department D 0200, Heidelberg, Germany.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

10666196

Citation

Schiedlmeier, B, et al. "Quantitative Assessment of Retroviral Transfer of the Human Multidrug Resistance 1 Gene to Human Mobilized Peripheral Blood Progenitor Cells Engrafted in Nonobese Diabetic/severe Combined Immunodeficient Mice." Blood, vol. 95, no. 4, 2000, pp. 1237-48.
Schiedlmeier B, Kühlcke K, Eckert HG, et al. Quantitative assessment of retroviral transfer of the human multidrug resistance 1 gene to human mobilized peripheral blood progenitor cells engrafted in nonobese diabetic/severe combined immunodeficient mice. Blood. 2000;95(4):1237-48.
Schiedlmeier, B., Kühlcke, K., Eckert, H. G., Baum, C., Zeller, W. J., & Fruehauf, S. (2000). Quantitative assessment of retroviral transfer of the human multidrug resistance 1 gene to human mobilized peripheral blood progenitor cells engrafted in nonobese diabetic/severe combined immunodeficient mice. Blood, 95(4), 1237-48.
Schiedlmeier B, et al. Quantitative Assessment of Retroviral Transfer of the Human Multidrug Resistance 1 Gene to Human Mobilized Peripheral Blood Progenitor Cells Engrafted in Nonobese Diabetic/severe Combined Immunodeficient Mice. Blood. 2000 Feb 15;95(4):1237-48. PubMed PMID: 10666196.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Quantitative assessment of retroviral transfer of the human multidrug resistance 1 gene to human mobilized peripheral blood progenitor cells engrafted in nonobese diabetic/severe combined immunodeficient mice. AU - Schiedlmeier,B, AU - Kühlcke,K, AU - Eckert,H G, AU - Baum,C, AU - Zeller,W J, AU - Fruehauf,S, PY - 2000/2/9/pubmed PY - 2000/3/18/medline PY - 2000/2/9/entrez SP - 1237 EP - 48 JF - Blood JO - Blood VL - 95 IS - 4 N2 - Mobilized peripheral blood progenitor cells (PBPC) are a potential target for the retrovirus-mediated transfer of cytostatic drug-resistance genes. We analyzed nonobese diabetic/severe combined immunodeficient (NOD/SCID) mouse-repopulating CD34+ PBPC from patients with cancer after retroviral transduction in various cytokine combinations with the hybrid vector SF-MDR, which is based on the Friend mink cell focus-forming/murine embryonic stem-cell virus and carries the human multidrug resistance 1 (MDR1) gene. Five to 13 weeks after transplantation of CD34+ PBPC into NOD/SCID mice (n = 84), a cell dose-dependent multilineage engraftment of human leukocytes up to an average of 33% was observed. The SF-MDR provirus was detected in the bone marrow (BM) and in its granulocyte fractions in 96% and 72%, respectively, of chimeric NOD/SCID mice. SF-MDR provirus integration assessed by quantitative real-time polymerase chain reaction (PCR) was optimal in the presence of Flt-3 ligand/thrombopoietin/stem-cell factor, resulting in a 6-fold (24% +/- 5% [mean +/- SE]) higher average proportion of gene-marked human cells in NOD/SCID mice than that achieved with IL-3 alone (P <.01). A population of clearly rhodamine-123(dull) human myeloid progeny cells could be isolated from BM samples from chimeric NOD/SCID mice. On the basis of PCR and rhodamine-123 efflux data, up to 18% +/- 4% of transduced cells were calculated to express the transgene. Our data suggest that the NOD/SCID model provides a valid assay for estimating the gene-transfer efficiency to repopulating human PBPC that may be achievable in clinical autologous transplantation. P-glycoprotein expression sufficient to prevent marrow aplasia in vivo may be obtained with this SF-MDR vector and an optimized transduction protocol. (Blood. 2000;95:1237-1248) SN - 0006-4971 UR - https://www.unboundmedicine.com/medline/citation/10666196/Quantitative_assessment_of_retroviral_transfer_of_the_human_multidrug_resistance_1_gene_to_human_mobilized_peripheral_blood_progenitor_cells_engrafted_in_nonobese_diabetic/severe_combined_immunodeficient_mice_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0006-4971(20)67069-0 DB - PRIME DP - Unbound Medicine ER -