Use of cell-free retroviral vector preparations for transduction of cells from the marrow of chronic phase and blast crisis chronic myelogenous leukemia patients and from normal individuals

Marc Etkin, Marylynne Filaccio, Debra Ellerson, Soon Pal Suh, David Claxton, Eugenia Gaozza, Malcolm Brenner, Robert Moen, John Belmont, Kateri A. Moore, A. M. Moseley, Christopher Reading, Issa Khouri, Moshe Talpaz, Hagop Kantarjian, Albert Deisseroth

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Marrow cells were exposed to the LNL6 or GIN safety-modified variants of the N2 retrovirus, which contain the G418 bacterial resistance gene neo. The frequency of acquisition of the G418 resistance phenotype following exposure to LNL6 or GIN was compared among hematopoictic progenitor cells from the marrow of patients with chronic phase chronic myelogenous leukemia (CML), blast crisis CML, or from nonleukemic individuals. Under the conditions of our experiments, the myeloid committed progenitor cells from 3 of 6 nonleukemic individuals, 9 of 18 chronic-phase CML patients, and 2 of 4 blast crisis CML patients acquired resistance to at least 1 mg/ml G418 following incubation with cell-free supernatants from the PA317 LNL6 or PA317 GIN producer cell lines. Ten of the 32 colonies growing up in 0.8 mg/ml G418 from chronic-phase marrow exposed to LNL6 were shown to contain the neo gene by polymerase chain reaction (PCR) assay of DNA. These results were consistent with estimates of the transduction frequency based on acquisition of resistance to G418 as the number of colonies growing under G418 selection was always greater at 0.8 mg/ml G418 than at higher concentrations of G418 (1.0-1.4 mg/ml). The average transduction frequency at each G418 concentration (1.0, 1.2, and 1.4 mg/ml) in cells from blast crisis CML cells ranged from 2 to 14%, as measured by acquisition of G418 resistance. Chronic-phase CML showed slightly lower average frequencies of transduction (0.6-2.8% of the colonies are G418 resistant). The average transduction frequency of cells from nonleukemic marrow was as high as that seen from the marrow of chronic-phase CML individuals. These results have implications for retroviral marking strategies which are designed to follow the purging of blastic leukemia cells from the marrow of CML patients used for autologous bone marrow transplantation.

Original languageEnglish (US)
Pages (from-to)137-145
Number of pages9
JournalHuman Gene Therapy
Volume3
Issue number2
DOIs
StatePublished - Jan 1 1992

Fingerprint

Blast Crisis
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Bone Marrow
Leukemia, Myeloid, Chronic Phase
antibiotic G 418
Myeloid Progenitor Cells
Bacterial Genes
Autologous Transplantation
Retroviridae
Bone Marrow Transplantation
Leukemia
Stem Cells

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Etkin, Marc ; Filaccio, Marylynne ; Ellerson, Debra ; Suh, Soon Pal ; Claxton, David ; Gaozza, Eugenia ; Brenner, Malcolm ; Moen, Robert ; Belmont, John ; Moore, Kateri A. ; Moseley, A. M. ; Reading, Christopher ; Khouri, Issa ; Talpaz, Moshe ; Kantarjian, Hagop ; Deisseroth, Albert. / Use of cell-free retroviral vector preparations for transduction of cells from the marrow of chronic phase and blast crisis chronic myelogenous leukemia patients and from normal individuals. In: Human Gene Therapy. 1992 ; Vol. 3, No. 2. pp. 137-145.
@article{4fb2281b31ac4a69be2f762ed376236e,
title = "Use of cell-free retroviral vector preparations for transduction of cells from the marrow of chronic phase and blast crisis chronic myelogenous leukemia patients and from normal individuals",
abstract = "Marrow cells were exposed to the LNL6 or GIN safety-modified variants of the N2 retrovirus, which contain the G418 bacterial resistance gene neo. The frequency of acquisition of the G418 resistance phenotype following exposure to LNL6 or GIN was compared among hematopoictic progenitor cells from the marrow of patients with chronic phase chronic myelogenous leukemia (CML), blast crisis CML, or from nonleukemic individuals. Under the conditions of our experiments, the myeloid committed progenitor cells from 3 of 6 nonleukemic individuals, 9 of 18 chronic-phase CML patients, and 2 of 4 blast crisis CML patients acquired resistance to at least 1 mg/ml G418 following incubation with cell-free supernatants from the PA317 LNL6 or PA317 GIN producer cell lines. Ten of the 32 colonies growing up in 0.8 mg/ml G418 from chronic-phase marrow exposed to LNL6 were shown to contain the neo gene by polymerase chain reaction (PCR) assay of DNA. These results were consistent with estimates of the transduction frequency based on acquisition of resistance to G418 as the number of colonies growing under G418 selection was always greater at 0.8 mg/ml G418 than at higher concentrations of G418 (1.0-1.4 mg/ml). The average transduction frequency at each G418 concentration (1.0, 1.2, and 1.4 mg/ml) in cells from blast crisis CML cells ranged from 2 to 14{\%}, as measured by acquisition of G418 resistance. Chronic-phase CML showed slightly lower average frequencies of transduction (0.6-2.8{\%} of the colonies are G418 resistant). The average transduction frequency of cells from nonleukemic marrow was as high as that seen from the marrow of chronic-phase CML individuals. These results have implications for retroviral marking strategies which are designed to follow the purging of blastic leukemia cells from the marrow of CML patients used for autologous bone marrow transplantation.",
author = "Marc Etkin and Marylynne Filaccio and Debra Ellerson and Suh, {Soon Pal} and David Claxton and Eugenia Gaozza and Malcolm Brenner and Robert Moen and John Belmont and Moore, {Kateri A.} and Moseley, {A. M.} and Christopher Reading and Issa Khouri and Moshe Talpaz and Hagop Kantarjian and Albert Deisseroth",
year = "1992",
month = "1",
day = "1",
doi = "10.1089/hum.1992.3.2-137",
language = "English (US)",
volume = "3",
pages = "137--145",
journal = "Human Gene Therapy",
issn = "1043-0342",
publisher = "Mary Ann Liebert Inc.",
number = "2",

}

Etkin, M, Filaccio, M, Ellerson, D, Suh, SP, Claxton, D, Gaozza, E, Brenner, M, Moen, R, Belmont, J, Moore, KA, Moseley, AM, Reading, C, Khouri, I, Talpaz, M, Kantarjian, H & Deisseroth, A 1992, 'Use of cell-free retroviral vector preparations for transduction of cells from the marrow of chronic phase and blast crisis chronic myelogenous leukemia patients and from normal individuals', Human Gene Therapy, vol. 3, no. 2, pp. 137-145. https://doi.org/10.1089/hum.1992.3.2-137

Use of cell-free retroviral vector preparations for transduction of cells from the marrow of chronic phase and blast crisis chronic myelogenous leukemia patients and from normal individuals. / Etkin, Marc; Filaccio, Marylynne; Ellerson, Debra; Suh, Soon Pal; Claxton, David; Gaozza, Eugenia; Brenner, Malcolm; Moen, Robert; Belmont, John; Moore, Kateri A.; Moseley, A. M.; Reading, Christopher; Khouri, Issa; Talpaz, Moshe; Kantarjian, Hagop; Deisseroth, Albert.

In: Human Gene Therapy, Vol. 3, No. 2, 01.01.1992, p. 137-145.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Use of cell-free retroviral vector preparations for transduction of cells from the marrow of chronic phase and blast crisis chronic myelogenous leukemia patients and from normal individuals

AU - Etkin, Marc

AU - Filaccio, Marylynne

AU - Ellerson, Debra

AU - Suh, Soon Pal

AU - Claxton, David

AU - Gaozza, Eugenia

AU - Brenner, Malcolm

AU - Moen, Robert

AU - Belmont, John

AU - Moore, Kateri A.

AU - Moseley, A. M.

AU - Reading, Christopher

AU - Khouri, Issa

AU - Talpaz, Moshe

AU - Kantarjian, Hagop

AU - Deisseroth, Albert

PY - 1992/1/1

Y1 - 1992/1/1

N2 - Marrow cells were exposed to the LNL6 or GIN safety-modified variants of the N2 retrovirus, which contain the G418 bacterial resistance gene neo. The frequency of acquisition of the G418 resistance phenotype following exposure to LNL6 or GIN was compared among hematopoictic progenitor cells from the marrow of patients with chronic phase chronic myelogenous leukemia (CML), blast crisis CML, or from nonleukemic individuals. Under the conditions of our experiments, the myeloid committed progenitor cells from 3 of 6 nonleukemic individuals, 9 of 18 chronic-phase CML patients, and 2 of 4 blast crisis CML patients acquired resistance to at least 1 mg/ml G418 following incubation with cell-free supernatants from the PA317 LNL6 or PA317 GIN producer cell lines. Ten of the 32 colonies growing up in 0.8 mg/ml G418 from chronic-phase marrow exposed to LNL6 were shown to contain the neo gene by polymerase chain reaction (PCR) assay of DNA. These results were consistent with estimates of the transduction frequency based on acquisition of resistance to G418 as the number of colonies growing under G418 selection was always greater at 0.8 mg/ml G418 than at higher concentrations of G418 (1.0-1.4 mg/ml). The average transduction frequency at each G418 concentration (1.0, 1.2, and 1.4 mg/ml) in cells from blast crisis CML cells ranged from 2 to 14%, as measured by acquisition of G418 resistance. Chronic-phase CML showed slightly lower average frequencies of transduction (0.6-2.8% of the colonies are G418 resistant). The average transduction frequency of cells from nonleukemic marrow was as high as that seen from the marrow of chronic-phase CML individuals. These results have implications for retroviral marking strategies which are designed to follow the purging of blastic leukemia cells from the marrow of CML patients used for autologous bone marrow transplantation.

AB - Marrow cells were exposed to the LNL6 or GIN safety-modified variants of the N2 retrovirus, which contain the G418 bacterial resistance gene neo. The frequency of acquisition of the G418 resistance phenotype following exposure to LNL6 or GIN was compared among hematopoictic progenitor cells from the marrow of patients with chronic phase chronic myelogenous leukemia (CML), blast crisis CML, or from nonleukemic individuals. Under the conditions of our experiments, the myeloid committed progenitor cells from 3 of 6 nonleukemic individuals, 9 of 18 chronic-phase CML patients, and 2 of 4 blast crisis CML patients acquired resistance to at least 1 mg/ml G418 following incubation with cell-free supernatants from the PA317 LNL6 or PA317 GIN producer cell lines. Ten of the 32 colonies growing up in 0.8 mg/ml G418 from chronic-phase marrow exposed to LNL6 were shown to contain the neo gene by polymerase chain reaction (PCR) assay of DNA. These results were consistent with estimates of the transduction frequency based on acquisition of resistance to G418 as the number of colonies growing under G418 selection was always greater at 0.8 mg/ml G418 than at higher concentrations of G418 (1.0-1.4 mg/ml). The average transduction frequency at each G418 concentration (1.0, 1.2, and 1.4 mg/ml) in cells from blast crisis CML cells ranged from 2 to 14%, as measured by acquisition of G418 resistance. Chronic-phase CML showed slightly lower average frequencies of transduction (0.6-2.8% of the colonies are G418 resistant). The average transduction frequency of cells from nonleukemic marrow was as high as that seen from the marrow of chronic-phase CML individuals. These results have implications for retroviral marking strategies which are designed to follow the purging of blastic leukemia cells from the marrow of CML patients used for autologous bone marrow transplantation.

UR - http://www.scopus.com/inward/record.url?scp=0026846946&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026846946&partnerID=8YFLogxK

U2 - 10.1089/hum.1992.3.2-137

DO - 10.1089/hum.1992.3.2-137

M3 - Article

C2 - 1391033

AN - SCOPUS:0026846946

VL - 3

SP - 137

EP - 145

JO - Human Gene Therapy

JF - Human Gene Therapy

SN - 1043-0342

IS - 2

ER -