Leukemic burden in subpopulations of CD34+ cells isolated from the mobilized peripheral blood of α-interferon-resistant or -intolerant patients with chronic myeloid leukemia

David Van Den Berg, Maija Wessman, Lesley Murray, Jie Tong, Benjamin Chen, Shirley Chen, Don Simonetti, Julie King, Glenn Yamasaki, Rhonda DiGiusto, Angello Carella, Francesco Frassoni, Norbert Claude Gorin, David Snyder, Irena Sciecinski, Robert Negrin, Albert Deisseroth, Ann Tsukamoto, David Gearing, Christopher ReadingRon Hoffman

Research output: Contribution to journalArticle

Abstract

We attempted to determine the frequency of normal hematopoietic stem cells (HSC) and contaminating leukemic cells in mobilized peripheral blood (MPB) collected from chronic myeloid leukemia (CML) patients, intolerant of α- interferon or with interferon-resistant disease. A total of 14 MPB samples, six from patients in chronic phase (CP) and eight from patients in accelerated phase or blast crisis (AP/BC) were studied. Cytogenetic analysis of MPB collected from AP/BC patients showed that 100% of the cells were Ph+, whereas cells from four of five CP MPB were Ph-. By contrast, fluorescence in situ hybridization (FISH) analysis of CP MPB showed a mean frequency of 14.7% Ph+ cells, while AP/BC MPB contained 39.2% Ph+ cells. In an attempt to purify normal HSC, subpopulations of the MPB CD34+ cells were isolated based on expression of the Thy-1 antigen (CDw90). The mean Ph+ cell frequency as determined by FISH within the CD34+Thy-1+ Lin- and CD34+Thy- 1-Lin- populations from CP patients was 19.2% and 33.9%, respectively. In the AP/BC patients, levels of residual leukemic cells were significantly greater with mean Ph+ cell frequencies of 59.2% and 72.7% for the CD34+Thy- 1+Lin- and CD34+Thy-1-Lin- fractions, respectively. The frequency of cobblestone area forming cells (CAFC) was used as a means of quantitating the numbers of functional HSC within these cell subpopulations. The mean CAFC frequency was 1 of 19 for the CD34+Thy-1+Lin- cells as compared with 1 of 133 for the Thy-1- fraction indicating a higher frequency of primitive progenitor cells in the Thy-1+ subpopulation. CD34+ cell subsets from two patients were also injected into SCID-hu bone assays to determine the in vivo behavior of these cell populations. After 8 weeks, multilineage donor engraftment was observed in these grafts. FISH analysis of the donor cells within the grafts showed that 55.3% and 60.0% of the cells were Ph+. We conclude that unfractionated MPB from this patient population is not leukemia-free and that the CD34+Thy-1+Lin- cell subpopulation, although predominantly enriched for normal HSC, still contains substantial numbers of residual leukemic cells.

Original languageEnglish
Pages (from-to)4348-4357
Number of pages10
JournalBlood
Volume87
Issue number10
Publication statusPublished - May 15 1996

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Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Interferons
Blood
Stem cells
Blast Crisis
Fluorescence
Grafts
Hematopoietic Stem Cells
Thy-1 Antigens
Fluorescence In Situ Hybridization
Assays
Bone
Cells
Tissue Donors
Population
Transplants
Cytogenetic Analysis

ASJC Scopus subject areas

  • Hematology

Cite this

Van Den Berg, D., Wessman, M., Murray, L., Tong, J., Chen, B., Chen, S., ... Hoffman, R. (1996). Leukemic burden in subpopulations of CD34+ cells isolated from the mobilized peripheral blood of α-interferon-resistant or -intolerant patients with chronic myeloid leukemia. Blood, 87(10), 4348-4357.

Leukemic burden in subpopulations of CD34+ cells isolated from the mobilized peripheral blood of α-interferon-resistant or -intolerant patients with chronic myeloid leukemia. / Van Den Berg, David; Wessman, Maija; Murray, Lesley; Tong, Jie; Chen, Benjamin; Chen, Shirley; Simonetti, Don; King, Julie; Yamasaki, Glenn; DiGiusto, Rhonda; Carella, Angello; Frassoni, Francesco; Gorin, Norbert Claude; Snyder, David; Sciecinski, Irena; Negrin, Robert; Deisseroth, Albert; Tsukamoto, Ann; Gearing, David; Reading, Christopher; Hoffman, Ron.

In: Blood, Vol. 87, No. 10, 15.05.1996, p. 4348-4357.

Research output: Contribution to journalArticle

Van Den Berg, D, Wessman, M, Murray, L, Tong, J, Chen, B, Chen, S, Simonetti, D, King, J, Yamasaki, G, DiGiusto, R, Carella, A, Frassoni, F, Gorin, NC, Snyder, D, Sciecinski, I, Negrin, R, Deisseroth, A, Tsukamoto, A, Gearing, D, Reading, C & Hoffman, R 1996, 'Leukemic burden in subpopulations of CD34+ cells isolated from the mobilized peripheral blood of α-interferon-resistant or -intolerant patients with chronic myeloid leukemia', Blood, vol. 87, no. 10, pp. 4348-4357.
Van Den Berg, David ; Wessman, Maija ; Murray, Lesley ; Tong, Jie ; Chen, Benjamin ; Chen, Shirley ; Simonetti, Don ; King, Julie ; Yamasaki, Glenn ; DiGiusto, Rhonda ; Carella, Angello ; Frassoni, Francesco ; Gorin, Norbert Claude ; Snyder, David ; Sciecinski, Irena ; Negrin, Robert ; Deisseroth, Albert ; Tsukamoto, Ann ; Gearing, David ; Reading, Christopher ; Hoffman, Ron. / Leukemic burden in subpopulations of CD34+ cells isolated from the mobilized peripheral blood of α-interferon-resistant or -intolerant patients with chronic myeloid leukemia. In: Blood. 1996 ; Vol. 87, No. 10. pp. 4348-4357.
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abstract = "We attempted to determine the frequency of normal hematopoietic stem cells (HSC) and contaminating leukemic cells in mobilized peripheral blood (MPB) collected from chronic myeloid leukemia (CML) patients, intolerant of α- interferon or with interferon-resistant disease. A total of 14 MPB samples, six from patients in chronic phase (CP) and eight from patients in accelerated phase or blast crisis (AP/BC) were studied. Cytogenetic analysis of MPB collected from AP/BC patients showed that 100{\%} of the cells were Ph+, whereas cells from four of five CP MPB were Ph-. By contrast, fluorescence in situ hybridization (FISH) analysis of CP MPB showed a mean frequency of 14.7{\%} Ph+ cells, while AP/BC MPB contained 39.2{\%} Ph+ cells. In an attempt to purify normal HSC, subpopulations of the MPB CD34+ cells were isolated based on expression of the Thy-1 antigen (CDw90). The mean Ph+ cell frequency as determined by FISH within the CD34+Thy-1+ Lin- and CD34+Thy- 1-Lin- populations from CP patients was 19.2{\%} and 33.9{\%}, respectively. In the AP/BC patients, levels of residual leukemic cells were significantly greater with mean Ph+ cell frequencies of 59.2{\%} and 72.7{\%} for the CD34+Thy- 1+Lin- and CD34+Thy-1-Lin- fractions, respectively. The frequency of cobblestone area forming cells (CAFC) was used as a means of quantitating the numbers of functional HSC within these cell subpopulations. The mean CAFC frequency was 1 of 19 for the CD34+Thy-1+Lin- cells as compared with 1 of 133 for the Thy-1- fraction indicating a higher frequency of primitive progenitor cells in the Thy-1+ subpopulation. CD34+ cell subsets from two patients were also injected into SCID-hu bone assays to determine the in vivo behavior of these cell populations. After 8 weeks, multilineage donor engraftment was observed in these grafts. FISH analysis of the donor cells within the grafts showed that 55.3{\%} and 60.0{\%} of the cells were Ph+. We conclude that unfractionated MPB from this patient population is not leukemia-free and that the CD34+Thy-1+Lin- cell subpopulation, although predominantly enriched for normal HSC, still contains substantial numbers of residual leukemic cells.",
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T1 - Leukemic burden in subpopulations of CD34+ cells isolated from the mobilized peripheral blood of α-interferon-resistant or -intolerant patients with chronic myeloid leukemia

AU - Van Den Berg, David

AU - Wessman, Maija

AU - Murray, Lesley

AU - Tong, Jie

AU - Chen, Benjamin

AU - Chen, Shirley

AU - Simonetti, Don

AU - King, Julie

AU - Yamasaki, Glenn

AU - DiGiusto, Rhonda

AU - Carella, Angello

AU - Frassoni, Francesco

AU - Gorin, Norbert Claude

AU - Snyder, David

AU - Sciecinski, Irena

AU - Negrin, Robert

AU - Deisseroth, Albert

AU - Tsukamoto, Ann

AU - Gearing, David

AU - Reading, Christopher

AU - Hoffman, Ron

PY - 1996/5/15

Y1 - 1996/5/15

N2 - We attempted to determine the frequency of normal hematopoietic stem cells (HSC) and contaminating leukemic cells in mobilized peripheral blood (MPB) collected from chronic myeloid leukemia (CML) patients, intolerant of α- interferon or with interferon-resistant disease. A total of 14 MPB samples, six from patients in chronic phase (CP) and eight from patients in accelerated phase or blast crisis (AP/BC) were studied. Cytogenetic analysis of MPB collected from AP/BC patients showed that 100% of the cells were Ph+, whereas cells from four of five CP MPB were Ph-. By contrast, fluorescence in situ hybridization (FISH) analysis of CP MPB showed a mean frequency of 14.7% Ph+ cells, while AP/BC MPB contained 39.2% Ph+ cells. In an attempt to purify normal HSC, subpopulations of the MPB CD34+ cells were isolated based on expression of the Thy-1 antigen (CDw90). The mean Ph+ cell frequency as determined by FISH within the CD34+Thy-1+ Lin- and CD34+Thy- 1-Lin- populations from CP patients was 19.2% and 33.9%, respectively. In the AP/BC patients, levels of residual leukemic cells were significantly greater with mean Ph+ cell frequencies of 59.2% and 72.7% for the CD34+Thy- 1+Lin- and CD34+Thy-1-Lin- fractions, respectively. The frequency of cobblestone area forming cells (CAFC) was used as a means of quantitating the numbers of functional HSC within these cell subpopulations. The mean CAFC frequency was 1 of 19 for the CD34+Thy-1+Lin- cells as compared with 1 of 133 for the Thy-1- fraction indicating a higher frequency of primitive progenitor cells in the Thy-1+ subpopulation. CD34+ cell subsets from two patients were also injected into SCID-hu bone assays to determine the in vivo behavior of these cell populations. After 8 weeks, multilineage donor engraftment was observed in these grafts. FISH analysis of the donor cells within the grafts showed that 55.3% and 60.0% of the cells were Ph+. We conclude that unfractionated MPB from this patient population is not leukemia-free and that the CD34+Thy-1+Lin- cell subpopulation, although predominantly enriched for normal HSC, still contains substantial numbers of residual leukemic cells.

AB - We attempted to determine the frequency of normal hematopoietic stem cells (HSC) and contaminating leukemic cells in mobilized peripheral blood (MPB) collected from chronic myeloid leukemia (CML) patients, intolerant of α- interferon or with interferon-resistant disease. A total of 14 MPB samples, six from patients in chronic phase (CP) and eight from patients in accelerated phase or blast crisis (AP/BC) were studied. Cytogenetic analysis of MPB collected from AP/BC patients showed that 100% of the cells were Ph+, whereas cells from four of five CP MPB were Ph-. By contrast, fluorescence in situ hybridization (FISH) analysis of CP MPB showed a mean frequency of 14.7% Ph+ cells, while AP/BC MPB contained 39.2% Ph+ cells. In an attempt to purify normal HSC, subpopulations of the MPB CD34+ cells were isolated based on expression of the Thy-1 antigen (CDw90). The mean Ph+ cell frequency as determined by FISH within the CD34+Thy-1+ Lin- and CD34+Thy- 1-Lin- populations from CP patients was 19.2% and 33.9%, respectively. In the AP/BC patients, levels of residual leukemic cells were significantly greater with mean Ph+ cell frequencies of 59.2% and 72.7% for the CD34+Thy- 1+Lin- and CD34+Thy-1-Lin- fractions, respectively. The frequency of cobblestone area forming cells (CAFC) was used as a means of quantitating the numbers of functional HSC within these cell subpopulations. The mean CAFC frequency was 1 of 19 for the CD34+Thy-1+Lin- cells as compared with 1 of 133 for the Thy-1- fraction indicating a higher frequency of primitive progenitor cells in the Thy-1+ subpopulation. CD34+ cell subsets from two patients were also injected into SCID-hu bone assays to determine the in vivo behavior of these cell populations. After 8 weeks, multilineage donor engraftment was observed in these grafts. FISH analysis of the donor cells within the grafts showed that 55.3% and 60.0% of the cells were Ph+. We conclude that unfractionated MPB from this patient population is not leukemia-free and that the CD34+Thy-1+Lin- cell subpopulation, although predominantly enriched for normal HSC, still contains substantial numbers of residual leukemic cells.

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