Reciprocal changes in fas and bcl2 expression may be responsible for the increased apoptosis observed in dividing cd34+ cells

C. M. Travcpff, Edward Srour

Research output: Contribution to journalArticle

Abstract

We have recently shown by in vitra cell tracking with the fluorescent membrane dye, PKH2, that apoptosis among dividing CD34+ cells from human BM, CB, and MPB Increases In association with the number of in vitro cellular divisions and correlates with a loss in hematopoietic potential in cultured CD34+ cells. To investigate mechanisms behind this increase in apoptosis, we examined the potential roles of FAS (CD95) and BCL2 in mediating apoptosis in dividing CD34+ cells. CD34+ cells from normal human BM, CB, and MPB were stained with PKH2 and cultured for 7 days with SCF, IL-3, and IL-6. and then analyzed for FAS or BCL2 expression In various fractions of expanded CD34+ cells. While CD34+ cells from all 3 tissues expressed low FAS on do, this percentage gradually increased among CD34+ cells during short-term culture such that by d7 40-80% of CD34+ cells were CD95+. Conversely, the percentage of CD34+ cells expressing BCL2 decreased from 90-100% on 00 to <50% by d7. When CD34+ cells were analyzed based on the number of in vitro cellular divisions, it was found that the percentage of CD95+ cells increased concomitantly with the number of In vitro cellular divisions, suggesting that expression of FAS may play a role in the proliferation-associated apoptosis observed in cultured CD34+ cells. This supposition was further substantiated when d7 CD34+/PKH2B" [cylokine non-responsive (CNR)] cells were fractionated based on their expression of CD95 and examined in long-term cultures (LTC). CNR cells lacking CD95 (CNR/CD95-) produced 3-fold more cells and 17-fold more progenitors by d25 in LTC than CNR/CD95+ celb. Furthermore, a greater percentage of CNR/CD95cells expressed BCL2 at d25 and with greater fluorescence intensity than CNR/CD95+ cells (68% @channel 281 vs. 56% @channel 185, respectively) Taken together, these results suggest that FAS-mediated apoptosis may play a role in the diminished hematopoietic potential observed among proliferating human BM, CB, and MPB CD34+ cells, and that this effect may be in part mediated by decreased expression of BCL2.

Original languageEnglish
Pages (from-to)1083
Number of pages1
JournalExperimental Hematology
Volume24
Issue number9
StatePublished - 1996

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Apoptosis
Cultured Cells
Cell Tracking
Interleukin-3
Fluorescent Dyes
Interleukin-6
Fluorescence
Membranes

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Hematology
  • Oncology
  • Transplantation

Cite this

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title = "Reciprocal changes in fas and bcl2 expression may be responsible for the increased apoptosis observed in dividing cd34+ cells",
abstract = "We have recently shown by in vitra cell tracking with the fluorescent membrane dye, PKH2, that apoptosis among dividing CD34+ cells from human BM, CB, and MPB Increases In association with the number of in vitro cellular divisions and correlates with a loss in hematopoietic potential in cultured CD34+ cells. To investigate mechanisms behind this increase in apoptosis, we examined the potential roles of FAS (CD95) and BCL2 in mediating apoptosis in dividing CD34+ cells. CD34+ cells from normal human BM, CB, and MPB were stained with PKH2 and cultured for 7 days with SCF, IL-3, and IL-6. and then analyzed for FAS or BCL2 expression In various fractions of expanded CD34+ cells. While CD34+ cells from all 3 tissues expressed low FAS on do, this percentage gradually increased among CD34+ cells during short-term culture such that by d7 40-80{\%} of CD34+ cells were CD95+. Conversely, the percentage of CD34+ cells expressing BCL2 decreased from 90-100{\%} on 00 to <50{\%} by d7. When CD34+ cells were analyzed based on the number of in vitro cellular divisions, it was found that the percentage of CD95+ cells increased concomitantly with the number of In vitro cellular divisions, suggesting that expression of FAS may play a role in the proliferation-associated apoptosis observed in cultured CD34+ cells. This supposition was further substantiated when d7 CD34+/PKH2B{"} [cylokine non-responsive (CNR)] cells were fractionated based on their expression of CD95 and examined in long-term cultures (LTC). CNR cells lacking CD95 (CNR/CD95-) produced 3-fold more cells and 17-fold more progenitors by d25 in LTC than CNR/CD95+ celb. Furthermore, a greater percentage of CNR/CD95cells expressed BCL2 at d25 and with greater fluorescence intensity than CNR/CD95+ cells (68{\%} @channel 281 vs. 56{\%} @channel 185, respectively) Taken together, these results suggest that FAS-mediated apoptosis may play a role in the diminished hematopoietic potential observed among proliferating human BM, CB, and MPB CD34+ cells, and that this effect may be in part mediated by decreased expression of BCL2.",
author = "Travcpff, {C. M.} and Edward Srour",
year = "1996",
language = "English",
volume = "24",
pages = "1083",
journal = "Experimental Hematology",
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T1 - Reciprocal changes in fas and bcl2 expression may be responsible for the increased apoptosis observed in dividing cd34+ cells

AU - Travcpff, C. M.

AU - Srour, Edward

PY - 1996

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N2 - We have recently shown by in vitra cell tracking with the fluorescent membrane dye, PKH2, that apoptosis among dividing CD34+ cells from human BM, CB, and MPB Increases In association with the number of in vitro cellular divisions and correlates with a loss in hematopoietic potential in cultured CD34+ cells. To investigate mechanisms behind this increase in apoptosis, we examined the potential roles of FAS (CD95) and BCL2 in mediating apoptosis in dividing CD34+ cells. CD34+ cells from normal human BM, CB, and MPB were stained with PKH2 and cultured for 7 days with SCF, IL-3, and IL-6. and then analyzed for FAS or BCL2 expression In various fractions of expanded CD34+ cells. While CD34+ cells from all 3 tissues expressed low FAS on do, this percentage gradually increased among CD34+ cells during short-term culture such that by d7 40-80% of CD34+ cells were CD95+. Conversely, the percentage of CD34+ cells expressing BCL2 decreased from 90-100% on 00 to <50% by d7. When CD34+ cells were analyzed based on the number of in vitro cellular divisions, it was found that the percentage of CD95+ cells increased concomitantly with the number of In vitro cellular divisions, suggesting that expression of FAS may play a role in the proliferation-associated apoptosis observed in cultured CD34+ cells. This supposition was further substantiated when d7 CD34+/PKH2B" [cylokine non-responsive (CNR)] cells were fractionated based on their expression of CD95 and examined in long-term cultures (LTC). CNR cells lacking CD95 (CNR/CD95-) produced 3-fold more cells and 17-fold more progenitors by d25 in LTC than CNR/CD95+ celb. Furthermore, a greater percentage of CNR/CD95cells expressed BCL2 at d25 and with greater fluorescence intensity than CNR/CD95+ cells (68% @channel 281 vs. 56% @channel 185, respectively) Taken together, these results suggest that FAS-mediated apoptosis may play a role in the diminished hematopoietic potential observed among proliferating human BM, CB, and MPB CD34+ cells, and that this effect may be in part mediated by decreased expression of BCL2.

AB - We have recently shown by in vitra cell tracking with the fluorescent membrane dye, PKH2, that apoptosis among dividing CD34+ cells from human BM, CB, and MPB Increases In association with the number of in vitro cellular divisions and correlates with a loss in hematopoietic potential in cultured CD34+ cells. To investigate mechanisms behind this increase in apoptosis, we examined the potential roles of FAS (CD95) and BCL2 in mediating apoptosis in dividing CD34+ cells. CD34+ cells from normal human BM, CB, and MPB were stained with PKH2 and cultured for 7 days with SCF, IL-3, and IL-6. and then analyzed for FAS or BCL2 expression In various fractions of expanded CD34+ cells. While CD34+ cells from all 3 tissues expressed low FAS on do, this percentage gradually increased among CD34+ cells during short-term culture such that by d7 40-80% of CD34+ cells were CD95+. Conversely, the percentage of CD34+ cells expressing BCL2 decreased from 90-100% on 00 to <50% by d7. When CD34+ cells were analyzed based on the number of in vitro cellular divisions, it was found that the percentage of CD95+ cells increased concomitantly with the number of In vitro cellular divisions, suggesting that expression of FAS may play a role in the proliferation-associated apoptosis observed in cultured CD34+ cells. This supposition was further substantiated when d7 CD34+/PKH2B" [cylokine non-responsive (CNR)] cells were fractionated based on their expression of CD95 and examined in long-term cultures (LTC). CNR cells lacking CD95 (CNR/CD95-) produced 3-fold more cells and 17-fold more progenitors by d25 in LTC than CNR/CD95+ celb. Furthermore, a greater percentage of CNR/CD95cells expressed BCL2 at d25 and with greater fluorescence intensity than CNR/CD95+ cells (68% @channel 281 vs. 56% @channel 185, respectively) Taken together, these results suggest that FAS-mediated apoptosis may play a role in the diminished hematopoietic potential observed among proliferating human BM, CB, and MPB CD34+ cells, and that this effect may be in part mediated by decreased expression of BCL2.

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