Stem Cells http://www.peprotech.com/
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Reprints/Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Huang, S.
Right arrow Articles by Law, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Huang, S.
Right arrow Articles by Law, P.
Stem Cells, Vol. 17, No. 5, 265-272, September 1999
© 1999 AlphaMed Press

Correlation Between IL-3 Receptor Expression and Growth Potential of Human CD34+ Hematopoietic Cells from Different Tissues

Shiang Huanga, Zhang Chenb, Ji Feng Yub, Dennis Younga, Asad Basheya, Anthony D. Hoc, Ping Lawa

a University of California San Diego, Division of Blood & Marrow Transplantation, La Jolla, California, USA;
b PharMingen Corporation, San Diego, California, USA;
c University of Heidelberg, Department of Internal Medicine V, Heidelberg, Germany

Key Words. CD34+ cells • CD123 • IL-3 receptor • Hematopoietic cells • Committed progenitors • Erythroid progenitors

Dr. Ping Law, Division of Blood and Marrow Transplantation, University of California San Diego, 9300 Campus Point Drive Mailcode 7621, La Jolla, California 92037-7621, USA.


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
CD123 ({alpha}-subunit of IL-3 receptor) expression on primitive and committed human hematopoietic cells was studied by multicolor sorting and single-cell culture. The sources of cells included fetal liver (FLV), fetal bone marrow, umbilical cord blood, adult bone marrow and mobilized peripheral blood. Three subsets of CD34+ cells were defined by the levels of surface CD123: CD123negative, CD123low, and CD123bright. Coexpression of lineage markers showed that a majority of CD34+CD123bright cells were myeloid and B-lymphoid progenitors, while erythroid progenitors were mainly in the CD34+CD123negative subset. The CD34+CD123low subset contained a heterogeneous distribution of early and committed progenitor cells. Single CD34+ cells from the CD123 subsets were cultured in a cytokine cocktail of stem cell factor, interleukin 3 (IL-3), IL-6, GM-CSF, erythropoietin, insulin-like growth factor-1, and basic fibroblast growth factor. After 14 days of incubation, a higher cloning efficiency (CE) was observed in the CD34+CD123negative and CD34+CD123low fractions (37 ± 23% and 44 ± 23%, respectively) than in the CD34+CD123bright fraction (15 ± 21%). Using previously published criteria that colonies containing dispersed, translucent cells (dispersed growth pattern, DGP) were derived from primitive cells and that colonies composed solely of clusters were from committed cells, early precursors were distributed evenly in the CD34+CD123negative and CD34+CD123low subsets. When CD38 and CD90 (Thy-1) were used for further characterization of CD34+ cells from FLV, CE increased from 37 ± 23% in CD123negative to 70 ± 19% in CD123negativeCD38- and from 44 ± 23% in CD123low to 66 ± 19% in CD123lowCD38-. No significant increase in CE or DGP progenitors was observed when CD34+ cells were sorted by CD90 and CD123. We concluded that: A) high levels of CD123 were expressed on B-lymphoid and myeloid progenitors; B) early erythroid progenitors had little or no surface CD123, and C) primitive hematopoietic cells are characterized by CD123negative/low expression.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
Growth of human hematopoietic stem/progenitor cells depends upon a hierarchy of growth factors, among which interleukin 3 (IL-3) is a multipotent cytokine capable of stimulating primitive as well as lineage-committed progenitors. IL-3 alone or in combination with other early-acting hematopoietic growth factors, such as stem cell factor (SCF), IL-1, and IL-6, has been shown to support the multilineage colony formation and expansion to different lineages in liquid culture [1-8]. In addition, IL-3 has been used in culture to produce blood cells of various lineages, including neutrophils, monocytes, erythrocytes, megakaryocytes, basophils, eosinophils, mast cells, and B-lymphoid cells [2, 5, 9-13]. Administration of IL-3 to human and primate subjects has enhanced multilineage-hematopoiesis [14-17]. Autologous progeny cells after culture/expansion in cytokine combination including IL-3 have been successfully infused into patients after a nonmyeloablative conditioning regimen [18]. In another study, when patients were given a myeloablative conditioning regimen, cells expanded in IL-3-containing cytokines failed to produce engraftment [19]. More recent data indicated that culture in IL-3 led to a decrease or loss of primitive hematopoietic cells with reconstitution potential [20-23].

IL-3 exerts its biological activities through binding to specific cell-surface receptors. A single class of high-affinity receptor for IL-3 is detected on different human cells, including stem/progenitor cells, granulocytes, monocytes, megakaryocytes, and B lymphocytes [24-28]. The human IL-3 receptor consists of a 70kD {alpha}-subunit specific for IL-3 and with low binding affinity, and a 120-140 kD ß-subunit with high binding affinity when coexpressed with the {alpha}-subunit [11, 28]. The human IL-3 receptor {alpha}-subunit was designated CD123, and consists of three extracellular domains, a transmembrane domain, and a short intracellular domain [29]. The N-terminal domain of this molecule was found to play a significant role in its binding to IL-3 [29]. The ß-subunit is a common component of IL-3, IL-5, and GM-CSF receptors and is critical for signal transduction, proliferation, and differentiation [30-33].

Studies of CD123 expression on primitive and committed progenitor cells can provide information on the effect of IL-3 on primitive and lineage-committed progenitors. Different levels of CD123 expression were reported on CD34+ cells at different stages of differentiation [24, 34-37]. Wognum et al. [35] reported that early primate hematopoietic progenitor cells identified as CD34+HLA-DRdull cells had a low level of CD123, while CD34+ cells with high or negative CD123 expression were committed myeloid and erythroid progenitors. Sato et al. [36] showed that a portion of human marrow and cord blood CD34+ cells expressed CD123 and that culture of the CD34+ cells for a short period (two days) in different growth factors increased CD123 expression. The authors speculated that primitive hematopoietic cells resided in the CD34+CD123- population. In contrast, Wagner et al. [37], using counter-flow elutriation, demonstrated that the primitive cells located in the small cell fraction had a heterogeneous expression of CD123. It is possible that CD123 expression on primitive CD34+ cells is similar to that of CD117 (c-kit); i.e., low expression on CD34+ cells signified the more primitive compartment, while the maturing progenitor had high surface levels [38-40]. Some reports, however, suggest that immature hematopoietic cells express high levels of kit [35, 41]. In this study, we investigated the surface expression of CD123 on human CD34+ cells from fetal liver (FLV), fetal bone marrow (FBM), umbilical cord blood (UCB), adult bone marrow (ABM), and mobilized peripheral blood (MPB) using multicolor flow cytometry, and correlated the growth of single CD34+ and CD34+CD38- cells to the level of CD123 expression. We found that the primitive hematopoietic progenitor cells were evenly distributed among the CD123negative and CD123low subsets of CD34+CD38- cells.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Preparation of Cells
ABM was collected from the iliac crests of healthy individuals. UCB was either collected at the University of California San Diego (UCSD) delivery room using ACD-A (Baxter-Fenwal; Deerfield, IL) as anticoagulant or obtained from Advanced Bioscience Resources ([ABR]; Alameda, CA). MPB was obtained from leukapheresis products (LPs) of healthy donors mobilized using a combination of GM-CSF (Immunex; Seattle, WA) and G-CSF (Amgen; Thousand Oaks, CA) in combinations of 5 µg/kg/d each for five or six days or GM-CSF (10 µg/kg/d) for three to four days followed by G-CSF (10 µg/kg/d) for two days. LPs were collected by leukapheresis procedure on Cobe Spectra (processing 10 l of blood) 24 h after the last administration of growth factors. The mobilization and collection of MPB have been published elsewhere [42, 43]. FLV and FBM (between 17 and 24 weeks of gestation) were procured from ABR. Use of all tissues has been reviewed and approved by the human subjects committee of UCSD. Informed consent was obtained from all live donors. Single-cell suspension of FBM was prepared by flushing the bone marrow cells from the humerus and femurs using a syringe and a 22-gauge needle into RPMI 1640 with 2% fetal bovine serum (FBS) (Germini; Calabasas, CA). FLV cells were homogenized through a cell strainer (Becton Dickinson Labware; Franklin Lakes, NJ) and washed once by centrifugation in RPMI 1640 with 2% FBS. Mononuclear cells (MNCs) from all tissues were separated by centrifugation on Ficoll-Hypaque (Histopaque-1077, Sigma; St. Louis, MO) and washed twice before staining and sorting.

Flow Cytometry and Cell Sorting
Phycoerythrin (PE)-conjugated anti-CD123 and anti-CD90 monoclonal antibody (mAb) were provided by PharMingen Corporation (San Diego, CA). Anti-CD123 is a mouse IgG2a (7G3) directed against the {alpha}-subunit of the human IL-3 receptor. All other mAbs were obtained from Becton Dickinson Immunocytometry Systems ([BDIS]; San Jose, CA), including: CD3-PerCP, CD10-fluorescein isothiocyanate (FITC), CD15-FITC, CD19-PerCP, CD34 (HPAC-2)-FITC, CD34-PerCP, CD61-FITC, CD71 (anti-transferrin receptor)-FITC, and HLA-DR-FITC. Cells were stained with mAb for 30 min on ice and washed twice with RPMI 1640 containing 2% FBS. Acquisition for CD34+ subset analysis was performed on a FACScan using LYSYS II (BDIS) software. At least 30,000 events were acquired for each data file. For CD34+ subset analysis, a minimum of 10,000 cells from each sample was collected in list-mode with light scatter gates (forward and side-scatter) to exclude neutrophils and RBC, and a fluorescence gate to include only CD34+ events. The proportion in each subset was calculated using the Paint-A-Gateplus or LYSYS II. Single cells were sorted into 96-well dishes (Costar; Cambridge, MA) using FACStarplus with an automated cell deposition unit. The accuracy of single-cell sorting and deposition is estimated to be >99% [44]. A typical sort window for CD34+ cells with different levels of CD123 expression is shown in Figure 1Go.



View larger version (30K):
[in this window]
[in a new window]
  		Figure 1. Expression of CD123 on CD34+ cells in FLV (A, B), UCB (C, D), and MPB (E, F). Low-density MNCs from FLV, UCB, and MPB were stained with CD34-FITC and CD123 PE. 30,000 events were acquired in list mode on a FACScan and analyzed with the CUT-A-CLUSTER software (BDIS). Light scatter characteristics of the cells were shown in A, C, and E.

 
Single Cell Culture and Scoring
The culture conditions and colony scoring for the sorted cells were described [45, 46]. Briefly, each well contained 200 µl of myeloid long-term culture medium (H5100, Stem Cell Technologies, Inc.; Vancouver, Canada) with 12.5% horse serum and 12.5% FBS, and was supplemented with 2.5 U of human recombinant erythropoietin, 40 ng/ml of recombinant human SCF (Amgen), 10 ng/ml of GM-CSF (Immunex), 10 ng/ml of recombinant human IL-3, 500 U/ml of recombinant human IL-6, 2.5 ng/ml of recombinant human basic fibroblast growth factor, and 10 ng/ml of recombinant human insulin-like growth factor-1 (Collaborative Research; Bedford, MA). The combination had been shown to support the growth of primitive as well as committed progenitor cells [45-47]. All cultures were incubated in 5% CO2/95% air at 37°C in a fully humidified incubator. A colony was defined as a cluster of >40 cells in a well after 14 days. Cloning efficiency (CE) was defined as the number of wells with colonies divided by the number of wells. Whether a colony-forming hematopoietic cell came from a primitive or committed progenitor was defined by the growth pattern: the colonies consisting solely of dispersed, translucent cells (dispersed growth pattern, [DGP]) were derived from primitive progenitors which had been shown to be capable of growth after replating of up to four generations [45]; the colonies consisting solely of tight clusters (cluster growth pattern [CGP]) were derived from committed progenitors which had been shown to have little or no replating potential; and the colonies with mixed growth pattern ([MGP], containing both DGP and CGP) were intermediate progenitors with limited replating potential. Erythroid progenitors were identified by colonies containing RBC.

Statistical Analysis
Mean ± standard deviation was calculated and is shown in tables and figures unless otherwise stated. Statistical significance was determined by Student's t-test. Differences were considered significant if p < 0.05.


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
Expression of CD123 on CD34+ Cells
Surface CD123 can be observed on 1% to 3% of the MNCs in FLV, UCB, and ABM. Typical dot-plots of CD34/CD123-stained samples are shown in Figure 1Go. CD34+ cells can be arbitrarily divided into three subsets: CD123negative, CD123low, and CD123bright. A majority of CD34+ cells expressed surface CD123 (CD123low + CD123bright). As summarized in Table 1Go, 79% of CD34+ cells were derived from fetal tissues, and 91% from mature sources were CD123+. Most CD34+CD123+ cells were located in CD123low region (61.1 ± 8.6% in fetal tissues and 80.4 ± 6.0% in mature sources). These results were consistent with early reports of IL-3R (receptor) expression in primate and human subjects [35, 36].


View this table:
[in this window]
[in a new window]
  		Table 1. Expression of CD123 on CD34+ cells from fetal and mature hematopoietic tissues
 
Single-Cell Culture of CD34+CD123 Subsets
The growth potential of different CD34+CD123 subsets was measured by single-cell sorting and culture. The CE of the sorted cells from FLV, UCB, and MPB is shown in Figure 2Go. In FLV, the CE of CD34+CD123bright cells was significantly lower than that of the other two subsets (p <= 0.05); however, the CE of different CD123 subsets of CD34+ cells in UCB and MPB was not significantly different. The presence of primitive hematopoietic cells was evaluated by the growth pattern of the colonies. Typical colonies with DGP, MGP, and CGP are shown in Figure 3Go (A, C, and D). Also included is a Wright-Giemsa-stained cell preparation from a DGP colony (B).



View larger version (16K):
[in this window]
[in a new window]
  		Figure 2. Cloning efficiency of CD34+ cells with different levels of CD123. Single cells of three different CD34+CD123 subsets were sorted according to the gates as defined in Figure 1Go. After 14 days of incubation in a mixture of cytokines, all wells that contained colonies of more than 40 cells were counted and divided by the total number of wells with sorted cells.

 


View larger version (116K):
[in this window]
[in a new window]
  		Figure 3. Growth patterns of primitive and committed CD34+CD123low FLV cells. Each colony was evaluated after 14 days of culture. (A) A colony with dispersed growth pattern (DGP) developed from primitive cells; (B) Cells from the DGP colony of (A) were collected and stained with Wright-Giemsa and photographed at 1,000x; (C) A colony with mixed growth pattern (MGP); (D) A colony with cluster growth pattern (CGP).

 
Growth pattern data are summarized in Table 2Go. In FLV, most of the colonies derived from primitive hematopoietic cells (DGP + MGP) were found in the CD123low subset (13 ± 9% versus 5% in CD123negative and 2% in CD123bright), while in UCB and MPB, the primitive cells were evenly distributed in CD123negative and CD123low subsets. In all tissues, few or no primitive hematopoietic cells could be detected in the CD123bright subset.


View this table:
[in this window]
[in a new window]
  		Table 2. Single cell culture of CD34+CD123 subsets: % of cells that formed colonies with dispersed (DGP), mixed (MGP) or cluster (CGP) growth patterns
 
Erythroid progenitor cells can be identified by the presence of RBCs in the MGP and CGP colonies. Most of erythroid progenitor cells, as summarized in Figure 4Go, were found in the CD123negative fraction in all tissues (FLV: 63 ± 26%; UCB: 40 ± 16%; MPB: 68 ± 23%). The values were significantly (p < 0.05) higher than those of other CD123 subsets of the same tissue. To confirm that the erythroid progenitors have little or no expression of CD123, CD34+CD123negative cells from UCB were further sorted according to CD71 into the single-cell culture system. As shown in Table 3Go, CE was higher in the CD71+ subset, and a majority of colonies were from erythroid progenitors, while the myeloid colonies were predominantly in the CD71- fraction.



View larger version (15K):
[in this window]
[in a new window]
  		Figure 4. Growth of CD34+ erythroid progenitor cells with different levels of CD123. MGP or CGP colonies with RBC were scored as erythroid colonies. There were no erythroid progenitor cells in the colonies with DGP.

 

View this table:
[in this window]
[in a new window]
  		Table 3. Erythroid differentiation of CD34+CD123negative cells
 
Cloning Efficiencies and Growth Patterns of Primitive CD34+CD123negative/low Subsets
To identify and characterize the primitive hematopoietic cell, CE and growth patterns of CD34+CD123negative and CD34+CD123low subsets in FLV were examined after fractionation using CD34, CD123, and CD38 or CD90 (Thy-1), and the results are summarized in Table 4Go. The CE and growth patterns from CD34+CD38-HLA-DR+ cells were included for comparison. When lack of CD38 expression was introduced to the single-cell sorting, CE increased from 37% ± 23% (Fig. 2Go) to 70% ± 19% (p < 0.05) in the CD123negative subset and from 44% ± 18% to 66% ± 19% in the CD123low subset (p < 0.05). The increase in primitive colonies (colonies with DGP and MGP) was statistically significant in the CD123negative subset (from 5% ± 4% to 18% ± 7%, p < 0.05), but not in the CD123low subset (from 13% ± 8% to 18% ± 8%, p > 0.1). Adding CD90 (positiveness) to the sorting gate did not result in any significant increase in CE (37% to 44% in CD123negative and 44% to 28% in CD123low subsets) or colonies with DGP and CGP (5% to 10% in CD123negative and 13% to 10% in CD123low subsets). Lack of surface CD38 was a more useful distinction than the presence of surface CD90 for early primitive hematopoietic cells. Growth (CE and pattern) of CD123negative and CD123low subsets was comparable to CD34+CD38-HLA-DR+ cells.


View this table:
[in this window]
[in a new window]
  		Table 4. Single cell culture of CD34+CD123negative and CD34+CD123low FLV cells: sorting with CD38 and CD90 (Thy 1)
 

   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
IL-3 plays a critical functional role in hematopoiesis and exerts its effects through a specific IL-3 receptor (IL-3R or CD123). In this study, our results in flow cytometry staining showed that a majority (79.7% to 93.9%) of CD34+ cells expressed IL-3R (fetal and adult hematopoietic tissues), comparable to the findings in other published reports [35, 36]. Based on the level of surface CD123, we sorted three subsets of CD34+ cells and found that the early hematopoietic cells were mainly located in the CD123low region in FLV, but not in UCB or MPB (Table 2Go).

The result indicating that the growth of CD34+CD123 subsets was different in fetal compared to mature hematopoietic tissues is interesting. Our previous finding showed that in hematopoietic cells with the same surface markers (e.g., CD34+CD38-HLA-DR+), growth (CE and percentage of primitive colonies) in the fetal tissues was superior (FLV + FBM > UCB > MPB > ABM) [46], but the distribution pattern of primitive colonies among CD34+ subsets remained the same (e.g., CD38-HLA-DR+ containing the highest proportion of primitive cells) for all tissues. There were no differences in the CE and growth patterns of CD123negative versus CD123low subsets in UCB and MPB. It is possible that neither the CD38 nor HLA-DR markers were directly associated with erythroid differentiation [48], while the CD34+CD123negative subset contained a majority of the erythroid precursors (Table 3Go). FLV is known to be highly erythropoietic as compared with UCB and MPB. Different behavior of the same CD34+ cell subsets among hematopoietic cell sources of different ontogenic ages was also reported by others [49, 50].

Both lack of CD38 expression and positiveness for CD90 expression on human CD34+ cells had been used for identifying primitive hematopoietic cells [47, 51-54]. Our previous report showed that CD38- is a better marker for primitive hematopoietic cells [53]. Results from the current study support the same conclusion, as the addition of CD90 into the CD123 subset fractionation did not improve CE or the proportion of primitive colonies (Table 4Go).

Recently, CD34- primitive hematopoietic stem cells have been isolated [55-62]. The cells were first reported in mice [59, 60] and later identified in humans [61, 62]. Experimental evidence suggested that the CD34- cells were more primitive than the CD34+CD38-HLA-DR+ subset in a well-established human-to-sheep in utero transplantation system [62]. Cytokine receptor expression on these primitive hematopoietic cells has not been established.

In conclusion, we have shown that A) high levels of surface CD123 were associated with B-lymphoid and myeloid differentiation; B) CD34+ erythroid progenitors had little or no surface CD123, and C) early hematopoietic cells were distributed evenly among the CD123negative and CD123low subsets of CD34+ cells.


   Acknowledgments
 
Supported in part by NIH grant no. R01-496619 and NCI Core Grant CA 23100.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. Ogawa M. Differentiation and proliferation of hematopoietic stem cells. Blood 1993;81:2844-2853.[Abstract/Free Full Text]

  2. Moore MA. Review: Stratton Lecture 1990. Clinical implications of positive and negative hematopoietic stem cell regulators. Blood 1991;78:1-19.[Free Full Text]

  3. Sieff C, Niemeyer C, Nathan D et al. Stimulation of human hematopoietic colony formation by recombinant gibbon multi-colony-stimulating factor or interleukin-3. J Clin Invest 1987;380:818-823.

  4. Kimura T, Sakabe H, Tanimukai S et al. Simultaneous activation of signals through gp130, c-kit, and interleukin-3 receptor promotes a trilineage blood cell production in the absence of terminally acting lineage-specific factors. Blood 1997;90:4767-4778.[Abstract/Free Full Text]

  5. Miller JS, McCullar V, Verfaillie CM. Ex vivo culture of CD34+/Lin-/DR- cells in stroma-derived soluble factors, interleukin-3, and macrophage inflammatory protein-1 alpha maintains not only myeloid but also lymphoid progenitors in a novel switch culture assay. Blood 1998;91:4516-4522.[Abstract/Free Full Text]

  6. Kawada H, Ando K, Tsuji T et al. Rapid ex vivo expansion of human umbilical cord hematopoietic progenitors using a novel culture system. Exp Hematol 1999;27:904-915.[Medline]

  7. Ahmed N, Khokher MA, Hassan HT. Cytokine-induced expansion of human CD34+ stem/progenitor and CD34+CD41+ early megakaryocytic marrow cells cultured on normal osteoblasts. STEM CELLS 1999;17:92-99.[Abstract/Free Full Text]

  8. Pierelli L, Scambia G, Bonanno G et al. Expansion of granulocyte colony-stimulating factor/chemotherapy-mobilized CD34+ hematopoietic progenitors: role of granulocyte-macrophage colony-stimulating factor/erythropoietin hybrid protein (MEN11303) and interleukin-15. Exp Hematol 1999;27:416-424.[Medline]

  9. Lévêque C, Grafte S, Paysant J et al. Regulation of interleukin 3 receptor alpha chain (IL-3R alpha) on human monocytes by interleukin (IL)-4, IL-10, IL-13, and transforming growth factor beta (TGF-beta). Cytokine 1998;10:487-494.[Medline]

  10. McKinstry WJ, Li CL, Rasko JE et al. Cytokine receptor expression on hematopoietic stem and progenitor cells. Blood 1997;89:65-71.[Abstract/Free Full Text]

  11. Miyajima A, Hara T, Kitamura T. Common subunits of cytokine receptors and the functional redundancy of cytokines. Trends Biochem Sci 1992;17:378-382.[Medline]

  12. Shalit M, Sekhsaria S, Mauhorter S et al. Early commitment to the eosinophil lineage by cultured human peripheral blood CD34+ cells: messenger RNA analysis. J Allergy Clin Immunol 1996;98:344-354.[Medline]

  13. Testa U, Fossati C, Samoggia P et al. Expression of growth factor receptors in unilineage differentiation culture of purified hematopoietic progenitors. Blood 1996;88:3391-3406.[Abstract/Free Full Text]

  14. Donahue R, Seehra J, Metzger M et al. Human IL-3 and GM-CSF act synergistically in stimulating hematopoiesis in primates. Science 1988;241:1820-1823.[Abstract/Free Full Text]

  15. van Gils F, van Teeffelen M, Neelis K et al. Interleukin-3 treament of Rhesus monkeys leads to increased production of histamine-releasing cells that express interleukin-3 receptors at high levels. Blood 1995;86:592-597.[Abstract/Free Full Text]

  16. Ganser A, Lindemann A, Seipelt G et al. Effect of recombinant human interleukin-3 in patients with normal hematopoiesis and in patients with bone marrow failure. Blood 1990;76:666-676.[Abstract/Free Full Text]

  17. Wagemaker G, van Gils F, Burger H et al. Highly increased production of derived blood cells by adminstration of homologous IL-3 to rhesus monkeys. Blood 1990;76:2235-2241.[Abstract/Free Full Text]

  18. Brugger W, Heimfeld S, Berenson RJ et al. Reconstitution of hematopoiesis after high-dose chemotherapy by autologous progenitor cells generated ex vivo [see comments]. N Engl J Med 1995;333:283-287.[Abstract/Free Full Text]

  19. Holyoake TL, Alcorn MJ, Richmond L et al. CD34 positive PBPC expanded ex vivo may not provide durable engraftment following myeloablative chemoradiotherapy regimens. Bone Marrow Transplant 1997;19:1095-1101.[Medline]

  20. Genechea G, Segovia JC, Albella B et al. Delayed engraftment of nonobese diabetic/severe combined immunodeficient mice transplanted with ex vivo-expanded human CD34(+) cord blood cells. Blood 1999;93:1097-1105.[Abstract/Free Full Text]

  21. Yonemura Y, Ku H, Hirayama F et al. Interleukin-3 or interleukin-1 abrogates the reconstituting ability of hematopoietic stem cells. Proc Natl Acad Sci USA 1996;93:4040-4044.[Abstract/Free Full Text]

  22. Matsunaga T, Hirayama F, Yonemura Y et al. Negative regulation by interleukin-3 (IL-3) of mouse early B-cell progenitors and stem cells in culture: transduction of the negative signals by betac and betaIL-3 proteins of IL-3 receptor and absence of negative regulation by granulocyte-macrophage colony-stimulating factor. Blood 1998;92:901-907.[Abstract/Free Full Text]

  23. Tisdale JF, Hanazono Y, Sellers SE et al. Ex vivo expansion of genetically marked rhesus peripheral blood progenitor cells results in diminished long-term repopulating ability. Blood 1998;92:1131-1141.[Abstract/Free Full Text]

  24. Park L, Friend D, Price V et al. Heterogeneity in human interleukin-3 receptors. A subclass that binds human granulocyte/macrophage colony stimulating factor. J Biol Chem 1989;264:5420-5427.[Abstract/Free Full Text]

  25. Gesner TG, Mufson RA, Norton CR et al. Specific binding, internalization, and degradation of human recombinant interleukin-3 by cells of the acute myelogenous, leukemia line, KG-1. J Cell Physiol 1988;136:493-499.[Medline]

  26. Uckun FM, Gesner TG, Song CW et al. Leukemic B-cell precursors express functional receptors for human interleukin-3. Blood 1989;73:533-542.[Abstract/Free Full Text]

  27. Budel LM, Elbaz O, Hoogerbrugge H et al. Common binding structure for granulocyte macrophage colony-stimulating factor and interleukin-3 on human acute myeloid leukemia cells and monocytes. Blood 1990;75:1439-1445.[Abstract/Free Full Text]

  28. Kitamura T, Sato N, Arai K et al. Expression cloning of the human IL-3 receptor cDNA reveals a shared beta subunit for the human IL-3 and GM-CSF receptors. Cell 1991;66:1165-1174.[Medline]

  29. Barry SC, Korpelainen E, Sun Q et al. Roles of the N and C terminal domains of the interleukin-3 receptor alpha chain in receptor function. Blood 1997;89:842-852.[Abstract/Free Full Text]

  30. Nicola NA, Metcalf D. Subunit promiscuity among hemopoietic growth factor receptors. Cell 1991;67:1-4.[Medline]

  31. Lopez A, Elliott M, Woodcock J et al. GM-CSF, IL-3 and IL-5: cross-competition on human hematopoietic cells. Immunol Today 1992;18:495-500.

  32. Bagley CJ, Woodcock JM, Stomski FC et al. The structural and functional basis of cytokine receptor activation: lessons from the common beta subunit of the granulocyte-macrophage colony-stimulating factor, interleukin-3 (IL-3), and IL-5 receptors. Blood 1997;89:1471-1482.[Free Full Text]

  33. McCormack MP, Gonda TJ. Expression of activated mutants of the human interleukin-3/interleukin-5/granulocyte-macrophage colony-stimulating factor receptor common beta subunit in primary hematopoietic cells induces factorindependent proliferation and differentiation. Blood 1997;90:1471-1481.[Abstract/Free Full Text]

  34. van Gils F, Budel L, Burger H et al. Interleukin-3 (IL-3) receptor on rhesus monkey cell: species specificity of human IL-3, binding characteristics, and lack of competition with GM-CSF. Exp Hematol 1994;22:248-255.[Medline]

  35. Wognum AW, de Jong MO, Wagemaker G. Differential expression of receptors for hemopoietic growth factors on subsets of CD34+ hemopoietic cells. Leuk Lymphoma 1996;24:11-25.[Medline]

  36. Sato N, Caux C, Kitamura T et al. Expression and factor-dependent modulation of the interleukin-3 receptor subunits on human hematopoietic cells. Blood 1993;82:752-761.[Abstract/Free Full Text]

  37. Wagner J, Collins D, Fuller S et al. Isolation of small primitive human hematopoietic stem cells: distribution of cell surface cytokine receptors and growth in SCID-Hu mice. Blood 1995;86:512-523.[Abstract/Free Full Text]

  38. Kawashima I, Zanjani ED, Almaida-Porada G et al. CD34+ human marrow cells that express low levels of Kit protein are enriched for long-term marrow-engrafting cells. Blood 1996;87:4136-4142.[Abstract/Free Full Text]

  39. Sansilvestri P, Cardoso AA, Batard P et al. Early CD34high cells can be separated into KIThigh cells in which transforming growth factor-beta (TGF-beta) downmodulates c-kit and KITlow cells in which anti-TGF-beta upmodulates c-kit. Blood 1995;86:1729-1735.[Abstract/Free Full Text]

  40. Sakabe H, Yahata N, Kimura T et al. Human cord blood-derived primitive progenitors are enriched in CD34+ c-kit- cells: correlation between long-term culture-initiating cells and telomerase expression. Leukemia 1998;12:728-734.[Medline]

  41. Ratajczak MZ, Pletcher CH, Marlicz W et al. CD34+, kit+, rhodamine123(low) phenotype identifies a marrow cell population highly enriched for human hematopoietic stem cells. Leukemia 1998;12:942-950.[Medline]

  42. Lane T, Law P, Maruyama M et al. Harvesting and enrichment of hematopoietic progenitor cells mobilized into the peripheral blood of normal donors by granulocyte-macrophage colony-stimulating factor (GM-CSF) or G-CSF: potential role in allogeneic marrow transplantation. Blood 1995;85:272-282.

  43. Lane TA, Ho AD, Bashey A et al. Mobilization of blood-derived stem and progenitor cells in normal subjects by granulocyte-macrophage- and granulocyte-colony-stimulating factors. Transfusion 1999;39:39-47.[Medline]

  44. Huang S, Terstappen LW. Formation of haematopoietic microenvironment and haematopoietic stem cells from single human bone marrow stem cells [retraction of Huang S, Terstappen LW. In: Nature 1992;360:745-749]. Nature 1994;368:664.[Medline]

  45. Huang S, Terstappen LW. Lymphoid and myeloid differentiation of single human CD34+, HLA-DR+, CD38- hematopoietic stem cells. Blood 1994;83:1515-1526.[Abstract/Free Full Text]

  46. Huang S, Law P, Young D et al. Candidate hematopoietic stem cells from fetal tissues, umbilical cord blood versus adult bone marrow and mobilized peripheral blood. Exp Hematol 1998;26:1162-1171.[Medline]

  47. Terstappen LW, Huang S, Safford M et al. Sequential generations of hematopoietic colonies derived from single nonlineage-committed CD34+CD38- progenitor cells. Blood 1991;77:1218-1227.[Abstract/Free Full Text]

  48. Terstappen L, Gandour D, Huang S et al. Assessment of hematopoietic cell differentiation by multidimensional flow cytometry. J Hematother 1993;2:431-447.[Medline]

  49. Weekx SF, Van Bockstaele DR, Plum J et al. CD34++ CD38- and CD34+ CD38+ human hematopoietic progenitors from fetal liver, cord blood, and adult bone marrow respond differently to hematopoietic cytokines depending on the ontogenic source. Exp Hematol 1998;26:1034-1042.[Medline]

  50. Zandstra PW, Conneally E, Piret JM et al. Ontogeny-associated changes in the cytokine responses of primitive human haemopoietic cells. Br J Haematol 1998;101:770-778.[Medline]

  51. Baum CM, Weissman IL, Tsukamoto AS et al. Isolation of a candidate human hematopoietic stem-cell population. Proc Natl Acad Sci USA 1992;89:2804-2808.[Abstract/Free Full Text]

  52. Hao QL, Smogorzewska EM, Barsky LW et al. In vitro identification of single CD34+CD38- cells with both lymphoid and myeloid potential. Blood 1998;91:4145-4151.[Abstract/Free Full Text]

  53. Ho AD, Young D, Maruyama M et al. Pluripotent and lineage-committed CD34+ subsets in leukapheresis products mobilized by G-CSF, GM-CSF versus a combination of both. Exp Hematol 1996;24:1460-1468.[Medline]

  54. D'Arena G, Musto P, Cascavilla N et al. Thy-1 (CDw90) and c-kit receptor (CD117) expression on CD34+ hematopoietic progenitor cells: a five dimensional flow cytometric study. Haematologia 1998;83:587-592.

  55. Dick JE. Absence of CD34 on some human SCID-repopulating cells. Ann NY Acad Sci 1999;872:211-217; discussion 217-219.[Medline]

  56. Donnelly DS, Zelterman D, Sharkis S et al. Functional activity of murine CD34+ and CD34- hematopoietic stem cell populations. Exp Hematol 1999;27:788-796.[Medline]

  57. Huss R. CD34- stem cells as the earliest precursors of hematopoietic progeny [letter]. Exp Hematol 1998;26:1022-1023.[Medline]

  58. Nakauchi H. Hematopoietic stem cells: are they CD34- positive or CD34-negative? [news; comment]. Nat Med 1998;4:1009-1010.[Medline]

  59. Nakauchi H, Takano H, Ema H et al. Further characterization of CD34-low/negative mouse hematopoietic stem cells. Ann NY Acad Sci 1999;872:57-66; discussion 66-70.[Medline]

  60. Osawa M, Hanada K, Hamada H et al. Long-term lymphohematopoietic reconstitution by a single CD34-low/negative hematopoietic stem cell. Science 1996;273:242-245.[Abstract]

  61. Zanjani ED, Almeida-Porada G, Livingston AG et al. Human bone marrow CD34- cells engraft in vivo and undergo multilineage expression that includes giving rise to CD34+ cells. Exp Hematol 1998;26:353-360.[Medline]

  62. Zanjani ED, Almeida-Porada G, Livingston AG et al. Engraftment and multilineage expression of human bone marrow CD34– cells in vivo. Ann NY Acad Sci 1999;872:220-231; discussion 231-232.[Medline]

accepted for publication August 13, 1999.



This article has been cited by other articles:


Home page
 BloodHome page
D. Baksh, J. E. Davies, and P. W. Zandstra
Soluble factor cross-talk between human bone marrow-derived hematopoietic and mesenchymal cells enhances in vitro CFU-F and CFU-O growth and reveals heterogeneity in the mesenchymal progenitor cell compartment
Blood, November 1, 2005; 106(9): 3012 - 3019.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Pathol.Home page
S. Lehtonen, E. Lehtonen, K. Kudlicka, H. Holthofer, and M. G. Farquhar
Nephrin Forms a Complex with Adherens Junction Proteins and CASK in Podocytes and in Madin-Darby Canine Kidney Cells Expressing Nephrin
Am. J. Pathol., September 1, 2004; 165(3): 923 - 936.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
M. G. Manz, T. Miyamoto, K. Akashi, and I. L. Weissman
Prospective isolation of human clonogenic common myeloid progenitors
PNAS, September 3, 2002; 99(18): 11872 - 11877.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
V. J. Weston, C. M. McConville, J. R. Mann, P. J. Darbyshire, S. Lawson, J. Gordon, P. A. H. Moss, A. Malcolm, R. Taylor, and T. Stankovic
Molecular Analysis of Single Colonies Reveals a Diverse Origin of Initial Clonal Proliferation in B-Precursor Acute Lymphoblastic Leukemia that Can Precede the t(12;21) Translocation
Cancer Res., December 1, 2001; 61(23): 8547 - 8553.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Reprints/Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Huang, S.
Right arrow Articles by Law, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Huang, S.
Right arrow Articles by Law, P.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
STEM CELLS THE ONCOLOGIST CME ALPHAMED PRESS JOURNALS
http://www.peprotech.com/