Comparative effects in vivo of recombinant murine interleukin 3, natural murine colony-stimulating factor-1, and recombinant murine granulocyte-macrophage colony-stimulating factor on myelopoiesis in mice.

Broxmeyer, Hal E.; Williams, Douglas E.; Cooper, Scott; Shadduck, RK; Gillis, Steven; Waheed, Abdül; Urdal, David L.; Bicknell, David

doi:10.1172/jci112877

Cited by 97 publications

(45 citation statements)

References 64 publications

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“…The percentage of progenitors in S phase was estimated by the high-specific-activity [ 3 H]thymidine kill technique, which eliminates cells in cycle from dividing in culture to form colonies. Briefly, the cells are exposed to a 30-s pulse of 50 Ci/ml (20 Ci/mmol) tritiated thymidine (PerkinElmer, Wellesley, MA) followed by a cold thymidine chase, prior to plating (9,34). Synergy assays with SCF and granulocyte-macrophage colonystimulating factor (GM-CSF) and chemokine inhibition assays were performed as described previously (26).…”

Section: Methodsmentioning

confidence: 99%

Aberrant Regulation of Hematopoiesis by T Cells in BAZF-Deficient Mice

Broxmeyer

Sehra

Cooper

et al. 2007

Molecular and Cellular Biology

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The BAZF (BCL-6b) protein is highly similar to the BCL-6 transcriptional repressor. While BCL-6 has been characterized extensively, relatively little is known about the normal function of BAZF. In order to understand the physiological role of BAZF, we created BAZF-deficient mice. Unlike BCL-6-deficient mice, BAZF-deficient mice are healthy and normal in size. However, BAZF-deficient mice have a hematopoietic progenitor phenotype that is almost identical to that of BCL-6-deficient mice. Compared to wild-type mice, both BAZF-deficient and BCL-6-deficient mice have greatly reduced numbers of cycling hematopoietic progenitor cells (HPC) in the BM and greatly increased numbers of cycling HPC in the spleen. In contrast to HPC from wild-type mice, HPC from BAZF-deficient and BCL-6-deficient mice are resistant to chemokine-induced myelosuppression and do not show a synergistic growth response to granulocyte-macrophage colony-stimulating factor plus stem cell factor. Depletion of CD8 T cells in BAZF-deficient mice reverses several of the hematopoietic defects in these mice. Since both BAZF- and BCL-6-deficient mice have defects in CD8 T-cell differentiation, we hypothesize that both BCL-6 and BAZF regulate HPC homeostasis by an indirect pathway involving CD8 T cells.

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Section: Methodsmentioning

confidence: 99%

Aberrant Regulation of Hematopoiesis by T Cells in BAZF-Deficient Mice

Broxmeyer

Sehra

Cooper

et al. 2007

Molecular and Cellular Biology

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“…After a 20-min incubation, thymidine (3-4 mg) in x M 0-mercaptoethanol, 30% FCS (HyClone), 1% BSA cold a-MEM was added (10,19,20). The cells were then washed (Sigma Chemicals, St. Louis, MO), 10% pokeweed-mitogentwice, resuspended, counted, and plated in clonogenic assays, as stimulated spleen cell-conditioned media, 3 U erythro~oietin, described below.…”

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confidence: 99%

In Vivo Effect of Interleukin-6 on Cycling Status of Hematopoietic Progenitors from Adults and Neonates

Liechty

Christensen

1990

Pediatr Res

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ABSTRACT. In vitro, IL-6 can induce hematopoietic pro-progenitors, are not completely known. Studies by Ikebuchi et genitors to progress from Go into cycle, but a role for IL-6 al. (12,13) and Leary et a/. (14) demonstrated that, in vitro, IL-in regulating cycling status of progenitors in vivo has not 6 is capable of inducing cycling of hematopoietic progenitors been established. In our studies, groups of five to six adult from adult mice and adult humans. Similarly, studies by our and newborn rats received i.v. injections of either IL-6 (1 group (1 5) demonstrated that IL-6 can accelerate cycling of fetal ng/g body wt) or the vehicle (control), after which cycling hematopoietic progenitors in vitro. However, the action of IL-6 of hematopoietic progenitors was evaluated by tritiated upon cycling status in vivo has not been evaluated. Thus, we thymidine suicide. Progenitors from adult rats injected with injected groups of adult and newborn rats with recombinant ILthe control had thymidine suicide rates of 7 2 1% (mean 6 or a vehicle control and assessed the cycling status of their f SEM), compared with 23 f 7% in the IL-6 recipients hematopoietic progenitors. . Before the injections, the tails were < 0.001; adults,p < 0.01). Thus, the effects of IL-6 in vivo prepared by washing with a 10% povidine-iodine solution, folin newborn and adult rats include cycle induction of he-lowed by a wash with 70% isopropyl alcohol. Four h after the matopoietic progenitors and release of neutrophils from injections, the animals were killed by CO2 inhalation, after which the storage pool into the circulation. (Pediatr Res 28: 323-blood was drawn from the inferior vena cava and both tibias 326,1990) were removed. Timed-pregnant Sprague-Dawley rats were allowed to deliver Abbreviations at the University of Utah Vivarium. Twenty-four to 48 h after delivery, groups of five to six pups, each weighing 5-8 g, received "tdr, tritiated thymidine i.v. injections (tail vein) of either IL-6 (1 ng/g body wt, in 4 pL/ PMN, polymorphonuclear cell g body wt), or the same volume of the vehicle used to suspend MEM, minimum essential medium the IL-6 (16). Four h after the injections, pups were killed by C 0 2 inhalation, after which blood was drawn from the internal jugular vein and the spleen and both femurs were removed.

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“…Colony assays for granulocytemacrophage (CFU-GM), erythroid (BFU-E), and multipotential (CFU-GEMM) progenitors were as described. 6,7 Megakaryocyte (CFU-Meg) progenitor assays were carried out using MegaCult-C medium (Stem Cell Technologies, Vancouver, BC, Canada). …”

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confidence: 99%

Selective enhancement of multipotential hematopoietic progenitors in vitro and in vivo by IL-20

Liu

Ding

Zeng

et al. 2003

Blood

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In a search for novel growth factors, we discovered that human interleukin-20 (IL-20) enhanced colony formation by CD34 ؉ multipotential progenitors. IL-20 had no effect on erythroid, granulocyte-macrophage, or megakaryocyte progenitors. IL- IntroductionCytokines are important regulators of the growth and development of hematopoietic cells 1,2 ; some are currently in clinical use. In a search for novel factors that regulate hematopoiesis, colony assays were used to screen novel secreted proteins identified through bioinformatics. One protein identified was identical to . 3,4 We demonstrate that IL-20 specifically enhances the proliferation of multipotential progenitors in vitro and in vivo without effect on more lineage-restricted progenitor cells. Study design Protein productionRecombinant human , containing a C-terminal FLAG tag (Eastman Kodak, Rochester, NY) followed by 6 histidine residues (Flis), was produced in 293EBNA1 cells. 5 We captured rhIL-20-Flis on Pharmacia Chelating Sepharose FF (Amersham-Pharmacia, Piscataway, NJ). The endotoxin level was less than 5.3 EU/mg rhIL-20. Colony assays for human and murine progenitorsCD34 ϩ human bone marrow or cord blood cells were purchased from BioWhittaker (Walkersville, MD). Colony assays for granulocytemacrophage (CFU-GM), erythroid (BFU-E), and multipotential (CFU-GEMM) progenitors were as described. 6,7 Megakaryocyte (CFU-Meg) progenitor assays were carried out using MegaCult-C medium (Stem Cell Technologies, Vancouver, BC, Canada). IL-20 transgenic miceTransgenic (TG) mice were generated by established techniques. 8 Human IL-20 was overexpressed in TG mice using apolipoprotein E gene promoter. IL-20 levels in mouse serum were determined using enzyme-linked immunosorbent assay (ELISA) with antihuman rhIL-20. rhIL-20 administration to normal miceFemale BDF1 mice (8-10 weeks of age; Harlan, Indianapolis, IN) were administered rhIL-20 (5 g/mouse) subcutaneously twice a day for 10 days. At day 11, mice were killed, bone marrow and spleen cells were counted, cells were used for colony assays, and the proportion of progenitors in S-phase of the cell cycle was estimated. 7,9,10 Results and discussion IL-20 in vitroIL-20 did not stimulate colony formation, but it increased the numbers of larger-sized colonies in combination with recombinant human stem cell factor (SCF) and erythropoietin (EPO) ( Figure 1Ai-ii). Colonies cultured with IL-20 contained cells with and without hemoglobin expression. Microscopic examination of 22 individual large colonies stained with Wright-Giemsa revealed mainly erythroblasts mixed with megakaryocytes. Granulocytes and monocytes were also detected (original magnification, ϫ 400) within these colonies but were less prominent than erythroblasts and megakaryocytes . This suggested that IL-20 enhanced CFU-GEMM.Colony assays were performed with human bone marrow (BM) and cord blood (CB) CD34 ϩ cells using various cytokine combinations. IL-20 (200 ng/mL), in combination with EPO and SCF, significantly enhanced BM CFU-GEMM numbers approximate...

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Comparative effects in vivo of recombinant murine interleukin 3, natural murine colony-stimulating factor-1, and recombinant murine granulocyte-macrophage colony-stimulating factor on myelopoiesis in mice.

Abstract: Purified murine colony-stimulating factors (CSF) recombinant interleukin 3 (IL-3), natural CSF-1, and recombinant granulocyte-macrophage (GM) CSF were assessed in vivo for their ef-

Cited by 97 publications

References 64 publications

Aberrant Regulation of Hematopoiesis by T Cells in BAZF-Deficient Mice

Aberrant Regulation of Hematopoiesis by T Cells in BAZF-Deficient Mice

In Vivo Effect of Interleukin-6 on Cycling Status of Hematopoietic Progenitors from Adults and Neonates

Selective enhancement of multipotential hematopoietic progenitors in vitro and in vivo by IL-20

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