Human granulocyte-macrophage colony-stimulating factor (hGM-CSF)–dependent in vitro and in vivo proliferation and differentiation of all hematopoietic.

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Human granulocyte-macrophage colony-stimulating factor (hGM-CSF)–dependent in vitro and in vivo proliferation and differentiation of all hematopoietic progenitor cells in hGM-CSF receptor transgenic mice Ichiko Nishijima, MSc, Sumiko Watanabe, PhD, Tatsutoshi Nakahata, MD, PhD, Ken-ichi Arai, MD, PhD Journal of Allergy and Clinical Immunology Volume 100, Issue 6, Pages S79-S86 (December 1997) DOI: 10.1016/S0091-6749(97)70011-3 Copyright © 1997 Mosby, Inc. Terms and Conditions

FIG. 1 Colony-forming assay of bone marrow cells from transgenic mice and normal mice. The experiments not described in this figure showed that the effects of mouse GM-CSF and mouse GM-CSF plus erythropoietin on colony numbers and types among normal mice were the same as those among transgenic mice. G, granulocyte colonies, M, macrophage colonies; GM, granulocyte-macrophage colonies; Eo, eosinophil colonies; MK, megakaryocyte colonies; Mast, mast cell colonies; CFU-E, erythrocyte colonies; BFU-E, erythroid burst-forming units; Bl, blast cell colonies; Mix, mixed hematopoietic colonies; Epo, erythropoietin. Journal of Allergy and Clinical Immunology 1997 100, S79-S86DOI: (10.1016/S0091-6749(97)70011-3) Copyright © 1997 Mosby, Inc. Terms and Conditions

FIG. 2 Erythroid colony formation by bone marrow cells from transgenic mice in the absence of erythropoietin (Epo). A, Photomicrograph of BFU-E colonies derived from bone marrow cells of transgenic mice in the presence of hGM-CSF. B, Addition of anti-erythropoietin antibody to cultures with hGM-CSF did not affect the number of CFU-E colonies, whereas the same antibody completely suppressed formation of erythroid colonies induced by IL-3 and erythropoietin. Journal of Allergy and Clinical Immunology 1997 100, S79-S86DOI: (10.1016/S0091-6749(97)70011-3) Copyright © 1997 Mosby, Inc. Terms and Conditions

FIG. 3 RT-PCR analysis of the expression of erythroid-specific transcription factors in BFU-E colonies. Each BFU-E colony was picked up with a micropipette from cultures with hGM-CSF or IL-3 plus erythropoietin. The lane marked with a minus sign is the PCR product obtained with mock complementary DNA (no reverse transcriptase in the complementary DNA synthesis reaction). Journal of Allergy and Clinical Immunology 1997 100, S79-S86DOI: (10.1016/S0091-6749(97)70011-3) Copyright © 1997 Mosby, Inc. Terms and Conditions

FIG. 4 Dose-response effects of hGM-CSF on the number of constituent cells (A) and hematopoietic progenitors (B) in spleens of transgenic mice as assayed by means of methylcellulose clonal culture. Treatment with hGM-CSF increased the number of both immature and mature erythroid cells. (From Nishijima I et al. Blood 1997;90:1031-8.) See Fig. 1 for abbreviations. Journal of Allergy and Clinical Immunology 1997 100, S79-S86DOI: (10.1016/S0091-6749(97)70011-3) Copyright © 1997 Mosby, Inc. Terms and Conditions

FIG. 5 Findings from flow cytometric analysis of cells from peripheral blood, bone marrow, spleen, and liver of normal littermates (left) and transgenic mice (right) treated with 500 ng hGM-CSF. Cells were stained with fluorescein isothiocyanate (FITC)–labeled anti-CD3 and phycoerythrin (PE)–labeled anti-NK1.1. Number of CD3- NK1.1+ cells increased in peripheral blood, bone marrow, spleen, and liver of transgenic mice treated with hGM-CSF. (From Nishijima I et al. Blood 1997;90:1031-8.) Journal of Allergy and Clinical Immunology 1997 100, S79-S86DOI: (10.1016/S0091-6749(97)70011-3) Copyright © 1997 Mosby, Inc. Terms and Conditions

FIG. 6 Cytotoxity of spleen mononuclear cells from transgenic mice induced by hGM-CSF. Target cells used in the cytotoxity assay were NK-resistant P815 cells and NK-sensitive YAC-1 cells. The cytotoxity of peripheral NK cells was induced by hGM-CSF. Journal of Allergy and Clinical Immunology 1997 100, S79-S86DOI: (10.1016/S0091-6749(97)70011-3) Copyright © 1997 Mosby, Inc. Terms and Conditions

FIG. 7 Two models that illustrate the possible relation between receptor expression and differentiation of hematopoietic cells. The results of in vitro and in vivo studies indicate that the differentiation of hematopoietic progenitor cells is not induced by exogenous cytokine stimulation (instruction model) but rather is determined by the intrinsic cell program with cytokines simply selecting cells that express the appropriate receptor (selection model). HSC, hematopoietic stem cell; Tg, transgenic. Journal of Allergy and Clinical Immunology 1997 100, S79-S86DOI: (10.1016/S0091-6749(97)70011-3) Copyright © 1997 Mosby, Inc. Terms and Conditions