Production and functional properties of eosinophils from bone marrow cultures

1985 ◽  
Vol 74 (1) ◽  
pp. 207-217
Author(s):  
M. Strath ◽  
C.J. Sanderson
Keyword(s):  
Bone Marrow ◽  
Continuous Production ◽  
Specific Antigen ◽  
Pokeweed Mitogen ◽  
Normal Bone Marrow ◽  
Spleen Cells ◽  
Differentiation Antigen ◽  
Normal Bone ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

Bone marrow cultures have been established from mice infected with Mesocestoides corti and undergoing parasitic eosinophilia. In the absence of added conditioned medium, eosinophil differentiation ceases, and eosinophils are undetectable by 7 days, whereas neutrophil production continues over several weeks as with normal bone marrow. Eosinophil production can be induced by adding pokeweed mitogen-stimulated spleen supernatants (MSSS) or specific antigen-stimulated spleen supernatants (ASSS) produced from-spleen cells of M. corti-infected mice. In contrast to the continuous production of neutrophils, eosinophil production is transient, suggesting that there is no continued production of eosinophil progenitor cells in these cultures. More eosinophils are produced when MSSS is added at the initiation of cultures, compared to after a delay of 2 weeks, and establishing the cultures at 33 degrees C does not appear to enhance eosinophil production. The eosinophils produced are shown to express the eosinophil differentiation antigen defined by monoclonal antibody NIMP-R13, they produce eosinophil peroxidase in similar amounts to eosinophils taken from mice. They show normal phagocytic activity of antibody-coated erythrocytes and lyse red cells coated with antibodies of IgG1, IgG2a, IgG2b, but not IgM isotypes.

scholarly journals Biologic significance of constitutive and subliminal growth factor production by bone marrow stroma

Blood ◽  
1992 ◽  
Vol 79 (12) ◽  
pp. 3168-3178 ◽  
Author(s):  
EL Kittler ◽  
H McGrath ◽  
D Temeles ◽  
RB Crittenden ◽  
VK Kister ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Growth Factor ◽  
Stromal Cells ◽  
Pokeweed Mitogen ◽  
Gm Csf ◽  
Bone Marrow Cultures ◽  
Growth Factor Production ◽  
Constitutive Production ◽  
Marrow Cultures

Abstract The “stromal” or adherent cells of long-term murine Dexter explant bone marrow cultures provide the best in vitro model of the bone marrow microenvironment. Colony-stimulating factor-1 (CSF-1) is produced constitutively by these cells and is easily detected, but most investigators have not found constitutive production of the other hemolymphopoietic cytokines. We have previously reported the detection of granulocyte-macrophage-CSF (GM-CSF) in murine stromal cultures and its induction by the lectin Pokeweed mitogen. The present studies analyzing stromal cytokine messenger RNA (mRNA) production by standard Northern blot analysis show constitutive production of mRNAs for CSF-1, GM-CSF, granulocyte-CSF (G-CSF), c-kit ligand (KL), and interleukin-6 (IL-6), but not IL-3, IL-4, or IL-5 by 3-week irradiated or nonirradiated murine Dexter stromal cells. Exposure of stromal cells to Pokeweed mitogen or IL-1 16 hours before RNA harvest induces the messages for GM-CSF, G-CSF, KL, and IL-6, but not IL-3, IL-4, IL-5, or CSF-1. Polymerase chain reaction amplification of cDNA made with reverse transcriptase from stromal RNA using two separate sets of IL-3- specific primers shows the presence of IL-3 message in irradiated stromal cells, which is only detectable with this more sensitive technique. The factor-dependent cell lines FDC-P1 and 32D are supported by the stromal cells without the addition of exogenous growth factors, demonstrating a cytokine activity in these cultures that is inhibited by the addition of anti-IL-3 or anti-GM-CSF antibodies. These data indicate that murine Dexter stromal cells constitutively produce CSF-1, GM-CSF, G-CSF, IL-6, KL, and IL-3. This growth factor production could explain the support of granulocyte, macrophage, and megakaryocyte production and stem cell maintenance in Dexter-type long-term murine bone marrow cultures.

scholarly journals Quantitative Assessment of Myeloid Nuclear Differentiation Antigen Distinguishes Myelodysplastic Syndrome From Normal Bone Marrow

2011 ◽  
Vol 135 (3) ◽  
pp. 380-385 ◽  
Author(s):  
Sara A. McClintock-Treep ◽  
Robert C. Briggs ◽  
Keith E. Shults ◽  
Leanne A. Flye-Blakemore ◽  
Claudio A. Mosse ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Myelodysplastic Syndrome ◽  
Quantitative Assessment ◽  
Normal Bone Marrow ◽  
Differentiation Antigen ◽  
Normal Bone ◽  
Nuclear Differentiation

scholarly journals Biologic significance of constitutive and subliminal growth factor production by bone marrow stroma

Blood ◽  
1992 ◽  
Vol 79 (12) ◽  
pp. 3168-3178 ◽  
Author(s):  
EL Kittler ◽  
H McGrath ◽  
D Temeles ◽  
RB Crittenden ◽  
VK Kister ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Growth Factor ◽  
Stromal Cells ◽  
Pokeweed Mitogen ◽  
Gm Csf ◽  
Bone Marrow Cultures ◽  
Growth Factor Production ◽  
Constitutive Production ◽  
Marrow Cultures

The “stromal” or adherent cells of long-term murine Dexter explant bone marrow cultures provide the best in vitro model of the bone marrow microenvironment. Colony-stimulating factor-1 (CSF-1) is produced constitutively by these cells and is easily detected, but most investigators have not found constitutive production of the other hemolymphopoietic cytokines. We have previously reported the detection of granulocyte-macrophage-CSF (GM-CSF) in murine stromal cultures and its induction by the lectin Pokeweed mitogen. The present studies analyzing stromal cytokine messenger RNA (mRNA) production by standard Northern blot analysis show constitutive production of mRNAs for CSF-1, GM-CSF, granulocyte-CSF (G-CSF), c-kit ligand (KL), and interleukin-6 (IL-6), but not IL-3, IL-4, or IL-5 by 3-week irradiated or nonirradiated murine Dexter stromal cells. Exposure of stromal cells to Pokeweed mitogen or IL-1 16 hours before RNA harvest induces the messages for GM-CSF, G-CSF, KL, and IL-6, but not IL-3, IL-4, IL-5, or CSF-1. Polymerase chain reaction amplification of cDNA made with reverse transcriptase from stromal RNA using two separate sets of IL-3- specific primers shows the presence of IL-3 message in irradiated stromal cells, which is only detectable with this more sensitive technique. The factor-dependent cell lines FDC-P1 and 32D are supported by the stromal cells without the addition of exogenous growth factors, demonstrating a cytokine activity in these cultures that is inhibited by the addition of anti-IL-3 or anti-GM-CSF antibodies. These data indicate that murine Dexter stromal cells constitutively produce CSF-1, GM-CSF, G-CSF, IL-6, KL, and IL-3. This growth factor production could explain the support of granulocyte, macrophage, and megakaryocyte production and stem cell maintenance in Dexter-type long-term murine bone marrow cultures.

scholarly journals THYMUS-MARROW IMMUNOCOMPETENCE

1971 ◽  
Vol 133 (5) ◽  
pp. 1026-1034 ◽  
Author(s):  
J. John Cohen ◽  
Henry N. Claman
Keyword(s):  
Bone Marrow ◽  
Proliferative Response ◽  
Normal Bone Marrow ◽  
Hydrocortisone Acetate ◽  
Spleen Cells ◽  
Normal Bone ◽  
Helper Cells ◽  
Sensitive Phase ◽  
Thymus Cells ◽  
Number Of Cells

Corticosteroids suppress the humoral antibody response of mice to sheep erythrocytes. This response depends on interactions between thymus-derived helper cells and bone marrow-derived antibody-forming cell precursors (AFC precursors). Previous experiments had shown that spleen cells (a mixture of thymus-derived and marrow-derived cells) were sensitive to corticosteroids while AFC precursors in the bone marrow were resistant. The present experiments showed that the thymus of a mouse given 2.5 mg of hydrocortisone acetate, although containing only about 5% of the number of cells of a normal thymus, was as effective as a normal thymus in cooperating with bone marrow when transferred to irradiated syngeneic mice and stimulated with SRBC. The proliferative response of thymus helper cells to SRBC was also resistant to hydrocortisone. In this context, the majority of thymic cells are in the cortex, are rapidly dividing, are sensitive to corticosteroids and are not iminunocompetent. A small number of thymic cells, probably located in the medulla, are resistant to corticosteroids, but are immunocompetent since they can serve as helper cells. The hydrocortisone-sensitive phase of the splenic response to SRBC was found to be the bone marrow-derived AFC precursor since spleens from hydrocortisone-treated donors had immunocompetence restored by normal bone marrow but not by normal thymus cells.

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