Skip to main page content
U.S. flag

An official website of the United States government

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2003 Feb 4;100(3):1244-9.
doi: 10.1073/pnas.252756999. Epub 2003 Jan 27.

Crowding-dependent production of colony-stimulating factors by cultured syngeneic or allogeneic hematopoietic cells

Affiliations

Crowding-dependent production of colony-stimulating factors by cultured syngeneic or allogeneic hematopoietic cells

Donald Metcalf et al. Proc Natl Acad Sci U S A. .

Abstract

Mitogenic stimulation in vitro of mouse T lymphocytes induces the production of the hematopoietic cytokines granulocyte-macrophage colony-stimulating factor and IL-3. The present experiments showed that simple crowding of murine spleen or lymph node cells was a sufficient inducing stimulus. Crowding did not have this consequence for thymus or marrow cells or spleen cells from nu/nu or Rag-1-/- mice lacking T lymphocytes. Crowding for as short a period as 24 h was sufficient to allow subsequent cytokine production in sparse cultures. Purified T lymphocytes also exhibited low levels of crowding induction of cytokine production and cytokine production was enhanced by IL-2 and IFN-gamma. However, IFN-gamma-/- spleen cells exhibited similar crowding-induced colony-stimulating factor production to that of control spleen cells. Excess cell crowding inhibited cytokine production. This inhibition was not caused by receptor internalization of cytokines but may contribute to the failure to observe IL-3 production in lymphoid organs in vivo. Coculture of allogeneic spleen or peritoneal cells was a strong inducing signal for colony-stimulating factor production but this parameter was unable to detect autoreactivity of T lymphocytes in mice that lack suppressor of cytokine signaling 1 and exhibit T lymphocyte activation.

PubMed Disclaimer

Figures

Figure 1
Figure 1
(A) Concentration-dependent rise and fall of GM-CSF and IL-3 in cultures of C57BL spleen cells with or without 2-mercaptoethanol (2ME). Means ± standard deviations from four replicate cultures are shown. (B) Concentration-dependent rise and fall in GM-CSF and IL-3 in cultures of βc−/− spleen cells lacking GM-CSF receptors versus control βc+/+ spleen cells. Mean values of duplicate cultures are shown. (C) Failure of GM-CSF production in cultures of nu/nu or Rag-1−/− spleen cells lacking T lymphocytes and relative inactivity of spleen cell cultures from NOD/SCID mice. Mean values of duplicate cultures are shown.
Figure 2
Figure 2
Failure of C57BL spleen cells to produce GM-CSF or IL-3 at 0.5 or 1.0 × 106 cells per ml but activity of cells at these concentrations after initial incubation for 24 h at a concentration of 5 × 106 cells per ml. Each column represents assays from an independent culture.
Figure 3
Figure 3
Increased production of GM-CSF and IL-3 in cocultures of 1 × 106 C57BL and 1 × 106 BALB/c spleen cells versus cultures of 2 × 106 C57BL or BALB/c cells. Shown are mean values of duplicate cultures.

Similar articles

References

    1. Parker J W, Metcalf D. J Immunol. 1974;112:502–510. - PubMed
    1. Cline M J, Golde D W. Nature. 1974;248:703–704. - PubMed
    1. Staber F G, Hültner L, Marcucci F, Krammer P H. Nature. 1982;298:79–82. - PubMed
    1. Kelso A, Metcalf D. Adv Immunol. 1990;48:69–105. - PubMed
    1. Schreier M H, Iscove N N. Nature. 1980;287:228–230. - PubMed

Publication types

MeSH terms

Substances