Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UNIPROT:P04141 (granulocyte-macrophage colony-stimulating factor)
 6,790 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have recently demonstrated that 50% of primitive human long-term culture-initiating cells (LTC-IC) are maintained for up to 8 weeks in stroma-dependent cultures in which progenitor-stroma contact is prevented (stroma noncontact), or when progenitors are cultured in medium conditioned by stromal feeders. This indicates that factors responsible for LTC-IC maintenance are present in soluble form in stromal supernatant (SN). Although the picogram concentrations of cytokines present in stromal SN can induce the differentiation of CD34+/HLA-DR- (DR-) cells to clonogenic cells (colony forming cells; CFC), they maintain only 10% of LTC-IC for 5 weeks, suggesting that factors other than these cytokines are required for LTC-IC maintenance. To characterize the factor(s) in stromal SN responsible for LTC-IC maintenance, we purified glycoproteins and proteoglycans (PG) from the SN of the LTC-IC supportive murine marrow stromal fibroblast cell line M2-10B4 by ion exchange high performance liquid chromatography (HPLC). Culture of DR- cells in a combination of M2-10B4-derived PG, but not glycoproteins and picogram concentrations of recombinant human interleukin-6 (IL-6), granulocyte colony-stimulating factor (G-CSF), stem cell factor (SCF), leukemia inhibitory factor (LIF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and macrophage inflammatory protein-1alpha (MIP-1alpha) resulted in the recovery of 96% +/- 8% of LTC-IC maintained in cultures supplemented with unfractionated stromal SN. LTC-IC maintenance was largely retained after digestion of the PG-rich fraction with proteinase K and after dissociative gel filtration chromatography, but was completely abolished following treatment with nitrous acid, which digests heparan sulfate glycosaminoglycans (HS GAG). As for M2-10B4-derived HS GAG, high concentrations of bovine kidney HS GAG, but not bovine tracheal chondroitin sulfate, significantly improved cytokine-mediated LTC-IC maintenance. Maintenance of LTC-IC by these nonmarrow-derived HS GAG was, however, significantly lower than that seen with M2-10B4-derived HS. These studies demonstrate a role for marrow stroma-derived HS GAG in the long-term in vitro maintenance of human LTC-IC. Further structure-function analysis of these HS GAG may have important implications for ex vivo stem cell expansion and gene transfer into hematopoietic progenitors.
 ...
PMID:Stromal fibroblast heparan sulfate is required for cytokine-mediated ex vivo maintenance of human long-term culture-initiating cells. 860 38

Aim-To develop methods of messenger RNA (mRNA) in situ hybridisation (ISH) for use with routinely processed bone marrow trephine biopsy specimens, decalcified using formic acid, and long term cultures in order to demonstrate sites of synthesis of mRNA encoding monocyte colony stimulating factor (M-CSF).Methods-Biotinylated oligonucleotide probes, directed against target sequences within M-CSF mRNA, were hybridised with sections from bone marrow trephine biopsy specimens and detected using Streptavidin-biotin alkaline phosphatase complex formation. Validation of results included demonstration of total mRNA and unrelated mRNA species in adjacent sections, with appropriate negative controls. Minor technical modifications were required to perform ISH with long term bone marrow cultures.Results-M-CSF mRNA was demonstrated successfully in trephine biopsy specimens and long term cultures. Biopsy specimens varied in their requirement for predigestion with proteinase K and in the strength of the final reaction product, presumably due to variation in fixation. M-CSF mRNA was present in myelocytes and promonocytes. No stromal production of M-CSF mRNA was detected in biopsy specimens. ISH using long term bone marrow cultures confirmed production of M-CSF mRNA by developing monocytes and macrophages. Weak M-CSF mRNA expression was also seen in stromal fibroblasts.Conclusions-ISH can be performed successfully with formic acid decalcified bone marrow trephine biopsy specimens and long term cultures. The presence of M-CSF mRNA in myelomonocytic cells suggests that an autocrine mechanism contributes to monocyte differentiation. The absence of detectable M-CSF mRNA in biopsy stroma and its presence in stromal fibroblasts within bone marrow cultures probably reflects reduced sensitivity of ISH following tissue fixation and processing.
 ...
PMID:Sites of M-CSF messenger RNA production in bone marrow trephine biopsy specimens and long term cultures demonstrated by nonisotopic in situ hybridisation. 1669 73

The acute-phase protein serum amyloid A (SAA) is commonly considered a marker for inflammatory diseases; however, its precise role in inflammation and infection, which often result in neutrophilia, remains ambiguous. In this study, we demonstrate that SAA is a potent endogenous stimulator of granulocyte colony-stimulated factor (G-CSF), a principal cytokine-regulating granulocytosis. This effect of SAA is dependent on Toll-like receptor 2 (TLR2). Our data demonstrate that, in mouse macrophages, both G-CSF mRNA and protein were significantly increased after SAA stimulation. The induction of G-CSF was blocked by an anti-TLR2 antibody and markedly decreased in the TLR2-deficient macrophages. SAA stimulation results in the activation of nuclear factor-kappaB and binding activity to the CK-1 element of the G-CSF promoter region. In vitro reconstitution experiments also support that TLR2 mediates SAA-induced G-CSF expression. In addition, SAA-induced secretion of G-CSF was sensitive to heat and proteinase K treatment, yet insensitive to polymyxin B treatment, indicating that the induction is a direct effect of SAA. Finally, our in vivo studies confirmed that SAA treatment results in a significant increase in plasma G-CSF and neutrophilia, whereas these responses are ablated in G-CSF- or TLR2-deficient mice.
 ...
PMID:Serum amyloid A induces G-CSF expression and neutrophilia via Toll-like receptor 2. 1895 97