The human granulocyte-macrophage colony stimulation factor (hGM-CSF) gene was cloned in the pET 23a( +) expression vector under the control of strong bacteriophage T7 transcription and translation signals. The hGM-CSF gene was transferred into E. coli strainBL21 (DE3)pLysS andIPTG was used for induction of GM-CSF gene. Production of the target protein was obtained as revealed by ELISA and ...

background: immunotoxins are comprised of both the cell targeting and the cell killing moieties. we previously established a new immunotoxin, i.e. shiga toxin granulocyte macrophage-colony stimulating factor (stxa1-gm-csf), comprises of catalytic domain of stx, as a killing moiety and gm-csf, as a cell targeting moiety. in this study, the ability of the immunotoxin to induce apoptosis and doubl...

Although erythropoietin (EPO) and its receptor (EPOR) are crucial for the proliferation, survival, and terminal differentiation of erythroid progenitors, it remains to be elucidated whether EPOR-unique signaling is required for erythropoiesis. To address this issue, human granulocyte-macrophage colony-stimulating factor (hGM-CSF) receptor (hGMR)-transgenic mice and heterozygous EPOR mutant mice...

Thymocytes show differential cytokine responses, depending on the stage of differentiation. Whether these responses are due to preferential cytokine receptor expression or due to downstream signaling mechanisms is unknown. In this study, we examined the relationship between receptor expression and T-cell proliferation or differentiation using thymocytes from transgenic mice constitutively expre...

To increase transgenic production of granulocyte-macrophage colony-stimulating factor (GM-CSF), we mutated the mRNA's 3'-untranslated region, AUUUA instability elements. Expression vectors containing human or murine GM-CSF cDNAs coding for wild-type (GM-AUUUA) or mutant versions with reiterated AUGUA repeats (GM-AUGUA) were transfected into cells in culture or animals using particle-mediated ge...

We evaluated the effects of ectopic granulocyte/macrophage colony-stimulating factor (GM-CSF) signals on hematopoietic commitment and differentiation. Lineage-restricted progenitors purified from mice with the ubiquitous transgenic human GM-CSF receptor (hGM-CSFR) were used for the analysis. In cultures with hGM-CSF alone, hGM-CSFR-expressing (hGM-CSFR+) granulocyte/monocyte progenitors (GMPs) ...

A panel of monoclonal antibodies was raised against the low-affinity human granulocyte-macrophage colony-stimulating factor (hGM-CSF) receptor a-chain expressed as recombinant protein on murine FDC-P1 cells. All the selected antibodies were of the lgG2A isotype and bound to protein A. They each recognized both native and recombinant receptors by indirect surface immunofluorescence and by immuno...

Acute myeloid leukemia colony forming cells (AML-CFU) require the addition of colony stimulating factors (CSFs) for in vitro proliferation. Recently, we isolated a human recombinant multilineage CSF (hMulti-CSF). We investigated the ability of hMulti-CSF to stimulate AML clonogenic cells in seven patients in direct comparison with the effects of human granulocyte CSF (hG-CSF), human granulocyte...