Establishment of multipotential and antigen presenting cell lines derived from myeloid leukemias in GM-CSF transgenic mice

Rasko, John E.J.; Metcalf, Donald; Alexander, B. J.; Strasser, Andreas; Begley, C. Glenn

doi:10.1038/sj.leu.2400614

Cited by 10 publications

(5 citation statements)

References 10 publications

(10 reference statements)

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“…As shown for seven of the lines in the lower panel of Figure 2, every line was resistant to growth inhibition, comparable to the deleted RB22.2 subclones, and several of the lines were even stimulated by the treatment. Similarly, we confirmed the observation of Rasko et al 30 that growth of the chr2-deleted tumor cell line IGM36 is stimulated by LPS (data not shown).…”

Section: Lps-resistant Phenotype Is Present In Radiationinduced Leukesupporting

confidence: 80%

Phenotypic effect correlating with loss of a novel tumor suppressor gene: towards cloning by complementation

Cook

Alexander

1998

Leukemia

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We intend to use a gene complementation approach to clone a tumor suppressor gene on mouse chromosome 2, the loss of which contributes to myeloid leukemia. An in vitro model system has been generated using a clonal cell line, in which tumorigenic chromosomal lesions have been selected along with myeloid differentiation. Among these lesions are deletions of chromosome 2. Comparison of subclones with deleted vs intact chromosomes 2 has allowed the identification of a growth related phenotypic pattern which correlates with the deletion, viz the retention of a marker of immature cells, resistance to inhibition by lipopolysaccharide (LPS), even in the presence of markers of mature myeloid cells, such as resistance to killing by apoptosis-inducing agents. The phenotype is shared by chromosome 2-deleted cell lines derived from conventional tumors. We have begun to investigate the mechanism of the phenotype. The LPS resistance does not correlate with lack of mRNA for CD14, a known cell surface receptor for this agent, or with failure to induce TNF␣ or nitric oxide synthase in response to its binding. The system should allow cloning of the gene using complementation of this phenotype in transfected cell lines.

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Section: Lps-resistant Phenotype Is Present In Radiationinduced Leukesupporting

confidence: 80%

Phenotypic effect correlating with loss of a novel tumor suppressor gene: towards cloning by complementation

Cook

Alexander

1998

Leukemia

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“…This question has only been posed for GM-CSF. While lifelong excess GM-CSF levels in transgenic mice were not leukaemogenic 94 , these mice were more susceptible to leukaemic transformation by the Moloney leukaemia virus 95 . Repopulation of mice for a period of a few months with marrow cells engineered to produce excess levels of G-CSF or IL3 did not lead to leukaemia development 87,88 .…”

Section: Csfs and Myeloid Leukaemiamentioning

confidence: 90%

The colony-stimulating factors and cancer

Metcalf¹

2010

Nat Rev Cancer

136

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The four colony stimulating factors (CSFs) are glycoproteins regulating the generation and some functions of infection-protective granulocytes and macrophages. Recombinant G-CSF and GM-CSF have now been used to elevate dangerously low white blood cell levels in many millions of cancer patients following chemotherapy. These CSFs also release haematopoietic stem cells to the peripheral blood (PBSC) and these cells have now largely replaced bone marrow as more effective populations for transplantation to cancer patients with treatment-induced bone marrow damage.

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“…Among the cytokines used, SCF was used to promote hHSC proliferation and prevent apoptosis 38,39 , and IL-3 aided stem cell expansion 40 . Flt3-L [41][42][43][44] along with G-CSF 45,46 promoted colony formation and neutrophil differentiation. Other chemokines that we tested like TPO and GM-CSF reduced the formation of multi-lobed nucleated cells.…”

Section: Discussionmentioning

confidence: 96%

Knockout studies using CD34+ hematopoietic cells suggest that CD44 is a physiological human neutrophil E-selectin ligand

Zhu,

Neelamegham

2023

Preprint

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The recruitment of peripheral blood neutrophils at sites of inflammation involves a multistep cascade, starting with E- and P-selectin expressed on the inflamed vascular endothelium binding sialofucosylated glycans on leukocytes. As the glycoconjugate biosynthesis pathways in different cells are distinct, the precise carbohydrate ligands of selectins varies both across species, and between different immune cell populations in a given species. To study this aspect in human neutrophils, we developed a protocol to perform CRISPR/Cas9 gene-editing on CD34+ hHSCs (human hematopoietic stem/progenitor cells) as they are differentiated towards neutrophil lineage. This protocol initially uses a cocktail of SCF (stem-cell factor), IL-3 (interleukin-3) and FLT-3L (FMS-like tyrosine kinase 3 ligand) to expand the stem/progenitor cells followed by directed differentiation to neutrophils using G-CSF (granulocyte colony-stimulating factor). Microfluidics based assays were performed on a confocal microscope platform to characterize the rolling phenotype of each edited cell type in mixed populations. These studies demonstrated that CD44, but not CD43, is a major E-selectin ligand on human neutrophils. The loss of function results were validated by developing sialofucosylated recombinant CD44. This glycosylated protein supported both robust E-selectin binding in a cell-free assay, and it competitively blocked neutrophil adhesion to E-selectin on inflamed endothelial cells. Together, the study establishes important methods to study human neutrophil biology and determines that sialoflucosylated-CD44 is a physiological human E-selectin ligand.

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Establishment of multipotential and antigen presenting cell lines derived from myeloid leukemias in GM-CSF transgenic mice

Cited by 10 publications

References 10 publications

Phenotypic effect correlating with loss of a novel tumor suppressor gene: towards cloning by complementation

Phenotypic effect correlating with loss of a novel tumor suppressor gene: towards cloning by complementation

The colony-stimulating factors and cancer

Knockout studies using CD34+ hematopoietic cells suggest that CD44 is a physiological human neutrophil E-selectin ligand

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