Flow Cytometry Analysis of Murine B Cell Lymphoma Differentiation

Warner, Noel L.; Daley, Michael J.; Richey, Judith; Spellman, Craig W.

doi:10.1111/j.1600-065x.1979.tb00304.x

Cited by 100 publications

(35 citation statements)

References 72 publications

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“…Cell lines ABE-8.1/2 (Burchiel and Warner, 1980) and WEHI-279 (Warner et al, 1979) cells were grown in Dulbecco's modified Eagle's medium (DMEM) (Mediatech), 10% FCS, 0.584 g/L L-Gln, and 50 ,uM 2-mercaptoethanol. 2PK-3 (Warner et al, 1979) were grown in the same media supplemented with 4.5 g/L glucose.…”

Section: Isolation Of Total Rna From Mouse Tissues and Cultured Mousementioning

confidence: 99%

Members of the syndecan family of heparan sulfate proteoglycans are expressed in distinct cell-, tissue-, and development-specific patterns.

Kim

Goldberger

Gallo

et al. 1994

MBoC

383

294

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The syndecans are a gene family of four transmembrane heparan sulfate proteoglycans that bind, via their HS chains, diverse components of the cellular microenvironment. To evaluate the expression of the individual syndecans, we prepared cDNA probes to compare mRNA levels in various adult mouse tissues and cultured mouse cells representing various epithelial, fibroblastic, endothelial, and neural cell types and B cells at various stages of differentiation. We also prepared antibody probes to assess whether the extracellular domains of the individual syndecans are shed into the conditioned media of cultured cells. Our results show that all cells and tissues studied, except B-stem cells, express at least one syndecan family member; most cells and tissues express multiple syndecans. However, each syndecan family member is expressed selectively in cell-, tissue-, and developmentspecific patterns. The extracellular domain of all syndecan family members is shed as an intact proteoglycan. Thus, most, if not all, cells acquire a distinctive repertoire of the four syndecan family members as they differentiate, resulting in selective patterns of expression that likely reflect distinct functions.

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Section: Isolation Of Total Rna From Mouse Tissues and Cultured Mousementioning

confidence: 99%

Members of the syndecan family of heparan sulfate proteoglycans are expressed in distinct cell-, tissue-, and development-specific patterns.

Kim

Goldberger

Gallo

et al. 1994

MBoC

383

294

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“…WEHI-231 (19) and Bal 17 (ref. 20; our isolate does not express mIgD) cells were resuspended at 5 x 106 cells per ml in growth medium containing 20 mM Hepes.…”

Section: Methodsmentioning

confidence: 99%

Tyrosine phosphorylation of components of the B-cell antigen receptors following receptor crosslinking.

Gold

Matsuuchi

Kelly

et al. 1991

Proc. Natl. Acad. Sci. U.S.A.

206

114

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Crosslinking membrane immunoglobulin (mIg), the B-cell antigen receptor, stimulates tyrosine phosphorylation of a number of proteins. Since many receptors are phosphorylated after ligand binding, we asked ifcomponents of the mIg receptor complexes were tyrosine-phosphorylated after mIg crosslinking. Both mIgM and mIgD are noncovalently associated with at least two other proteins. mIgM Is associated with the MB-1 protein, which is disulfide-linked to a protein designated Ig-f3. mIgD is not associated with MB-1 but is with IgD-a, which is also disulfide-linked to Ig-fi. Using immunoprecipitation with a specific anti-MB-i antiserum followed by anti-phosphotyrosine immunoblotting, we found that crosslinking mIgM stimulated tyrosine phosphorylation of (5), were resuspended at 16.7 x 106 cells per ml in a modified Hepes-buffered saline solution (9). Cells were stimulated at 37°C, washed, and resuspended at 108 cells per ml in lysis buffer [20 mM Tris'HCI, pH 8/137 mM NaCl/10% (wt/vol) glycerol/1% Triton X-100/1 mM Na3VO4/2 mM EDTA/1 mM phenylmethylsulfonyl fluoride/20 ,uM leupeptin/0.15 unit of aprotinin per ml] as described (5). Detergent-insoluble material was removed by centrifugation, and protein concentrations were determined by the BCA (bicinchoninic acid) protein assay (Pierce).Anti-MB-i Antiserum. A 34-amino acid peptide from the putative cytoplasmic domain of the murine MB-1 protein [amino acids 187-220 (21) were collected on protein A-Sepharose for 1 hr. The beads were suspended in 0.5 ml of lysis buffer containing 50 ,uM Na3VO4, 0.4% deoxycholate, and 0.3% SDS (wash buffer), layered over 0.4 ml of 30%o sucrose in 0.5 x wash buffer, and centrifuged 5 min at 10,000 x g. The beads were then washed once each with wash buffer and 50 ,uM Na3VO4. Proteins were eluted by boiling in nonreducing SDS/PAGE sample Abbreviations: mlg, membrane immunoglobulin; anti-Tyr(P), antiphosphotyrosine.tTo whom reprint requests should be addressed.tThe first two authors contributed equally to the experiments in this paper. 3436The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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“…In any scheme of the sequential acquisition of surface antigens with differentiation, the influence of quantitative variation in the amount of antigen expressed and the lower limit of detection by the methods employed must be considered (16,17). The flow cytometer is the ideal instrument with which to quantitate monoclonal antibody binding to specific surface markers expressed by leukemic cells in concert with other important cellular features.…”

Section: Resultsmentioning

confidence: 99%

“…We reasoned from studies with other differentiation antigens (16)(17)(18)(19)(20) that quantitation of CALLA on leukemic blasts might disclose substantial differences in its expression, which could be related to the predicted pattern of differentiation within the B precursor pathway (15). Additionally, since about one-third…”

Section: Introductionmentioning

confidence: 99%

Quantitative variation of the common acute lymphoblastic leukemia antigen (gp100) on leukemic marrow blasts.

Look¹,

Melvin²,

Brown³

et al. 1984

J. Clin. Invest.

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Flow Cytometry Analysis of Murine B Cell Lymphoma Differentiation

Cited by 100 publications

References 72 publications

Members of the syndecan family of heparan sulfate proteoglycans are expressed in distinct cell-, tissue-, and development-specific patterns.

Members of the syndecan family of heparan sulfate proteoglycans are expressed in distinct cell-, tissue-, and development-specific patterns.

Tyrosine phosphorylation of components of the B-cell antigen receptors following receptor crosslinking.

Quantitative variation of the common acute lymphoblastic leukemia antigen (gp100) on leukemic marrow blasts.

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