Current Search: B cells (x)
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Title
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Detection and purification of lupus B cells.
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Creator
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Saccocio, Seth., Florida Atlantic University, Hartmann, James X.
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Abstract/Description
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This study developed a method for positive selection of anti-single stranded (ss)DNA B cells from peripheral blood mononuclear cells (PBMCs) using a twenty nucleotide oligomer of deoxythymidine (oligo(dT) 20mer), coupled to microscopie magnetic beads. Oligo(dT) 20mer specificity for plasma anti-ssDNA antibody (Ab) was established through indirect enzyme linked immunosorbent assay (ELISA) and flow cytometry. A novel method for semi-quantitative detection of ssDNA specific, B cells from PBMC...
Show moreThis study developed a method for positive selection of anti-single stranded (ss)DNA B cells from peripheral blood mononuclear cells (PBMCs) using a twenty nucleotide oligomer of deoxythymidine (oligo(dT) 20mer), coupled to microscopie magnetic beads. Oligo(dT) 20mer specificity for plasma anti-ssDNA antibody (Ab) was established through indirect enzyme linked immunosorbent assay (ELISA) and flow cytometry. A novel method for semi-quantitative detection of ssDNA specific, B cells from PBMC purified peripheral blood that utilized flow cytometric analysis of oligo(dT) 20mer Texas Red labeled cells was developed. Qualitative purification of ssDNA specific, B cells using oligo(dT) coupled magnetic beads was determined through light microscopy and flow cytometric analysis of positive sclected cell populations. Cross-reaetivity of oligo(dT) 20mer with receptors distinct from membrane Ab, resulted in the use of oligo(dC) 20mer as a useful blocking agent. Results show anti-ssDNA Ab titer does not correlate with numbers of peripheral blood ssDNA specific, B cells.
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Date Issued
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2003
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PURL
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http://purl.flvc.org/fcla/dt/13086
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Subject Headings
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Systemic lupus erythematosus, B cells, DNA antibodies
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Format
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Document (PDF)
