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Detection and purification of lupus B cells

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Date Issued:
2003
Summary:
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 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.
Title: Detection and purification of lupus B cells.
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Name(s): Saccocio, Seth.
Florida Atlantic University, Degree grantor
Hartmann, James X., Thesis advisor
Type of Resource: text
Genre: Electronic Thesis Or Dissertation
Issuance: monographic
Date Issued: 2003
Publisher: Florida Atlantic University
Place of Publication: Boca Raton, Fla.
Physical Form: application/pdf
Extent: 76 p.
Language(s): English
Summary: 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 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.
Identifier: 9780496219247 (isbn), 13086 (digitool), FADT13086 (IID), fau:9950 (fedora)
Collection: FAU Electronic Theses and Dissertations Collection
Note(s): Thesis (M.S.)--Florida Atlantic University, 2003.
Charles E. Schmidt College of Science
Subject(s): Systemic lupus erythematosus
B cells
DNA antibodies
Held by: Florida Atlantic University Libraries
Persistent Link to This Record: http://purl.flvc.org/fcla/dt/13086
Sublocation: Digital Library
Use and Reproduction: Copyright © is held by the author with permission granted to Florida Atlantic University to digitize, archive and distribute this item for non-profit research and educational purposes. Any reuse of this item in excess of fair use or other copyright exemptions requires permission of the copyright holder.
Use and Reproduction: http://rightsstatements.org/vocab/InC/1.0/
Host Institution: FAU
Is Part of Series: Florida Atlantic University Digital Library Collections.