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The role of bacterial lipopolysaccharide in the production of the lupus B cell phenotype
Title
The role of bacterial lipopolysaccharide in the production of the lupus B cell phenotype.
Creator
Nikolic, Veljko., Florida Atlantic University, Hartmann, James X.
Abstract/Description
The purpose of this study was to determine the effect of bacterial lipopolysaccharide (LPS) on the expression of RP105 (CD180) in human B lymphocytes from normal, leukemic, and lupus patients. The percentage of cells that express RP105 on their surface increased following a 24 hour exposure to LPS. However, continued exposure for a total of four days resulted in a marked decrease in the expression of this receptor molecule. Human B cells were purified by a combination of density gradient and...
Show moreThe purpose of this study was to determine the effect of bacterial lipopolysaccharide (LPS) on the expression of RP105 (CD180) in human B lymphocytes from normal, leukemic, and lupus patients. The percentage of cells that express RP105 on their surface increased following a 24 hour exposure to LPS. However, continued exposure for a total of four days resulted in a marked decrease in the expression of this receptor molecule. Human B cells were purified by a combination of density gradient and negative magnetic bead selection and maintained in culture with and without LPS. Enzyme linked immunoassay for the detection of anti-dsDNA antibodies following LPS treatment of isolated B cells was negative. The percentage of RP105 positive or negative B cells from lupus patients could not be accurately determined because too few B cells were available from these lymphopenic patients following negative selection.
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Date Issued
2004
PURL
http://purl.flvc.org/fcla/dt/13177
Subject Headings
Microbial polysaccharides, Bacterial genetics, Systemic lupus erythematosus--Etiology, Systemic lupus erythematosus--Molecular aspects
Format
Document (PDF)
Systemic Lupus Erythematosus: A.Studies on an ldiotype Specific Dendritic Cell Vaccine. B.Association of Lupus Calcinosis with Calcifying Nanoparticles.
Title
Systemic Lupus Erythematosus: A.Studies on an ldiotype Specific Dendritic Cell Vaccine. B.Association of Lupus Calcinosis with Calcifying Nanoparticles.
Creator
Keating, Patricia, Florida Atlantic University, Hartmann, James X., Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description
Systemic lupus erythrematosus (SLE) is an autoimmune disease characterized by the production of anti-DNA antibodies. The primary goal of this study was to activate T cells with specificity toward lupus B cells presenting anti-DNA antibody idiotopes on their surface. Monocyte derived dendritic cells were obtained from peripheral blood of healthy donors and lupus patients. Affinity purified anti-DNA antibodies were obtained from lupus patients' plasmas. The efficacy of different carrier...
Show moreSystemic lupus erythrematosus (SLE) is an autoimmune disease characterized by the production of anti-DNA antibodies. The primary goal of this study was to activate T cells with specificity toward lupus B cells presenting anti-DNA antibody idiotopes on their surface. Monocyte derived dendritic cells were obtained from peripheral blood of healthy donors and lupus patients. Affinity purified anti-DNA antibodies were obtained from lupus patients' plasmas. The efficacy of different carrier proteins, conjugated to lgG, was evaluated, and KLH found to be the most efficient for antigen uptake. The cognate dendritic cells were evaluated for their capacity to activate autologous T cells, and generate a Th1 mediated response which was evaluated by proliferation assays and interferony secretion. During the vaccine study a patient presented with panniculitis, CREST syndrome and a calcinotic exudate. A secondary goal of my study was to analyze this exudate. Calcifying nanoparticles were isolated in lymphocyte culture medium. They were characterized by Von Kassa staining for hydroxyapatite, solubilization by the calcium chelating agent EDT A, and light and scanning electron microscopy. A novel method was developed, using a specific monoclonal antibody, to analyze the calcifying nanoparticles. This method allowed for an approximate quantification of the particles. These particles increased in numbers when incubated for different time periods, and their numbers were decreased when incubated in the presence of minocyclin. Concomitantly, the panniculi in the patient underwent remission with long term antibiotic therapy. CNPs were also obtained from fetal bovine serum and human plasma samples from both lupus patients and healthy donors. Peripheral blood mononuclear cells from healthy donors and lupus patients were analyzed in vitro for their reactivity when incubated in the presence of a biofilm, generated by the calcifying nanoparticles. Viability, proliferation, and co-stimulatory marker up-regulation were determined in the presence or absence of the particles. Osteopontin was found highly expressed in the supernatants of the cells grown with CNPs. Microarray of the mononuclear cells of a healthy donor and a lupus patient incubated in the presence or absence of CNPs was performed, and the results coincided with those determined in vitro.
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Date Issued
2007
PURL
http://purl.flvc.org/fau/fd/FA00000865
Subject Headings
Systemic lupus erythematosus--Molecular aspects, Systemic lupus erythematosus--Immunological aspects, Cell surface antigens, Autoimmune diseases--Research, Monoclonal antibodies--Diagnostic use
Format
Document (PDF)