Toll-Like Receptor Mediated Activation of Natural Autoantibody Producing B Cell Subpopulations in an Autoimmune Disease Model
Abstract
:1. Introduction
2. Results
2.1. CD180 Expression Is Decreased in dcSSc B Cells
2.2. TLR Ligation Results in Reduced CD180 mRNA and Protein Expression of B Cells
2.3. The Frequency of CD180+ Cells Is the Highest in the Non-Switched Memory B Cell Subset
2.4. Anti-CD180 Stimulation Resulted in Activation of B Cell Subsets
2.5. TLR Stimulation Differentially Affects IL-6 and IL-10 Production of B Cells
2.6. CD180 Stimulation Induces Natural Autoantibody Production
3. Discussion
4. Materials and Methods
4.1. Patients
4.2. Mononuclear Cell Isolation, B Cell Separation
4.3. Cell Stimulation
4.4. RNA Isolation, cDNA Synthesis, and qPCR
4.5. Flow Cytometric Analysis
4.6. IL-10 and IL-6 ELISA
4.7. Measurement of Natural and Pathologic Autoantibodies
4.8. Statistical Analysis
Author Contributions
Funding
Conflicts of Interest
Abbreviations
ANA | Antinuclear antibodies |
Breg | Regulatory B cell |
CpG | 5’—C—phosphate—G—3’ dinucleotide |
CS | Citrate synthase |
dcSSc | Diffuse cutaneous systemic sclerosis |
DN B | Double negative B cell |
ELISA | Enzyme-linked immunosorbent assay |
FSC | Forward scatter |
GAPDH | Glyceraldehyde 3-phosphate dehydrogenase |
HC | Healthy control |
Hu | Human |
LEAF | Low endotoxin, azide-free |
MFI | Mean fluorescence intensity |
Ms | Mouse |
MZ | Marginal zone |
NS B | Non-switched B cell |
PBMC | Peripheral blood mononuclear cells |
PFA | Paraformaldehyde |
qPCR | Quantitative polymerase chain reaction |
RQ | Relative quantification |
S B | Switched B cell |
SLE | Systemic lupus erythematosus |
SSc | Systemic sclerosis |
SSC | Side scatter |
TLR | Toll-like receptor |
TMB | 3,3′,5,5′-Tetramethylbenzidine |
Topo I | Topoisomerase I |
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Erdő-Bonyár, S.; Rapp, J.; Minier, T.; Ráth, G.; Najbauer, J.; Czirják, L.; Németh, P.; Berki, T.; Simon, D. Toll-Like Receptor Mediated Activation of Natural Autoantibody Producing B Cell Subpopulations in an Autoimmune Disease Model. Int. J. Mol. Sci. 2019, 20, 6152. https://doi.org/10.3390/ijms20246152
Erdő-Bonyár S, Rapp J, Minier T, Ráth G, Najbauer J, Czirják L, Németh P, Berki T, Simon D. Toll-Like Receptor Mediated Activation of Natural Autoantibody Producing B Cell Subpopulations in an Autoimmune Disease Model. International Journal of Molecular Sciences. 2019; 20(24):6152. https://doi.org/10.3390/ijms20246152
Chicago/Turabian StyleErdő-Bonyár, Szabina, Judit Rapp, Tünde Minier, Gábor Ráth, József Najbauer, László Czirják, Péter Németh, Timea Berki, and Diána Simon. 2019. "Toll-Like Receptor Mediated Activation of Natural Autoantibody Producing B Cell Subpopulations in an Autoimmune Disease Model" International Journal of Molecular Sciences 20, no. 24: 6152. https://doi.org/10.3390/ijms20246152