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Expression of Toll-like receptor -7 and -9 in B cell subsets from patients with primary Sjögren's syndrome.

Karlsen M, Hansen T, Nordal HH, Brun JG, Jonsson R, Appel S - PLoS ONE (2015)

Bottom Line: An increased number of naïve B cells, as well as fewer pre-switched memory B cells were found in pSS patients.This study shows that pSS patients have an alteration in the B cell subpopulation composition compared to controls, with less pre-switched memory B cells and more naïve B cells.We did not detect any significant disparities in TLR-7 and -9 expression between the two groups.

View Article: PubMed Central - PubMed

Affiliation: Broegelmann Research Laboratory, Department of Clinical Science, University of Bergen, Bergen, Norway.

ABSTRACT

Introduction: Sjögren's syndrome (SS) is a rheumatic autoimmune disease characterized by inflammation of exocrine glands. As autoantibodies are present in a majority of patients, B cells have been suggested to play an important role in onset and development of the disease. Toll-like receptors (TLRs) are pattern recognition receptors triggering innate immune responses. Since an increased expression of TLRs has been detected in other rheumatic diseases the purpose of this study was to explore TLRs in B cells of SS patients.

Methods: The expression of TLR-7 and -9 in B cell subsets of 25 patients with primary SS (pSS) and 25 healthy controls was analysed in peripheral blood using flow cytometry and real time quantitative PCR.

Results: We detected similar levels of CD19+ B cells in pSS patients and healthy controls. An increased number of naïve B cells, as well as fewer pre-switched memory B cells were found in pSS patients. No significant differences were observed in TLR-7 and -9 expression in B cells between pSS patients and healthy controls.

Conclusion: This study shows that pSS patients have an alteration in the B cell subpopulation composition compared to controls, with less pre-switched memory B cells and more naïve B cells. We did not detect any significant disparities in TLR-7 and -9 expression between the two groups.

No MeSH data available.


Related in: MedlinePlus

Similar TLR-7 and -9 mRNA level in pSS patients and controls.No significant disparities were detected in mRNA level, however a slightly lower level of TLR-9 was noticed in pSS. RNA was isolated from negatively isolated B cells, and cDNA was synthesized with High-Capacity RNA-to-cDNA Kit. Taqman gene expression assays were used to detect TLR-7 and -9. GAPDH was used for normalization. Median is indicated in the figure.
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pone.0120383.g005: Similar TLR-7 and -9 mRNA level in pSS patients and controls.No significant disparities were detected in mRNA level, however a slightly lower level of TLR-9 was noticed in pSS. RNA was isolated from negatively isolated B cells, and cDNA was synthesized with High-Capacity RNA-to-cDNA Kit. Taqman gene expression assays were used to detect TLR-7 and -9. GAPDH was used for normalization. Median is indicated in the figure.

Mentions: In order to analyse TLR expression in B cell subsets, PBMC were stained with surface markers to identify different B cell populations before cells were fixed, permeabilized and stained for TLR-7 and -9. We used CD27 and IgD to distinguish the B cell subsets, and the gating strategy is shown in Fig. 3. No significant differences in TLR-7 and -9 expression were detected between pSS patients and controls (Fig. 4), though different expression levels of the TLRs were observed in the various B cells subsets. Naïve B cells expressed less TLR-7 and -9 than the memory B cell subsets in both, patients and controls (Fig. 4B). To explore if there were differences between protein expression and amount of transcripts, we analysed the mRNA levels of TLR-7 and -9 in isolated B cells from most of the pSS patients and healthy controls (n = 21 and 18, respectively). No significant changes were detected (Fig. 5).


Expression of Toll-like receptor -7 and -9 in B cell subsets from patients with primary Sjögren's syndrome.

Karlsen M, Hansen T, Nordal HH, Brun JG, Jonsson R, Appel S - PLoS ONE (2015)

Similar TLR-7 and -9 mRNA level in pSS patients and controls.No significant disparities were detected in mRNA level, however a slightly lower level of TLR-9 was noticed in pSS. RNA was isolated from negatively isolated B cells, and cDNA was synthesized with High-Capacity RNA-to-cDNA Kit. Taqman gene expression assays were used to detect TLR-7 and -9. GAPDH was used for normalization. Median is indicated in the figure.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4366264&req=5

pone.0120383.g005: Similar TLR-7 and -9 mRNA level in pSS patients and controls.No significant disparities were detected in mRNA level, however a slightly lower level of TLR-9 was noticed in pSS. RNA was isolated from negatively isolated B cells, and cDNA was synthesized with High-Capacity RNA-to-cDNA Kit. Taqman gene expression assays were used to detect TLR-7 and -9. GAPDH was used for normalization. Median is indicated in the figure.
Mentions: In order to analyse TLR expression in B cell subsets, PBMC were stained with surface markers to identify different B cell populations before cells were fixed, permeabilized and stained for TLR-7 and -9. We used CD27 and IgD to distinguish the B cell subsets, and the gating strategy is shown in Fig. 3. No significant differences in TLR-7 and -9 expression were detected between pSS patients and controls (Fig. 4), though different expression levels of the TLRs were observed in the various B cells subsets. Naïve B cells expressed less TLR-7 and -9 than the memory B cell subsets in both, patients and controls (Fig. 4B). To explore if there were differences between protein expression and amount of transcripts, we analysed the mRNA levels of TLR-7 and -9 in isolated B cells from most of the pSS patients and healthy controls (n = 21 and 18, respectively). No significant changes were detected (Fig. 5).

Bottom Line: An increased number of naïve B cells, as well as fewer pre-switched memory B cells were found in pSS patients.This study shows that pSS patients have an alteration in the B cell subpopulation composition compared to controls, with less pre-switched memory B cells and more naïve B cells.We did not detect any significant disparities in TLR-7 and -9 expression between the two groups.

View Article: PubMed Central - PubMed

Affiliation: Broegelmann Research Laboratory, Department of Clinical Science, University of Bergen, Bergen, Norway.

ABSTRACT

Introduction: Sjögren's syndrome (SS) is a rheumatic autoimmune disease characterized by inflammation of exocrine glands. As autoantibodies are present in a majority of patients, B cells have been suggested to play an important role in onset and development of the disease. Toll-like receptors (TLRs) are pattern recognition receptors triggering innate immune responses. Since an increased expression of TLRs has been detected in other rheumatic diseases the purpose of this study was to explore TLRs in B cells of SS patients.

Methods: The expression of TLR-7 and -9 in B cell subsets of 25 patients with primary SS (pSS) and 25 healthy controls was analysed in peripheral blood using flow cytometry and real time quantitative PCR.

Results: We detected similar levels of CD19+ B cells in pSS patients and healthy controls. An increased number of naïve B cells, as well as fewer pre-switched memory B cells were found in pSS patients. No significant differences were observed in TLR-7 and -9 expression in B cells between pSS patients and healthy controls.

Conclusion: This study shows that pSS patients have an alteration in the B cell subpopulation composition compared to controls, with less pre-switched memory B cells and more naïve B cells. We did not detect any significant disparities in TLR-7 and -9 expression between the two groups.

No MeSH data available.


Related in: MedlinePlus