Skip to main content
main-content

25.07.2017 | Original Article | Ausgabe 5/2017

Immunologic Research 5/2017

Interleukin-10 production and T cell-suppressive capacity in B cell subsets from atherosclerotic apoE−/− mice

Zeitschrift:
Immunologic Research > Ausgabe 5/2017
Autoren:
Héctor Rincón-Arévalo, Janny Villa-Pulgarín, Jorge Tabares, Mauricio Rojas, Gloria Vásquez, José R. Ramírez-Pineda, Diana Castaño, Lina M. Yassin
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1007/​s12026-017-8939-6) contains supplementary material, which is available to authorized users.
Diana Castaño and Lina M. Yassin contributed equally to the present work.

Abstract

The evidence regarding the role of regulatory B cells (Breg) in atherosclerosis are scarce, and there are contradictory data about their atheroprotective properties. Due to the demonstrated protective function of Breg in different inflammatory diseases mainly through interleukin-10 (IL-10) production, the knowledge of their participation in atherosclerosis immunopathology would be very valuable. To further study which B cell subsets participate in IL-10 production and their regulatory role, splenocytes from apolipoprotein-E-deficient mice were evaluated by ex vivo and in vitro cultures. Atherosclerotic mice had increased frequency of IL-10+ B cells, which presented high CD1d, CD19, and IgM, but variable CD5, CD21, and CD23 expression. IL-10+ B cells were not enriched in B cell subsets previously reported as Breg. Increased frequency of IL-10+ B cells with transitional 1-like (T1-like) and follicular (FO) and reduced CD5+ and marginal zone (MZ) phenotypes were observed ex vivo. Increased frequency of IL-10+ B cells with T1-like and MZ, and decreased IL-10+ FO and T2 phenotypes were also observed in vitro. To determine regulatory capacity of B cells in the atherosclerotic model, each subset were co-cultured with CD4+CD25 T cells. CD5+, FO, MZ, and T1-like cells from atherosclerotic mice exhibited regulation in an IL-10-dependent manner. However, only FO cells decreased both frequency of interferon gamma (IFN-γ)+ and tumor necrosis factor alpha (TNF-α)+ and proliferation of T cells. Finally, splenocytes showed increased frequency of IFN-γ+ and TNF-α+ cells only when FO-depleted B cells were evaluated. These results suggest that mainly FO B cells can modulate in some level the inflammatory responses observed in atherosclerosis.

Bitte loggen Sie sich ein, um Zugang zu diesem Inhalt zu erhalten

★ PREMIUM-INHALT
e.Med Interdisziplinär

Mit e.Med Interdisziplinär erhalten Sie Zugang zu allen CME-Fortbildungen und Fachzeitschriften auf SpringerMedizin.de. Zusätzlich können Sie eine Zeitschrift Ihrer Wahl in gedruckter Form beziehen – ohne Aufpreis.

Bis zum 22.10. bestellen und 100 € sparen!

Weitere Produktempfehlungen anzeigen
Zusatzmaterial
ESM S1 Frequency of B cells secreting IL-10 in atherosclerotic mice. Percentage of IL-10 secreting splenic B cells from WT-HFD and apoE −/− HFD mice after A. ex vivo and B. in vitro cultures. Briefly, isolated B cells were cultured as previously described, incubated for the last 45 min with an anti-IL-10 conjugated to an anti-CD45 antibody to capture secreted IL-10 by each B cell, and finally extracellularly labeled with anti-IL-10. Median and interquartile range of IL-10+ B cells from three to seven mice per group are shown from two independent experiments. Mann-Whitney test. *p < 0.05. (GIF 9 kb)
12026_2017_8939_Fig6_ESM.gif
High-resolution image (TIFF 28572 kb)
12026_2017_8939_MOESM1_ESM.tif
ESM S2Frequency of aortic IL-10+ B cells after ex vivo culture. Aortic B cells were cultured ex vivo as explained in the “Materials and methods.” Median and interquartile range from nine to ten mice per group are shown from two independent experiments. Mann-Whitney test. (GIF 3 kb)
High-resolution image (TIFF 10392 kb)
12026_2017_8939_MOESM2_ESM.tif
ESM S3 Frequency of Foxp3+ cells in IL-10+ B cells region. A. Representative pseudocolor plots of Foxp3 expression in splenic IL-10+ (left) and IL-10− B cells (right) after ex vivo culture. B-C. Frequency of Foxp3+ in IL-10+ and IL-10-splenic B cells after B. ex vivo and C. in vitro cultures. Median and interquartile range from nine mice (WT-HFD and apoE −/− ) are shown. D-E. Frequency of Foxp3+ cells in splenic IL-10+ B cells from WT-HFD and apoE −/− HFD mice after D. ex vivo and E. in vitro cultures. Median and interquartile range from four to five mice per group are shown. Mann-Whitney test. ∞ p < 0.0001; Φ p < 0.001; *p < 0.05. (GIF 27 kb)
12026_2017_8939_Fig8_ESM.gif
High-resolution image (TIFF 70302 kb)
12026_2017_8939_MOESM3_ESM.tif
ESM S4 B cell subsets analysis. Representative pseudocolor plots of CD5+, follicular (FO, CD5−CD21lowCD23+CD24low), transitional 1-like (T1-like, CD5−CD21lowCD23−CD24low), transitional 2 (T2, CD5−CD21hiCD23+CD24hi), and marginal zone (MZ, CD5−CD21hiCD23−CD24hi) B cell subsets from WT-HFD mouse after A ex vivo and B in vitro cultures. (GIF 18 kb)
12026_2017_8939_Fig9_ESM.gif
High-resolution image (TIFF 30809 kb)
12026_2017_8939_MOESM4_ESM.tif
High-resolution image (TIFF 172021 kb)
12026_2017_8939_MOESM5_ESM.tif
ESM S6 Representative pseudocolor plots of enriched B cell subsets from a WT-HFD mouse, according to CD5, CD21, and CD23 markers for the sorting of B cells subsets. A. B cells before sorting. B-F. B cell subsets after sorting. B. CD5+, C. follicular (FO, CD5−CD21lowCD23+CD24low), D. transitional 1-like (T1-like, CD5−CD21lowCD23−CD24low), E. transitional 2 (T2, CD5−CD21hiCD23+CD24hi), and F. marginal zone (MZ, CD5−CD21hiCD23−CD24hi) B cell subsets. (GIF 22 kb)
12026_2017_8939_Fig11_ESM.gif
High-resolution image (TIFF 54429 kb)
12026_2017_8939_MOESM6_ESM.tif
ESM S7 Proliferation of CD4+CD25− T cells co-cultured with each B cell subsets from atherosclerotic mice. A. Fold change in the proliferation index (total number of divisions per the number of cells that went into division) of CD4+CD25− T cells co-cultured with each B cell subset (CD5+, follicular (FO), marginal zone (MZ), transitional 1-like (T1-like), and transitional 2 (T2)) from apoE −/− HFD mice. B. Fold change in the division index (average number of cell divisions that a cell in the original population has undergone) of T cells co-cultured with B cell subsets from apoE −/− HFD mice. Median and interquartile range from 8 apoE −/− HFD mice for each co-culture are shown from two independent experiments. Two-way ANOVA test with Šídák post-test. Differences between co-cultures and cultures without B cells were indicated as follows: ∞ p < 0.0001; Φ p < 0.001; # p < 0.01; *p < 0.05. Symbol on the bracket indicates difference between co-cultures at a ratio of 1:1 and 2:1. The threshold on the y-axis was stablished based on T cells cultured without B cells. (GIF 27 kb)
12026_2017_8939_Fig12_ESM.gif
High-resolution image (TIFF 49166 kb)
12026_2017_8939_MOESM7_ESM.tif
ESM S8 Frequencies of B cell subsets after culture. A. Percentage of B cell subsets after A. ex vivo and B. in vitro cultures. Median and interquartile range from 13 to 14 mice per group are shown from two independent experiments. Kruskal-Wallis test with Dunn’s post-test. ∞ p < 0.0001; Φ p < 0.001; # p < 0.01; *p < 0.05. (GIF 48 kb)
12026_2017_8939_Fig13_ESM.gif
High-resolution image (TIFF 122342 kb)
12026_2017_8939_MOESM8_ESM.tif
Literatur
Über diesen Artikel

Weitere Artikel der Ausgabe 5/2017

Immunologic Research 5/2017Zur Ausgabe
  1. Das kostenlose Testabonnement läuft nach 14 Tagen automatisch und formlos aus. Dieses Abonnement kann nur einmal getestet werden.

Neu im Fachgebiet HNO

 

 

 
 

Mail Icon II Newsletter

Bestellen Sie unseren kostenlosen Newsletter Update HNO und bleiben Sie gut informiert – ganz bequem per eMail.

Bildnachweise