Limits...
Characterization of Regulatory B Cells in Graves' Disease and Hashimoto's Thyroiditis.

Kristensen B, Hegedüs L, Lundy SK, Brimnes MK, Smith TJ, Nielsen CH - PLoS ONE (2015)

Bottom Line: B-cell expression of two surface marker combinations previously associated with regulatory B-cell functions, CD24hiCD38hi and CD27+CD43+, did not differ between patients and healthy donors.In conclusion, our findings indicate that autoimmune thyroiditis is not associated with reduced frequency of IL-10+ B cells.These results do not rule out regulatory B-cell dysfunction, however.

View Article: PubMed Central - PubMed

Affiliation: Institute for Inflammation Research, Department of Infectious Diseases and Rheumatology, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark; Department of Endocrinology and Metabolism, Odense University Hospital, Odense, Denmark.

ABSTRACT
A hallmark of regulatory B cells is IL-10 production, hence their designation as IL-10+ B cells. Little is known about the ability of self-antigens to induce IL-10+ B cells in Graves' disease (GD), Hashimoto's thyroiditis (HT), or other autoimmune disease. Here we pulsed purified B cells from 12 HT patients, 12 GD patients, and 12 healthy donors with the thyroid self-antigen, thyroglobulin (TG) and added the B cells back to the remaining peripheral blood mononuclear cells (PBMCs). This procedure induced IL-10+ B-cell differentiation in GD. A similar tendency was observed in healthy donors, but not in cells from patients with HT. In GD, B cells primed with TG induced IL-10-producing CD4+ T cells. To assess the maximal frequency of inducible IL-10+ B cells in the three donor groups PBMCs were stimulated with PMA/ionomycin. The resulting IL-10+ B-cell frequency was similar in the three groups and correlated with free T3 levels in GD patients. IL-10+ B cells from both patient groups displayed CD25 or TIM-1 more frequently than did those from healthy donors. B-cell expression of two surface marker combinations previously associated with regulatory B-cell functions, CD24hiCD38hi and CD27+CD43+, did not differ between patients and healthy donors. In conclusion, our findings indicate that autoimmune thyroiditis is not associated with reduced frequency of IL-10+ B cells. These results do not rule out regulatory B-cell dysfunction, however. The observed phenotypic differences between IL-10+ B cells from patients and healthy donors are discussed.

No MeSH data available.


Related in: MedlinePlus

Frequencies of IL-10+ B cells expressing CD5, CD25 and TIM-1.PBMCs from healthy donors (HD; N = 12), patients with Graves’ disease (GD; N = 12) or patients with Hashimoto’s thyroiditis (HT; N = 12) were stimulated with PMA/ionomycin and stained with antibodies against CD19, IL-10, CD5, CD25, and TIM-1. The proportion of (A) CD5+, (B) CD25+ and (C) TIM-1+ cells within the IL-10+CD19+ B cell subset was analyzed. The frequencies of IL-10+ B cells (light grey) and the remaining B cells (IL-10-; dark grey) expressing (D) CD5, (E) CD25 and (F) TIM-1 are shown for each donor category. The box plots indicate median, interquartile range (box) and range (whiskers). Brackets show differences between groups with the corresponding raw P-values (Mann-Whitney U test). ** P<0.01; *** P<0.001 between IL-10+ cells and IL-10- B cells within each group (Wilcoxon matched-pairs signed rank test).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4446335&req=5

pone.0127949.g004: Frequencies of IL-10+ B cells expressing CD5, CD25 and TIM-1.PBMCs from healthy donors (HD; N = 12), patients with Graves’ disease (GD; N = 12) or patients with Hashimoto’s thyroiditis (HT; N = 12) were stimulated with PMA/ionomycin and stained with antibodies against CD19, IL-10, CD5, CD25, and TIM-1. The proportion of (A) CD5+, (B) CD25+ and (C) TIM-1+ cells within the IL-10+CD19+ B cell subset was analyzed. The frequencies of IL-10+ B cells (light grey) and the remaining B cells (IL-10-; dark grey) expressing (D) CD5, (E) CD25 and (F) TIM-1 are shown for each donor category. The box plots indicate median, interquartile range (box) and range (whiskers). Brackets show differences between groups with the corresponding raw P-values (Mann-Whitney U test). ** P<0.01; *** P<0.001 between IL-10+ cells and IL-10- B cells within each group (Wilcoxon matched-pairs signed rank test).

Mentions: Representative dot plots of CD5, CD25 and TIM-1 surface expression on IL-10+ B cells from a healthy donor are shown in Fig 4A–4C. As expected from earlier studies, IL-10+ B cells were enriched with all three markers, compared to the rest of the B-cell pool, in all three donor groups (Fig 4D–4F). The median proportion of CD25+IL-10+ B cells constituted 30% in both GD and HT, compared to only 17% in healthy donors (P = 0.039 and P = 0.0009, respectively; Fig 4E).


Characterization of Regulatory B Cells in Graves' Disease and Hashimoto's Thyroiditis.

Kristensen B, Hegedüs L, Lundy SK, Brimnes MK, Smith TJ, Nielsen CH - PLoS ONE (2015)

Frequencies of IL-10+ B cells expressing CD5, CD25 and TIM-1.PBMCs from healthy donors (HD; N = 12), patients with Graves’ disease (GD; N = 12) or patients with Hashimoto’s thyroiditis (HT; N = 12) were stimulated with PMA/ionomycin and stained with antibodies against CD19, IL-10, CD5, CD25, and TIM-1. The proportion of (A) CD5+, (B) CD25+ and (C) TIM-1+ cells within the IL-10+CD19+ B cell subset was analyzed. The frequencies of IL-10+ B cells (light grey) and the remaining B cells (IL-10-; dark grey) expressing (D) CD5, (E) CD25 and (F) TIM-1 are shown for each donor category. The box plots indicate median, interquartile range (box) and range (whiskers). Brackets show differences between groups with the corresponding raw P-values (Mann-Whitney U test). ** P<0.01; *** P<0.001 between IL-10+ cells and IL-10- B cells within each group (Wilcoxon matched-pairs signed rank test).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4446335&req=5

pone.0127949.g004: Frequencies of IL-10+ B cells expressing CD5, CD25 and TIM-1.PBMCs from healthy donors (HD; N = 12), patients with Graves’ disease (GD; N = 12) or patients with Hashimoto’s thyroiditis (HT; N = 12) were stimulated with PMA/ionomycin and stained with antibodies against CD19, IL-10, CD5, CD25, and TIM-1. The proportion of (A) CD5+, (B) CD25+ and (C) TIM-1+ cells within the IL-10+CD19+ B cell subset was analyzed. The frequencies of IL-10+ B cells (light grey) and the remaining B cells (IL-10-; dark grey) expressing (D) CD5, (E) CD25 and (F) TIM-1 are shown for each donor category. The box plots indicate median, interquartile range (box) and range (whiskers). Brackets show differences between groups with the corresponding raw P-values (Mann-Whitney U test). ** P<0.01; *** P<0.001 between IL-10+ cells and IL-10- B cells within each group (Wilcoxon matched-pairs signed rank test).
Mentions: Representative dot plots of CD5, CD25 and TIM-1 surface expression on IL-10+ B cells from a healthy donor are shown in Fig 4A–4C. As expected from earlier studies, IL-10+ B cells were enriched with all three markers, compared to the rest of the B-cell pool, in all three donor groups (Fig 4D–4F). The median proportion of CD25+IL-10+ B cells constituted 30% in both GD and HT, compared to only 17% in healthy donors (P = 0.039 and P = 0.0009, respectively; Fig 4E).

Bottom Line: B-cell expression of two surface marker combinations previously associated with regulatory B-cell functions, CD24hiCD38hi and CD27+CD43+, did not differ between patients and healthy donors.In conclusion, our findings indicate that autoimmune thyroiditis is not associated with reduced frequency of IL-10+ B cells.These results do not rule out regulatory B-cell dysfunction, however.

View Article: PubMed Central - PubMed

Affiliation: Institute for Inflammation Research, Department of Infectious Diseases and Rheumatology, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark; Department of Endocrinology and Metabolism, Odense University Hospital, Odense, Denmark.

ABSTRACT
A hallmark of regulatory B cells is IL-10 production, hence their designation as IL-10+ B cells. Little is known about the ability of self-antigens to induce IL-10+ B cells in Graves' disease (GD), Hashimoto's thyroiditis (HT), or other autoimmune disease. Here we pulsed purified B cells from 12 HT patients, 12 GD patients, and 12 healthy donors with the thyroid self-antigen, thyroglobulin (TG) and added the B cells back to the remaining peripheral blood mononuclear cells (PBMCs). This procedure induced IL-10+ B-cell differentiation in GD. A similar tendency was observed in healthy donors, but not in cells from patients with HT. In GD, B cells primed with TG induced IL-10-producing CD4+ T cells. To assess the maximal frequency of inducible IL-10+ B cells in the three donor groups PBMCs were stimulated with PMA/ionomycin. The resulting IL-10+ B-cell frequency was similar in the three groups and correlated with free T3 levels in GD patients. IL-10+ B cells from both patient groups displayed CD25 or TIM-1 more frequently than did those from healthy donors. B-cell expression of two surface marker combinations previously associated with regulatory B-cell functions, CD24hiCD38hi and CD27+CD43+, did not differ between patients and healthy donors. In conclusion, our findings indicate that autoimmune thyroiditis is not associated with reduced frequency of IL-10+ B cells. These results do not rule out regulatory B-cell dysfunction, however. The observed phenotypic differences between IL-10+ B cells from patients and healthy donors are discussed.

No MeSH data available.


Related in: MedlinePlus