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Expansion of FasL-Expressing CD5 + B Cells in Type 1 Diabetes Patients

View Article: PubMed Central - PubMed

ABSTRACT

Fas ligand drives insulitis in the non-obese diabetic mouse model of type 1 diabetes (T1D) and negatively regulates IL-10-producing (IL-10pos) CD5+ B cells in pancreata. Relevance of these phenomena to the human disease is poorly understood. Here, using splenocytes from T1D, autoantibody (Ab+), and non-diabetic (ND) human subjects, we show that a subpopulation of CD5+ B cells that is characterized by expression of FasL (FasLhiCD5+) was significantly elevated in T1D subjects, many of whom had significantly reduced frequency of IL-10posCD5+ B cells compared to Ab+ subjects. The majority of FasLhiCD5+ B cells did not produce cytokines and were more highly resistant to activation-induced cell death than their IL-10posCD5+ counterparts. These results associate expansion of FasL-expressing CD5+ B cells with T1D and lay the groundwork for future mechanistic studies to understand specific role in disease pathogenesis.

No MeSH data available.


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FasLhi CD5+, but not of FasLhi CD5– B cells, are selectively enriched in type 1 diabetes (T1D) as compared to Ab+ and ND subjects. Cryopreserved splenocytes from T1D, Ab+, and ND subjects were thawed and surface-stained, and indicated subsets identified as described in Figure 1A were gated and analyzed for surface expression of FasL. (A) Representative dot plots show FasL expression by gated CD5+ and CD5− B cells and T cells in different subjects. (B) Graphs show cumulative data pooled from at least 14 independent experiments. In each experiment, one T1D subject is paired with Ab+ and/or an ND subject. Each symbol represents one subject. Data were analyzed using the Mann–Whitney test and expressed as mean ± SEM; p < 0.05 is statistically significant.
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Figure 2: FasLhi CD5+, but not of FasLhi CD5– B cells, are selectively enriched in type 1 diabetes (T1D) as compared to Ab+ and ND subjects. Cryopreserved splenocytes from T1D, Ab+, and ND subjects were thawed and surface-stained, and indicated subsets identified as described in Figure 1A were gated and analyzed for surface expression of FasL. (A) Representative dot plots show FasL expression by gated CD5+ and CD5− B cells and T cells in different subjects. (B) Graphs show cumulative data pooled from at least 14 independent experiments. In each experiment, one T1D subject is paired with Ab+ and/or an ND subject. Each symbol represents one subject. Data were analyzed using the Mann–Whitney test and expressed as mean ± SEM; p < 0.05 is statistically significant.

Mentions: Next, we determined whether FasL expression is dysregulated in B or T cells in any of the three subject groups. We detected significant differences that, as above, were limited to the CD5+ B cell subpopulation, albeit in the opposite direction, as the frequency of FasL-expressing CD5+ B cells (FasLhi CD5+) was significantly higher in T1D as compared to Ab+ and ND subjects (Figure 2A). The frequency of FasL-expressing CD5− B cells (FasLhi CD5−) was generally low and slightly higher in T1D, but the difference among the groups did not reach statistical significance (Figure 2B). Likewise, the frequency of FasL-expressing T cells was comparable in the three groups, thereby excluding a generally dysregulated expression of FasL in T1D subjects (Figure 2B). In addition, CD5+ B cells in T1D, Ab+, and ND subjects had comparable surface expression of CD86 and CD40, confirming that the detected changes in IL-10 and FasL expression by these cells was specific and not due to generalized modulation of their surface profile (Figure S5 in Supplementary Material). Taken together, our results show IL-10pos CD5+ B cells are selectively expanded in Ab+ subjects as opposed to selective expansion of FasLhi CD5+ B cells T1D subjects.


Expansion of FasL-Expressing CD5 + B Cells in Type 1 Diabetes Patients
FasLhi CD5+, but not of FasLhi CD5– B cells, are selectively enriched in type 1 diabetes (T1D) as compared to Ab+ and ND subjects. Cryopreserved splenocytes from T1D, Ab+, and ND subjects were thawed and surface-stained, and indicated subsets identified as described in Figure 1A were gated and analyzed for surface expression of FasL. (A) Representative dot plots show FasL expression by gated CD5+ and CD5− B cells and T cells in different subjects. (B) Graphs show cumulative data pooled from at least 14 independent experiments. In each experiment, one T1D subject is paired with Ab+ and/or an ND subject. Each symbol represents one subject. Data were analyzed using the Mann–Whitney test and expressed as mean ± SEM; p < 0.05 is statistically significant.
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Related In: Results  -  Collection

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Figure 2: FasLhi CD5+, but not of FasLhi CD5– B cells, are selectively enriched in type 1 diabetes (T1D) as compared to Ab+ and ND subjects. Cryopreserved splenocytes from T1D, Ab+, and ND subjects were thawed and surface-stained, and indicated subsets identified as described in Figure 1A were gated and analyzed for surface expression of FasL. (A) Representative dot plots show FasL expression by gated CD5+ and CD5− B cells and T cells in different subjects. (B) Graphs show cumulative data pooled from at least 14 independent experiments. In each experiment, one T1D subject is paired with Ab+ and/or an ND subject. Each symbol represents one subject. Data were analyzed using the Mann–Whitney test and expressed as mean ± SEM; p < 0.05 is statistically significant.
Mentions: Next, we determined whether FasL expression is dysregulated in B or T cells in any of the three subject groups. We detected significant differences that, as above, were limited to the CD5+ B cell subpopulation, albeit in the opposite direction, as the frequency of FasL-expressing CD5+ B cells (FasLhi CD5+) was significantly higher in T1D as compared to Ab+ and ND subjects (Figure 2A). The frequency of FasL-expressing CD5− B cells (FasLhi CD5−) was generally low and slightly higher in T1D, but the difference among the groups did not reach statistical significance (Figure 2B). Likewise, the frequency of FasL-expressing T cells was comparable in the three groups, thereby excluding a generally dysregulated expression of FasL in T1D subjects (Figure 2B). In addition, CD5+ B cells in T1D, Ab+, and ND subjects had comparable surface expression of CD86 and CD40, confirming that the detected changes in IL-10 and FasL expression by these cells was specific and not due to generalized modulation of their surface profile (Figure S5 in Supplementary Material). Taken together, our results show IL-10pos CD5+ B cells are selectively expanded in Ab+ subjects as opposed to selective expansion of FasLhi CD5+ B cells T1D subjects.

View Article: PubMed Central - PubMed

ABSTRACT

Fas ligand drives insulitis in the non-obese diabetic mouse model of type 1 diabetes (T1D) and negatively regulates IL-10-producing (IL-10pos) CD5+ B cells in pancreata. Relevance of these phenomena to the human disease is poorly understood. Here, using splenocytes from T1D, autoantibody (Ab+), and non-diabetic (ND) human subjects, we show that a subpopulation of CD5+ B cells that is characterized by expression of FasL (FasLhiCD5+) was significantly elevated in T1D subjects, many of whom had significantly reduced frequency of IL-10posCD5+ B cells compared to Ab+ subjects. The majority of FasLhiCD5+ B cells did not produce cytokines and were more highly resistant to activation-induced cell death than their IL-10posCD5+ counterparts. These results associate expansion of FasL-expressing CD5+ B cells with T1D and lay the groundwork for future mechanistic studies to understand specific role in disease pathogenesis.

No MeSH data available.


Related in: MedlinePlus