Short-Chain Fatty Acids Promote Immunotherapy by Modulating Immune Regulatory Property in B Cells

Figure 4

TTP forms complexes with IL-10 mRNA in human B cells. (a–c) CD5⁺ and CD5^- B cells were isolated from blood samples collected from HC subjects and AR patients. Violin plots show TTP mRNA (a), and immunoblots show TTP protein (TTP) levels in B cells. Boxplots show the integrated density of TTP immunoblots (c). (d, e) IP products with an anti-TTP Ab as a bait. Violin plots show IL-10 mRNA in the IP products, indicating that TTP forms a complex with IL-10 mRNA. (f–h) B cells were prepared from HC subject blood samples and treated with the methods denoted below the panels. (f) Boxplots show IL-10 mRNA levels in B cells. 1^st stage: B cells were cultured overnight in the presence of CpG; 2^nd stage: B cells were collected from the 1^st stage culture, washed, and cultured in fresh medium for 1 h; TTP RNAi: B cells were treated with TTP RNAi; Control RNAi: B cells were treated with control RNAi; TTP overexp: B cells were transfected with TTP expressing plasmids; Overexp control: B cells were treated with empty plasmids. (g, h) B cells were cultured overnight with the treatment denoted below the plots. PA: 10 μM in the culture; BA: 2 μM in the culture. Boxplots show TTP mRNA levels in B cells. Immunoblots show TTP protein levels in B cells. , , , compared with the HC group (a, c, e), the CpG group (f), or the saline group (g). ^## and ^###, compared with the medium group. Statistical methods: Student test (e) and ANOVA+Dunnett’s test (a, c, f, g).

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