Journal: Journal of immunology (Baltimore, Md. : 1950)
Article Title: LLT1 and CD161 Expression in Human Germinal Centers Promotes B Cell Activation and CXCR4 Downregulation.
doi: 10.4049/jimmunol.1502462
Figure Lengend Snippet: FIGURE 5. LLT1 promotes B cell activation. (A) Purified B cells from PBMCs were stimulated with anti-BCRs only or with anti-BCRs with and without rCD161 or IC. Geometric mean fluorescence intensities (GeoMFI) of CD83 and CD38 were measured by flow cytometry. B cell stimulation with rCD161 resulted in enhanced levels of CD83 and CD38; data were pooled from three independent experiments (n = 7, **p , 0.01). (B) Tonsillar cells were cultured for 3 d with CpG or anti-CD40 plus IL-4. Blocking LLT1–CD161 interaction resulted in a decreased expression of CD83 and CD38 on B cells; data were pooled from four independent experiments (n = 12, ****p , 0.0001). (C) Depletion of CD161+ cells from tonsillar cells resulted in decreased expression of CD83 (anti-CD40 plus IL-4 stimulation) and CD38 (CpG stimulation) on B cells after 3 d; data were pooled from two independent experiments (n = 8; *p , 0.05, ****p , 0.0001, paired t test). (D) Overnight stimulation of tonsillar B cells with rCD161 or 2H7 anti-LLT1 Ab did not change the expression of CD83 on GC B cells (n = 5). (E) However, a decrease in CXCR4 expression levels was observed GC B cells (n = 10; **p , 0.01, ****p , 0.0001). (F) Repeated experiments on sorted DZ B cells revealed a downregulation of CXCR4 mRNA. All statistics were two-way ANOVA using a Bonferroni multiple comparisons test, unless stated otherwise.
Article Snippet: For LLT1 crosslinking, recombinant CD161 (R&D Systems) or IgG1 isotype control (R&D Systems) were bound to a 96-well ELISA plate (Greiner Bio-One, Stonehouse, U.K.) overnight prior to the addition of B cells and BCR stimulus as described above.
Techniques: Activation Assay, Cytometry, Cell Stimulation, Cell Culture, Blocking Assay, Expressing
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