Journal: PLoS Pathogens
Article Title: Hantavirus infection-induced B cell activation elevates free light chains levels in circulation
doi: 10.1371/journal.ppat.1009843
Figure Lengend Snippet: B cells were enriched from blood of 3 healthy volunteers and infected with live or UV-inactivated purified PUUV, together with Sendai and Sindbis viruses, for comparison (multiplicity of infections for all viruses equals 1). The TLR9 agonist CpG and no treatment were used as a positive and negative controls for B cell activation, respectively. After culturing for 8 days, kappa and lambda FLCs in (A) and IgA, IgM and IgGs in (B) were measured from cell supernatants. Remaining cells were subjected to IgA, IgM and IgG-specific antibody secreting cell (ASC) quantification using Elispot. IgA + and IgM + ASCs were simultaneously detected in one well, using AP-conjugated anti-IgA and HRP-conjugated anti-IgM Abs, and IgG + ASCs in a separate well using HRP-conjugated anti-IgG Ab. Representative Elispot images from one donor are shown in (C), and ASC quantification per input cell number measured from all donors in (D). Significant differences of each treatment group, as compared to non-treated cells, were assessed using two-way ANOVA and Dunnett’s multiple comparison tests. *p < 0.05, **p < 0.01, ***p < 0.001.
Article Snippet: The following mAbs were used: anti-kappa light chain PercP-Cy5.5 (G20-193), anti-lambda light chain BV605 (JDC-12), anti-IgM APC (G20-127), anti-IgG AF700 (G18-145; all from BD) and anti-IgA PE-Vio770 (IS11-8E10; Miltenyi).
Techniques: Infection, Purification, Comparison, Activation Assay, Enzyme-linked Immunospot