Journal: Cells

Article Title: Bone Marrow-Derived IL-1Ra Increases TNF Levels Poststroke

doi: 10.3390/cells10040956

Figure Lengend Snippet: Effect of bone marrow (BM) cells on ischemia-induced proteins in brain and serum. ( A ) BM cell treatment 30 min after tMCAo reduces infarct volumes (IFV) in LM–LM and Tg–LM mice as well as brain IL-1β levels (pg/mg) in Tg–LM mice 24 h after tMCAo, compared to LM tMCAo mice (Table with extracted data from ) (the experimental overview was created with BioRender.com). ( B – K ) Comparison of TNF ( C ), IL-4 ( D ), IL-10 ( E ), CXCL1 ( F ), IL-12p70 ( G ), IL-5 ( H ), IL-6 ( I ), IFNγ ( J ) and IL-2 ( K ), levels in the brain (pg/mg) and serum (pg/mL) of unlesioned controls and in LM, LM–LM, and Tg–LM mice 24 h after tMCAo. Data are presented as mean ± SEM ( n = 10–12/group). One-way ANOVA, followed by Sidak’s post hoc tests, was performed. Significance indicated as * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. N.D, not detected.

Article Snippet: Electrochemiluminescence analysis was performed on brain, serum, cells, and media using the MSD Mouse Proinflammatory V-Plex Plus Kit (IFNγ, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p70, CXCL1, TNF; K15012C, Mesoscale Discovery), the U-Plex Kit (TNFR1; K15069L, Mesoscale), and the R-Plex kit (TNFR2; K150ZSR-2, Mesoscale Discovery) according to the manufacturer’s instructions.

Techniques: Comparison

Journal: Frontiers in Immunology

Article Title: IgA2 immune complexes selectively promote inflammation by human CD103 + dendritic cells

doi: 10.3389/fimmu.2023.1116435

Figure Lengend Snippet: IgA2 immune complexes selectively increase cytokine production by intestinal CD103 + DCs. Monocytes, macrophages and CD103 + dendritic cells (DCs; 40,000 cells/well) were stimulated with TLR2/1 agonist Pam3CSK4 (Pam3) and serum isolated total IgA (A) , IgA1, or IgA2 (B) , and analyzed for pro-inflammatory cytokine production after 24 hours. TNF production by CD103 + DCs after (co)-stimulation with different concentrations of IgA subclasses was analyzed (C) . Production of different cytokines by CD103 + DCs upon (co-) stimulation was measured by Meso Scale Discovery multiplex assay (D) . TNF production upon alternative PRR stimulation with PGN or LPS was tested by ELISA (E) . Data of multiple donors (A, B, D, E) or a representative example measured in triplicate (C) . *p < 0.05; **p<0.01; ns – not significant.

Article Snippet: Multiplexing human cytokine assay kit U-PLEX (IL-1β, IL-6, IL-8, IL-10, TNF, and CXCL10) were purchased from Meso Scale Discovery (MSD).

Techniques: Isolation, Multiplex Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: IgA2 immune complexes selectively promote inflammation by human CD103 + dendritic cells

doi: 10.3389/fimmu.2023.1116435

Figure Lengend Snippet: Increased cytokine production by IgA2 is at least partially mediated by enhanced gene transcription. mRNA expression of cytokines by CD103 + DCs (40,000 cells/well) upon stimulation by Pam3CSK4 (Pam3), or co-stimulation with IgA1 or IgA2 was measured at different time points. Expression levels of indicated genes were normalized to the mean expression of RACK1 , HPRT1 , and UBB . Data shown of four donors (mean + SD). *p < 0.05.

Article Snippet: Multiplexing human cytokine assay kit U-PLEX (IL-1β, IL-6, IL-8, IL-10, TNF, and CXCL10) were purchased from Meso Scale Discovery (MSD).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: IgA2 immune complexes selectively promote inflammation by human CD103 + dendritic cells

doi: 10.3389/fimmu.2023.1116435

Figure Lengend Snippet: IgA2-induced cytokine production is only partially dependent on FcαRI. Surface expression of FCαRI (CD89) and CD103 was analyzed on in vitro differentiated CD103 + DCs (A) . Expression was compared to unstained cells (grey). Data shown of a representative example of four donors. FCAR mRNA expression after (co-) stimulation with Pam3CSK4 (Pam3) and IgA1 or IgA2 was measured at indicated time points and normalized to the mean expression of RACK1 , HPRT1 , and UBB (data of four donors shown as mean + SD) (B) . Pro-inflammatory cytokine production upon (co-) stimulation (40,000 cells/well) after pre-treatment with anti-CD89 antibodies (FcαRI block) was analyzed (representative example, measured in triplicate) (C) . The effect of anti-CD89 pre-treatment on TNF production was compared to co-stimulation of non-treated CD103 + DCs (D) . Data of six independent experiments. Effects of N-glycosylation on cytokine production was tested for multiple donors using de-glycosylated IgA1 (IgA1-dg) in stimulation experiments with Pam3 (E) . *p < 0.05.

Article Snippet: Multiplexing human cytokine assay kit U-PLEX (IL-1β, IL-6, IL-8, IL-10, TNF, and CXCL10) were purchased from Meso Scale Discovery (MSD).

Techniques: Expressing, In Vitro, Blocking Assay, Glycoproteomics

Journal: Frontiers in Immunology

Article Title: IgA2 immune complexes selectively promote inflammation by human CD103 + dendritic cells

doi: 10.3389/fimmu.2023.1116435

Figure Lengend Snippet: IgA-induced cytokine production can be abolished by small molecule inhibitors targeting downstream signaling molecules for FcαRI. Cytokine production by CD103 + DCs (40,000 cells/well) upon co-stimulation with Pam3CSK4 and IgA1 or IgA2 after treatment with small molecule inhibitors R406 (A, B) , alpelisib, or BX795 (B) specific for Syk, PI3K and TBK1/IKKϵ, respectively, was measured by ELISA. TNF production of cells pre-treated with inhibitors was normalized to that of DMSO controls. Data of a representative example measured in triplicate (mean + SD) (A) or multiple donors (B) . *p < 0.05.

Article Snippet: Multiplexing human cytokine assay kit U-PLEX (IL-1β, IL-6, IL-8, IL-10, TNF, and CXCL10) were purchased from Meso Scale Discovery (MSD).

Techniques: Enzyme-linked Immunosorbent Assay