Journal: Mucosal immunology

Article Title: IL-13-mediated immunological control of enterochromaffin cell hyperplasia and serotonin production in the gut.

doi: 10.1038/mi.2012.58

Figure Lengend Snippet: Figure 7 Phorbol ester (phorbol-12-myristate-13-acetate (PMA)) induces the release of 5-hydroxytryptamine (5-HT) from BON cells after 1 h in a dose-dependent manner, but recombinant human interleukin- 13 (rhIL-13) at three different concentrations does not. BON cells were treated with serum-free media (Control) containing either PMA or rhIL- 13 for 1 h at 37 ° C. Media were collected and 5-HT concentration was determined by enzyme-linked immunosorbent assay (ELISA). Each bar represents mean ± s.e.m. of six biological replicates (wells). a, b and c represent statistical significance. a is significantly different from b and c, b is significantly different than a and c, and all the conditions with c are not significantly different from each other.

Article Snippet: Colon samples were analyzed using a mouse IL-13 and IL-4 ELISA kit (R & D Systems, Minneapolis, MN) according to the manufacturer ’ s instructions.

Techniques: Recombinant, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of the American Society of Nephrology

Article Title: Complement Activation Regulates the Capacity of Proximal Tubular Epithelial Cell to Stimulate Alloreactive T Cell Response

doi: 10.1097/01.asn.0000135974.06478.7b

Figure Lengend Snippet: Figure 6. Detection of IL-2 and IFN- by ELISA. IFN-–stimulated PTEC were treated with NS or HIS for 30 min and subsequently co-cultured with T cells. (A and B) Supernatants were collected from co-culture of 2 105 PTEC with 2 105 T cells at various time points and assayed for IL-2 and IFN- by sandwich ELISA. P values are for comparisons between NS and HIS treatment. *P 0.05; **P 0.005; ***P 0.0003.

Article Snippet: Sandwich ELISA was performed using ELISA Kit for mouse IFN- or IL-2 (R&D Systems, Oxford, UK) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Co-Culture Assay, Sandwich ELISA

Journal: Journal of the American Society of Nephrology

Article Title: Complement Activation Regulates the Capacity of Proximal Tubular Epithelial Cell to Stimulate Alloreactive T Cell Response

doi: 10.1097/01.asn.0000135974.06478.7b

Figure Lengend Snippet: Figure 7. Relative involvement of CD4 and CD8 T cells in complement-enhanced alloreactive T cell response. IFN-–stimulated and NS- or HIS-treated PTEC were co-cultured with purified allo- reactive CD4 or CD8 T cells. Supernatants were collected from co-culture of PTEC with T cells after 72 h of co-culturing and assayed for IFN- by sandwich ELISA. P values are for comparisons between NS and HIS treatment. **P 0.005.

Article Snippet: Sandwich ELISA was performed using ELISA Kit for mouse IFN- or IL-2 (R&D Systems, Oxford, UK) according to the manufacturer’s instructions.

Techniques: Cell Culture, Purification, Co-Culture Assay, Sandwich ELISA

Journal: Journal of the American Society of Nephrology

Article Title: Complement Activation Regulates the Capacity of Proximal Tubular Epithelial Cell to Stimulate Alloreactive T Cell Response

doi: 10.1097/01.asn.0000135974.06478.7b

Figure Lengend Snippet: Figure 8. Complement-mediated PTEC stimulation of alloreactive T cell response can be inhibited with CTLA4-Ig. IFN-–stimulated and NS- or HIS-treated PTEC were co-cultured with purified alloreactive CD4 T cells in medium alone or in the presence of CTLA4-Ig (5 g/ml) or control Ig (human IgG1, 5 g/ml). Supernatants were collected from co-culture of PTEC with T cells after 72 h of co- culturing and assayed for IFN- by sandwich ELISA. P values are for comparisons between CTLA4-Ig treatment and medium alone or control Ig treatment. **P 0.005.

Article Snippet: Sandwich ELISA was performed using ELISA Kit for mouse IFN- or IL-2 (R&D Systems, Oxford, UK) according to the manufacturer’s instructions.

Techniques: Cell Culture, Purification, Control, Co-Culture Assay, Sandwich ELISA

Journal: Frontiers in Microbiology

Article Title: Staphylococcus aureus increases Prostaglandin E 2 secretion in cow neutrophils by activating TLR2, TLR4, and NLRP3 inflammasome signaling pathways

doi: 10.3389/fmicb.2023.1163261

Figure Lengend Snippet: Staphylococcus aureus lipoprotein is an important protein that can induce the secretion of PGE 2 and cytokines by neutrophils in dairy cows and can also activate TLR2, TLR4, and NLRP3 receptors and their mediated MAPK and Caspase-1 signaling pathways. ELISA results showed the secretion levels of cytokines IL-1β (A) , IL-6 (B) , IL-10 (C) , IL-8 (D) , and PGE 2 (E) in the uninfected, SA113, and Δ lgt groups after neutrophil infection with MOI 3:1 and MOI 10:1. Western blot analysis revealed the activation of the MAPK (F–H) and Caspase-1 (I, J) signaling pathways. TLR2, TLR4, and NLRP3 receptor genes (K–M) and protein (N–Q) expressions were detected by qRT-PCR and Western blot. ns P > 0.05, * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Fetal bovine serum (FBS), RPMI1640 medium, and PBS (Hyclon, Logen, UT); dairy cow peripheral blood neutrophil isolation kit (Hao Yang, Tian Jing, China); MH broth (Oxoid, Hampshire, UK); TLR2 receptor inhibitor (C29, MCE, China), TLR4 receptor inhibitor (TAK-242, MCE, China), NLRP3 receptor inhibitor (MCC950, MCE, China); COX-2 inhibitor (CAY10404), mPGES-1 inhibitor (MF63) (Cayman Chemical, MI, United States); gentamicin sulfate(Promega, Wisconsin, United States); Bovine IL-1β, IL-10, IL-8 ELISA Kits (Kingfisher, Biotech); Bovine IL-6 ELISA Kit (R&D Systems, California, United States); PGE 2 ELISA Kit (Cayman Chemical, MI, United States); M-PER mammalian protein extraction reagent, Halt protease inhibitor, pre-stained protein ladders, blocking buffer, starting block T20 (TBS), LIVE/DEAD Bac Light Bacterial Viability Kits (Thermo Fisher Scientific); SDS-PAGE loading buffer (Takara, Shiga, Japan); Pierce BCA Protein Assay Kit (Rockford, United States); SDS-PAGE gel electrophoresis kit (Solarbio, Beijing, China); Axy Prep Multisource Total mRNA Miniprep Kit (Axygen Scientific, Union City, United States); Primer Script RT Master Mix (Takara); SYBR Green Master (Rox) (Roche, Basel, Switzerland).

Techniques: Protein-Protein interactions, Enzyme-linked Immunosorbent Assay, Infection, Western Blot, Activation Assay, Quantitative RT-PCR

Journal: Frontiers in Microbiology

Article Title: Staphylococcus aureus increases Prostaglandin E 2 secretion in cow neutrophils by activating TLR2, TLR4, and NLRP3 inflammasome signaling pathways

doi: 10.3389/fmicb.2023.1163261

Figure Lengend Snippet: The secretion of PGE 2 and cytokines mediated by neutrophils depends on TLR2, TLR4, and NLRP3 receptors. ELISA results showed the secretion levels of cytokines IL-1β (A) , IL-6 (B) , IL-8 (C) , IL-10 (D) , and PGE2 (E) in the SA113, C29, TAK-242, and MCC950 groups after neutrophils infection with MOI 3:1 and MOI 10:1. Western blot analysis revealed the activation of the MAPK (F, G, I) and Caspase-1 (H, J) signaling pathways. ns P > 0.05, * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Fetal bovine serum (FBS), RPMI1640 medium, and PBS (Hyclon, Logen, UT); dairy cow peripheral blood neutrophil isolation kit (Hao Yang, Tian Jing, China); MH broth (Oxoid, Hampshire, UK); TLR2 receptor inhibitor (C29, MCE, China), TLR4 receptor inhibitor (TAK-242, MCE, China), NLRP3 receptor inhibitor (MCC950, MCE, China); COX-2 inhibitor (CAY10404), mPGES-1 inhibitor (MF63) (Cayman Chemical, MI, United States); gentamicin sulfate(Promega, Wisconsin, United States); Bovine IL-1β, IL-10, IL-8 ELISA Kits (Kingfisher, Biotech); Bovine IL-6 ELISA Kit (R&D Systems, California, United States); PGE 2 ELISA Kit (Cayman Chemical, MI, United States); M-PER mammalian protein extraction reagent, Halt protease inhibitor, pre-stained protein ladders, blocking buffer, starting block T20 (TBS), LIVE/DEAD Bac Light Bacterial Viability Kits (Thermo Fisher Scientific); SDS-PAGE loading buffer (Takara, Shiga, Japan); Pierce BCA Protein Assay Kit (Rockford, United States); SDS-PAGE gel electrophoresis kit (Solarbio, Beijing, China); Axy Prep Multisource Total mRNA Miniprep Kit (Axygen Scientific, Union City, United States); Primer Script RT Master Mix (Takara); SYBR Green Master (Rox) (Roche, Basel, Switzerland).

Techniques: Enzyme-linked Immunosorbent Assay, Infection, Western Blot, Activation Assay, Protein-Protein interactions

Journal: Frontiers in Microbiology

Article Title: Staphylococcus aureus increases Prostaglandin E 2 secretion in cow neutrophils by activating TLR2, TLR4, and NLRP3 inflammasome signaling pathways

doi: 10.3389/fmicb.2023.1163261

Figure Lengend Snippet: PGE 2 can affect the secretion of pro-inflammatory factors, anti-inflammatory factors, and chemokines in neutrophils of cows infected with S. aureus . ELISA results showed that IL-1β (A) , IL-6 (B) , IL-8 (C) , IL-10 (D) , and PGE 2 (E) were secreted in the SA113 group, the uninfected group, the CAY10404 group, and the MF63 group. ns P > 0.05, * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Fetal bovine serum (FBS), RPMI1640 medium, and PBS (Hyclon, Logen, UT); dairy cow peripheral blood neutrophil isolation kit (Hao Yang, Tian Jing, China); MH broth (Oxoid, Hampshire, UK); TLR2 receptor inhibitor (C29, MCE, China), TLR4 receptor inhibitor (TAK-242, MCE, China), NLRP3 receptor inhibitor (MCC950, MCE, China); COX-2 inhibitor (CAY10404), mPGES-1 inhibitor (MF63) (Cayman Chemical, MI, United States); gentamicin sulfate(Promega, Wisconsin, United States); Bovine IL-1β, IL-10, IL-8 ELISA Kits (Kingfisher, Biotech); Bovine IL-6 ELISA Kit (R&D Systems, California, United States); PGE 2 ELISA Kit (Cayman Chemical, MI, United States); M-PER mammalian protein extraction reagent, Halt protease inhibitor, pre-stained protein ladders, blocking buffer, starting block T20 (TBS), LIVE/DEAD Bac Light Bacterial Viability Kits (Thermo Fisher Scientific); SDS-PAGE loading buffer (Takara, Shiga, Japan); Pierce BCA Protein Assay Kit (Rockford, United States); SDS-PAGE gel electrophoresis kit (Solarbio, Beijing, China); Axy Prep Multisource Total mRNA Miniprep Kit (Axygen Scientific, Union City, United States); Primer Script RT Master Mix (Takara); SYBR Green Master (Rox) (Roche, Basel, Switzerland).

Techniques: Infection, Enzyme-linked Immunosorbent Assay