Journal: Journal of Immunotherapy (Hagerstown, Md. : 1997)

Article Title: NUSAP1 Promotes Immunity and Apoptosis by the SHCBP1/JAK2/STAT3 Phosphorylation Pathway to Induce Dendritic Cell Generation in Hepatocellular Carcinoma

doi: 10.1097/CJI.0000000000000531

Figure Lengend Snippet: NUSAP1 intervene in the differentiation of PBMCs around HCC into DC. (A) Elisa assay test IL-6 levels in the cell supernatant; (B and C) After co-culture, the expression levels of the surface factors CD1a and CD86 of PBMCs were determined by flow cytometry; (D and E) After LMT-28 intervene, the expression levels of the surface factors CD1a and CD86 of PBMCs were determined by flow cytometry; (F and G) After Coumermycin A1 intervene, the expression levels of the surface factors CD1a and CD86 of PBMCs were determined by flow cytometry; (H) After co-culture, WB detect Apoptosis-related proteins in HCC.

Article Snippet: According to the instructions of IL-6 ELISA kits purchased from Boster Biological Co. (Cat# MEK2004; Boster, USA), IL-6 level in the cultured cell supernatants of the different groups were detected, respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Expressing, Flow Cytometry

Journal: BMC Veterinary Research

Article Title: Quick and improved immune responses to inactivated H9N2 avian influenza vaccine by purified active fraction of Albizia julibrissin saponins

doi: 10.1186/s12917-020-02648-1

Figure Lengend Snippet: Effect of AJSAF on H9–Ag-specific IgG and its isotype antibody titers in the immunized mice. Mice were s.c. injected with 0.2 ml of IH9V (10 7 TCID 50 /0.1 ml) alone or containing Quil A or AJSAF, or 0.2 ml of CH9V (10 7 TCID 50 /0.1 ml) on days 1 and 15. Sera were collected 14 days after the boosting immunization, and serum H9–Ag-specific IgG, IgG1, IgG2a, and IgG2b antibody titers were measured by ELISA. The values are presented as means ± SD ( n = 6). Significant differences with IH9V alone group were designated as * P < 0.05, † P < 0.01, and ‡ P < 0.001; those with CH9V group as § P < 0.05, # P < 0.01, and ¶ P < 0.001

Article Snippet: Assoc., Birmingham, AL, USA; rabbit anti-chicken IgY horseradish peroxidase conjugate was from Promega Corporation, Madison, WI, USA; mouse cytokine detecting ELISA kits were from Wuhan Boster Bio-Tech.

Techniques: Injection, Enzyme-linked Immunosorbent Assay

Journal: BMC Veterinary Research

Article Title: Quick and improved immune responses to inactivated H9N2 avian influenza vaccine by purified active fraction of Albizia julibrissin saponins

doi: 10.1186/s12917-020-02648-1

Figure Lengend Snippet: Effect of AJSAF on haemagglutination inhibition (HI) and H9–Ag-specific IgY antibody levels in the immunized chicken. The chickens were prime- and boost-immunized with 0.4 ml of IH9V (10 7 TCID 50 /0.1 ml) alone or containing AJSAF, or 0.4 ml of CH9V (10 7 TCID 50 /0.1 ml) at 3-week interval. Sera were collected on designated days post-immunization, and the serum HI antibody titers ( a ) and H9–Ag-specific IgY antibody ( b ) levels were measured by HI assay and ELISA, respectively. The values are presented as means ± SD ( n = 30). Significant differences with IH9V alone group were designated as * P < 0.05, † P < 0.01, and ‡ P < 0.001

Article Snippet: Assoc., Birmingham, AL, USA; rabbit anti-chicken IgY horseradish peroxidase conjugate was from Promega Corporation, Madison, WI, USA; mouse cytokine detecting ELISA kits were from Wuhan Boster Bio-Tech.

Techniques: Inhibition, HI Assay, Enzyme-linked Immunosorbent Assay

Journal: BMC Veterinary Research

Article Title: Quick and improved immune responses to inactivated H9N2 avian influenza vaccine by purified active fraction of Albizia julibrissin saponins

doi: 10.1186/s12917-020-02648-1

Figure Lengend Snippet: Effect of AJSAF on cytokine secretion from splenocytes in the immunized mice. Splenocytes were incubated with Con A (48 h) or H9–Ag (72 h), and the supernatants were collected for detecting IL-2, IFN-γ, and IL-10 levels using ELISA kits. The values are presented as means ± SD ( n = 6). Significant differences with IH9V alone group were designated as * P < 0.05, † P < 0.01, and ‡ P < 0.001; those with CH9V group as # P < 0.01 and ¶ P < 0.001

Article Snippet: Assoc., Birmingham, AL, USA; rabbit anti-chicken IgY horseradish peroxidase conjugate was from Promega Corporation, Madison, WI, USA; mouse cytokine detecting ELISA kits were from Wuhan Boster Bio-Tech.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: Molecular Therapy. Nucleic Acids

Article Title: Modulation of experimental acute lung injury by exosomal miR-7704 from mesenchymal stromal cells acts through M2 macrophage polarization

doi: 10.1016/j.omtn.2023.102102

Figure Lengend Snippet: Effects on macrophage polarization of IT-sMSC-exos Twenty-four hours after the addition of MSC-exo and I/T-MSC-exo, macrophages were harvested and measured using real-time PCR (A) ( Cd206 , Cd86 , Arg1 , Ifn-γ , Il-6 , Tnf , and Il-1β ) (each group n = 6). (B) Inflammatory cytokine expression (IFN-γ, IL-6, TNF, and IL-1β) of macrophages was measured using ELISA (each group n = 6). Results are presented as mean ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001; ns, not significant, one-way ANOVA.

Article Snippet: Cell culture CM or BALF from mice were collected and analyzed using the Multiplex ELISA Kit for Mouse Cytokine Panel 1 (6-Plex, MEK1011, BOSTER) following the manufacturer’s instructions.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Molecular Therapy. Nucleic Acids

Article Title: Modulation of experimental acute lung injury by exosomal miR-7704 from mesenchymal stromal cells acts through M2 macrophage polarization

doi: 10.1016/j.omtn.2023.102102

Figure Lengend Snippet: Analysis of effects on macrophage polarity by miR-7704 (A) miR-7704 expression in different phenotypes of macrophages (each group n = 6). The transfection efficiency of miR-7704 in M1 macrophages (M1+miR-7704) and NC (M1+NC) was analyzed using qPCR (B) and immunofluorescence staining conjugated with Cy5 (C) Scale bar, 90 μm (each group n = 6). (D) Macrophage surface makers (CD86 and CD206) and their quantified percentage (E) were analyzed using flow cytometry (each group n = 6). (F) Gene expression of M1, M1+NC, M1+miR-7704, and M2 macrophages. M1 marker genes ( Cd86 , Cd80 , Il-1β , TNF , IFN-γ , Il-6 , Il-18 , and inos ); M2 related genes ( Cd206 , Cd163 , Arg1 , and Il-1 0) (each group n = 6). (G) Cytokine expression of M1, M1+NC, M1+miR-7704, and M2 through ELISA (IL1-β, IL-6, IFN-γ, and TNF; each group n = 6). Results are presented as mean ± SD. Statistical analyses were performed using one-way ANOVA. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001; ns, not significant.

Article Snippet: Cell culture CM or BALF from mice were collected and analyzed using the Multiplex ELISA Kit for Mouse Cytokine Panel 1 (6-Plex, MEK1011, BOSTER) following the manufacturer’s instructions.

Techniques: Expressing, Transfection, Immunofluorescence, Staining, Flow Cytometry, Gene Expression, Marker, Enzyme-linked Immunosorbent Assay

Journal: Molecular Therapy. Nucleic Acids

Article Title: Modulation of experimental acute lung injury by exosomal miR-7704 from mesenchymal stromal cells acts through M2 macrophage polarization

doi: 10.1016/j.omtn.2023.102102

Figure Lengend Snippet: Amelioration of LPS-induced ALI by miR-7704 (A) Schematic showing the experimental design of ALI, control (PBS/PBS), LPS (LPS/PBS), miR-7704 (LPS/miR-7704), IT-sMSC-exo (LPS/IT-sMSC-exo), IT-sMSC-exo plus anti-miR-7704 (LPS/IT-sMSC-exo+αmiR) administration, and analysis (each group n = 6). (B) miR-7704 expression in different groups (each group n = 6). (H) BALF protein concentration in different groups (each group n = 6). (C) Representative H&E staining in different groups. Scale bar, 200 μm. (D) Lung injury score in different groups. Representative IHC staining for CD86 (E), CD206 (F), and F4/80 (G) expression in different groups under 200× magnification. Brown: positively stained cells. Scale bar, 200 μm. (H) Protein concentration of BALF in different groups. (I) Cytokine expression of BALF measured using ELISA (IL1-β, IL-6, IFN-γ, TNF, and IL-10). (J) Inflammatory-related gene expression of lung tissue through real-time PCR in different groups (each group n = 6). (K) Immunoblotting of lung tissue protein (MyD88, STAT1, and pSTAT1). Results are presented as mean ± SD. Statistical analyses were performed using one-way ANOVA. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001; ns, not significant.

Article Snippet: Cell culture CM or BALF from mice were collected and analyzed using the Multiplex ELISA Kit for Mouse Cytokine Panel 1 (6-Plex, MEK1011, BOSTER) following the manufacturer’s instructions.

Techniques: Control, Expressing, Protein Concentration, Staining, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Gene Expression, Real-time Polymerase Chain Reaction, Western Blot