Journal: Science Advances

Article Title: Sympathetic nerve aggravates autoimmune skin disease via NE–adrenergic receptor axis: Neuroimmune cross-talk insights from vitiligo

doi: 10.1126/sciadv.aea7017

Figure Lengend Snippet: ( A and B ) Flow cytometry and statistical analysis of CD8 + CD45 + T cells in the mouse tail epidermis and dermis of vehicle, 6-OHDA, and 6-OHDA + NE groups ( n =3 per group). ( C and D ) Flow cytometry and statistical analysis of IFN-γ + CD8 + T cells in the mouse tail epidermis and dermis of vehicle, 6-OHDA, and 6-OHDA + NE groups ( n =3 per group). ( E and F ) Flow cytometry and statistical analysis of Granzyme B + CD8 + T cells in the mouse tail epidermis and dermis of vehicle, 6-OHDA, and 6-OHDA + NE groups ( n =3 per group). ( G ) The secretion levels of CXCL9, CXCL10, IL-6, and IL-15 in the tail skin of mice in the vehicle, 6-OHDA, and 6-OHDA + NE groups were detected by ELISA ( n =4 per group). All vehicle mice were only treated with 0.1% ascorbic acid in 0.9% sterile NaCl. Error bars represent mean ± SD from three independent experiments. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: ELISA analysis on serum samples, skin samples, and cell culture supernatants were performed using the NE ELISA Kit (E-EL-0047c, Elabscience, Wuhan, China), Human CXCL9 ELISA Kit (EHC114.96, Neobioscience Technology Co, Ltd., China), Human CXCL10 ELISA Kit (EHC157.96, Neobioscience Technology Co, Ltd., China), Human IL-6 ELISA Kit (EHC007.96, Neobioscience Technology Co, Ltd., China), Human IL-15 ELISA Kit (EHC013.96, Neobioscience Technology Co, Ltd., China), Mouse CXCL9 ELISA Kit (E-EL-M3077, Elabscience, Wuhan, China), Mouse CXCL10 ELISA Kit (E-EL-M0021, Elabscience, Wuhan, China), Mouse IL-6 ELISA Kit (EMC004.96, Neobioscience Technology Co, Ltd., China), and Mouse IL-15 ELISA Kit (EMC126.96, Neobioscience Technology Co, Ltd., China) following the manufacturer’s instructions.

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay, Sterility

Journal: Science Advances

Article Title: Sympathetic nerve aggravates autoimmune skin disease via NE–adrenergic receptor axis: Neuroimmune cross-talk insights from vitiligo

doi: 10.1126/sciadv.aea7017

Figure Lengend Snippet: ( A ) Volcano plots illustrated the expression of adrenergic receptors on fibroblasts in human skin, including ADRA1A, ADRA1B, ADRA1D, ADRA2A, ADRA2B, ADRA2C, ADRB1, ADRB2, and ADRB3. ( B ) Bubble map illustrated the up-regulation of ADRA2A expression on fibroblasts in the lesional skin of patients with vitiligo and the normal skin of healthy controls. ( C ) Representative immunofluorescence images of fibroblasts, ADRA2A, CXCL9, CXCL10, and CD8 + T cells from the normal skin of the healthy control and the lesional skin of the patient with vitiligo ( n =3 per group). Nuclei were counterstained with DAPI (blue). Scale bars, 50 μm. ( D ) Immunofluorescence analysis of ADRA2A (red) expression in BJ cells after treatment with 5 μM NE for 48 hours. Cell nuclei were stained with DAPI (blue). Scale bar, 50 μm. ( E ) Secretion levels of CXCL9, CXCL10, IL-6, and IL-15 in BJ cells were detected by ELISA with treatment with aposcopolamine (Apos) or NE ( n =3 per group). ( F ) Secretion levels of CXCL9, CXCL10, IL-6, and IL-15 in BJ cells with ADRA2A siRNA or control siRNA were detected by ELISA ( n =3 per group). Error bars represent mean ± SD from three independent experiments. * P < 0.05, ** P < 0.01, and *** P < 0.001; ns, not significant.

Article Snippet: ELISA analysis on serum samples, skin samples, and cell culture supernatants were performed using the NE ELISA Kit (E-EL-0047c, Elabscience, Wuhan, China), Human CXCL9 ELISA Kit (EHC114.96, Neobioscience Technology Co, Ltd., China), Human CXCL10 ELISA Kit (EHC157.96, Neobioscience Technology Co, Ltd., China), Human IL-6 ELISA Kit (EHC007.96, Neobioscience Technology Co, Ltd., China), Human IL-15 ELISA Kit (EHC013.96, Neobioscience Technology Co, Ltd., China), Mouse CXCL9 ELISA Kit (E-EL-M3077, Elabscience, Wuhan, China), Mouse CXCL10 ELISA Kit (E-EL-M0021, Elabscience, Wuhan, China), Mouse IL-6 ELISA Kit (EMC004.96, Neobioscience Technology Co, Ltd., China), and Mouse IL-15 ELISA Kit (EMC126.96, Neobioscience Technology Co, Ltd., China) following the manufacturer’s instructions.

Techniques: Expressing, Immunofluorescence, Control, Staining, Enzyme-linked Immunosorbent Assay

Journal: Science Advances

Article Title: Sympathetic nerve aggravates autoimmune skin disease via NE–adrenergic receptor axis: Neuroimmune cross-talk insights from vitiligo

doi: 10.1126/sciadv.aea7017

Figure Lengend Snippet: ( A ) Volcano plots illustrated the expression of adrenergic receptors on keratinocytes in human skin, including ADRA1A, ADRA1B, ADRA1D, ADRA2A, ADRA2B, ADRA2C, ADRB1, ADRB2, and ADRB3. ( B ) Bubble map illustrated the up-regulation of ADRB2 expression on keratinocytes in the lesional skin of patients with vitiligo and the normal skin of healthy controls. ( C ) Representative immunofluorescence images of keratinocytes, ADRB2, CXCL9, CXCL10, and CD8 + T cells from the normal skin of the healthy control and the lesional skin of the patient with vitiligo ( n =3 per group). Nuclei were counterstained with DAPI (blue). Scale bars, 50 μm. ( D ) Immunofluorescence analysis of ADRB2 (red) expression in keratinocytes after treatment with 5 μM NE for 48 hours. Cell nuclei were stained with DAPI (blue). Scale bar, 50 μm. ( E ) Secretion levels of CXCL9, CXCL10, IL-6, and IL-15 in keratinocytes were detected by ELISA with treatment with NE or ICI ( n =3 per group). ( F ) Secretion levels of CXCL9, CXCL10, IL-6, and IL-15 in keratinocytes with ADRB2 siRNA or control siRNA were detected by ELISA ( n =3 per group). Error bars represent mean ± SD from three independent experiments. * P < 0.05, ** P < 0.01, and *** P < 0.001; ns, not significant.

Article Snippet: ELISA analysis on serum samples, skin samples, and cell culture supernatants were performed using the NE ELISA Kit (E-EL-0047c, Elabscience, Wuhan, China), Human CXCL9 ELISA Kit (EHC114.96, Neobioscience Technology Co, Ltd., China), Human CXCL10 ELISA Kit (EHC157.96, Neobioscience Technology Co, Ltd., China), Human IL-6 ELISA Kit (EHC007.96, Neobioscience Technology Co, Ltd., China), Human IL-15 ELISA Kit (EHC013.96, Neobioscience Technology Co, Ltd., China), Mouse CXCL9 ELISA Kit (E-EL-M3077, Elabscience, Wuhan, China), Mouse CXCL10 ELISA Kit (E-EL-M0021, Elabscience, Wuhan, China), Mouse IL-6 ELISA Kit (EMC004.96, Neobioscience Technology Co, Ltd., China), and Mouse IL-15 ELISA Kit (EMC126.96, Neobioscience Technology Co, Ltd., China) following the manufacturer’s instructions.

Techniques: Expressing, Immunofluorescence, Control, Staining, Enzyme-linked Immunosorbent Assay

Journal: Journal of Veterinary Science

Article Title: Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine

doi: 10.4142/jvs.2014.15.1.99

Figure Lengend Snippet: Humoral immune responses in vaccinated piglets from the different groups detected with a commercial ELISA kit.

Article Snippet: To further evaluate cellular immune responses, IFN-γ levels in peripheral blood isolated from the vaccinated piglets at 35 dpi were measured using a commercial ELISA kit (Pig Interferon-γ, IFN-γ ELISA Kit; CUSABIO, USA) according to the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: The veterinary quarterly

Article Title: Holstein × Montbéliarde-sired F1 generation crossbred female calves have an increased cellular immune response potential compared with purebred Holsteins.

doi: 10.1080/01652176.2024.2435982

Figure Lengend Snippet: Figure 2. Gating strategies to define bovine T cell subsets and their IFN-γ expression. Mononuclear cells were isolated from blood of 3 months-old female calf and surface stained with antibody cocktails (CD4, CD8, CD44, IFN-γ) after PMA/ionomycin mixture stimulation. The lymphocytes are chosen with FSC versus SSC (A) and then singlets are gated using FSC-a and FSC-H (B). By outputting CD4 versus CD8, CD4+ CD8-, CD8+ CD4- are identified, respectively (C). The CD4+ CD8- T cells expressing IFN-γ (D). The CD8+ CD4- T cells expressing IFN-γ (G). CD4+ CD8- T cells and CD4+ CD8- T cells can be further subdivided into CD44+ and CD44- subsets (E and F), respectively. The lymphocytes expressing IFN-γ (H).

Article Snippet: Monoclonal antibodies (mAb) specific for bovine TCRγδ (WC1), CD8, CD4 and IFN-γ with different fluorochrome conjugates were purchased from BIO-RAD (BIO-RAD, Shanghai, China) and mAb to human CD27, mouse CD44 were purchased from Biolegend (Biolegend, San Diego, USA) (Table 1).

Techniques: Expressing, Isolation, Staining

Journal: The veterinary quarterly

Article Title: Holstein × Montbéliarde-sired F1 generation crossbred female calves have an increased cellular immune response potential compared with purebred Holsteins.

doi: 10.1080/01652176.2024.2435982

Figure Lengend Snippet: Figure 5. Differences of IFN-γ expression in lymphocytes and their T cells of MH and HO calves. Representative dot-plots depict IFN-γ+ T cell subsets in CD4+ (a), CD8+ (B) T cells and total lymphocytes (C). Representative histograms showed the expressions of IFN-γ in lymphocytes (D), CD4+ (E) and CD8+ (F) T cells in the whole blood of MH and HO calves. Data were shown in LSM ± SEM (n = 25). *p < 0.05, **p < 0.01.

Article Snippet: Monoclonal antibodies (mAb) specific for bovine TCRγδ (WC1), CD8, CD4 and IFN-γ with different fluorochrome conjugates were purchased from BIO-RAD (BIO-RAD, Shanghai, China) and mAb to human CD27, mouse CD44 were purchased from Biolegend (Biolegend, San Diego, USA) (Table 1).

Techniques: Expressing

Journal: The veterinary quarterly

Article Title: Holstein × Montbéliarde-sired F1 generation crossbred female calves have an increased cellular immune response potential compared with purebred Holsteins.

doi: 10.1080/01652176.2024.2435982

Figure Lengend Snippet: Figure 6. The expression of IFN-γ was derived from subsets of CD4+ and CD8− T cells, characterized by CD44+ cells rather than CD44− cells. Representative dot-plots depict CD4+ (A) and CD8- (B) T cells subsets of IFN-γ and CD44 from MH (left panel) and HO (right panel) calves. Numbers represent the percentages of cells in each quadrant. Bar graphs showed the expressions of IFN-γ by CD44+ or CD44− subsets in CD4+ (C) and CD8+ (D) T cells of MH and HO calves. Data were shown in LSM ± SEM (n = 25). Labels (a, b, c and d) of different letters indicated a significant difference between any two different sets of data (p < 0.01).

Article Snippet: Monoclonal antibodies (mAb) specific for bovine TCRγδ (WC1), CD8, CD4 and IFN-γ with different fluorochrome conjugates were purchased from BIO-RAD (BIO-RAD, Shanghai, China) and mAb to human CD27, mouse CD44 were purchased from Biolegend (Biolegend, San Diego, USA) (Table 1).

Techniques: Expressing, Derivative Assay

Journal: Vaccines

Article Title: Mannose-Modified Chitosan-Nanoparticle-Based Salmonella Subunit OralVaccine-Induced Immune Response and Efficacy in a Challenge Trial in Broilers

doi: 10.3390/vaccines8020299

Figure Lengend Snippet: Antibodies used for lymphocyte cell surface and intracellular IFNγ immunostaining.

Article Snippet: Rabbit anti-chicken IFNγ , Cat# AHP945Z; Bio-Rad; Hercules, CA, USA.

Techniques: Immunostaining