mab against human tnf alpha  (Hycult Biotech)

Journal: International Journal of Molecular Sciences

Article Title: Human Monocytes Are Suitable Carriers for the Delivery of Oncolytic Herpes Simplex Virus Type 1 In Vitro and in a Chicken Embryo Chorioallantoic Membrane Model of Cancer

doi: 10.3390/ijms24119255

Figure Lengend Snippet: CD14 + - and EGFP-oHSV-1-infected (ICP4 + ) cells can be observed in UM-SCC-11B-derived tumors grown on eggs injected with EGFP-oHSV-1-infected THP-1 cells. The 10× immunohistochemistry (IHC) pictures of UM-SCC-11B tumors grown on the chorioallantoic membrane (CAM) of embryonated chicken eggs. ( a ) Image of a section of an UM-SCC-11B tumor from an egg injected with intravascular EGFP-oHSV-1-infected THP-1 cells (treated) stained with primary anti-CD14 antibody and secondary horseradish peroxidase (HRP)-conjugated antibody; ( b ) image of a section of an UM-SCC-11B tumor from a treated egg stained with primary anti-ICP4 antibody and secondary horseradish peroxidase (HRP)-conjugated antibody; ( c ) 10× IHC picture of a section of an UM-SCC-11B tumor from an egg injected intravascularly with PBS (untreated), stained with primary anti-CD14 antibody and secondary horseradish peroxidase (HRP)-conjugated antibody; ( d ) 10× IHC picture of a section of an UM-SCC-11B tumor from an untreated egg, stained with primary anti-ICP4 antibody and secondary horseradish peroxidase (HRP)-conjugated antibody; ( e , f ) selected untreated ( e ) and treated ( f ) tumor samples were analyzed via fluorescence microscopy upon incubation with a rabbit polyclonal antibody against pan-cytokeratin wide spectrum screening (Agilent Technologies, Santa Clara, CA, USA) followed by a secondary Alexa Fluor 594-conjugated antibody (ThermoFisher Scientific, Waltham, MA, USA). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI, Vector Laboratories, Newark, CA, USA). Pictures were taken with an Eclipse E800 microscope equipped with DS-U1 cooled digital camera (all from Nikon Instruments, Nikon Europe B.V., Amstelveen, The Netherlands). Filters with the following characteristics were used: for EGFP excitation (EX) 480/30, dichroic mirror (DM) 505, barrier (BA) 535/45; for Alexa Fluor 594 fluorophore EX 560/40, DM 595, BA 630/60. Merged imagines are displayed. Scale bars are reported.

Article Snippet: Selected tumor sections, fixed and cut as above described, were immunostained with a mouse monoclonal antibody against CD14 (Abcam, ab181470) and/or a rabbit polyclonal antibody against pan-cytokeratin wide spectrum screening (Agilent, Z0622) as a tumor cell marker.

Techniques: Infection, Derivative Assay, Injection, Immunohistochemistry, Staining, Fluorescence, Microscopy, Incubation, Plasmid Preparation

Journal: International Journal of Molecular Sciences

Article Title: Human Monocytes Are Suitable Carriers for the Delivery of Oncolytic Herpes Simplex Virus Type 1 In Vitro and in a Chicken Embryo Chorioallantoic Membrane Model of Cancer

doi: 10.3390/ijms24119255

Figure Lengend Snippet: CD14 + - and EGFP-oHSV-1-infected (ICP4 + ) cells were minimally observed in chicken embryos’ livers and kidneys following the injection of EGFP-oHSV-1-infected THP-cells. ( a – d ) Panels display 10× IHC pictures of representative liver ( a , b ) or kidney ( c , d ) sections obtained from EGFP-oHSV-1-injected eggs (treated) stained with primary anti-CD14 ( a , c ) or anti-ICP4 ( b , d ) antibody and secondary horseradish peroxidase (HRP)-conjugated antibody. ( e – h ) The same procedure was performed with control (untreated) liver ( e , f ) and kidney ( g , h ) samples. Scale bar is shown.

Article Snippet: Selected tumor sections, fixed and cut as above described, were immunostained with a mouse monoclonal antibody against CD14 (Abcam, ab181470) and/or a rabbit polyclonal antibody against pan-cytokeratin wide spectrum screening (Agilent, Z0622) as a tumor cell marker.

Techniques: Infection, Injection, Staining

Journal: International Journal of Molecular Sciences

Article Title: Human Monocytes Are Suitable Carriers for the Delivery of Oncolytic Herpes Simplex Virus Type 1 In Vitro and in a Chicken Embryo Chorioallantoic Membrane Model of Cancer

doi: 10.3390/ijms24119255

Figure Lengend Snippet: CD14 + - and EGFP-oHSV-1-infected (ICP4 + ) cells are more frequently found in tumors than in the livers and kidneys of treated chicken embryos. Microscopy pictures of UM-SCC-11B tumors, livers and kidneys of chicken embryos were analyzed using LuciaG 5.0 software (Nikon Instruments). The difference in staging grade between tumor and liver or tumor and kidney was statistically significant (*** p < 0.001 and ** p < 0.01, Student’s t -test) for both CD14 ( left panel) and ICP4 ( right panel). Semiquantitative staging was defined as 0 = no positivity; 1 = sparse positive cells; 2 = clumps of positive cells; 3 = widespread cell positivity.

Article Snippet: Selected tumor sections, fixed and cut as above described, were immunostained with a mouse monoclonal antibody against CD14 (Abcam, ab181470) and/or a rabbit polyclonal antibody against pan-cytokeratin wide spectrum screening (Agilent, Z0622) as a tumor cell marker.

Techniques: Infection, Microscopy, Software

Journal: Frontiers in Immunology

Article Title: γδ T Cells May Aggravate Acute Graft-Versus-Host Disease Through CXCR4 Signaling After Allogeneic Hematopoietic Transplantation

doi: 10.3389/fimmu.2021.687961

Figure Lengend Snippet: Assay for the reactivity of γδ T cells in response to stimulation of allogeneic αβ T cells. Mixed lymphocyte reactions (MLRs) were performed by co-culture of purified allogenic γδ T cells with PBMCs depleted of γδ T cells (PBMCΔγδ T cells) isolated from healthy donors, at a ratio of 1:3. Cells were collected on day 5 after co-culture and stained with antibodies against γδ TCR (A–J) , HLA-DR (A, F) , CD38 (B, G) , TNF-α (C, H) , IFN-γ (D, I) and IL-17 (E, J) . Data are expressed as the mean ± SD of triplicate cultures. P values are shown on the graphs.

Article Snippet: Immunophenotypic analyses were conducted on FFPE sections using antibodies against human TNF-α (Boster, China), TCRγδ (Biolegend, USA), CD4 (Affinity Biosciences, China) and CXCR4 (Biosynthesis Biotechnology, China).

Techniques: Co-Culture Assay, Purification, Isolation, Staining

Journal: Frontiers in Immunology

Article Title: γδ T Cells May Aggravate Acute Graft-Versus-Host Disease Through CXCR4 Signaling After Allogeneic Hematopoietic Transplantation

doi: 10.3389/fimmu.2021.687961

Figure Lengend Snippet: Assay for the reactivity of CD4 T cells in response to stimulation of allogeneic γδ T cells. MLRs were performed by co-culture of purified allogenic γδ T cells with PBMCΔγδ T cells isolated from healthy donors, at a ratio of 1:3. Cells were collected on day 5 after co-culture and stained with antibodies against CD4 (A–J) , HLA-DR (A, F) , CD38 (B, G) , TNF-α (C, H) , IFN-γ (D, I) and IL-17 (E, J) . Data are expressed as the mean ± SD of triplicate cultures. P values are shown on the graphs.

Article Snippet: Immunophenotypic analyses were conducted on FFPE sections using antibodies against human TNF-α (Boster, China), TCRγδ (Biolegend, USA), CD4 (Affinity Biosciences, China) and CXCR4 (Biosynthesis Biotechnology, China).

Techniques: Co-Culture Assay, Purification, Isolation, Staining

Journal: Frontiers in Immunology

Article Title: γδ T Cells May Aggravate Acute Graft-Versus-Host Disease Through CXCR4 Signaling After Allogeneic Hematopoietic Transplantation

doi: 10.3389/fimmu.2021.687961

Figure Lengend Snippet: Assay for the reactivity of CD8 T cells in response to stimulation of allogeneic γδ T cells. MLRs were performed by co-culture of purified allogenic γδ T cells with PBMCΔγδ T cells isolated from healthy donors, at a ratio of 1:3. Cells were collected on day 5 after co-culture and stained with antibodies against CD8 (A–J) , HLA-DR (A, F) , CD38 (B, G) , TNF-α (C, H) , IFN-γ (D, I) and IL-17 (E, J) . Data are expressed as the mean ± SD of triplicate cultures. P values are shown on the graphs.

Article Snippet: Immunophenotypic analyses were conducted on FFPE sections using antibodies against human TNF-α (Boster, China), TCRγδ (Biolegend, USA), CD4 (Affinity Biosciences, China) and CXCR4 (Biosynthesis Biotechnology, China).

Techniques: Co-Culture Assay, Purification, Isolation, Staining

Journal: Frontiers in Immunology

Article Title: γδ T Cells May Aggravate Acute Graft-Versus-Host Disease Through CXCR4 Signaling After Allogeneic Hematopoietic Transplantation

doi: 10.3389/fimmu.2021.687961

Figure Lengend Snippet: Migrations of human γδ T cells and CD4 T cells expressing CXCR4 in mice with or without aGVHD. Immunodeficient mice were transplanted with human PBMCs as described. The proportion of human CD45-positive cells was detected in mouse peripheral blood by flow cytometry at 2 and 3 weeks after injection (A) . Histopathological examination was performed on biopsy specimens of liver obtained from transplanted mice. Histopathological feature and expression of TNF-α in liver were detected by hematoxylin and eosin (H&E, ×100) staining and immunohistochemistry analyses, ×200 (B) . Three adjacent sections of liver biopsies were stained with antibodies against CXCR4, TCRγδ and CD4, ×200 (C) . CXCR4-expressing γδ T and CD4 T cells were detected by flow cytometry in the skin (D, E) and liver (F, G) of mice with or without aGVHD. n = 4, P values are shown on the graphs.

Article Snippet: Immunophenotypic analyses were conducted on FFPE sections using antibodies against human TNF-α (Boster, China), TCRγδ (Biolegend, USA), CD4 (Affinity Biosciences, China) and CXCR4 (Biosynthesis Biotechnology, China).

Techniques: Expressing, Flow Cytometry, Injection, Staining, Immunohistochemistry

Journal: International Ophthalmology

Article Title: New pharmacotherapy options for noninfectious posterior uveitis

doi: 10.1007/s10792-021-01763-8

Figure Lengend Snippet: Intravitreally administered compounds in clinical use or in development for posterior segment uveitis

Article Snippet: Adalimumab , Human monoclonal IgG antibody against TNF-alpha , Case series (off-label for intravitreal use) (Ref…….) , AbbVie.

Techniques: Permeability, Migration, Activation Assay, Cell Differentiation, Inhibition, Recombinant