Journal: FASEB bioAdvances

Article Title: Manipulation of the tumor microenvironment by cytokine gene transfection enhances dendritic cell‐based immunotherapy

doi: 10.1096/fba.2019-00052

Figure Lengend Snippet: Lung metastasis in the LM8‐inoculated mice

Article Snippet: Cells infiltrating tumor tissues were evaluated by immunohistochemistry using following primary Abs: for IFNγ, Bioss Cat# bs‐0480R‐PE‐Cy7, RRID:AB_11092092; for CD40L, Biorbyt Cat# orb10326, RRID:AB_10746923; for CD8, clone 53‐6.7, Thermo Fisher Scientific Cat# 14‐0081‐81, RRID:AB_467086; for Foxp3, clone FJK‐16s, Thermo Fisher Scientific Cat# 14‐5773‐80, RRID:AB_467575; for Iba1, Wako Pure Chemical Industries Inc, Cat# 019‐19741, RRID:AB_839504; for granzyme B, Spring Bioscience Cat# E2580, RRID:AB_1661202.

Techniques:

Journal: FASEB bioAdvances

Article Title: Manipulation of the tumor microenvironment by cytokine gene transfection enhances dendritic cell‐based immunotherapy

doi: 10.1096/fba.2019-00052

Figure Lengend Snippet: Effect of IFNγ and CD40L (Comb) gene‐transfection on DC‐based therapy. (A) Survival of mice with the indicated treatments. (B) The relative tumor volume (with the volume on day 0 set at 1) of each mouse in the control groups. Black solid lines indicate the relative tumor volumes of the [Untreated] group; black dotted lines indicate those of the [Cont (IT)] group; gray solid lines indicate those of the [Cont (IV)] group. (C) The relative tumor volume of each mouse in the intratumoral (IT) gene‐administrated group. Black solid lines indicate relative tumor volumes of the [Comb (IT) +DC] group. Black dotted lines indicate those of the [Comb (IT)] group. Gray solid lines indicate those of the [Cont (IT) + DC] group. (D) The relative tumor volume of each mouse in the intravenous (IV) gene‐administrated group. Black solid lines indicate relative tumor volumes of the [Comb (IV) +DC] group. Black dotted lines indicate those of the [Comb (IV)] group. Gray solid lines indicate those of the [Cont (IV) + DC] group. Experiments were performed independently three times using a total of six mice in each group. All mice were humanely euthanized according to the criteria in Section . * P < .05, vs the [Cont (IT)] group. ** P < .05 vs the [Untreated] group, and P < .01, vs the [Cont (IV)] group. *** P < .05 vs the [Untreated] group, and P < .005, vs the [Cont (IT) +DC] group or the [Cont (IV) +DC] group, respectively

Article Snippet: Cells infiltrating tumor tissues were evaluated by immunohistochemistry using following primary Abs: for IFNγ, Bioss Cat# bs‐0480R‐PE‐Cy7, RRID:AB_11092092; for CD40L, Biorbyt Cat# orb10326, RRID:AB_10746923; for CD8, clone 53‐6.7, Thermo Fisher Scientific Cat# 14‐0081‐81, RRID:AB_467086; for Foxp3, clone FJK‐16s, Thermo Fisher Scientific Cat# 14‐5773‐80, RRID:AB_467575; for Iba1, Wako Pure Chemical Industries Inc, Cat# 019‐19741, RRID:AB_839504; for granzyme B, Spring Bioscience Cat# E2580, RRID:AB_1661202.

Techniques: Transfection

Journal: FASEB bioAdvances

Article Title: Manipulation of the tumor microenvironment by cytokine gene transfection enhances dendritic cell‐based immunotherapy

doi: 10.1096/fba.2019-00052

Figure Lengend Snippet: Effect of IFNγ and CD40L (Comb) gene‐transfection therapy on the infiltration of CD8 + cells into LM8 tumors. Tumor tissue was collected from mice with the indicated treatments at 7 d after the second treatment. Immunohistochemistry was performed to detect CD8 + cells present in the tumor tissue. (A) Typical photos of tumors collected from the indicated treatment groups are shown. Expression of CD8 is shown as brown color products. (B) Counts of CD8 + cells in 1000 cells in tumors are shown. Results are expressed as mean ± SE. The experiment was performed using three mice in each treatment group. All mice were humanely euthanized according to the criteria in Section . * P < .05, ** P < .01 by the Tukey‐Kramer test

Article Snippet: Cells infiltrating tumor tissues were evaluated by immunohistochemistry using following primary Abs: for IFNγ, Bioss Cat# bs‐0480R‐PE‐Cy7, RRID:AB_11092092; for CD40L, Biorbyt Cat# orb10326, RRID:AB_10746923; for CD8, clone 53‐6.7, Thermo Fisher Scientific Cat# 14‐0081‐81, RRID:AB_467086; for Foxp3, clone FJK‐16s, Thermo Fisher Scientific Cat# 14‐5773‐80, RRID:AB_467575; for Iba1, Wako Pure Chemical Industries Inc, Cat# 019‐19741, RRID:AB_839504; for granzyme B, Spring Bioscience Cat# E2580, RRID:AB_1661202.

Techniques: Transfection, Immunohistochemistry, Expressing

Journal: FASEB bioAdvances

Article Title: Manipulation of the tumor microenvironment by cytokine gene transfection enhances dendritic cell‐based immunotherapy

doi: 10.1096/fba.2019-00052

Figure Lengend Snippet: Effect of the IFNγ and CD40L (Comb) gene‐transfection therapy on tumor‐infiltrating cells. Tumor tissue was collected from mice with the indicated treatments at 7 d after the second treatment. The Iba 1 + activated macrophage and granzyme B + killer cells in the tumor tissue were detected by immunohistochemistry. Results are expressed as mean ± SE. The experiment was performed using three mice in each treatment group. All mice were humanely euthanized according to the criteria in Section . * P < .05, ** P < .01 by the Tukey‐Kramer test

Article Snippet: Cells infiltrating tumor tissues were evaluated by immunohistochemistry using following primary Abs: for IFNγ, Bioss Cat# bs‐0480R‐PE‐Cy7, RRID:AB_11092092; for CD40L, Biorbyt Cat# orb10326, RRID:AB_10746923; for CD8, clone 53‐6.7, Thermo Fisher Scientific Cat# 14‐0081‐81, RRID:AB_467086; for Foxp3, clone FJK‐16s, Thermo Fisher Scientific Cat# 14‐5773‐80, RRID:AB_467575; for Iba1, Wako Pure Chemical Industries Inc, Cat# 019‐19741, RRID:AB_839504; for granzyme B, Spring Bioscience Cat# E2580, RRID:AB_1661202.

Techniques: Transfection, Immunohistochemistry

Journal: FASEB bioAdvances

Article Title: Manipulation of the tumor microenvironment by cytokine gene transfection enhances dendritic cell‐based immunotherapy

doi: 10.1096/fba.2019-00052

Figure Lengend Snippet: Effect of IFNγ and CD40L (Comb) gene‐transfection therapy on tumor‐infiltrating cells. Tumor tissue was collected from mice with the indicated treatments at 7 d after the second treatment. Mature DCs (MHC II high CD83 + cells), MDSCs (CD11b + , Gr1 + cells), NK (NK1.1 + ) cells and PD‐L1 + cells in the tumor tissues were detected in FCM. Results are expressed as mean ± SE. The experiment was performed using three mice in each treatment group. All mice were humanely euthanized according to the criteria in Section . * P < .05, ** P < .01 by the Tukey‐Kramer test

Article Snippet: Cells infiltrating tumor tissues were evaluated by immunohistochemistry using following primary Abs: for IFNγ, Bioss Cat# bs‐0480R‐PE‐Cy7, RRID:AB_11092092; for CD40L, Biorbyt Cat# orb10326, RRID:AB_10746923; for CD8, clone 53‐6.7, Thermo Fisher Scientific Cat# 14‐0081‐81, RRID:AB_467086; for Foxp3, clone FJK‐16s, Thermo Fisher Scientific Cat# 14‐5773‐80, RRID:AB_467575; for Iba1, Wako Pure Chemical Industries Inc, Cat# 019‐19741, RRID:AB_839504; for granzyme B, Spring Bioscience Cat# E2580, RRID:AB_1661202.

Techniques: Transfection

Journal: FASEB bioAdvances

Article Title: Manipulation of the tumor microenvironment by cytokine gene transfection enhances dendritic cell‐based immunotherapy

doi: 10.1096/fba.2019-00052

Figure Lengend Snippet: Effect of IFNγ and CD40L (Comb) gene‐transfection therapy on systemic immune responses. Spleen cells were collected from the LM8 tumor‐bearing mice with the indicated treatments at 7 d after the second treatment. Cytotoxic activity of the cells against LM8 (A), BW5147 (B), and YAC‐1 (C) was evaluated in vitro. The experiments were repeated twice. Each experiment was performed using three mice in each treatment group. A typical result is shown. All mice were humanely euthanized according to the criteria in Section . Results are expressed as mean ± SE. * P < .05, ** P < .01 vs the [Untreated] group, † P < .05 vs the [Comb (IT) + DC] and [Comb (IV)] group at the E/T ratio

Article Snippet: Cells infiltrating tumor tissues were evaluated by immunohistochemistry using following primary Abs: for IFNγ, Bioss Cat# bs‐0480R‐PE‐Cy7, RRID:AB_11092092; for CD40L, Biorbyt Cat# orb10326, RRID:AB_10746923; for CD8, clone 53‐6.7, Thermo Fisher Scientific Cat# 14‐0081‐81, RRID:AB_467086; for Foxp3, clone FJK‐16s, Thermo Fisher Scientific Cat# 14‐5773‐80, RRID:AB_467575; for Iba1, Wako Pure Chemical Industries Inc, Cat# 019‐19741, RRID:AB_839504; for granzyme B, Spring Bioscience Cat# E2580, RRID:AB_1661202.

Techniques: Transfection, Activity Assay, In Vitro

Journal: FASEB bioAdvances

Article Title: Manipulation of the tumor microenvironment by cytokine gene transfection enhances dendritic cell‐based immunotherapy

doi: 10.1096/fba.2019-00052

Figure Lengend Snippet: Effect of IFNγ and CD40L (Comb) gene‐transfection therapy on spleen cells. Spleen was collected from mice with the indicated treatments at 7 days after the second treatment. Mature DCs (MHC II high CD83 + cells), MDSCs (CD11b + , Gr1 + cells), NK (NK1.1 + ) cells and PDL‐1 + cells in the tumor tissues were detected in FCM. Results are expressed as mean ± SE. The experiment was performed using three mice in each treatment group. All mice were humanely euthanized according to the criteria in Section . * P < .05, ** P < .01 by the Tukey‐Kramer test

Article Snippet: Cells infiltrating tumor tissues were evaluated by immunohistochemistry using following primary Abs: for IFNγ, Bioss Cat# bs‐0480R‐PE‐Cy7, RRID:AB_11092092; for CD40L, Biorbyt Cat# orb10326, RRID:AB_10746923; for CD8, clone 53‐6.7, Thermo Fisher Scientific Cat# 14‐0081‐81, RRID:AB_467086; for Foxp3, clone FJK‐16s, Thermo Fisher Scientific Cat# 14‐5773‐80, RRID:AB_467575; for Iba1, Wako Pure Chemical Industries Inc, Cat# 019‐19741, RRID:AB_839504; for granzyme B, Spring Bioscience Cat# E2580, RRID:AB_1661202.

Techniques: Transfection

Journal: Infection and Immunity

Article Title: Mucosal Vaccination against Tuberculosis Using Inert Bioparticles

doi: 10.1128/IAI.00786-13

Figure Lengend Snippet: IFN-γ responses. IFN-γ was determined from splenocytes stimulated using an ELISA or ELISPOT assay. (a and b) ELISA detection of IFN-γ from splenocytes stimulated with rMPT64 (a) or with rAcr or rAg85B (b). (c) ELISPOT assay determination of the number of spot-forming units (SFU) in splenocytes stimulated with rMPT64 or rAcr-Ag85B. Spleens were taken from mice (n = 2) immunized using dosing regimen 1. The BCG group received one dose of BCG.

Article Snippet: Free binding sites were blocked with 10% FCS in PBS at RT for 1 h. Culture supernatants were tested at RT for 2 h, and IFN-γ was detected with a biotinylated anti-mouse IFN-γ–streptavidin–horseradish peroxidase conjugate (BD Biosciences) at RT for 1 h. Recombinant mouse IFN-γ (BD Biosciences) was used as a standard.

Techniques: Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

Journal: Infection and Immunity

Article Title: Mucosal Vaccination against Tuberculosis Using Inert Bioparticles

doi: 10.1128/IAI.00786-13

Figure Lengend Snippet: Gating strategy for polyfunctional T cell analysis. Representative FlowJo scatter plots of a naive mouse splenocyte sample are presented to show the gating strategy. Samples were first gated by forward scatter (FSC-A)/side scatter (SSC-A) to select lymphocytes and were then gated by FSC-H/FSC-A to gate for single cells. This population was then gated for CD3+ CD4+ or CD3+ CD8+ staining. These two populations were then analyzed for IFN-γ, IL-2, and TNF-α by staining.

Article Snippet: Free binding sites were blocked with 10% FCS in PBS at RT for 1 h. Culture supernatants were tested at RT for 2 h, and IFN-γ was detected with a biotinylated anti-mouse IFN-γ–streptavidin–horseradish peroxidase conjugate (BD Biosciences) at RT for 1 h. Recombinant mouse IFN-γ (BD Biosciences) was used as a standard.

Techniques: Staining

Journal: Scientific Reports

Article Title: CD11b deficiency suppresses intestinal tumor growth by reducing myeloid cell recruitment

doi: 10.1038/srep15948

Figure Lengend Snippet: The accumulation of Gr-1 + CD11b + MDSC cells in the tumor microenvironment of the Apc Min/ + mice compared with the normal intestine tissues from the C57 mice; the Gr-1 + cells were decreased in the tumor microenvironment of the Apc Min/ + ;CD11b −/− mice ( a ). The number of the Ly6G + Ly6C low subset of MDSCs (gated on CD45 + cells) in the bone marrow of C57, CD11b −/− , Apc Min/ + and Apc Min/ + ;CD11b −/− mice ( b ) and in the peripheral blood, spleen and tumor tissues of the Apc Min/ + and Apc Min/ + ;CD11b −/− mice ( c ) was measured by FACS. The results of the IF staining and FACS represent of 11 independent mice (all mice were 16–weeks-old). The proteins expression of IFN-γ and CXCL9 in the tumor tissues from the Apc Min/ + and Apc Min/ + ;CD11b −/− mice was examined by immunoblotting ( d ). The protein bands intensities were quantitated and normalized to those of GAPDH. The statistical data are expressed as the means ± S.D. * P <0.05, ** P < 0.01 and *** P < 0.001. Scale bars: 50 μm ( a ).

Article Snippet: The primary antibodies were used for immunohistochemical (IHC), immunofluorescence (IF) analysis and Western blotting (WB) assays: and mouse anti-BrdU antibody (RPN202, diluted at 1:150 for IHC) was obtained from GE Healthcare (Little Chalfond, UK); rabbit anti-CyclinD1 (BM0771, diluted at 1:100 for IHC and 1:400 for WB), rabbit anti-CD34 (BA0532, diluted at 1:100 for IHC), rabbit anti-CD45 (BA3371, diluted at 1:100 for IHC) and rabbit anti-p65 (BA0610, diluted at 1:100 for IHC) were obtained from Boster (Wuhan, China); mouse anti-TNF-α (ab10863, diluted at 1:5000 for WB) was obtained from Abcam (Cambridge, UK); rabbit anti-IFN-γ (bs-0480R), rabbit anti-CXCL-9 (bs-2551R) (both diluted at 1:500 for WB), rabbit anti-CD11b (bs-1014R) and rabbit anti-Gr-1 (bs-2576R, diluted at 1:100 for IF) were obtained from Bioss (Beijing, China); mouse anti-Cytokeratin 8 (CK8, ZM-0310, diluted at 1:100 for IF) was purchased from ZSGB-BIO (Beijing, China); mouse anti-CD11b (Santa Cruz, USA, diluted at 1:100 for IF); mouse anti-E-cadherin (#610181, diluted at 1:100 for IHC and 1:5000 for WB) and mouse anti-β-catenin (#610154, diluted at 1:100 for IHC, and 1:2000 for WB) were purchased from BD Transduction Laboratories (Franklin Lakes, NJ); rabbit anti-GAPDH (#2118s, diluted at 1:2000 for WB) was purchased from CST (USA); rabbit anti-pp65 (Ser276, sc-101749, diluted at 1:500 for IHC) was obtained from Santa Cruz Biotechnology Inc. (USA).

Techniques: Staining, Expressing, Western Blot