cd11c primary antibody  (Proteintech)

Journal: Journal for immunotherapy of cancer

Article Title: In situ endoscopic photodynamic therapy combined with immature DC vaccination induces a robust T cell response against peritoneal carcinomatosis.

doi: 10.1136/jitc-2024-009752

Figure Lengend Snippet: Figure 3 PDT-induced ferroptosis promotes iDC infiltration in three-dimensional (3D) spheroids together with an increase in MHCI and MHCII expression. (A) Protocol of iDC transfer to CT26 spheroids prechallenged with photoactivated OR141 or RSL3. CT26 3D spheroids were pretreated with 10 µM ferrostatin-1 and incubated 48 hours later with PS OR141 in the dark for 4 hours, followed by a washing step. Photoactivation was performed using an LED light source for 90 min and incubation was continued after the renewal of ferrostatin-1. A similar protocol was applied for RSL3 treatment with 48 hours pretreatment and renewal for final incubation. Exogenous iDCs (5×103 per spheroid) were added 48 hours post-PDT or after the last RSL3 addition. (B, D) Representative IF pictures depicting the extent of CD11c+ cell infiltration after spheroid exposition to (B) PDT or (D) 5 µM RSL3. Scale bar: 100 µm. (C, E) Representative modeling of CD11c+ cell infiltration (red) derived from light sheet microscopy- based reconstruction (Imaris program). Scale bar: 50 µm. (F) Quantification of wholemount IF-based infiltrated CD11c+ cells and (G) impact on spheroid growth for the indicated conditions. (H) Flow cytometry-based quantification of infiltrated CD11c+ cells and associated changes in (I) iDC volume, (J) MHCI and (K) MHCII expression. Data are plotted as the means±SEM (*p≤0.05; **p≤0.01; ***p≤0.001; ****p≤0.0001; ns, p>0.05) from 3 independent experiments performed with ≥3 technical replicates (minimum of 6 spheroids were pooled together per condition). Significance was determined by one-way ANOVA with Tukey’s multiple comparison test. ANOVA, analysis of variance; iDC, immature dendritic cell; ns, not significant; PDT, photodynamic therapy.

Article Snippet: Frozen sections (5 μm) were permeabilized 10 min in PBS- triton 0.5% at RT and then blocked with PBS- triton 0.05% (PBST)- 5% BSA during 1 hour at RT, incubated overnight at 4°C with anti- pimonidazole primary antibody (#Pab2627, 1/400, Hyopxyprobe) or anti- CD11c primary antibody (#97585, 1/150, Cell Signaling) in PBST- 1% BSA.

Techniques: Expressing, Incubation, Derivative Assay, Microscopy, Flow Cytometry, Comparison

Journal: Journal for immunotherapy of cancer

Article Title: In situ endoscopic photodynamic therapy combined with immature DC vaccination induces a robust T cell response against peritoneal carcinomatosis.

doi: 10.1136/jitc-2024-009752

Figure Lengend Snippet: Figure 5 Laparoscopic PDT administration promotes iDC-based (memory) immune response against peritoneal tumors. Balb/ CByJ mice were i.p. injected with 1×106 luciferase-positive AB1 tumor cells and exposed 7 days later to PDT. PS-OR141 (0.4 or 4 mg/kg) was injected i.p. 90 min before laparoscopy and photo-activated for 15 min on endoscopal light exposure; sham animals were used as controls. (A) Experimental set-up depicting laparoscopy procedure to introduce the endoscope LED light source in the inflated peritoneal cavity. (B) Representative multiplex immunofluorescence pictures depicting immune cell infiltration (CD11c+ cells in red, CD3 + T cells light blue and CD8+ T cells in green) 6 days post in situ PDT. Scale bar: 200 µm. (C) Protocol of in situ PDT-based iDC vaccination against mouse peritoneal tumors. iDCs (2×106) were injected i.p. 60 min after in situ PDT. (D–F) Tumor burden monitored by bioluminescence (expressed as photons/s/cm2/sr) and (G–I) corresponding survival curves. n=6–8 mice per group. Significance was determined by comparing curves presented in G–I with log-rank (Mantel-Cox) test (*p≤0.05). Experimental endpoint was determined when mice reached the ethical limits (ie, bioluminescence signal over 50,000 ph/s/cm2/sr or euthanasia (†) on major ascite accumulation). In (I), arrow at day 69 indicates rechallenge by i.p. injection of 1×106 luciferase-positive AB1 tumor cells. (J, K) Immune cell activation analysis carried out by flow cytometry from mesenteric lymph nodes collected 10 days after treatment. Bar graphs represent (J) CD40+, CD80 +, CD86 + MHCII + and OX40L + MHCI + CD11+ cells and (K) CD62L+ CD69+ early activated CD8+ and CD4+ T cells in PDT 4 mg/kg+iDC experimental condition (vs sham mice). Data are plotted as the means±SEM (*p≤0.05; ns, p>0.05) from 3 to 4 mice per group. Significance was determined by Student’s t-test. iDC, immature dendritic cell; i.p., intraperitoneal; MLN, mesenteric lymph nodes; PDT, photodynamic therapy.

Article Snippet: Frozen sections (5 μm) were permeabilized 10 min in PBS- triton 0.5% at RT and then blocked with PBS- triton 0.05% (PBST)- 5% BSA during 1 hour at RT, incubated overnight at 4°C with anti- pimonidazole primary antibody (#Pab2627, 1/400, Hyopxyprobe) or anti- CD11c primary antibody (#97585, 1/150, Cell Signaling) in PBST- 1% BSA.

Techniques: Injection, Luciferase, Introduce, Multiplex Assay, Immunofluorescence, In Situ, Activation Assay, Flow Cytometry