il12p70 levels  (R&D Systems)

Journal: Journal of Nanobiotechnology

Article Title: Cu-doped TiO 2 nanoparticles improve local antitumor immune activation and optimize dendritic cell vaccine strategies

doi: 10.1186/s12951-023-01844-z

Figure Lengend Snippet: A Histogram displaying the ratio of CD8 + T cells over CD4 + T cells obtained upon in vitro stimulation by OVA-peptide loaded DCs that had either been left untreated (control) or were activated classically, or upon exposure to 33% Cu-doped TiO 2 NPs. B , C Histograms displaying the relative level (control: 100%) of B) granzyme B release, or C) perforin release from T cells obtained from part A that had been exposed to the OVA-derived SIINFEKL peptide. D , E Histograms displaying the level of D) IL12p70 release or E) relative Ccr7 surface expression (control: 100%) on DCs that had either been left untreated (control) or were activated classically, or exposed to 33% Cu-doped TiO 2 NPs. F , G) Histograms displaying the F) number of migrated DCs or G) the level of IL12p70 secreted from migratory DCs obtained from DCs that had either been left untreated (control) or were activated classically, or exposed to 33% Cu-doped TiO 2 NPs and subsequently exposed to the CCR7 ligand 6C-kine. H , I Histograms displaying the in vivo antibody titer for H) OVA-specific IgG1 or I) OVA-specific IgG2α obtained upon intravenous administration of OVA-pulsed DCs that had either been left untreated (control) or were activated classically, or exposed to 33% Cu-doped TiO 2 NPs. J Histogram displaying the level of IFNγ obtained from ex vivo splenocytes obtained from animals receiving the different types of OVA-pulsed DCs and incubated with OVA ex vivo. K Tumor volumes obtained from OVA-expressing E0771 tumors grafted subcutaneously in C57Bl6 mice receiving the different OVA-pulsed DCs that had either been left untreated (control) or were activated classically, or exposed to 33% Cu-doped TiO 2 NPs. L , M Histograms displaying the relative level (control: 100%) of L) granzyme B release, or M) perforin release from CD8 + T cells isolated from OVA-E0771 tumor-bearing animals having received OVA-pulsed DC grafts of the different DC types and then exposed to the OVA-derived SIINFEKL peptide ex vivo. N Histogram displaying the level of OVA-specific CD8 + T cells isolated from the spleen of OVA-E0771 tumor-bearing animals having received OVA-pulsed DC grafts of the different DC types

Article Snippet: Media were then removed and fresh media (5 ml) was given with either no additions (controls), 33% Cu-doped TiO 2 NPs at 40 μg/ml (NP condition) or addition of maturation factors IL1β (25 ng/ml) TNFα (50 ng/ml) and IFNγ (1000 units/ml) (all from PeproTech; this is the “classical” activation scheme) for 24 h. Media were then removed and cells were incubated with soluble recombinant CD40L (BioTechne) at 16 μg/ml for 24 h after which the supernatants was collected and used to determine IL12p70 levels by ELISA (M1270, R&D Systems).

Techniques: In Vitro, Control, Derivative Assay, Expressing, In Vivo, Ex Vivo, Incubation, Isolation

Journal: PLoS ONE

Article Title: Endoplasmic Reticulum Aminopeptidase-1 Functions Regulate Key Aspects of the Innate Immune Response

doi: 10.1371/journal.pone.0069539

Figure Lengend Snippet: CD11c+ dendritic cells were isolated from C57BL/6 WT (n = 4) or ERAP1-KO mice (n = 6) and in vitro stimulated with specified concentrations of rEA protein. Cultured media was used to perform an IL12p70 ELISA as described in Materials and Methods. The bars represent Mean ± SEM. 0.1 ng/ml of rEA is an optimal stimulation dose. Statistical analysis was completed using two-tailed homoscedastic Student’s t-tests; *, - indicate values, statistically different between WT_rEA and ERAP1-KO_rEA groups, p<0.05. Representative of three independent experiments is shown.

Article Snippet: Following incubation, culture medium was analyzed for mouse IL12p70 levels using an ELISA kit and following its enclosed instructions (R&D Systems, Minneapolis, MN).

Techniques: Isolation, In Vitro, Cell Culture, Enzyme-linked Immunosorbent Assay, Two Tailed Test