nb110 89474 cd80 rabbit polyclonal  (Novus Biologicals)

Journal: Cell Reports Medicine

Article Title: Potentiating immunotherapy in “immune-cold” solid tumors through orchestrating T cell immunity via tumor-specific genetic engineering

doi: 10.1016/j.xcrm.2025.102510

Figure Lengend Snippet: P αCD3&LIGHT improved anti-melanoma efficacy of CAR-T cells without obvious systemic toxicity (A) Therapeutic scheme of P αCD3&LIGHT in combination with CAR-T cells. hCD19-B16 melanoma-bearing mice were treated with four intravenous injections of P αCD3&LIGHT or other controls and two intravenous injections of CAR-T cells at different doses (0–4 × 10 6 cells per mouse). (B) Experimental timeline of blood serum collection for evaluating severe cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS). (C) Flow cytometry analysis of CAR-T cells in tumor tissues of hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + CAR-T cells or other controls. n = 8. (D) Tumor weights of hCD19-B16 melanoma-bearing mice treated with 0–4 × 10 6 CAR-T cells. n = 9 or 10. (E–G) ELISA of SAA (E), IL-6 (F), and IL-1β (G) in blood serum of hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + CAR-T cells or other controls. n = 8. (H and I) Body temperature (H) and body weight (I) of hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + CAR-T cells or other controls. n = 10. (J) Radar map of fold changes of five CRS-related markers. The larger the area enclosed by the five markers, the more severe the CRS-related symptoms. (K) Representative immunohistochemistry images of CD11b-positive staining to indicate vascular leakage in lung, spleen, and liver tissues from hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + CAR-T cells or other controls. Scale bar: 100 μm. (L) Representative immunohistochemistry images of H&E staining to show the thickness of meninges in brain tissues from hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + CAR-T cells or other controls. Scale bar: 100 μm. (M) Therapeutic scheme of P αCD3&LIGHT in combination with 1.5 × 10 6 CAR-T cells. hCD19-B16 melanoma-bearing mice were treated with four intravenous injections of P αCD3&LIGHT or other controls and two intravenous injections of PBS or total 1.5 × 10 6 CAR-T cells. (N) Representative IVIS spectrum images of DiR-labeled CAR-T cells at tumor sites in hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + 1.5 × 10 6 CAR-T cells or other controls. (O–Q) Flow cytometry analysis of CD69-positive activated CAR-T cells (O), TCF-1-positive stem cell-like (P), and Ki67-positive self-renewing CAR-T cells (Q) in tumor tissues from hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + 1.5 × 10 6 CAR-T cells or other controls. n = 8. (R–T) ELISA of IFN-γ (R), TNF-α (S), and Gzm-B (T) in tumor tissues from hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + 1.5 × 10 6 CAR-T cells or other controls. n = 8. (U) Flow cytometry analysis of cleaved caspase-3-positive apoptotic hCD19-B16 tumor cells in tumor tissues from hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + 1.5 × 10 6 CAR-T cells or other controls. n = 8. (V–X) Individual (V) and average (W) tumor growth curves and survival curves (X) of hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + 1.5 × 10 6 CAR-T cells or other controls. n = 8–10. (Y) Magnetic resonance imaging (MRI) images of complete cured melanoma-bearing mice treated with P αCD3&LIGHT + 1.5 × 10 6 CAR-T cells. Data are represented as mean ± SD (error bars) from biological replicates. p values were determined by unpaired two-tailed Student’s t test for (C) and one-way ANOVA with Tukey’s test for (D), (O)–(U), and (W). n.s., not significant; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. See also .

Article Snippet: Rabbit Anti-mouse CD11b Polyclonal Antibody , Servicebio , Cat# GB115689.

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining, Labeling, Magnetic Resonance Imaging, Two Tailed Test

Journal: Theranostics

Article Title: A new nano-encapsulated TSPO ligand reduces neuroinflammation and improves cognitive functions in Alzheimer's disease model

doi: 10.7150/thno.106083

Figure Lengend Snippet: Expression of the pro- and anti-inflammatory markers (iNOS, CD86 and IL-1Ra) in LPS-stimulated BV2 microglia exposed to SANP-TEMNAP (100 nM). (A) Representative confocal images displaying the expression of Iba1 (red) and iNOS (green) in BV2 microglia cell cultures in control condition (a-c) and in LPS-stimulated microglia co-exposed to the following conditions: (i) self-assembling nanoparticle (SANP) (d-f); (ii) nanoencapsulated-TEMNAP (100 nM) (SANP-TEMNAP) (g-i); (iii) TEMNAP (100 nM) non-nanoencapsulated (j-l). Nuclei were stained by the nuclear dye Hoechst-33258. Scale bars: 10 µm. (B) Representative confocal images displaying the expression of Iba1 (red) and CD86 (green) in BV2 microglia cell cultures in control condition (a-c) and in LPS-stimulated microglia co-exposed to the following conditions: (i) self-assembling nanoparticle (SANP) (d-f); (ii) nanoencapsulated-TEMNAP (100 nM) (SANP-TEMNAP) (g-i); (iii) TEMNAP (100 nM) non-nanoencapsulated (j-l). Nuclei were stained by the nuclear dye Hoechst-33258. Scale bars: 10 µm. (C) Quantitative analysis of iNOS (a) and CD86 (b) fluorescence intensities. The values represent the mean±SEM; (n = 3 independent experimental replicates); *p < 0.05 versus control. (D) Representative confocal images displaying the expression of CD11b (red) and IL-1Ra (green) in BV2 microglia cell cultures in LPS-stimulated microglia co-exposed to the following conditions: (i) nanoencapsulated-TEMNAP (100 nM) (SANP-TEMNAP) (a-c); (ii) transferrin-conjugated nanoparticles containing TEMNAP (100 nM) (SANP-TF-TEMNAP) (d-f); Nuclei were stained by the nuclear dye Hoechst-33258. Scale bars: 20 µm.

Article Snippet: The primary antibodies used were the following: mouse monoclonal anti-Iba1 (NCNP24) (Wako, Cat#016-26721, RRID:AB_2811160); rabbit anti-Iba1 (Wako, Cat#019-19741, RRID:AB_839504); rabbit anti-iNOS (EPR16635) (Abcam, Cambridge, UK, Cat#ab178945, RRID:AB_2861417); monoclonal mouse anti-iNOS (NOS-IN) (Sigma-Aldrich, St. Louis, Missouri, USA, Cat#N9657, RRID:AB_260818); mouse monoclonal anti-CD86 (B7-2 (D-6)) (Santa Cruz Biotechnology, Dallas, Texas, USA, Cat#sc-28347, RRID:AB_627200), rabbit polyclonal CD11b (Novus, Italy, Cat#NB110-89474, RRID:AB_1216361), mouse monoclonal IL1-Ra (A4) (Santa Cruz Biotechnology, Dallas, Texas, USA, Cat#sc-374084, RRID: AB_10917936).

Techniques: Expressing, Control, Staining, Fluorescence