Journal: Journal of neuroinflammation

Article Title: Brain endothelial CXCL12 attracts protective natural killer cells during ischemic stroke.

doi: 10.1186/s12974-023-02689-x

Figure Lengend Snippet: Fig. 5 NK cells positively regulate behavioral deficits after PT stroke induction. A Coordinates of the lesion on the brain of photothromobotic (PT) mouse model used for beam-walk sensorimotor test analysis. The center of the lesion is 0.3 mm in front of bregma and 2.0 mm lateral to midline, with the 2 mm of diameter. B Nissl staining on the vibratome section of stroke brain at day 2 after PT. Data represent n = 7 mice. C The beam-walk sensorimotor test analysis was performed by calculating percentage of contralateral hindlimb faults. Anti-NK1.1 or IgG2a was injected at day 9 (P-9) and day 2 (P-2) before PT induction and the efficiency of depletion in blood was tested at P-7, P-1, P6, P12 and P18. The videos were recorded just before PT induction (P0) and every second day afterwards. (n = 3 mice for IgG2a, n = 4 mice for anti-NK1.1). D Lesion sizes of brains from the isotype control (IgG2a) (blue) vs. anti-NK1.1 (red) injected mice at P2 and P18 (n = 3 mice for IgG2a, n = 4 mice for anti-NK1.1). E Control (n = 4) and KO (n = 4) mice after PT induction by calculating the percentage of contralateral hindlimb faults.. P2 **p = 0.0058, P4 **P = 0.0053, P6 **P = 0.0096

Article Snippet: For in vivo depletion of NK cells, mice were injected 100 μg of InVivoPlus anti-mouse Nk1.1 (Clone PK136, Bio X Cell) or InVivoPlus mouse IgG2a (Clone C1.18.4, Bio X Cell) resolved in PBS at P-9 and P-2, respectively.

Techniques: Staining, Injection, Control

Journal: Neoplasia (New York, N.Y.)

Article Title: An antibody targeting an immune checkpoint molecule BTN2A2 enhances anti-tumor immunity

doi: 10.1016/j.neo.2025.101161

Figure Lengend Snippet: hBTN2A2-Ig preparation and functional validation. (A) The extracellular segment of hBTN2A2 nucleotide sequence was identified by ordinary PCR and agarose gel electrophoresis. The purified hBTN2A2-Ig protein was verified by Coomassie blue-stained SDS- PAGE, Western blot with anti-IgG2a antibody and anti-hBTN2A2 antibody. (B–G) The effects of hBTN2A2-Ig protein on murine T cell activation and proliferation in vitro. Splenocytes from C57BL/6 mice were incubated on 96-well plate precoated with 0.5 μg/ml of anti-CD3 antibody and the indicated dose of hBTN2A2-Ig or control Ig. The splenocytes were stained with anti-CD4, CD8, CD44, CD62L, Ki67 antibodies, followed by analyzed for the expression of CD69 18 h later (B–C) and the percentages of CD44 lo CD62L hi naive and CD44 hi CD62L lo effective memory CD4+ and CD8+ T cells 72 h later (D–E) , and T cell proliferation by CFSE dilution assay and Ki67 staining 5 days later (F–G) . The data are representative of three independent experiments with similar results. ( n = 3 each experiment) * P < 0.05, ** P < 0.01 and *** P < 0.001, compared with control Ig.

Article Snippet: The recombinant mouse IgG2a Fc protein (control Ig) was purchased from BioXCell (West Lebanon, NH) [ , ].

Techniques: Functional Assay, Biomarker Discovery, Sequencing, Agarose Gel Electrophoresis, Purification, Staining, SDS Page, Western Blot, Activation Assay, In Vitro, Incubation, Control, Expressing, Dilution Assay

Journal: Chinese Medical Journal

Article Title: Neutralization of Interleukin-9 Decreasing Mast Cells Infiltration in Experimental Autoimmune Encephalomyelitis

doi: 10.4103/0366-6999.204110

Figure Lengend Snippet: Anti-IL-9 antibody treatment decreased mast cell infiltration of the CNS. Purified cells (mainly composed of mast cells) were harvested from the CNS on days 0, and 5, 15, and 20 after EAE immunization. CD45 + CD117 + mast cells were detected by flow cytometry. CNS mast cells maintained a high level throughout the disease course in EAE group. However, mast cells were significantly reduced from day 5, and remained low in anti-IL-9 Abs group, compared with IgG group. Data are shown as mean ± SE. * P < 0.01, anti-IL-9 Abs group versus IgG group. CNS: Central nervous system; EAE: Experimental autoimmune encephalomyelitis; SE: Standard error; IL: Interleukin.

Article Snippet: The following antibodies were used in this study: FITC-anti-mouse CD45 (eBioscience, USA), PE-Cyanine5-anti-mouse CD117 (eBioscience), anti-mouse IL-9 (BE0181; BioXCell, USA), anti-mouse IgG2a isotype control (BE0085; BioXCell), anti-mouse IL-9 receptor (IL-9R) (SC699; Santa Cruz, USA), anti-mouse IL-2Rγ (SC668; Santa Cruz), anti-mouse IgG isotype control (GTX35009; Santa Cruz), and donkey pAb to Rb IgG Alexa Flour 555 (Ab150074; Abcam, USA).

Techniques: Purification, Flow Cytometry

Journal: Chinese Medical Journal

Article Title: Neutralization of Interleukin-9 Decreasing Mast Cells Infiltration in Experimental Autoimmune Encephalomyelitis

doi: 10.4103/0366-6999.204110

Figure Lengend Snippet: IL-9 blockade reduced production of chemokine recruiting mast cells in the CNS. Five days after immunization, mRNA expressions of Pecam1, SCF, Vcam-1, CCL2, and CCL5 in CNS tissue of EAE mice were detected by RT-PCR. After IL-9 neutralization, mRNA expressions of CCL5 and Vcam-1 were significantly decreased in anti-IL-9 Abs group, compared with IgG group. * P < 0.01. Pecam1: Platelet and endothelial cell adhesion molecule 1; SCF: Supercoiling factor; Vcam-1: Vascular cell adhesion molecule 1; CCL2: C-C motif chemokine ligand 2; CCL5: C-C motif chemokine ligand 5; CNS: Central nervous system; EAE: Experimental autoimmune encephalomyelitis; IL: Interleukin; RT-PCR: Reverse transcription-polymerase chain reaction; mRNA: Messenger RNA.

Article Snippet: The following antibodies were used in this study: FITC-anti-mouse CD45 (eBioscience, USA), PE-Cyanine5-anti-mouse CD117 (eBioscience), anti-mouse IL-9 (BE0181; BioXCell, USA), anti-mouse IgG2a isotype control (BE0085; BioXCell), anti-mouse IL-9 receptor (IL-9R) (SC699; Santa Cruz, USA), anti-mouse IL-2Rγ (SC668; Santa Cruz), anti-mouse IgG isotype control (GTX35009; Santa Cruz), and donkey pAb to Rb IgG Alexa Flour 555 (Ab150074; Abcam, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Neutralization

Journal: Chinese Medical Journal

Article Title: Neutralization of Interleukin-9 Decreasing Mast Cells Infiltration in Experimental Autoimmune Encephalomyelitis

doi: 10.4103/0366-6999.204110

Figure Lengend Snippet: In vitro , the effect of anti-IL-9 antibody on splenic mast cells. Splenocytes were harvested from experimental autoimmune encephalomyelitis mice 5 days after MOG immunization. After co-culture with anti-IL-9 antibody or anti-mouse IgG for 7 h, mast cell number was counted by flow cytometry. Splenic mast cells cultured with anti-IL-9 antibody showed significantly lower levels in a dose-dependent manner. This trend was particularly evident with anti-IL-9 antibody concentrations up to 20 μg/ml. * P < 0.05; † P < 0.01. IL: Interleukin; MOG: Myelin oligodendrocyte glycoprotein.

Article Snippet: The following antibodies were used in this study: FITC-anti-mouse CD45 (eBioscience, USA), PE-Cyanine5-anti-mouse CD117 (eBioscience), anti-mouse IL-9 (BE0181; BioXCell, USA), anti-mouse IgG2a isotype control (BE0085; BioXCell), anti-mouse IL-9 receptor (IL-9R) (SC699; Santa Cruz, USA), anti-mouse IL-2Rγ (SC668; Santa Cruz), anti-mouse IgG isotype control (GTX35009; Santa Cruz), and donkey pAb to Rb IgG Alexa Flour 555 (Ab150074; Abcam, USA).

Techniques: In Vitro, Co-Culture Assay, Flow Cytometry, Cell Culture