Journal: Experimental and Therapeutic Medicine

Article Title: IL-35 improves T reg -mediated immune suppression in atherosclerotic mice

doi: 10.3892/etm.2016.3649

Figure Lengend Snippet: Plasma levels of Foxp3. Peripheral blood mononuclear cells were collected from each group, and the proportions of CD4 + CD25 + Foxp3 + T reg /CD4 + T-cells were analyzed by flow cytometry, and quantified using FlowJo 7.6.1 software. (A) Histogram. Data are presented as the mean ± standard deviation. *P<0.01, vs. the AS group. (B) Negative group, (C) AS group, (D) atorvastatin group and (E) IL-35 group. Foxp3, forkhead box protein 3; AS, atherosclerosis; IL-35, interleukin-35; APC, allophycocyanin; PE, phycoerythrin.

Article Snippet: Anti-Foxp3 antibody was purchased from Wuhan Boster Biological Technology, Ltd. (Wuhan, China).

Techniques: Clinical Proteomics, Flow Cytometry, Software, Standard Deviation

Journal: Experimental and Therapeutic Medicine

Article Title: IL-35 improves T reg -mediated immune suppression in atherosclerotic mice

doi: 10.3892/etm.2016.3649

Figure Lengend Snippet: Levels of Foxp3 in atherosclerotic lesions. The deposition of Foxp3 in arteries from various groups was detected by immunohistochemistry (magnification, ×400). Positive Foxp3 was shown as brown nuclei. In the (A) negative and (B) AS groups, the deposition of Foxp3 in the lesions was minimal. Conversely, Foxp3 deposition was increased in the lesions of the (C) atorvastatin and (D) IL-35 groups, as compared with the AS group. (E) This was shown to be significant following quantification. There was no significant difference in the levels of Foxp3 between the atorvastatin and IL-35 groups. *P<0.01, vs. the negative control. Foxp3, forkhead box protein 3; AS, atherosclerosis; IL-35, interleukin-35.

Article Snippet: Anti-Foxp3 antibody was purchased from Wuhan Boster Biological Technology, Ltd. (Wuhan, China).

Techniques: Immunohistochemistry, Negative Control

Journal: International Journal of Nanomedicine

Article Title: A Mansonone Derivative Coupled with Monoclonal Antibody 4D5-Modified Chitosan Inhibit AKR1C3 to Treat Castration-Resistant Prostate Cancer

doi: 10.2147/IJN.S241324

Figure Lengend Snippet: Affinity of CS-4D5 and CS-4D5/6e for 22Rv1 and LNCaP cells. ( A ) Western blot analysis relative expression of HER2 receptor in 22RV1 and LNCaP cells. Data are expressed as the mean SEM (n = 3). * P < 0.05 vs α-tubulin. ( B ) Immunofluorescence double co-localization assay was used to compare the affinity of CS-4D5 and CS-4D5/6e in 22RV1 and LNCAP cells. Red fluorescence (Alexa Fluor 555) represents the location of the HER2 receptor, whereas green fluorescence (Alexa Fluor 488) represents the location of CS-4D5 and CS-4D5/6e. Abbreviations: HER2, human epidermal growth factor receptor 2; CS, chemically chitosan; 4D5, single chain antibody fragment 4D5; 6e, a derivative of mansonone F; DAPI, 4ʹ,6-diamidino-2-phenylindole; SEM, standard error of mean.

Article Snippet: Cell membranes were probed with primary antibodies overnight against AR (rabbit monoclonal antibody (mAb); 5153S; Cell Signaling Technology), PSA (rabbit mAb; 5877S; Cell Signaling Technology), AKR1C3 (rabbit mAb; ab203834; Abcam, Cambridge, UK), HER2 (mouse mAb; bsm-33051M; Bioss) or α-tubulin (rabbit mAb; A01080; Abbkine, Wuhan, China) followed by incubation with anti-mouse (AA75181; Bioworld Technology, Saint Louis Park, MN, USA) or anti-rabbit (AA01191; Bioworld Technology) horseradish peroxidase-conjugated secondary antibody (1:5000 dilution) for 1 h, followed by chemiluminescence detection.

Techniques: Western Blot, Expressing, Immunofluorescence, Fluorescence

Journal: bioRxiv

Article Title: Decoupling individual host response and immune cell engager cytotoxic potency

doi: 10.1101/2024.06.22.600188

Figure Lengend Snippet: ADCC assay and dependence of potency (EC 50 ) on bispecific antibody (bsAb) and NK cell donor. A. Schematic representation of co-culture assay mixing adherent tumour target cells (MCF7-WT, SKBR3 or MCF7-HER2+) and primary human NK cells from 15 donors in the presence of bsAb. Six bsAb anti-HER2 × CD16 constructs are tested, based on 3 anti-HER2 (Nanobodies CE4, CA5 or C7b) and 2 anti-CD16 (Nanobodies C21 or C28). B. Example of lysis fraction vs bsAb concentration measured on the MCF7-HER2+ target cell line and bispecific Ab C7b-21, donor A . Data was fitted with Eq. (1) (black line). C. Result of Hill fit for all conditions. Data are normalized using the fitting parameters Min, Max and EC 50 and compared to the normalized Hill function c/ (1 + c ) with c in nM units (black line). Raw residuals are shown below. D. EC 50 for each target cell line and bsAb. Each point is the median on the donors, with the bar representing 95% percentile interval. bsAbs are ranked according to the median value for SKBR3 cell line. E. EC 50 for each target cell line and donor. Each point is the median on the bsAbs, with the bar representing 95% percentile interval. Donors are ranked according to the median value for SKBR3 cell line.

Article Snippet: To obtain them, MCF7 cells were electroporated with 1 µ g of DNA plasmid HER2-GFP from Sino biologicals (ref HG10004-ACG) with Nucle-ofector 2b device (Lonza), and selected by antibiotic hygromycine.

Techniques: ADCC Assay, Co-culture Assay, Construct, Lysis, Concentration Assay

Journal: bioRxiv

Article Title: Decoupling individual host response and immune cell engager cytotoxic potency

doi: 10.1101/2024.06.22.600188

Figure Lengend Snippet: Physical model for bsAb dependent cell-mediated cytotoxicity. A. Reaction scheme for the two-step binding of bispecific antibody L at the immune synapse on membrane receptors R 1 (tumour side, HER2) and R 2 (effector side, CD16). K 1 , K 2 , K S 1 , K S 2 are dissociation constants. B-C. The equilibrium density σ of bridging bsAbs as a function of bulk bsAb concentration c given by the solution of . Each colored curve has been obtained for a different value of the cooperativity parameter α varied from 0.01 to 10. Other parameters K 1 , K 2 , ∑ 1 , ∑ 2 correspond to Target MCF7-WT and antibody C7b-21 (B) or Target SKBR3 and antibody CE4-28 (C) (values in Tabs. S1 and S2). The horizontal line in (B,C) represents an example of σ threshold value for the NK cell cytotoxic response (here σ 50 = 0.1 molec/ µ m 2 ). Together with the value of the cooperativity (here α = 0.01), it sets the value of c 50 , such that σ ( c 50 ) = σ 50 . The maximum density σ * is found for .

Article Snippet: To obtain them, MCF7 cells were electroporated with 1 µ g of DNA plasmid HER2-GFP from Sino biologicals (ref HG10004-ACG) with Nucle-ofector 2b device (Lonza), and selected by antibiotic hygromycine.

Techniques: Binding Assay, Membrane, Concentration Assay

Journal: bioRxiv

Article Title: Decoupling individual host response and immune cell engager cytotoxic potency

doi: 10.1101/2024.06.22.600188

Figure Lengend Snippet: Correlation of best-fit parameters of the multiscale model between cell lines. A. MCF7-WT and SKBR3. B. MCF7-WT and MCF7-HER2+. C. SKBR3 and MCF7-HER2+. P: Pearson coefficient. S: Spearman coefficient. n: number of points. Dashed lines indicate x = y .

Article Snippet: To obtain them, MCF7 cells were electroporated with 1 µ g of DNA plasmid HER2-GFP from Sino biologicals (ref HG10004-ACG) with Nucle-ofector 2b device (Lonza), and selected by antibiotic hygromycine.

Techniques: