Journal: Gene therapy

Article Title: Synergistic antileukemia effect of combinational gene therapy using murine beta-defensin 2 and IL-18 in L1210 murine leukemia model.

doi: 10.1038/sj.gt.3302966

Figure Lengend Snippet: Figure 1 Leukemogenicity of genetically modified L1210 cells. Groups of DBA/2 mice (n ¼ 10) were injected i.p. with live L1210- MBD2, L1210-pro-IL-18-ICE and a mixture of equal numbers of the two cells. Pro-IL-18, ICE and pIRES empty plasmid transduced and wild-type L1210 cells were injected as controls. Cells were diluted in 200 ml of PBS before injection. (a) The dose of cells injected was 1 105. (b) The dose of cells injected was 1 106. Experiments were repeated three times. The overall survival rates of mice vaccinated with L1210-MBD2 or L1210-pro-IL-18-ICE or the mix of the two cells were significantly higher than those of the other groups (Po0.001). The survival rates of the mice vaccinated with the mix of the L1210-MBD2 and L1210-pro-IL-18-ICE were significantly higher than that with either of the cells alone (Po0.05). MBD2, murine b-defensin 2.

Article Snippet: Transfection of L1210 cells and IL-18 bioassay The pro-IL-18-ICE, pro-IL-18 and ICE expression plasmid and empty plasmid vector were transduced into L1210 cells by using Lipofectamine 2000 reagent (Life Technologies, Grand Island, NY, USA).

Techniques: Injection, Plasmid Preparation

Journal: Gene therapy

Article Title: Synergistic antileukemia effect of combinational gene therapy using murine beta-defensin 2 and IL-18 in L1210 murine leukemia model.

doi: 10.1038/sj.gt.3302966

Figure Lengend Snippet: Figure 2 Therapeutic effect of the irradiated genetically modified L1210 cells. DBA/2 mice (n ¼ 6) were first injected i.p. with 104 live parental L1210 cells, and were treated on the same day with eight subsequent subcutaneous injections of irradiated (10 Gy) L1210-MBD2 or L1210-pro-IL-18-ICE or a mixture of equal numbers of the two cell lines, delivered two times a week. The same amount of irradiated L1210-pro-IL-18, L1210-ICE, L1210-pIRES and parental L1210 cells were used as controls. Experiments were repeated three times. Compared with the four controls, L1210-MBD2 and the mixture of L1210-MBD2 and L1210-pro-IL-18-ICE cells signifi- cantly prolonged the survival time (Po0.05), and the mixture of L1210-MBD2 and L1210-pro-IL-18-ICE elicited more marked therapeutic effect than either of cells alone (Po0.05). MBD2, murine b-defensin 2.

Article Snippet: Transfection of L1210 cells and IL-18 bioassay The pro-IL-18-ICE, pro-IL-18 and ICE expression plasmid and empty plasmid vector were transduced into L1210 cells by using Lipofectamine 2000 reagent (Life Technologies, Grand Island, NY, USA).

Techniques: Irradiation, Injection

Journal: Gene therapy

Article Title: Synergistic antileukemia effect of combinational gene therapy using murine beta-defensin 2 and IL-18 in L1210 murine leukemia model.

doi: 10.1038/sj.gt.3302966

Figure Lengend Snippet: Figure 3 NK cells and CTL cytotoxicity and IFN-g production induced by genetically modified L1210 cells. Naı¨ve DBA/2 mice (n ¼ 3) were injected with 1 105 live L1210-MBD2 or L1210-pro-IL- 18-ICE or a mixture of equal numbers of the two cells. Pro-IL-18, ICE and pIRES empty plasmid transduced and wild-type L1210 cells were injected as controls. Seven days after injection, spleen cells were obtained from each mouse. (a) NK activity. NK activity was assessed by a standard lactate dehydrogenase (LDH) released assay, using the spleen cells as effector cells and YAC-1 cells as target cells. Values were presented as mean7s.d. of three mice per group. Experiments were repeated two times with similar results. The cytotoxicity assay showed significantly higher NK cell mediated lysis of YAC-1 in groups of mice vaccinated with MBD2, pro-IL-18-ICE, and MBD2 + pro-IL-18-ICE transduced cells, compared with the four control groups (Po0.05), and there were no significant differences among the three test groups (P>0.05). (b) CTL activity. Spleen cells were restimulated with irradiated parental L1210 cells for 5 days. CTL activity of the resultant effector cells was measured against parental L1210 targets in a standard LDH released assay. mean7s.d. were shown for three mice per group. Similar results were obtained in two independent experiments. CTL activity of mice inoculated with MBD2, pro-IL-18-ICE, and a mix of MBD2 and pro-IL-18-ICE transduced L1210 cells were significantly higher than that of the four control groups (Po0.05). MBD2+pro-IL-18- ICE group induced significantly stronger CTL activity than either MBD2 or pro-IL-18-ICE alone (Po0.05). (c) IFN-g production. Spleen cells were restimulated with irradiated L1210 cells in an IFN-g ELISPOT plate for 1 day, and spots were counted with background being subtracted. The spot numbers were shown as mean7s.d. from three mice per group. Similar results were obtained in two independent experiments. Mice inoculated with L1210-MBD2 and L1210-MBD2+L1210-pro-IL-18-ICE generated significantly increased IFN-g, as compared with the four control groups (Po0.05). Among all the groups, the combinatorial delivery of MBD2 and pro-IL-18-ICE transduced L1210 cells resulted in the highest production of IFN-g (Po0.05). 1, L1210-MBD2+L1210- proIL18-ICE; 2, L1210-MBD2; 3, L1210-proIL-18-ICE; 4, L1210- proIL-18; 5, L1210-ICE; 6, L1210-pIRES; 7, parental L1210; 8, saline.

Article Snippet: Transfection of L1210 cells and IL-18 bioassay The pro-IL-18-ICE, pro-IL-18 and ICE expression plasmid and empty plasmid vector were transduced into L1210 cells by using Lipofectamine 2000 reagent (Life Technologies, Grand Island, NY, USA).

Techniques: Injection, Plasmid Preparation, Activity Assay, Cytotoxicity Assay, Lysis, Control, Irradiation, Enzyme-linked Immunospot, Generated, Saline

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Transgenic Overexpression of Tissue‐Nonspecific Alkaline Phosphatase ( TNAP ) in Vascular Endothelium Results in Generalized Arterial Calcification

doi: 10.1161/JAHA.115.002499

Figure Lengend Snippet: Expression of eTNAP. A, AP activity staining of the aorta, heart, lung, kidney, liver, small intestine, spleen, thymus, pancreas, and mesenteric LN from WT and eTNAP mice aged 8 weeks. B, Colocalization of AP activity with endothelial‐specific lectin ( IB 4) staining in the consecutive sections of myocardium counterstained with DAPI ( DNA ). C, Whole‐body x‐ray images of WT and eTNAP mice aged 8 weeks. AP indicates alkaline phosphatase; eTNAP, endothelial tissue‐nonspecific alkaline phosphatase; IB4, isolectin B4; LN, lymph node; WT, wild type.

Article Snippet: ECs were detected with FITC‐labeled isolectin B4 (IB4) from Bandeiraea simplicifolia (item FL1201; Vector Laboratories).

Techniques: Expressing, Activity Assay, Staining

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Transgenic Overexpression of Tissue‐Nonspecific Alkaline Phosphatase ( TNAP ) in Vascular Endothelium Results in Generalized Arterial Calcification

doi: 10.1161/JAHA.115.002499

Figure Lengend Snippet: Structural characteristics of arterial lesions in eTNAP mice. A, Detection of Alizarin Red–positive lesions in flat‐mounted preparations of the aortas from eTNAP mice aged 8 weeks. B, Calcification in coronary artery sinus (top) and coronary arteries (bottom) in eTNAP mice aged 8 weeks visualized by Alizarin Red staining. C, Subendothelial localization of coronary lesions (highlighted by arrows) in eTNAP mice aged 13 weeks detected by AP and endothelial‐specific ( IB 4) staining in consecutive sections counterstained with DAPI ( DNA ). D, von Kossa–positive calcium deposits in mesenteric arteries of eTNAP mice aged 13 weeks. *, lumen. E, Mesenteric arteries from eTNAP mice aged 13 weeks, with hematoxylin and eosin staining; (double arrow) neointima. *, lumen. F, Masson's trichrome staining of coronary, renal, mesenteric, and spenic arteries from eTNAP mice aged 13 weeks. *, lumen. AP indicates alkaline phosphatase; eTNAP, endothelial tissue‐nonspecific alkaline phosphatase; IB4, isolectin B4; WT, wild type.

Article Snippet: ECs were detected with FITC‐labeled isolectin B4 (IB4) from Bandeiraea simplicifolia (item FL1201; Vector Laboratories).

Techniques: Staining

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Transgenic Overexpression of Tissue‐Nonspecific Alkaline Phosphatase ( TNAP ) in Vascular Endothelium Results in Generalized Arterial Calcification

doi: 10.1161/JAHA.115.002499

Figure Lengend Snippet: Expression of Runx2, collagen II , osteopontin, and sialic acid in coronary lesions of eTNAP mice. A through D, coronary lesions were identified by DIC imaging. A, Consecutive sections of the hearts from eTNAP mice aged 8 weeks were stained with Runx2 antibody or secondary antibody only, DNA binding dye DAPI was used as counterstain. B, Lesions from eTNAP mice aged 8 weeks were stained with collagen II antibody or secondary antibody alone and counterstained with DAPI. C, Osteopontin expression was detected in the lesions from eTNAP mice aged 13 weeks by direct immunofluorescent staining; IB 4 was used as counterstain. D, The presence of sialic acid was detected in the coronary lesions from eTNAP mice aged 13 weeks with fluorescein‐labeled SNA lectin; DAPI was used as nuclear counterstain. Affected (left panels) and unaffected (right panels) coronary arteries from the same tissue section are shown. DIC indicates differential interference contrast; eTNAP, endothelial tissue‐nonspecific alkaline phosphatase; IB4, isolectin B4; SNA, Sambucus nigra .

Article Snippet: ECs were detected with FITC‐labeled isolectin B4 (IB4) from Bandeiraea simplicifolia (item FL1201; Vector Laboratories).

Techniques: Expressing, Imaging, Staining, Binding Assay, Labeling

Journal: PLoS ONE

Article Title: Plaque Size Is Decreased but M1 Macrophage Polarization and Rupture Related Metalloproteinase Expression Are Maintained after Deleting T-Bet in ApoE Null Mice

doi: 10.1371/journal.pone.0148873

Figure Lengend Snippet: Primary (A) and secondary (B) antibodies and Griffonia simplicifolia Lectin I (GSL).

Article Snippet: , GSL I , VectorLabs, USA B1215 , Lectin , 1:100.

Techniques: Concentration Assay