The work described the synthesis and evaluation of PEI-g-comb-PEG-transferrin as a potential system for gene therapy in vitro. The MW of PEG was 10KDa, and PEI was 2KDa. Its structure was identified by NMR, FT-IR and TGA spectroscopy. MTT assay found that at concentration up to 4000 n mol/L of the polymer, cell viability was over 85%. The bio-character of polymer/DNA complex was characterized by agarose gel electrophoresis, ethidium bromide exclusion and zeta-potential assay. The polymer could retardate DNA at N/P ratio 3.0-3.5 （mol/mol）. The particle size of the polymer/DNA complex was less than 300 nm. Transfection efficiency of the complex was studied in COS7 and NT2 cell lines.
Here,we investigated the antitumor effect of adenovirus-mediated gene transfer of LIGHT,the tumor-necrosis factor(TNF)superfamily member also known as TNFSF14,in the murine A20 B-cell lymphoma.LIGHT gene modification resulted in upregulated expression of Fas and the accessory molecule-intercellular adhesion molecule-1(ICAM-1)on A20 cells and led to enhanced A20 cell apoptosis.LIGHT-modified A20 cells effectively stimulated the proliferation of T lymphocytes and interferon(IFN)-c production in vitro.Immunization of BALB/c mice with a LIGHT-modified A20 cell vaccine efficiently elicited protective immunity against challenge with the parental tumor cell line.Adenovirus-mediated gene transfer of LIGHT by intratumoral injection exerted a very potent antitumor effect against pre-existing A20 cell lymphoma in BALB/c mice.This adenovirus-mediated LIGHT therapy induced substantial splenic natural killer(NK)and cytotoxic T lymphocyte(CTL)activity,enhanced tumor infiltration by inflammatory cells and increased chemokine expression of CC chemokine ligand 21(CCL21),IFN-inducible protein-10(IP-10)and monokine induced by IFN-c(Mig)from tumor tissues.Thus,adenovirus-mediated LIGHT therapy might have potential utility for the prevention and treatment of B-cell lymphoma.
Guili HuYang LiuHong LiDekuang ZhaoLiuqing YangJiangen ShenXuejun HongXuetao CaoQingqing Wang
