Enhanced transduction efficiency of fiber-substituted adenovirus vectors by the incorporation of RGD peptides in two distinct regions of the adenovirus serotype 35 fiber knob

3区 · 医学

Article

作者: Katsuhisa Tashiro ; Kenji Kawabata ; Fuminori Sakurai ; Hayato Matsui ; Kazufumi Katayama ; Kumiko Sugio ; Hiroyuki Mizuguchi ; Shinnosuke Kurachi

Fiber-substituted Ad serotype 5 vectors containing the fiber protein from Ad serotype 35 (Ad5F35) exhibit properties that render them suitable as a platform for targeted Ad vectors. Ad5F35 vectors do not show apparent tropism in certain organs, including the liver, and they elicit less innate immunity than other vectors after intravenous administration. In order to develop a targeted Ad vector, we previously developed fiber-mutant Ad5F35 vectors containing the integrin binding Arg-Gly-Asn (RGD) motif in the FG or HI loop of the Ad35 fiber knob. Mutant Ad5F35 vectors containing the RGD peptide in the FG or HI loop transduced CD46-negative cells more efficiently in an RGD-dependent manner, as compared to the efficiency achieved with unmodified Ad5F35 vectors (Matsui et al., 2009. Gene Therapy 16, 1050-1057). However, the transduction efficiency of the mutant Ad5F35 vectors in CD46-negative cells remained lower than had been expected. Ad5F35 vectors containing the RGD peptide in the HI or FG loop enabled a 6-fold higher transduction efficiency than that achieved with unmodified Ad5F35 vectors in CD46-negative cells, although this cell type abundantly expresses α(v)-integrins. In the present study, we aimed to enhance the transduction efficiency of fiber-mutant Ad5F35 vectors. To this end, we developed an Ad5F35-vector system in which foreign peptides could be incorporated into regions of FG and HI loops of the Ad35 fiber knob by means of in vitro ligation. Using this Ad5F35-vector system, firefly luciferase-expressing mutant Ad5F35 vectors containing the RGD peptides in both loops (Ad5F35-2xRGD-L2) were constructed. In CD46-negative cells, Ad5F35-2xRGD-L2 showed 12-fold and 3-fold greater transduction efficiency than unmodified Ad5F35 vectors and mutant Ad5F35 vectors containing only one copy of the RGD peptide in the FG or the HI loop. In addition, transduction with Ad5F35-2xRGD-L2 in CD46-negative cells was RGD peptide-dependent. These results indicate that fiber-mutant Ad5F35 vectors, by which foreign peptides can be simultaneously incorporated into both the FG and the HI loops of the Ad35 fiber knob, could be a promising gene delivery vehicle for various gene therapies, and could facilitate basic research efforts such as analyses of gene function.

2008-08-01·Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology

[Dendritic cells transfected with recombinant adenovirus Ad5F35-LMP2 induces LMP2 specific immunity mediated by cytotoxic T lymphocytes in vitro].

Article

作者: Zeng, Yi ; Tang, An-Zhou ; Huang, Guang-Wu ; Mo, Wu-Ning ; Zhou, Ling ; Wang, Zhan

OBJECTIVETo observe whether dendritic cells (DCs) transfected with recombinant adenovirus Ad5F35-LMP2 induces LMP2 specific immunity mediated by cytotoxic T lymphocytes in vitro.METHODSDendritic cells have been generated in vitro, and cocultured with autologous T cell after the DCs were infected with Ad5F35-LMP2, then the proliferation of the induced T cells and their cytotoxic activity against CNE-2 tumor cells which express EBV-LMP2 protein on membrane were assessed by MTT method.RESULTSThe dendritic cells could be transfected with Ad5F35-LMP2 and the CTL activated by Ad5F35-LMP2-DC could effectively suppress the proliferation of CNE-2 cells compared with control groups.CONCLUSIONThe dendritic cells transfected with recombinant adenovirus Ad5F35-LMP2 showed cytotoxicity effect by activating T lymphocytes.

2007-09-01·Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology

[Immune responses induced by recombinant adenovirus Ad5F35-LMP2 in rhesus monkeys].

Article

作者: Du, Hai-Jun ; Wang, Qi ; Yu, Shuang-Qing ; Ye, Shu-Qing ; Huang, Guang-Wu ; Wu, Xiao-Bing ; Zeng, Yi ; Mo, Wu-Ning ; Zhou, Ling ; Tang, An-Zhou ; Wang, Zhan

OBJECTIVETo observe the specific cellular and humoral immune responses after immunization with recombinant adenovirus Ad5F35-LMP2 in rhesus monkeys.METHODSSixteen rhesuses were immunized with Ad5F35-LMP2 through intra-muscular injection in three groups: high dosage group (1.5 x 10(10) TCID(50)/rhesus), medium dosage group (1.5 x 10(9)TCID(50)/rhesus), low dosage group (1.5 x 10(8)TCID50/rhesus) and the last group was control (PBS 4 ml/rhesus). They were totally immunized three times at intervals of one month. The EBV-LMP2 specific cellular immune responses were tested during the 0, 4, 8, 12 weeks by Elispot after immunization respectively. And the titers of anti-LMP2 antibody were tested by EIA at the same time.RESULTSEBV-LMP2 specific cellular and humoral immune responses which were induced by recombinant adenovirus Ad5F35-LMP2 can be found in all the three dosage groups. The potency of immune responses was related with the dosage of immunization. Higher dosage elicited more potent immune response.CONCLUSIONThe recombinant adenovirus Ad5F35-LMP2 could elicit LMP2 specific cellular and humoral immune responses in rhesus.

100 项与 Ad5f35-LMPd1-2-transduced autologous dendritic cells(National Cancer Institute) 相关的药物交易

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