Suppression effect of secreting-trail adenovirus on the prostate cancer PC-3 cells in the tumor bearing mice / 实用医学杂志

Objective To investigate the suppression effect of secreting-trail adenovirus on the prostate cancer PC-3 cells in the tumor bearing mice. Methods Apoptosis of PC-3 cells was measured by DAPI staining in vitro . Invasion of PC-3 cells was measured by transwell chamber assay. The tumor bearing mice were prepared and used to test the inhibition rates in different groups. The protein expression of TRAIL in different groups tumor bearing mice was determined by immunohistochemistry. The variation of cell apoptosis in different groups of tumor bearing mice was detected by TUNEL assay. Results DAPI staining showed that the apoptosis level in Ad-sTRAIL group was significantly higher than that in the control group and that in the Ad-LacZ group ,respectively. The number of PC-3 cells invading the inferior chamber in the Ad-sTRAIL group(24.8 ± 3.70) was significantly decreased compared with that in the control group(47.6 ± 4.28,q=10.68,P<0.01)and that in the Ad-LacZ group (49.6 ± 4.39,q=11.61,P<0.01). Growth inhibition assays in vivo showed the inhibition rate of Ad-sTRAIL was 3 d 8.15%,6 d 11.55%,9 d 17.23%,12 d 20.05%,15 d 27.18%,18 d 34.27%,and 21 d 33.08%,respective-ly. Immunohistochemistry results showed that the expression level of TRAIL in tumors of Ad-sTRAIL group tumor-bearing mice was significantly higher than that in the control group and that in the Ad-LacZ group. TUNEL assay showed that the apoptosis level of tumor bearing mice tumor cells in the Ad-sTRAIL group was significantly higher than that in the control group and that in the Ad-LacZ group. Conclusion The secreting-trail adenoviral vector could inhibit the growth of tumor bearing mice tumor cells,furthermore it could induce the apoptosis of PC-3 cells in the tumor bearing mice.

A novel rabies vaccine based on the chimpanzee adenoviral vector AdC68 / 中华微生物学和免疫学杂志

Objective To investigate the genetic stability, immunogenicity and protective efficacy of AdC68-rab. gp, a novel rabies vaccine based on the replication-defective chimpanzee adenoviral vector AdC68-ept. Methods The recombinant adenovirus AdC68-rab. gp expressing the glycoprotein of rabies vi-rus ERA strain was constructed. Genomes of the AdC68-rab. gp of different generations were extracted and analyzed. HEK293 and Huh7 cells were infected with the AdC68-rab. gp of different generations. ICR mice were immunized with the AdC68-rab. gp and blood samples were collected 4 weeks or 6 months after immuni-zation. Rapid fluorescent focus inhibition test ( RFFIT) was performed to detect the neutralizing antibody against rabies virus in mice serum samples. ICR mice were challenged with lethal dose of rabies virus 4 weeks after the immunization with AdC68-rab. gp to evaluate the protective efficacy of AdC68-rab. gp. Re-sults The genome of AdC68-rab. gp was stable after 15 passages, which was identical to that of the 5th and 1st generations. High levels of neutralizing antibody against rabies virus in serum samples were detected in mice immunized with AdC68-rab. gp and maintained for a long period of time. Immunization mice with one dose of AdC68-rab. gp could protect all mice from the lethal dose challenge of rabies virus. Conclusion The novel AdC68-rab. gp was characterized by good genetic stability and ideal protective effi-cacy. The adenoviral vector based vaccine could be further developed as a potential candidate for the substi-tute of current rabies vaccine.

Effect of Heat Shock Protein 72 Expression on Etoposide-induced Cell Death of Rat Retinal Ganglion Cells

PURPOSE: To assess whether the expression of heat shock protein 72 (Hsp72) protects rat retinal ganglion cells (RGC-5) from apoptotic cell death. METHODS: Hsp72 expression in RGC-5 cells transduced with replication-deficient recombinant adenovirus was analyzed by Western blot analysis and immunofluorescence. The effect of Hsp72 expression on etoposide-induced apoptotic cell death was examined by microscopic analysis and confirmed by cell proliferation assay. RESULTS: Western blot analysis and immunofluorescence clearly showed adenovirus-mediated Hsp72 expression in RGC-5 cells. Treatment with etoposide resulted in the death of a proportion of the cells by apoptosis. However, this apoptotic cell death was significantly reduced in cells expressing Hsp72, with the reduction in cell death correlating to the level of Hsp72 expression. CONCLUSIONS: Over-expression of Hsp72 alone is sufficient to rescue neuronal cells from apoptotic cell death, suggesting that fine-tuning its expression may be an effective neuroprotective approach in retinal degenerative disease.

Effect of blocking the signal pathway of TGFβ on presenting antigen of dendritic cells / 肿瘤研究与临床

Objective To explore the dysfunction of dendritic cells (DC) related to TGFβ reversed after blocking the TGFβ signal pathway by recombinant adenovirus vector encoding for Smad7.Methods Smad7 by recombinant adenovirus vector was transfected into dendritic cells.Expression of immunologic phenotypes was detected by FCM,and CTL activity induced by DC was compared.Results The DC modified with Smad7 still expressed high adhesiveness factor related to maturation even if existing exogenous TGFβ1,which was significant statistically compared with DC transfected with control adenoviral vector (P ＜0.01).Even if existing exogenous TGFβ1,the DC modified with Smad7 pulsed with soluble antigen associated with Lewis pulmonary carcinoma could still induce potent CTL activity against Lewis pulmonary carcinoma,which showed significant difference with DC-Ad-c (P ＜0.01).Conclusion The inhibitory effects on function of DC of TGFβ may be reversed by blocking the Smad signal of TGFβ pathway.

The growth-inhibition of osteosarcoma xenografts by adenovirus-mediated ING4 gene in nude mice / 中国免疫学杂志

Objective:To study the inhibitory effect and anti-cancer mechanisms of adenovirus-mediated ING4 gene on the MG-63 osteosarcoma xenografts in nude mice.Methods:Ad-ING4 was transfected into QBI-293 cells and harvested.15 nude mice of the subcutaneous tumor models were established with MG-63 osteosarcoma cells and were randomly divided into PBS,Ad-GFP and Ad-ING4 groups.Then PBS(100 μl),Ad-GFP(100 μl,10~9pfu/ml) and Ad-ING4 (100 μl,10~9pfu/ml) for each one were given respectively QOD for 5 times,with intratumor injections.Tumor volume changes were monitored;and the 15 mice were sacrificed 2 weeks after treatment,the tumors were removed,weighed and ratios of tumor-suppression were calculated.The morphological changes of apoptotic tumor cells were observed under microscope.Bcl-2,Bax,Caspase-3,VEGF,CD34 expression was tested by immumohistochemistry.Results:High titer(10~9pfu/ml)adenoviral vector of ING4 gene were obtained.In nude mice bearing MG-63 osteosarcoma xenografts,the growth of MG-63 tumors treated by intratumoral injecting of Ad-ING4 was significantly suppressed,compared with PBS group and Ad-GFP group.The ratios of tumor weight-suppression of Ad-ING4 group was 59.3%(P＜0.05).Immumohistochemistry displayed that the expression of Bax,Caspase-3 was up-regulated and the expression of Bcl-2,VEGF,CD34 was down-regulated by Ad-ING4.Conclusion:Ad-ING4 can inhibit the growth of MG-63 osteosarcoma xenografts in nude mice,which may be via activating the apoptosis pathway and inhibiting tumor angiogenesis.

Construction and expression of secretary recombinant adenovis vectors carrying mPSMA / 基础医学与临床

Objective To construct secretary recombinant adenoviral vector carrying the mouse prostate-specific mem-brane antigen gene through AdEasy vector system and the immunological outcome was assessed. Methods mPSMA was amplified from plasmid pCR-BluntⅡ-TOPO by PCR and subcloned into transfer vector pAdeno Vator CMV5, The signal peptide DNA sequence of hIL-2 was fused to 5'terminal of mPSMA gene to construct a secretary Ad-mPSMA. pAdv-mPSMA was co-transformed with pAdeno Vator △E1/E3 through homologous recombination. The recombinant adenoviruses were packaged, amplified and purified in HEK293 cells. HeLa cell was infected by recombinant ade-novirus Ad-mPSMA and the expression of mouse prostate-specific membrane antigen gene was detected by RT-PCR and Western blot. The recombinant adenovirus had been immuned mice, sera antibody against mPSMA from immu-nized mice was detected by ELISA. Results The secretary pAd-mPSMA was constructed successfully and typical cytopathic effect (CPE) was observed. The titer of the recombinant adenovirus was 1.32 × 10~(11)IU/mL and expres-sion of mPSMA was confirmed by RT-PCR and Western blot. The specific antibody against mPSMA had been found in serum of the immunized mice. Conclusion mPSMA gene recombinant adenovirus was constructed successfully, which provide a basis for further study on the anti-tumor immunotherapy role of PSMA.

Production and Quality Control of Adenoviral Vectors for Clinical Trials

The importance of recombinant adenoviral vectors for the development of gene therapy and prophylactic and therapeutic vaccines has led to efforts for process development of large scale production of clinically safe adenoviral vectors. First of all, cell lines producing replication incompetent adenoviral vectors required for clinical application have been developed and the concept of banking and characterization of cell lines and adenoviral vectors has been established. In order to meet the need of amount of adenoviral vectors for clinical trials, various large scale suspension culture methods using serum-free media have been developed along with development of large scale purification methods using chromatography instead of cesium chloride method. In addition, methods for the quality control of adenoviral vectors have been established and applied for the clinical lots.

B Cells Transduced with HPV16 E6/E7-expressing Adenoviral Vector Can Efficiently Induce CTL-dependent Anti-Tumor Immunity

BACKGROUND: Human papillomavirus (HPV) infection is responsible for cervical cancer, a common cancer in women. Since HPV infection and cancer development are controlled by the host immune system, immunotherapy against HPV can be helpful in preventing or treating HPV-associated cervical cancer. Two oncoproteins of HPV16, E6 and E7, are promising targets for immunotherapy against cervical cancer, because they are constitutively expressed in cervical cancer. METHODS: Since cellular vaccines using B cells as well as dendritic cells offer an efficient approach to cancer immunotherapy, we opted to use B cells. We evaluated the immunogenicity and anti-tumor effects of a B cell vaccine transduced with HPV16 E6/E7-expressing adenovirus. RESULTS: Vaccination with HPV16 E6/E7-transduced B cells induced E6/E7-specific CD8+ T cell-dependent immune responses and generated anti-tumor effects against E6/E7-expressing TC-1 tumor. The anti-tumor effect induced by this B cell vaccine was similar to that elicited by DC vaccine, showing that B cells can be used as an alternative to dendritic cells for cellular vaccines. CONCLUSION: Thisstudy has shown the feasibility of using B cells as immunogenic APCs and the potential for developing prophylactic and therapeutic vaccines against HPV-associated cervical cancer using a B cell vaccine transduced with adenovirus expressing HPV16 E6/E7.

Construction and expression of secretary recombinant adenovis vectors carrying mPSMA / 基础医学与临床

Objective To construct secretary recombinant adenoviral vector carrying the mouse prostate-specific membrane antigen gene through AdEasy vector system and the immunological outcome was assessed.Methods mPSMA was amplified from plasmid pCR-BluntII-TOPO by PCR and subcloned into transfer vector pAdenoVator CMV5,The signal peptide DNA sequence of hIL-2 was fused to 5′terminal of mPSMA gene to construct a secretary Ad-mPSMA.pAdv-mPSMA was co-transformed with pAdenoVator ?E1/E3 through homologous recombination.The recombinant adenoviruses were packaged,amplified and purified in HEK293 cells.HeLa cell was infected by recombinant adenovirus Ad-mPSMA and the expression of mouse prostate-specific membrane antigen gene was detected by RT-PCR and Western blot.The recombinant adenovirus had been immuned mice,sera antibody against mPSMA from immunized mice was detected by ELISA.Results The secretary pAd-mPSMA was constructed successfully and typical cytopathic effect(CPE) was observed.The titer of the recombinant adenovirus was 1.32?1011IU/mL and expression of mPSMA was confirmed by RT-PCR and Western blot.The specific antibody against mPSMA had been foundin serum of the immunized mice.Conclusion mPSMA gene recombinant adenovirus was constructed successfully,which provide a basis for further study on the anti-tumor immunotherapy role of PSMA.

Construction of recombinant adenoviral vector carrying the LEDGFp52 gene by homologous recombination in bacteria and its expression in vitro / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To construct recombinant adenoviral vector carrying the LEDGFp52 gene by homologous recombination in bacteria and to detect its expression in vitro.METHODS: The LEDGFp52 gene was cloned to adenoviral shuttle plasmid pAdTrack-CMV. Then, the resultant pAdTrack-CMV-LEDGFp52 was cotransfected into BJ5 183 bacteria with the adenoviral backbone plasmid pAdeasy-1. The adenoviral plasmid carrying LEDGFp52 was generated with homologous recombination in bacteria, and the adenoviruses were produced in 293 cells. These 293 cells were then infected with adenoviruses, and the expression of LEDGFp52 was detected by CPE (cytopathic effect) and western blot.RESULTS: The titer of Ad-LEDGFp52 adenoviruses was up to 5×1012 pfu/L after proliferation in 293 cells. LEDGFp52 was expressed efficiently in 293 cells after infection.CONCLUSION: The recombinant adenoviruses vector expressing LEDGFp52 was constructed successfully and can be used in further gene transfection experiments.

Construction of hNGF-betarecombinant adenovirus and screening of its expression after transfection into various cell lines

Antitumor Activity of TRAIL Recombinant Adenovirus in Human Malignant Glioma Cells

Tumor necrosis factor-Related Apoptosis-Inducing Ligand (TRAIL) has been reported to specifically kill malignant cells but to be relatively nontoxic to normal cells. One of disadvantages to previous in vivo protocols was the need for large quantities of TRAIL recombinant protein to suppress tumor growth. To evaluate the antitumor activity and therapeutic value of the TRAIL gene, we constructed adenoviral vectors expressing the human TRAIL gene (Ad.hTRAIL) and transferred them into malignant glioma cells in vitro and tumors in vivo, as an alternative to recombinant soluble TRAIL protein. The results show that TRAIL-sensitive glioma cells infected Ad.hTRAIL undergo apoptosis through the production and expression of TRAIL protein. The in vitro transfer elicited apoptosis, as demonstrated by the quantification of viable or apoptotic cells and by the analysis of cleavage of poly (ADP-ribose) polymerase. Furthermore, in vivo administration of Ad.hTRAIL at the site of tumor implantation suppressed the outgrowth of human glioma xenografts in SCID mice. These results further define Ad.hTRAIL as an anti-tumor therapeutic and demonstrate its potential use as an alternative approach to treatment for malignant glioma.

Expression of Wild Type and Truncated Myocilins in Trabecular Meshwork Cells: their Subcellular Localization

PURPOSE: To investigate subcellular localizations of wild type myocilin and trunctated (Q368X) myocilin in cultured human trabecular meshwork (TM) cells. METHODS: GFP and GFP tagged truncated myocilin, full-length myocilin, and stromelysin were expressed in TM cells using adenoviral vectors, and their secretory properties were examined by western blotting. To determine the subcellular localizations of myocilins, cellular organelles of the infected TM cells were stained with antibodies or orgenelle specific fluorescent indicators and examined under confocal microscope. RESULTS: Wild type myocilin was expressed as discrete fine vesicles in the perinuclear region of TM cells as well as in ER, but not in microtubules or mitochondria. Colocalization of wild type and truncated myocilin, indicative of in vivo interaction of the proteins, was also observed in cells co-expressing the proteins. Truncated myocilin was found to be accumulated as aggregates in ER, and inhibited the secretion of normal myocilin. CONCLUSIONS: Our result suggests that intracellular accumulation of truncated myocilin may cause a dysfunction of the cells, resulting in alterations in structural compartmentalization of trabecular ECM and obstruction of aqueous outflow.

Growth Suppression and Induction of Chemosensitivity in Human Gallbladder Epithelial Carcinoma Cells (GBCE) by Adenovirus-Mediated Transfer of the Wild-type p53 Gene / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Mutations in the p53 gene are reported in 50~90% of gallbladder and bile duct cancer, and have been implicated in chemoresistance. We undertook this study to determine whether the introduction of the wild type p53 gene into GBCE (human gallbladder cancer cell line with a heterozygous p53 mutation) by an adenoviral vector could increase the sensitivity of the cell to 5-FU, a commonly used drug in the treatment of gallbladder cancer. MATERIALS AND METHODS: GBCE cells were transfected with either Ad/p53 or Ad/E1 in the presence of 5-FU. Gene expression was confirmed by western blotting. Nude mice were injected subcutaneously with GBCE cells. When tumors formed, intratumoral injection of Ad/p53 was performed. Reduction of tumor size was compared in two weeks of Ad/p53 gene transfection. RESULTS: Ad/53 transfection induced a dose-dependent inhibition of tumor growth. Tumor colony formation was more inhibited with p53 gene transfection than with mock transfection in the presence of 5-FU. The reduction in tumor size was more pronounced with p53 transfection than with mock infection. CONCLUSION: These treatment modalities could be utilized in the treatment of p53 mutant human gallbladder cancers.

Attenuation of the Expression of Fibrogenic Molecules by Transforming Growth Factor-beta1 Antisense in Cultured Rat Mesangial Cells / 대한신장학회잡지

BACKGROUND: The diabetic milieu augments the expression of transforming growth factor-beta1 (TGF-beta1) in mesangial cells, which, in turn, stimulates the accumulation of extracellular matrix in glomeruli. Accordingly, mesangial cells have been shown to produce increased amount of TGF-beta1 when exposed to high glucose concentrations in vitro. METHODS: In the present study, we examined the effects of antisense TGF-beta1 gene transfer on the expressions of several cytokines implicated in kidney fibrosis. The DNA fragment containing bases -162 to +168 of rat TGF-beta1 sequence was isolated by reverse transcription-polymerase chain reaction (RT- PCR) from rat kidney tissue. This PCR product was ligated in the xbaI-KpnI restriction sites of a shuttle vector in forward and reverse orientations and replication defective recombinant adenoviral vectors containing either the sense or antisense TGF-beta1 genes under the control of a cytomegalovirus promoter - rAdTGFbeta1S and rAdTGFbeta1AS, respectively - were generated. Rat mesangial cells infected with recombinant adenoviruses were grown in normal (NG, 100 mg/dL) or high glucose (HG, 450 mg/dL) media for six days, and mRNA expressions of the monolayers were evaluated by RT-PCR. RESULTS: An adenoviral vector containing the antisense TGF-beta1 gene significantly downregulated the gene expressions of TGF-beta1, collagen, fibronectin and PDGF-B in cultured rat mesangial cells. CONCLUSION: The expression of fibrogenic molecules were significantly attenuated by adenoviral vectors with antisense TGF-beta1, and such results should be confirmed in the future study of animal models of diabetic nephropathy or chronic kidney diseases.

The Cytotoxicity of Truncated (Q368X) Myocilin in Trabecular Meshwork (TM) Cells

PURPOSE: To investigate the cytotoxicities of wild type myocilin and trunctated (Q368X) myocilin in cultured human trabecular meshwork (TM) cells. METHODS: GFP tagged truncated myocilin and DsRED1 tagged wild type myocilin were expressed in TM cells using adenoviral vectors and observed colocalization by confocal microscope. Cytopathic effects in the cells were examined by light microscope and WST-1 cell proliferation assay. RESULTS: Colocalization of wild type and truncated myocilin was observed in cells co-expressing the proteins. Truncated myocilin was found to be toxic to cells, leading to deformed cellular morphology and diminished cell proliferation. CONCLUSIONS: The intracellular accumulation of truncated myocilin exhibited cytotoxicity in trabecular meshwork cells, and eventually resulted in diminished number and dysfunction of trabecular meshwork cells, which might be involved in the pathogenesis of glaucoma.

Studies of killing and bystander effect of tumor necrosis factor related apoptosis-inducing ligand gene on hepatic carcinoma cell / 中华消化杂志

Objective To study the bystander effect of tumor necrosis factor related apoptosis inducing ligand(TRAIL) gene and it's mechanism. Methods Full length cDNA of human TRAIL was transfected into SMMC7721 with binary adenoviral vectors system. RT PCR was used to determine the expression of TRAIL gene. By MTT assay the effects of the TRAIL gene on the proliferation of SMMC7721 was studied; The apoptosis inducing ability of TRAIL gene on SMMC7721 was tested by fluorescence activated cell sorting (FACS). Bystander effect by testing the proliferation of the mixed cells of SMMC7721/TRAIL and SMMC7721 with different ratios, and the mechanism of bystander effect by testing the proliferation of SMMC7721 cultured with media of SMMC7721/TRAIL without cell components were also studied. Results TRAIL gene expressed by binary adenoviral vector system was able to inhibit proliferation(91.2%) and induce apoptosis(29.07%) of SMMC7721, significant difference between TRAIL gene and the other three controls were observed(PBS, LacZ, Bax)( P

The Growth-Suppressive Effect of Adenovirus-mediated Wild-type p53 Transfection on Gastric Cancer Cell Line / 中国医师杂志

Objective To study the inhibitory effects of wild-type p53 on gastric cancer cell lines BGC-7901 and to explore the importance of wild-type p53 in gene therapy for gastric cancer. Methods Adenoviral vector with exogenous wild-type p53 gene was transfected into the gastric cancer cell line BGC-7901 which harbours a mutation in codon 204 of the p53 gene.The transfer efficiency and expression effect of p53 were examined by immunohistochemistry and in situ hybridization. The growth of BGC-7901 cells in vitro was determined by cell counting and MTT assay. Flow cytometric analysis was used for observing cell cycle.Results Transfer efficiency reached 100% in BGC-7901 cells when adenovirus infection ability was over 100MOI. 3 days after transfection, the introduction of exogenous wild-type p53 into the tumor cells resulted in decreased expression of the protein of mutant p53 and increased level of mRNA of wild-type p53 gene. The growth of the BGC-7901 cells was significantly slower after transfection with wild-type p53 gene than that before transfection. The percentage of G 1/G 0 phase of the cells with exogenous wild-type p53 gene was much higher than that of the cells without exogenous wild-type p53 gene.It was 56 47% and 79 40% respectively(P

Restoration of Wild-Type p53 by Adenovirus-Mediated Gene Transfer May Enhance the Therapeutic Efficacy of Chemotherapy in Human Ovarian Cancer Cells / 대한부인종양콜포스코피학회잡지

OBJECTIVE: In an effort to develop a more effective therapeutic strategy for ovarian cancer, we examined whether the restoration of the wild-type p53 gene can enhance the therapeutic effect of chemotherapy. METHODS: In this study, Ov-ca-2774 cells, which are known to have p53 point mutation and cisplatin-resistance, were selected and currently used chemotherapeutic agents including cisplatin, carboplatin, paclitaxel, etoposide, topotecan, and doxorubicin were added concurrently or sequentially with adenovirus-mediated p53 gene transfer (Ad5CMV-p53). RESULTS: Transfer of the wild-type p53 cDNA gene into Ov-ca-2774 cells showed 55% cell killing in vitro at a multiplicity of infection (MOI) of 40. Although the combination of carboplatin or paclitaxel followed by p53 gene transfer with an interval of 48 h manifested no enhanced cell killing compared with cells infected with Ad5CMV-p53 alone, the other combinations of chemotherapeutic agents and p53 gene transfer resulted in 15% to 37% further cell killing (P<0.05). Furthermore, p53 gene transfer followed by doxorubicin with an interval of 24 h and concurrent combination of etoposide with p53 gene transfer showed significant difference in cell killing in contrast to the other combination strategies in the respective chemotherapeutic agent exposure groups (P<0.05). CONCLUSION: Our data demonstrated that combination of p53 gene transfer and chemotherapeutic agents had higher cell killing than either of these two modality alone.

Subject The Effect of Apolipoprotein E Overexpression on Plasma Lipoprotein Profile in Mice Fed on Long-term High Cholesterol Diet

BACKGROUND AND OBJECTIVES: Apolipoprotein E (apoE), a 34-kD plasma glycoapolipoprotein, plays a key role in lipoprotein metabolism by facilitating cellular uptake of remnants of triglyceride-rich chylomicrons and VLDL and may have other important biological functions. Various studies using apoE-knockout mice have elucidated the role of apoE in lipolysis, remnant clearance, and atherogenesis. Despite the growing evidence of the protective role exerted by apoE against atherosclerosis, the direct in vivo effects of the apoE overexpression on lipoprotein metabolism in the presence of endogenous mouse apoE are not yet fully understood. In this study, the technique of adenovirus-mediated gene transfer was employed to investigate the in vivo effect of apoE overexpression on lipid level and lipoprotein profile in mice fed on normal chow or high cholesterol diet. MATERIALS AND METHODS: Recombinant adenovirus (rAd.mApoE) containing mouse apoE cDNA driven by a cytomegalovirus promoter was generated and infused via tail vein in mice fed on normal chow or high cholesterol diet. Recombinant adenoviruses have emerged as the most efficient vectors for transient delivery of functional genes to the mammalian liver. RESULTS: rAd.mApoE in the various mouse tissues one week after injection was expressed mainly in the liver. ApoE overexpression decreased the cholesterol and triglyceride concentration in mice fed on normal chow. In mice fed on high cholesterol diet, apoE overexpression resulted in decrease in triglyceride concentration and increase in cholesterol. VLDL and LDL fraction were decreased, HDL was increased by apoE overexpression in both mice fed on normal chow and high cholesterol diet. CONCLUSION: These data suggest that overexpression of mouse apoE in mice with endogenous apoE may exert antiatherogenic effect by inducing favorable change in the lipoprotein profile, regardless of diet and consequent plasma lipid level. In the future, the studies regarding the effect of human apoE overexpression on the lipid and lipoprotein profile in mice fed on normal chow and high cholesterol diet will be helpful to understand the species differences or similarities in apoE activity.