809 Time for TEA T Cells Expanded on Artificial Antigen Presenting Cells? Molecular Therapy Volume 18, Supplement 1, May 2010 Copyright © The American Society of Gene & Cell Therapy S312 CANCER IMMUNO[.]
CANCER - IMMUNOTHERAPY III Jurkat clones, we demonstrate that this drug combination does not signicantly alter the expression of endogenous genes nearest to the proviral integration site 806 Highly Efcient Expression and Purication of Hepatitis C Virus Core Protein in E.coli Yixin Bian,1 Guozhen Qiu,1 Tingting Li,1 Zixuan Chen,1 Wenjing Wang,1 Chengyao Li.1 Southern Medical University, Guangzhou, Guangdong, China Hepatitis C virus (HCV) infection is major cause of chronic liver diease and hepatocellular carcinoma, but currently there are no prophylactic HCV vaccines available Therefore, the early diagnosis and the early treatment are critical to hepatitis control Although there are numerous commercial HCV diagnostic antigens available, their specicity and sensitivity need to be improved HCV core protein-encoding sequence is among the most conservative genes in the HCV genome The core protein localizes to the endoplasmic reticulum (ER) through a C-terminal hydrophobic region that is cotranslationally inserted into the ER membrane In this study, a 360bp cDNA encoding 120 amino acid of HCV type-1b core protein, which lacked the C-terminal hydrophobic region, was amplied by PCR, and then cloned into an expression plasmid pET-28a(+) The recombinant plasmid pET-28a-core120 was transduced into E.coli BL21 The expression of the recombinant core protein was induced by IPTG Then the recombinant protein was puried by Ni+ -afnity column and renatured by G-25 molecular sieve Finally, the expressed HCV core protein, accounting for 30% of the total protein was identied using SDS-PAGE, western blot and ELISA The results revealed its specic immunoreactivity with serum from patients with hepatitis C In conclusion, high level of the truncated HCV core protein was expressed in E.coli with high HCV specicity and good antigenicity It may suggest the application on studying and developing of HCV diagnostic antigens 807 Molecular Epidemiology of HCV Infection among Recovery and Chronic Blood Donors in Guangdong, China Tingting Li,1 Jinfeng Zeng,2 Zixuan Chen,1 Lifang Shuai,1 Anqi Wang,1 Chengyao Li.1 School of Biotechnology, Southern Medical University, Guangzhou, Guangdong, China; 2Shenzhen Blood Center, Shenzhen, Guangdong, China Hepatitis C virus (HCV) infection is gaining importance in Asian countries in recent years But epidemiological studies conducted on recovery and chronic blood donors with HCV infection in China are little known A study was carried out to determine the ratio between HCV chronic and recovery infection in blood donors from Guangdong, a province in southern China HCV genotyping and phylogenetic analyses were also performed A total of 122 antibody positive plasmas detected by both Lizhu and Ortho anti-HCV EIAs were tested for viral load, conrmed by Nested-PCR, and then classied into two statuses, chronic or recovery infection Approximately 37.5% of conrmed anti-HCV carriers had no detectable viral RNA and were considered to have cleared the virus and recovered from the infection Chronic samples were amplied by 5’-NCR PCR and sequenced for genotyping (215-218bp) The result showed genotype (29.3%), 2(8.3%), (29.3%), (33.3%) was present in chronic blood donors This initial study will help us to understand HCV infection among recovery and chronic blood donors in China S312 808 Occult Hepatitis B Virus Infection in Shenzhen, China Xin Zheng,1 Yixin Bian,2 Ling Zhang,1 Wenjing Wang,1 Lifang Shuai,3 Anqi Wang,1 Jean-Pierre Allain,4 Daniel Candotti,4 Chengyao Li.1 School of Biotechnology, Southern Medical University, Guangzhou, Guangdong, China; 2Shenzhen Blood Centre, Shenzhen, China; 3Guangzhou Centre of Disease Control of PLA, Guangzhou, China; 4Department of Haematology, London, United Kingdom Characterization of occult HBV infection(OBI) in blood donors in China remains far unknown This study is to explore the molecular characterization of occult hepatitis B virus (HBV) infection in blood donors in Shenzhen, China The HBsAg negative donors plasmas collected for HBV/HCV/HIV screening from 2003 by the ELISA were detected for HBV DNA by Nucleic Acid Testing (NAT) in blood donors from Shenzhen blood center All samples of China origin OBIs were quantied for HBV DNA loads by real-time PCR(QPCR) The amplied basic core promoter/precore, pre-S/S, and whole genome were cloned and sequenced, phylogenetic analysis were performed based on one to three sequences of HBV clones Twenty-nine samples of OBI screened from 165371 donors were conrmed and genotyped, according to sequences of Pre-S/S region Genotype C was more frequent (8 strains) than genotype B (5 strains) OBI donors were obtained for females (27.6%) and 21males(72.4%) Viral load ranged between unquantiable and 7321IU/ml (median 17.4IU/ml) Sequence analyses of full length genome showed occult isolates were variants from clones The amino acid substitutions of core and pre-s/s open reading frame in OBI were more frequent than HBsAg+ sample in genotype B(P< 0.03), but not in the core region of genotype C(P>0.05) Frequency of diversity of core regular element in OBIs was signicantly higher than that in wildtype HBV(P