The Impact Of Different Species Sources Of Anthocyanins On The Biological Activity Of DF-1Cell Line Which Infected By Avian Leukemia Virus Subgroup A

Avian leukemia (Avian leukosis AL) refers to a variety of transmissible benign andmalignant tumors in the chickens aroused by the members of the genus of the RetrovirusBranch of avian retrovirus avian leukosis/sarcoma virus group of viruses (ALV/RSV).According to clinical symptoms, the diseases are divided into Lymphoblastic leukemia,Erythroblast leukemia, myeloid leukemia, myelocytomatosis, connective tissue tumors andosteopetrosis.1. Identification and quantification of avian leukemia virus subgroup A (ALV-A)A PCR method for identification of ALV-A was established, by amplifying the ALV-P27and ALV-A fragments with special primers respectively. The sequence homology of ALV-P27with Avian leukosis virus-RSA (M37980.1) and Myeloblastosis-associated virus (L10923.1)strains of ALV-A was97%and96.6%respectively, while the sequence homology of ALV-Awith Avian leukosis virus-RSA (M37980.1) and Myeloblastosis-associated virus (L10923.1)strains of ALV-A was93.6%and96.8%respectively. According to the env gene sequence ofALV-A, a pair of primers and Taqman probe were designed. A Real-time fluorescentquantitative method was established and applied for quantitative detection of ALV-A, fornoobvious CPE in the cells infected by ALV-A. The result showed that there was104.67copies/μL of ALV-A in the sample, which was used in the later researches.2. The impact of different species sources of anthocyanins on the DF-1cellsTo investigate the impact and the final concentration of different species sources ofanthocyanin on DF-1cell line, the different concentrations of ten sorts of different speciessources of anthocyanins were used to study by direct observation of the adherence and growthof DF-1cell. The results showed that different concentrations of Blueberry anthocyanin,Radish anthocyanin, Purple cabbage anthocyanin, purple sweet potato anthocyanin,Elderberry anthocyanin and Purple corn anthocyanin had a significant impact on promting theadherence and growth of DF-1cell, while black soybean anthocyanin, blackcurrantanthocyanin, Vaccinium vitis-idaea L anthocyanin and mountain ash anthocyanin hamperedDF-1cell growth and leaded to cell death. By MTT assay, Blueberry anthocyanin, Radishanthocyanin, Purple cabbage anthocyanin, purple sweet potato anthocyanin, Elderberry anthocyanin and Purple corn anthocyanin also had a significant impact on promting thebiological activity and viability of DF-1cell.3. Biological activity of the DF-1cells infected with ALV-A by MTTTo investigate the impact of the six anthocyanins on the biological activity of the DF-1cellsinfected with ALV-A, the different concentrations (50μg/mL,75μg/mL and100μg/mL) ofBlueberry anthocyanin, Radish anthocyanin, Purple cabbage anthocyanin, purple sweet potatoanthocyanin, Elderberry anthocyanin and Purple corn anthocyanin were added into the DF-1cells at the mean time (Protection) or1hour postinfection (Treatment) of ALV-A. The resultsindicated that different concentrations of Radish anthocyanin had a significant impact onpromting the biological activity of the DF-1cells in Protection and Treatment groups, whilethe other five anthocyanins had no significant impact.4. The detection of NF-κBAt12h,24h and48h postinfection of the DF-1cells infected by ALV-A, the nucleoproteinswere extracted to detect NF-κB. Compared with control cells, the amount of nucleoproteinwas significantly increased at12h and24h postinfection, but there was no significant changeat48h postinfection. When detection of NF-κB by ELISA, there were no significant changesat12h,24h and48h postinfection.