మాలిక్యులర్ మరియు జెనెటిక్ మెడిసిన్

Typical reverse genetics systems for generating influenza viruses require the insertion of each genome segments by DNA ligation into vectors for genome synthesis and expression. Herein is described the construction and use of a novel pair of plasmid vectors for cloning all eight genome segments of influenza A virus by homologous recombination for influenza virus reverse genetics. Plasmids, pLLBA and pLLBG, were constructed to possess opposing RNA polymerase I and RNA polymerase II transcription units for generating influenza genomic and messenger RNAs, respectively. In addition these promoters flanked a recombination cassette which comprised the conserved 5’ (13bp) and 3’ (12bp) terminal promoters of influenza virus. These vectors differed due to the presence of an A or a G (plus sense) to correspond to differences at nucleotide position 4 among negative-sense influenza virus promoters. The cloning approach involved homologous recombination of each influenza gene segment and the appropriate linearized pLLBA or pLLBG vectors in E. coli. Direct cloning by recombination was simpler and faster than conventional restriction digestion and ligation methods. This new vector system was successfully used to clone and rescue various influenza viruses and thus has the potential to promote the rapid analysis and vaccine development of novel influenza strains.

With antigenically novel epidemic and pandemic influenza strains persistently on the horizon it is of fundamental importance that we understand whether heterosubtypic antibodies gained from exposures to circulating human influenzas exist and can protect against emerging novel strains. Our studies of IVIG obtained from an infection-naive population (Australian) enabled us to reveal heterosubtypic influenza antibodies that cross react with H5N1. We now expand those findings for an Australian donor population to include IVIG formulations from a variety of northern hemisphere populations. Examination of IVIGs from European and South East-Asian (Malaysian) blood donor populations further reveal heterosubtypic antibodies to H5N1 in humans from different global regions. Importantly these protect against highly pathogenic avian H5N1 infection in vitro, albeit at low titres of inhibition. Although there were qualitative and quantitative differences in binding and protection between globally different formulations, the heterosubtypic antibody activities for the respective IVIGs were in general quite similar. Of particular note because of the relative geographic proximity to the epicentre of H5N1 and the majority of human infections, was the similarity in the antibody binding responses between IVIGs from the Malayan peninsula, Europe and Australia. These findings highlight the value of employing IVIGs for the study of herd immunity, and particularly heterosubtypic antibody responses to viral antigens such as those conserved between circulating human influenzas and emerging influenza strains such as H5N1. They also open a window into a somewhat ill defined arena of antibody immunity, namely heterosubtypic immunity.

? ??????? ????? ??????? ???????‌?? ???????? ????? ????? ??????????? ???????? ?????? (GPV) ????? ????? ???????????? ????? ?????. ?????? GPV ??????? ?????? VP2 ????? VP3 ?????? ????????‌? ?????????????? ????? ??????? ??????????? ??????????‌??? ????? ?????, ????????, ??????, ???? ????? ??????‌??? ??????????? ??? ??????? ????????. ?????? ????? ????????? ??????????? ?????????? ????? ???????? ????? ???????? ???????? ???? ????? ?????? ?????????????? ??????? 5%?? ?????????. ? ????????? ?????????????? ?????? ?????? GPV ?????? ????? ???????? ??? ??????????? ???? MDPV FM ??????? ??????????? ??????????‌?? ???????? ?????? ?????????????. ??????????? ????????? ?????? ????? ????????‌??? ????? ??????????? ?????? GPV ?????? ????? ?????? ?????? ???????? ???????????? ???? ?????????????.

????????? ??????????? ????? ???????????????? (HA) ????? ?????????????? (NA) ????? ?????‌?????????, ???????????????? ????‌??? ????????? ????? ????????? ?????????????? ????????????? ????????? ????????????? ???? ????? ?????????????. ??? ??????????? ??????? ????? ???? HA ????? NA ?????????‌? ??????? ????????????? ??????????‌??? ????????????, ? ?????? ?????? ????? ???? ?????? ????????? ??????????? ?????? ?????????????? ????????? ??????? ??????????, ????? ???????????? ?????????? ???? ???????? ??????????‌??? ??????????? ????? ??????? ??????????. ?????? ?????????? ?????????‌?????? ????????? ??? ???????? ?????? ????????? ??????????? ????? ??????? ????. ??? ????? ???? ??????? ????????????? ????????? ??????? ???????? ???????‌??? ????? ????????? ????????? ????????? ????????????? ???????????. ???????????, ??????? ????????????? ????????? ??????? ???? ?????? ????????? ?????????? ????? ??????, ??? ??????????? ???????‌??? ???????? ?????? ????? ???? ?????????? ????? ????? ????? ???????? ??????? ???????????. ????????? ??????????????? ????????? ????? ???? ??????? ?????????, ????? ???????????? ???????? ? ????? ????????? ????????? ??????? ???? ??????? ???????. ????? ??????? ?????????? ????????????? ????‌??? ?????? ???????? ????? ?????? ???????????? ????? ????????, ????????? ????? ???????????? ?????? ?????????? H5N1 ???? ????????????? ??? ????? ????? ???????? ??????? ??????? ??????????.

?????? ??????????? (AI)?? ??????????? ????? ?????????? ?????? ????? 2003 ????? ????????????? ????? 2006 ????? ????????? H5N1 ?????? ?????????? ?????? ??????????? (HPAI) ????????????? ??????????? ????? ?????? ??????????????. ???????? ????? ???????? ????????? ??? ?????? ??????? ???????? ????? ????? ??????? ??????? ??????? ????????? HPAI?? ??????????? ???? ?????? ???????????? ??????? ??????? ?????? ????????????. ???????????, ???? ????????? ?????, ????????? AI ???? ????? ?????? ?????????? ??????? ???? ????????? ????. HPAI?? ??????????? ?????????? ?????? ?????? ????????????, ???????? ???????? ????? ???? ?????? ????? ????????? ??????????????. ?????? ?????? ??????? ???????? ?????????? ???????????? ???????? ????????? ????? ??????? ?????????? ???? ?????????? ????? ????????? ??????? ??????? ???????????? ?????? ???????. ???? ?????? ????? ?????? ???????? ?????? ????? ???????????? ????? ?????? ?????? ????????, ??????????? ?????????? ??????? ???????? ????? ?????? ???????????‌??? ??????? ?????? ??????? ??????????. ????-??????? ?????? ????? ??? ???????? ??????? ????????? ????? ??????? ??????? ??????, ???????????, ??????? ???????? ?????????? ????? ???????? ?????? ?????? ?????? ??????????? ??????????? ?????? ?????????? ????????????. ????, ???????? ???????, ?????? ??????? ????????? ????? ???-?????????? ????? ????? ??????? ?????? ?????????? ??????? ?????? ???????? ???????????? AI ?????? ????? ?? ???????? ??????? ???????????. ??????? ???? ??????????????, AI ???? ?????? ???????? ?????? ?????? ??????????? ????? ???????. ???????????, ???? ?????????? ?????? ??????? ???-?????????? ?????????? ???????, ?????????? ?????? ????????? ??????? ????????? ????? ?????????? ??????? ???????? ????? ???????? ????? ????????? ???????? ???????? ????? ???????? ???????????? ????????? ?????? ??????? ????????.

1918 ?????????? ???????? ???‌???????? ????? ????? ????????? ????????? ??????? ?????, ????? 1918 ????? ????? ?????? ??? ???? ?????‌?? ???????, ??? ????? ?????? ???????? ??????? ????????? ?????????? ?????? ????? ????. ??????? ????????? ????? ???????? ?????????? ???????? ??????? ??????????? ????? ????????? ?????????? ????? ??????????????? ?????? ???????. ?????? ???? ?????? ???????????, ???: ??? ??????? ?????? ???????? ????? ?????????? ??????? ?????? ?????, ??????? ???? ?????? ???? ???????? ?????, ???????, ???????, ???, ????? ???????? ???????? ????????? ???????????????. ???? ????? ??????????????? ??????, ???????? ???????????? ??????, ?????? ????????, ??? ????? ???????, ?????? ????? ??? ?????????, ???? ????? ??????? ????????? ??????. ????, ? ?????????? ????? ???? ????? ?????? ??????????? ???? ?????? ????????????? ?????? ???? ????? ???????? 1918 ?????????? ???????? ????????????? ???????????? ?????????‌?? ????????? ?????? ????.