A Self-Biomineralized Novel Adenovirus Vectored COVID-19 Vaccine for Boosting Immunization of Mice

SARS-CoV-2 has caused more than 3.8 million deaths worldwide,and several types of COVID-19 vaccines are urgently approved for use,including adenovirus vectored vaccines.However,the thermal instability and pre-existing immunity have limited its wide applications.To circumvent these obstacles,we constructed a self-biomineralized adenovirus vectored COVID-19 vaccine(Sad23L-n Co V-S-Ca P)by generating a calcium phosphate mineral exterior(Ca P)based on Sad23L vector carrying the full-length gene of SARS-Co V-2 spike protein(S)under physiological condition.This Sad23L-n Co V-S-Ca P vaccine was examined for its characteristics of structure,thermostability,immunogenicity and avoiding the problem of preexisting immunity.In thermostability test,Sad23L-n Co V-S-Ca P could be stored at 4°C for over 45 days,26°C for more than 8 days and 37°C for approximately 2 days.Furthermore,Sad23L-n Co V-S-Ca P induced higher level of S-specific antibody and T cell responses,and was not affected by the pre-existing anti-Sad23L immunity,suggesting it could be used as boosting immunization on Sad23L-n Co V-S priming vaccination.The boosting with Sad23L-n Co V-S-Ca P vaccine induced high titers of 10^(5.01)anti-S1,10^(4.77)anti-S2 binding antibody,10^(3.04) pseudovirus neutralizing antibody(IC_(50)),and robust T-cell response of IFN-c(1466.16 SFCs/10^(6)cells)to S peptides,respectively.In summary,the selfbiomineralization of the COVID-19 vaccine Sad23L-n Co V-S-Ca P improved vaccine efficacy,which could be used in prime-boost regimen for prevention of SARS-Co V-2 infection in humans.

rAAV2-CTGF和TIMP1双基因联合转染恒河猴退变腰椎间盘的生物学效应

目的研究腺相关病毒（adeno—associated virus2，AAV2）介导的结缔组织生长冈子（connective tissue growth factor，CTGF）和基质金属蛋白酶组织抑制因子1（tissue inhibitor of metalloproteinasesl，TIMP1）双基因体内转染退变的恒河猴腰椎间盘的生物学效应。方法恒河猴9只，雌性4只，雄性5只；年龄4-7岁，体重4．5-7kg。CT引导下经皮穿刺纤维环诱导恒河猴腰椎间盘退变，微创经皮注入携带CTGF和TlMPI双基因腺相关病毒颗粒。应用Rrr_PCR测定细胞因子的表达，RT—PCR、35S整合法观察双基因联合转染对退变椎间盘的生物学作用。结果双基因转染后8、16、24周时CTGFmRNA表达量分别为PBS对照组的10．02、2．39、0．91倍；TIMPImRNA表达量分别为PBS对照组的6．08、3．8l、2．67倍；Ⅱ型胶mRNA表达量分别为PBS对照组的145．51％、174．72％、113．73％；蛋白多糖mRNA表达量分别为PBS对照组的461．19％、191．46％、301．39％；蛋白多糖合成效率为分别PBS对照组的455．06％、285．97％、165．58％。结论CTGF和TIMPI双基因联合转染后可以在体内较长期表达，并能促进体内蛋白多糖和Ⅱ型胶原的合成，延缓椎间盘的退变。