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A novel live attenuated vaccine candidate protects chickens against subtype B avian metapneumovirus

Journal of Integrative Agriculture 2024年第5期23卷 1658-1670页

作者： Lingzhai Meng Mengmeng Yu Suyan Wang Yuntong Chen Yuanling Bao Peng Liu Xiaoyan Feng Tana He Ru Guo Tao Zhang Mingxue Hu Changjun Liu Xiaole Qi Kai Li Li Gao Yanping Zhang Hongyu Cui Yulong Gao Avian Immunosuppressive Diseases Division State Key Laboratory for Animal Disease Control and PreventionHarbin Veterinary Research InstituteChineseAcademy of Agricultural SciencesHarbin150069China Avian ImmunosuppressiveDiseases DivisionStateKey LaboratoryforAnimal Disease Cont National Poultry Laboratory Animal Resource Center Harbin 150069China

Avian metapneumovirus(aMPV) is a highly contagious pathogen that causes acute upper respiratory tract diseases in chickens and turkeys, resulting in serious economic losses. Subtype B aMPV has recently become the dominant epidemic strain in China. We developed an attenuated aMPV subtype B strain by serial passaging in Vero cells and evaluated its safety and efficacy as a vaccine candidate. The safety test showed that after the 30th passage, the LN16-A strain was fully attenuated, as clinical signs of infection and histological lesions were absent after inoculation.The LN16-A strain did not revert to a virulent strain after five serial passages in chickens. The genomic sequence of LN16-A differed from that of the parent wild-type LN16(wtLN16) strain and had nine amino acid mutations. In chickens, a single immunization with LN16-A induced robust humoral and cellular immune responses, including the abundant production of neutralizing antibodies, CD4^(+) T lymphocytes, and the Th1(IFN-γ) and Th2(IL-4 and IL-6)cytokines. We also confirmed that LN16-A provided 100% protection against subtype B aMPV and significantly reduced viral shedding and turbinate inflammation. Our findings suggest that the LN16-A strain is a promising live attenuated vaccine candidate that can prevent infection with subtype B aMPV.

关键词： avian metapneumovirus attenuated protection vaccine candidate chickens

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A highly immunogenic live-attenuated vaccine candidate prevents SARS-CoV-2 infection and transmission in hamsters

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The Innovation 2022年第2期3卷 126-135页

作者： Xiao-Feng Li Zhen Cui Hang Fan Qi Chen Lei Cao Hong-Ying Qiu Na-Na Zhang Yan-Peng Xu Rong-Rong Zhang Chao Zhou Qing Ye Yong-Qiang Deng Yan Guo Si Qin Kaiyue Fan Lei Wang Zijing Jia Yujun Cui Xiangxi Wang Cheng-Feng Qin Department of Virology State Key Laboratory of Pathogen and BiosecurityBeijing Institute of Microbiology and EpidemiologyAcademy of Military Medical SciencesBeijing 100071China CAS Key Laboratory of Infection and Immunity National Laboratory of MacromoleculesInstitute of BiophysicsChinese Academy of SciencesBeijing 100101China School of Medicine Tsinghua UniversityBeijing 100084China Research Unit of Discovery and Tracing of Natural Focus Diseases Chinese Academy of Medical SciencesBeijing 100071China

The highly pathogenic and readily transmissible SARS-CoV-2 has caused a global coronavirus pandemic,urgently requiring effective countermeasures against its rapid expansion.All available vaccine platforms are being used to generate safe and effective COVID-19 vaccines.Here,we generated a live-attenuated candidate vaccine strain by serial passaging of a SARSCoV-2 clinical isolate in Vero cells.

关键词： vaccine attenuated infection

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Efficacy of live attenuated vaccines after two doses of intravenous immunoglobulin for Kawasaki disease

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World Journal of Pediatrics 2022年第10期18卷 706-709页

作者： Yoshihiko Morikawa Hiroshi Sakakibara Masaru Miura Clinical Research Support Center Tokyo Metropolitan Children’s Medical Center2-8-29 MusashidaiFuchuTokyo183-8561Japan Department of General Pediatrics Tokyo Metropolitan Children’s Medical CenterTokyoJapan Department of Cardiology Tokyo Metropolitan Children’s Medical CenterTokyoJapan

Kawasaki disease(KD)is a vasculitis of unknown etiology occurring predominantly in children under five years old[1].Coronary artery aneurysm,its most serious complica-tion,occurs in about 25%of untreated patients[2].High-dose intravenous immunoglobulin(IVIG)2 g/kg is used to prevent coronary artery aneurysms[1,3,4],but about 20%of patients are nonresponsive and require a second dose for a total of 4 g/kg.The appropriate timing of live attenuated vaccine(LAV)administration in such cases is unclear.

关键词： doses attenuated intravenous

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attenuated plasmodium sporozoite expressing MAGE-A3 induces antigen-specific CD8+ T cell response against lung cancer in mice

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Cancer Biology & Medicine 2019年第2期16卷 288-298页

作者： Dong Zhou Hong Zheng Quanxing Liu Xiao Lu Xufeng Deng Li Jiang Bing Hou Yong Fu Feng Zhu Yan Ding Wenyue Xu Jigang Dai Department of Thoracic Surgery Xinqiao Hospital Third Military Medical University Chongqing 400037 China Department of Cardiothoracic Surgery The First People's Hospital of Zunyi Zunyi 563000 China Department of Pathogenic Biology Third Military Medical University Chongqing 400037 China

Objective: Cancer vaccines that rely on tumor antigen-specific CD8^+ T cell responses, are promising anti-cancer adjuvant immunotherapies. This study investigated whether genetically attenuated Plasmodium sporozoite(GAS) could be used as a novel vector to induce antigen-specific CD8^+ T cell responses against lung cancer.Methods: We constructed GAS/MAGE-A3, a recombinant GAS engineered to express the lung cancer-specific antigen, melanomaassociated antigen 3(MAGE-A3), and assessed its therapeutic effects against lung cancer.Results: Robust parasite-specific CD8αlow CD11ahigh and CD49dhigh CD11ahigh CD4^+ T cell responses as well as a MAGE-A3-specific CD8^+ T cell response were induced in GAS/MAGE-A3-immunized mice. Adoptive transfer of GAS/MAGE-A3-induced CD8^+ T cells from HLA-A2 transgenic mice into lung cancer-bearing nude mice inhibited tumor growth and prolonged survival.Conclusions: These findings demonstrate that GAS/MAGE-A3 induces a strong MAGE-A3-specific CD8^+ T cell response against lung cancer in vivo, and indicate that GAS is a novel and efficacious antigen delivery vector for antitumor immunotherapy.

关键词： Vaccine lung cancer genetically attenuated sporozoites MAGE-A3 malaria

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Induction of deletion mutation on ompR gene of Salmonella enterica serovar Typhi isolates from asymptomatic typhoid carriers to evolve attenuated strains for vaccine development

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Asian Pacific Journal of Tropical Medicine 2014年第12期7卷 933-939页

作者： Senthilkumar B Anbarasu K Senbagam D Rajasekarapandian M Centre for Biotechnology Muthayammal College of Arts & SeienceRasipuram-637408Tamil NaduIndia Department of Marine Biotechnology Bharathidasan UniversityTiruchirappalli-620024.Tamil NaduIndia Department of Biotechnology Virekanandha College of Engineering for WomenTiruchengode-637205.Tamil NaduIndia Department of Zoology Arignar Anna Government Arts CollegeNamakkal-637001Tamil NaduIndia

Objective:To develop allenualed slrains of Salmonella enterica serorar Typhi(S.typhi) for the candidate vaccine by osmolar stress.Mothods:S.typhi SS3 and SS5 strains were isolated from asymptomatic typhoid carriers in Mamakkal,Tamil Nadu.India.Both strains were grown in LB(Luria Bertani) medium supplemented with various concentration of NaCl(0.1- 0.7M) respectively.The effecl of osmolar stress was determined at molecular level by PCR using MCR 06 and MCR07 primers corresponding to ompR with chromosomal DNA of S.typhi SS3 and SS5 strains.Attenuation by osmolar stress results in deletion mutation of the.S.typhi slrains was determined by agglutination assays,precipitation method.SDS PAGE analysis and by animal models.Results:The 799 bp amplified ompR gene product from wild type S.typhi SS3 and SS5 illustrate the presence of virulent gene.Interestingly,there was only a 282 bp amplified product from S.typhi SS3 and SS5 grown in the presence of 0.***.0.6 and 0.7 M NaCl.This illustrates the occurrence of deletion mutation in ompR gene al high concentration of NaCl.Furthermore,both the wildtype and mutant S.typhi outer membrane SDS-PAGF.profile reveals the differences in the expression of ompF.ompC and ompA proteins.In mice,wild type and mutant strains lethal dose(LD_(50)) were determined.The mice died within 72 h when both the wild type strains were injected intraperitoneally with 3 log CFU-mL^(-1).When the mice were injected with the mutants in same dosage,no clinical symptoms were observed;whereas the serum antibodv litre was elicited within two weeks indicated that the mutants have the ability to induce protective humoral immune response.These results suggest that S.typhi SS3 and SS5 may bo used as good candidate strains for the development of live attenuated vaccine against salmonellosis.Conclusions:This study demonstrates that the S.typhi strains were allenualed and could be good vaccine candidates in future.

关键词： attenuated live vaccine Salmonella enterica serovar Typhi Mutants

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Evaluating the efficacy of an attenuated Streptococcus equi ssp. zooepidemicus vaccine produced by multi-gene deletion in pathogenicity island SeseCisland_4

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Journal of Integrative Agriculture 2019年第5期18卷 1093-1102页

作者： MA Fang WANG Guang-yu ZHOU Hong MA Zhe LIN Hui-xing FAN Hong-jie MOE Joint International Research Laboratory of Animal Health and Food Safety College of Veterinary Medicine Nanjing Agricultural University National Center of Meat Quality and Safety Control Nanjing Agricultural University Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses Yangzhou University

Streptococcus equi ssp. zooepidemicus(SEZ) is a pathogen associated with a wild range of animal species. Frequent outbreaks have occurred in recent years in pigs, horses, goats and dogs which is liable to infect humans. There is a lack of efficient vaccines against this disease and the occurrence of antibiotic resistance may render drug therapies ineffective. In this study, gene deletion mutant(ΔSEZ) in pathogenicity islands SeseCisland_4 was constructed. The mutant ΔSEZ had a 52-fold decrease in 50% lethal dose(LD_(50)) and had less capacity to adhere epithelial cells. Importantly, immunization of mice with attenuated vaccine ΔSEZ at the dose of 10~2 colony-forming units(CFU) mL^(–1) elicited a significant humoral antibody response, with an antibody titer of 1:12 800. Therefore, 10~2 CFU mL^(–1) might be used as the appropriate immune dose for the attenuated vaccine ΔSEZ, which provided mice with efficient protection against virulent SEZ. In addition, the hyperimmune sera against 10~2 CFU m L–1 attenuated vaccine ΔSEZ could confer significant protection against virulent SEZ infection in the passive immunization experiment and exhibited efficient bactericidal activity in the whole blood assay. Meanwhile, no viable bacteria was detected in blood when mice were immunized with ΔSEZ at the dose of 10~2 CFU mL^(–1) via hypodermic injection. Thereafter, the mutant ΔSEZ at the dose of 10~2 CFU mL^(–1) could confer significant protection in mice and had less negative effects on host, which could be an effective attenuated vaccine candidate for the prevention of SEZ.

关键词： Streptococcus equi ssp.zooepidemicus zoonosis attenuated vaccine immune dose

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Current status of the genetic susceptibility in attenuated adenomatous polyposis

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World Journal of Gastrointestinal Oncology 2019年第12期11卷 1101-1114页

作者： Víctor Lorca Pilar Garre Laboratorio de Oncología Molecular Grupo de Investigación Clínica y Traslacional en OncologíaHospital Clínico San CarlosMadrid 28040Spain Laboratorio de Oncología Molecular Servicio de OncologíaHospital Clínico San CarlosMadrid 28040Spain

Adenomatous polyposis(AP)is classified according to cumulative adenoma number in classical AP(CAP)and attenuated AP(AAP).Genetic susceptibility is the major risk factor in CAP due to mutations in the known high predisposition genes APC and MUTYH.However,the contribution of genetic susceptibility to AAP is lower and less understood.New predisposition genes have been recently proposed,and some of them have been validated,but their scarcity hinders accurate risk estimations and prevalence calculations.AAP is a heterogeneous condition in terms of severity,clinical features and heritability.Therefore,clinicians do not have strong discriminating criteria for the recommendation of the genetic study of known predisposition genes,and the detection rate is low.Elucidation and knowledge of new AAP high predisposition genes are of great importance to offer accurate genetic counseling to the patient and family members.This review aims to update the genetic knowledge of AAP,and to expound the difficulties involved in the genetic analysis of a highly heterogeneous condition such as AAP.

关键词： attenuated adenomatous polyposis Genetic susceptibility High predisposition gene Genetic heterogeneity Colorectal cancer

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Recombinant constructions and infectivity analysis of tobacco mosaic virus and attenuated tomato mosaic virus N14 genomes

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Chinese Science Bulletin 2000年第15期45卷 1399-1404,1442页

作者： Gong Yang Xiangguo Liu Bingsheng Qiu Chinese Acad Sci Inst Microbiol Dept Mol Virol & Bioengn Beijing 100080 Peoples R China

The recombinant clones of pTN and pNT have been constructed by exchanging the coding regions of the movement proteins (MP), coat proteins (CP) and 3’ noncoding regions between the cDNAs of the tobacco mosaic virus (Chinese Isolate, TMV-Cv) and the attenuated tomato mosaic virus N14 genomes, and used as templates for in vitro runoff transcription. Their transcripts have been used for tobacco infection assays. The infection results show that the transcripts of pTN and pNT are infectious. Local lesions were observed in the leaves of Nicotiana tabacum cv. Samsun NN inoculated with pTN transcript, but were fewer than those in the same kind of plant induced by pTMV-Cv transcript. Systemic symptoms were also observed in N. tabacum cv. Huangmiaoyu induced by pTN transcript, but were slighter than those on the same kind of tobacco induced by pTMV-Cv transcript. Local lesions were shown in N. tabacum cv. Samsun NN inoculated with pNT transcript, but were more than those in the same kind of plant induced by pN14

关键词： tobacco mosaic virus (TMV) attenuated tomato mosaic virus (ToMV) N14 genomes infectivity attenuation mechanism.

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Prophylaxis of tumor through oral administration of IL-12 GM-CSF gene carried by live attenuated salmonella

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Chinese Science Bulletin 2001年第13期46卷 1107-1112页

作者： LI Yuhua CHEN Hui QUO Kunyuan XIE Yongmei WANG Feng XIE Kuangcheng HUANG Jiangsheng REN Darning Department of Hematology Zhujiang HospitalFirst Military Medical UniversityGuangzhou 510282China Institute of Genetics School of Life ScienceSlate Key Laboratory of Genetic EngineeringFudan UniversityShangshai 200433China Central Research Lab of First Hospital of Shanghai Shangshai 200000China

A live attenuated AraA autotrophic mutant of Salmonella typhimurium (SL3261) was used as carrier for eukaryotic expression vectors EGFPN1, pCMVmIL-12, pCMVWL-12, pCMVmGM-CSF and PCMVhGM-CSF and was administered orally to BALB/c and C57BL/6 mice. After 6 weeks, these mice were challenged with 4T1 and Lewis tumor cells respectively. GFP expression and gene integration could be detected in mice’s livers, spleens, intestines, kidneys and tumors. The serum level of cytokines increasedsignificantly in treated mice, so did the ratio of CD8+/CD4+,which resulted in the tumor regression and prolongation of the survival time of those mice. These researches laid an experimental foundation for the tumor gene therapy using live attenuated salmonella.

关键词： oral administration gene therapy attenuated salmonella.

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MOLECULAR CLONING OF attenuated HEPATITIS A VIRAL cDNA

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Chinese Science Bulletin 1991年第22期36卷 1916-1920页

作者：杨能宇陈勇柴少爱洪艳毛江森 Institute of Viral Disease Research Zhejiang Academy of Medical Sciences Hangzhou 310013 PRC

An attenuated hepatitis A virus (HAV) strain (H2 strain) has been developed successfully by Mao et al. in our laboratory, which has been proven to have a good immanogenicity in animal and human trials. The molecular biological research for H2 strain has been carried out in order to darify the attenuation mechanism of H2 strain on the molecular level.

关键词： hepatitis A virus attenuated strain cDNA molecular cloning

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