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Establishment of a Chinese street rabies virus library and its application for detecting neutralizing activity

Infectious Diseases of Poverty 2018年第1期7卷 1217-1223页

作者： Peng-Cheng Yu Xiao-Yan Tao Li-Hua Wang Qing Tang Li-Yun Fan Shu-Xia Zhang Shu-Qing Liu Xue-Xin Lu Gui-Zhen Wu Wu-Yang Zhu Key Laboratory of Medical Virology Ministry of HealthNational Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China

Background:The injection of rabies immune globulin(RIG)is of the utmost importance in the management of category III exposures to rabies-suspect *** of the high cost and limited availability of existing RIG,one possible replacement for RIG is monoclonal antibodies(MAbs)against the rabies virus(RABV).Consequently,it is necessary to determine the neutralizing activity of the MAbs against rabies viruses,especially street rabies ***,the method to detect the neutralizing activity of MAbs against street rabies virus remains ***:To establish a method for detecting the neutralizing activity of MAbs against street rabies virus,we constructed a library consisting of 12 strains of street RABV from 11 provinces in *** this street RABV library and the Reed-Muench formula,we established a method for detecting the neutralizing titer of the *** reliability and repeatability of the method were evaluated by repeatedly measuring the neutralizing activity of a MAb and a post vaccination ***:A total of 12 strains of street RABV were chosen for inclusion in the street RABV library,which covered six Chinese lineages(China I-China VI)and grew to high titers in N2A cells(>105 FFD50/ml).On the basis of the library,we constructed the method to detect the neutralizing activity of the *** results of repeatedly measuring the MAbs and positive serum showed excellent reliability and repeatability of the method established in this ***:This study established a street RABV library reflecting the epidemiological features of Chinese rabies viruses,which provides a platform for detecting the neutralizing activity of MAbs against rabies viruses circulating in China.

关键词： Street RABV MAbs neutralizing activity

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“Unconventional”neutralizing activity of Antibodies Against HIV

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Virologica Sinica 2007年第6期22卷 462-470页

作者： Michael WEN Paul ZHOU Laboratory of Anti-viral Immunity and Genetic Therapy Institute Pasteur of ShanghaiChinese Academy of SciencesShanghai 200025China

neutralizing antibodies are recognized to be one of the essential elements of the adaptive immune response that must be induced by an effective vaccine against HIV. However, only a limited number of antibodies have been identified to neutralize a broad range of primary isolates of HIV-1 and attempts to induce such antibodies by inununization were unsuccessful. The difficulties to generate such antibodies are mainly due to intrinsic properties of HIV-1 envelope spikes, such as high sequence diversity, heavy glycosylation, and inducible and transient nature of certain epitopes. In vitro neutralizing antibodies are identified using ＂conventional＂ neutralization assay which uses phytohemagglutinin （PHA）-stimulated human PBMCs as target cells. Thus, in essence the assay evaluates HIV-1 replication in CD4^＋T cells. Recently, several laboratories including us demonstrated that some monoclonal antibodies and HIV-1-specific polyclonal IgG purified from patient sera, although they do not have neutralizing activity when tested by the ＂conventional＂ neutralization assay, do exhibit potent and broad neutralizing activity in ＂unconventional＂ ways. The neutralizing activity of these antibodies and IgG fractions is acquired through post-translational modifications, through opsonization of virus particles into macrophages and inunature dendritic cells （iDCs）, or through expression of antibodies on the surface of HIV-1-susceptible cells. This review will focus on recent findings of this area and point out their potential applications in the development of preventive strategies against HIV.

关键词： Antibody neutralizing activity Human immunodeficiency virus (HIV)

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Preparation and Characterization of Monoclonal Antibodies against VP1 Protein of Foot-and-mouth Disease Virus O/China99

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Virologica Sinica 2009年第6期24卷 566-572页

作者： Shuai SONG Tong LIN Jun-jun SHAO Shan-dian GAO Guo-zheng CONG Jun-zheng DU Hui-yun CHANG National FMD Reference Laboratory Key Laboratory of Animal Virology of Ministry of Agriculture State Key Laboratory of veterinary Etiological Biology Lanzhou Veterinary Research Institute Chinese Academy of Agricutral Science Lanzhou 730046 China Guangdong Open Laboratory of Veterinary Public Healty Institute of Veterinary Medicine Guangdong Academy of Agricultural Science Guangzhou 510640 China

Monoclonal antibodies (McAbs) 1A9 and 9F12 against Foot-and-mouth disease virus (FMDV) serotype O were produced by fusing SP2/0 myeloma cells with splenocyte from the mouse immunized with O/China99. Both McAbs reacted with O/China99 but not with Asia 1, as determined by immunohistochemistry assay. The microneutralization titer of the McAbs 1A9 and 9F12 were 640 and 1 280, respectively. Both McAbs contain kappa light chains, but the McAbs 1A9 and 9F12 were IgG1 and IgM, respectively. In order to define the McAbs binding epitopes, the reactivity of these McAbs against VP1, P20 and P14 were examined using indirect ELISA, the result showed that both McAbs reacted with VP1 and P20. McAbs may be used for further studies of vaccine, diagnostic methods, prophylaxis, etiological and immunological researches on FMDV.

关键词： Foot-and-mouth disease virus (FMDV) Monoclonal antibody neutralizing activity VP1 protein

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Develope Monoclonal Antibody against Foot-and-mouth Disease Virus A Type

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Virologica Sinica 2011年第4期26卷 273-278页

作者： Tong Lin Jing Li Jun-jun Shao Guo-zheng Cong Jun-zheng Du Shan-dian Gao Hui-yun Chang State Key Laboratory of Veterinary Etiological Biology National Foot and Mouth Disease Reference Laboratory Lanzhou Veterinary Research Institute Chinese Academy of Agricultural Sciences Lanzhou 730046 China

In order to develop an anti-FMDV A Type monoclonal antibody （mAb）, BABL/c mice were immunized with FMDV A type. Monoclonal antibodies （mAbs） 7B 11 and 8H4 against Foot-and-mouth disease virus （FMDV） serotype A were produced by fusing SP2/0 myeloma cells with splenocyte from the mouse immunized with A/AV88. The microneutralization titer of the mAbs 7Bll and 8H4 were 1024 and 512, respectively. Both mAbs contain kappa light chainS, the mAbs were IgG1. In order to define the mAbs binding epitopes, the reactivity of these mAbs against A Type FMDV, were examined using indirect ELISA, the result showed that both mAbs reacted with A Type FMDV. These mAbs may be used for further vaccine studies, diagnostic methods, prophylaxis, etiological and immunological research on FMDV. Characterization of these ncindicated that prepared anti-FMDV A mAbs had no cross-reactivity with Swine Vesicular Disease （SVD） or FMDV O, Asial and C Type antigens. Their titers in abdomen liquor were 1：5×10^6 and 1：2×10^6, respectively. 7B 11 was found to be of subtype IgGb 8H4 was classified as IgG2b subtype. The mAbs prepared in this study, are specific for detection of FMDV serotype A, and is potentially useful for pen-side diagnosis.

关键词： Foot-and-mouth disease virus (FMDV) A Type Monoclonal antibody (mAb) neutralizing activity

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A broadly neutralizing human monoclonal antibody against the hemagglutinin of avian influenza virus H7N9

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Chinese Medical Journal 2022年第7期135卷 799-805页

作者： Jingxin Li Li Zhang Linlin Bao Yuxiao Wang Lin Qiu Jialei Hu Rong Tang Huiyan Yu Jun Shan Yan Li Chuan Qin Fengcai Zhu Department for Vaccine Clinical Evaluation NHC Key Laboratory of Enteric Pathogenic MicrobiologyJlangsu Provincial Center for Diseases Control and PreventionNanjingJiangsu 210009China Institute of Laboratory Animal Science Chinese Academy of Medical Sciences(CAMS)&Comparative Medicine CenterPeking Union Medical Collage(PUMC)Key Laboratory of Human Disease Comparative MedicineMinistry of HealthBeijing Key Laboratory for Animal Models of Emerging and Reemerging Infectious DiseasesBeijing 100101China Department of Public Health Southeast UniversityNanjingJiangsu 210009China Shanghai Institute for Biological Sciences Chinese Academy of SciencesShanghai 200031China

Background:The new emerging avian influenza A H7N9 virus,causing severe human infection with a mortality rate of around 41%.This study aims to provide a novel treatment option for the prevention and control of ***:H7 hemagglutinin(HA)-specific B cells were isolated from peripheral blood plasma cells of the patients previously infected by H7N9 in Jiangsu Province,*** human monoclonal antibodies(mAbs)were generated by amplification and cloning of these HA-specific B ***,all human mAbs were screened for binding activity by enzyme-linked immunosorbent ***,those mAbs,exhibiting potent affinity to recognize H7 HAs were further evaluated by hemagglutination-inhibiting(HAI)and microneutralizationin vitro ***,the lead mAb candidate was selected and tested against the lethal challenge of the H7N9 virus using murine ***:The mAb 6-137 was able to recognize a panel of H7 HAs with high affinity but not HA of other subtypes,including H1N1 and *** mAb 6-137 can efficiently inhibit the HA activity in the inactivated H7N9 virus and neutralize 100 tissue culture infectious dose 50(TCID_(50))of H7N9 virus(influenza A/Nanjing/1/2013)invitro,with neutralizing activity as low as 78 ng/*** addition,the mAb 6-137 protected the mice against the lethal challenge of H7N9 prophylactically and ***:The mAb 6-137 could be an effective antibody as a prophylactic or therapeutic biological treatment for the H7N9 exposure or infection.

关键词： Avian influenza H7N9 Monoclonal antibody neutralizing activity

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