Objective To investigate the anti-tumor effects in vitro and in vivo distribution of the human/murine chimeric antibody (D2C). Methods The CD71 positive target cells (K562, GEM and SMMC7721) and the effector cells, fr...Objective To investigate the anti-tumor effects in vitro and in vivo distribution of the human/murine chimeric antibody (D2C). Methods The CD71 positive target cells (K562, GEM and SMMC7721) and the effector cells, freshly isolated human PBMC, with the ratio of target cells to effector cells 1:50, were incubated in various dilutions of D2C antibody ( Ab) . Antibody dependent cytotoxicity (AD-CC) was tested by using an LDH-release assay. Instead of effector cells, complement was added to the target cells (GEM, SMMC-7721) with various dilutions of D2C Ab. A method of counting death cells was used in complement dependent cytotoxicity (CDC) assay. Tumor localization and distribution of the chimeric antibody (D2C) were observed by labeling the chimeric Ab with radioiodine(131I) and injecting it into nude mice (Balb/c nu/nu) transplanted with human hepatocellular carcinoma cells (SMMC-7721).Results A significant ADCC was observed with the increased concentration of the D2C Ab. Cytolysis of CD71-positive target cells by the D2C Ab was found in the presence of fresh rabbit complement. Labeled D2C administered by intraperitoneal as well as tumor regional injection, was visualized by SPECT. The distribution of D2C Ab in murine organs and tissues showed that non-specific binding was lower following tumor regional administration than when the antibody was administered by an intraperitoneal injection. The human/murine chimeric antibody (D2C) has in vitro anti-tumor effects and can exert its effects in specific tumor localization. Its distribution and local effects in vivo can be detected by radioimmunoimaging.Conclusion CD71 human/murine chimeric antibody showed marked killing of tumor cells in vitro, and specific recognition and high affinity binding to tumor tissue in vivo展开更多
P74 is a per os infectivity factor of baculovirus.Here,we report the production of three monoclonal antibodies (mAbs),denoted as 20D9,20F9 and 21E1,raised against P74 of Helicoverpa armigera nucleopolyhedrovirus (Hear...P74 is a per os infectivity factor of baculovirus.Here,we report the production of three monoclonal antibodies (mAbs),denoted as 20D9,20F9 and 21E1,raised against P74 of Helicoverpa armigera nucleopolyhedrovirus (HearNPV),and the identification of their recognition epitopes.The full-length P74,without the transmembrane domains at the C-terminus,was first divided into three segments (N,M and C,respectively),based on the proposed cleavage model for the protein,which were then expressed individually.Western blot analyses revealed specific cross-reactions with the N fragment,for both 20D9 and 21E1.Extensive truncation,followed by prokaryotic expression,of the P74 N fragment was then performed in order to screen for linear epitopes of P74.The recognition regions of 20D9 and 21E1 were revealed to be localized at R144-T153 and T199-C219,respectively.In addition,immunofluorescence microscopy indicated that 20D9 and 20F9 could recognize native P74 in HearNPV-infected cells.These findings will facilitate further investigations of the proteolytic processing of HearNPV P74,and of its involvement in virus-host interactions.展开更多
Wheat powdery mildew (Pro) is a major disease of wheat worldwide. During the past years, numerous studies have been published on molecular mapping of Pm resistance gene(s) in wheat. We summarized the relevant find...Wheat powdery mildew (Pro) is a major disease of wheat worldwide. During the past years, numerous studies have been published on molecular mapping of Pm resistance gene(s) in wheat. We summarized the relevant findings of 89 major re- sistance gene mapping studies and 25 quantitative trait loci (QTL) mapping studies. Major Pm resistance genes and QTLs were found on all wheat chromosomes, but the Pm resistance genes/QTLs were not randomly distributed on each chromosome of wheat. The summarized data showed that the A or B genome has more major Pm resistance genes than the D genome and chromosomes 1A, 2A, 2B, 5B, 5D, 6B, 7A and 7B harbor more major Pm resistance genes than the other chromosomes. For adult plant resistance (APR) genes/QTLs, B genome of wheat harbors more APR genes than A and D genomes, and chromo- somes 2A, 4A, 5A, 1B, 2B, 3B, 5B, 6B, 7B, 2D, 5D and 7D harbor more Pm resistance QTLs than the other chromosomes, suggesting that A genome except 1A, 3A and 6A, B genome except 4B, D genome except 1D, 3D, 4D, and 6D play an impor- tant role in wheat combating against powdery mildew. Furthermore, Pm resistance genes are derived from wheat and its rela- tives, which suggested that the resistance sources are diverse and Pm resistance genes are diverse and useful in combating against the powdery mildew isolates. In this review, four APR genes, Pm38/Lr34/Yr18/Sr57, Pm46/Lr67/Yr46/Sr55, Pm?/Lr27/Yr30/ SY2 and Pm39/Lr46/Yr29, are not only resistant to powdery mildew but also effective for rust diseases in the field, indicating that such genes are stable and useful in wheat breeding programmes. The summarized data also provide chromosome locations or linked markers for Pm resistance genes/QTLs. Markers linked to these genes can also be utilized to pyramid diverse Pm resis- tance genes/QTLs more efficiently by marker-assisted selection.展开更多
基金National Sciences Foundation of China(No.39970693)
文摘Objective To investigate the anti-tumor effects in vitro and in vivo distribution of the human/murine chimeric antibody (D2C). Methods The CD71 positive target cells (K562, GEM and SMMC7721) and the effector cells, freshly isolated human PBMC, with the ratio of target cells to effector cells 1:50, were incubated in various dilutions of D2C antibody ( Ab) . Antibody dependent cytotoxicity (AD-CC) was tested by using an LDH-release assay. Instead of effector cells, complement was added to the target cells (GEM, SMMC-7721) with various dilutions of D2C Ab. A method of counting death cells was used in complement dependent cytotoxicity (CDC) assay. Tumor localization and distribution of the chimeric antibody (D2C) were observed by labeling the chimeric Ab with radioiodine(131I) and injecting it into nude mice (Balb/c nu/nu) transplanted with human hepatocellular carcinoma cells (SMMC-7721).Results A significant ADCC was observed with the increased concentration of the D2C Ab. Cytolysis of CD71-positive target cells by the D2C Ab was found in the presence of fresh rabbit complement. Labeled D2C administered by intraperitoneal as well as tumor regional injection, was visualized by SPECT. The distribution of D2C Ab in murine organs and tissues showed that non-specific binding was lower following tumor regional administration than when the antibody was administered by an intraperitoneal injection. The human/murine chimeric antibody (D2C) has in vitro anti-tumor effects and can exert its effects in specific tumor localization. Its distribution and local effects in vivo can be detected by radioimmunoimaging.Conclusion CD71 human/murine chimeric antibody showed marked killing of tumor cells in vitro, and specific recognition and high affinity binding to tumor tissue in vivo
基金supported by the National Science Foundation of China(31130058 to Z.H.).Monoclonal Antibodies against HearNPV P74
文摘P74 is a per os infectivity factor of baculovirus.Here,we report the production of three monoclonal antibodies (mAbs),denoted as 20D9,20F9 and 21E1,raised against P74 of Helicoverpa armigera nucleopolyhedrovirus (HearNPV),and the identification of their recognition epitopes.The full-length P74,without the transmembrane domains at the C-terminus,was first divided into three segments (N,M and C,respectively),based on the proposed cleavage model for the protein,which were then expressed individually.Western blot analyses revealed specific cross-reactions with the N fragment,for both 20D9 and 21E1.Extensive truncation,followed by prokaryotic expression,of the P74 N fragment was then performed in order to screen for linear epitopes of P74.The recognition regions of 20D9 and 21E1 were revealed to be localized at R144-T153 and T199-C219,respectively.In addition,immunofluorescence microscopy indicated that 20D9 and 20F9 could recognize native P74 in HearNPV-infected cells.These findings will facilitate further investigations of the proteolytic processing of HearNPV P74,and of its involvement in virus-host interactions.
基金Supported by the NSF of China(Grant no.31471488)State Key Laboratory of Crop Biology(2017KF03)+3 种基金Shandong Province Key Technology Innovation Project(2014GJJS0201-1)Transgenic Special Item(2016ZX08002003)National Modern Agricultural Industry System Construction Project(CARS-03-1-8)The Scholars of Taishan Seed Industry Project(2014-2019)
文摘Wheat powdery mildew (Pro) is a major disease of wheat worldwide. During the past years, numerous studies have been published on molecular mapping of Pm resistance gene(s) in wheat. We summarized the relevant findings of 89 major re- sistance gene mapping studies and 25 quantitative trait loci (QTL) mapping studies. Major Pm resistance genes and QTLs were found on all wheat chromosomes, but the Pm resistance genes/QTLs were not randomly distributed on each chromosome of wheat. The summarized data showed that the A or B genome has more major Pm resistance genes than the D genome and chromosomes 1A, 2A, 2B, 5B, 5D, 6B, 7A and 7B harbor more major Pm resistance genes than the other chromosomes. For adult plant resistance (APR) genes/QTLs, B genome of wheat harbors more APR genes than A and D genomes, and chromo- somes 2A, 4A, 5A, 1B, 2B, 3B, 5B, 6B, 7B, 2D, 5D and 7D harbor more Pm resistance QTLs than the other chromosomes, suggesting that A genome except 1A, 3A and 6A, B genome except 4B, D genome except 1D, 3D, 4D, and 6D play an impor- tant role in wheat combating against powdery mildew. Furthermore, Pm resistance genes are derived from wheat and its rela- tives, which suggested that the resistance sources are diverse and Pm resistance genes are diverse and useful in combating against the powdery mildew isolates. In this review, four APR genes, Pm38/Lr34/Yr18/Sr57, Pm46/Lr67/Yr46/Sr55, Pm?/Lr27/Yr30/ SY2 and Pm39/Lr46/Yr29, are not only resistant to powdery mildew but also effective for rust diseases in the field, indicating that such genes are stable and useful in wheat breeding programmes. The summarized data also provide chromosome locations or linked markers for Pm resistance genes/QTLs. Markers linked to these genes can also be utilized to pyramid diverse Pm resis- tance genes/QTLs more efficiently by marker-assisted selection.