Oncolysate, a debris of tumor cells, has been provento be effective in tumor active immunotherapy, it wasreported that the vaccinia virus, especially recombinantvaccinia viruses encoding human IL-2 (rVV-IL-2 ),enhance...Oncolysate, a debris of tumor cells, has been provento be effective in tumor active immunotherapy, it wasreported that the vaccinia virus, especially recombinantvaccinia viruses encoding human IL-2 (rVV-IL-2 ),enhanced the immunogenicity of transfected tumor cells.In this experiment, the murine melanoma cell B16-F10oncolysates trans fected by rVV-IL-2 (IL-2VBO) wereused as vaccine. The IL-2VBO or TK-VBO was preparedby incubating B16-F10 cells with rVV-IL-2 or rVV-TK展开更多
Direct gene transfer into somatic tissue in vivo is adeveloping technology with potential application forcancer gene therapy. Retrovirus vector, which was aneffective vehicle, still has some disadvantages ingenerating...Direct gene transfer into somatic tissue in vivo is adeveloping technology with potential application forcancer gene therapy. Retrovirus vector, which was aneffective vehicle, still has some disadvantages ingenerating high titer recombinant vectors andmanipulating to mediate in viro gene transfer. In thispaper, recombinant vaccinia virus vector encoding展开更多
The adaptive immune system produces a large and diverse set of antibodies,each with an individual evolutionary and clonal history.This so called"antibody repertoire"protects each individual against insults s...The adaptive immune system produces a large and diverse set of antibodies,each with an individual evolutionary and clonal history.This so called"antibody repertoire"protects each individual against insults such as infection and cancer,and responds to vaccination with B cell proliferation in response to the antigenic stimulation.Hybridomas and antigen-specific FACSbased analysis have given us much insight on how the immune system generates the complex and diverse immune response required to protect the body from the wide variety of potential pathogens.However,these methods have not been sufficient to make global and unbiased characterizations of the clonal structure of the immune system of a particular individual。展开更多
Objective The malaria parasites remodel the host erythrocyte structure by exporting parasite proteins that interact with the membrane skeleton proteins of red blood cells(RBCs),facilitating their intracellular surviva...Objective The malaria parasites remodel the host erythrocyte structure by exporting parasite proteins that interact with the membrane skeleton proteins of red blood cells(RBCs),facilitating their intracellular survival and pathogenicity.Skeletonbinding protein 1(SBP1)is a conserved exported protein across Plasmodium species.In Plasmodium falciparum,SBP1 has been reported to interact with erythrocyte membrane skeleton proteins 4.1R and spectrin,while its contribution to erythrocyte remodeling and parasite virulence in Plasmodium berghei(Pb)remains unclear.This study aims to determine whether PbSBP1 associates with the host cytoskeletal protein 4.1R and to investigate its role in the remodeling of host RBCs and the pathogenicity of Plasmodium berghei.Methods In Plasmodium berghei,the relationship between PbSBP1 and the erythrocyte cytoskeletal protein 4.1R was examined using co-immunoprecipitation.A Pbsbp1 gene knockout mutant of Plasmodium berghei(Pbsbp1Δ)was generated based on the principle of double crossover homologous recombination.The deformability of erythrocytes infected with Pbsbp1Δparasites was assessed using microfluidic methods.Microchannels with an array of cylindrical pillars were used to detect modifications in infected RBC deformability.The infected RBCs were squashed between the rows and recovered between the columns and the transit velocity(μm/s)of infected RBCs travelling through the microchannel was recorded.The component of the erythrocyte membrane skeleton junctional complex,tropomodulin(TMOD),was fluorescently labeled,and the cytoskeletal network of infected erythrocytes was imaged using super-resolution stochastic optical reconstruction microscopy(STORM)to analyze ultrastructural changes in the cytoskeleton of wild-type(WT)and Pbsbp1Δ-infected erythrocytes.Actin-based junctional complexes were displayed as individual clusters by the labeled TMOD in the STORM images,and the cluster densities and distances between adjacent clusters of infected RBCs were calculated.Additionally,rodent malaria models(BALB/c mice)and experimental cerebral malaria models(C57BL/6 mice)were employed to monitor the growth of Pbsbp1Δand WT parasites during the intraerythrocytic stage and their capacity to induce cerebral malaria in mice.Results PbSBP1 may participate in the remodeling of infected erythrocytes through direct or indirect interaction with the erythrocyte cytoskeletal protein 4.1R.Microfluidic assays revealed that the deformability of erythrocytes infected with Pbsbp1Δparasites was significantly enhanced compared to those infected with WT parasites.STORM imaging further demonstrated that the ultrastructure of the erythrocyte cytoskeleton in Pbsbp1Δ-infected cells was altered relative to that in WTinfected erythrocytes.The distances between nearest neighbors of clusters had a tendency to increase while the cluster densities were decreased in Pbsbp1Δ-infected RBCs compared to WT-infected RBCs.Subsequent phenotypic analysis indicated that the growth rate of Pbsbp1Δparasites during the intraerythrocytic stage was significantly slower than that of WT parasites,and their ability to induce cerebral malaria in mice was also attenuated.These findings suggest that PbSBP1 is involved in the remodeling of the erythrocyte membrane skeleton,likely through its direct or indirect interaction with protein 4.1R,thereby regulating the deformability of infected erythrocytes and influencing the pathogenicity of the blood-stage parasites.Conclusion This study establishes a role for PbSBP1 in host erythrocyte remodeling and parasite virulence,providing new research strategies for the prevention and treatment of malaria.展开更多
文摘Oncolysate, a debris of tumor cells, has been provento be effective in tumor active immunotherapy, it wasreported that the vaccinia virus, especially recombinantvaccinia viruses encoding human IL-2 (rVV-IL-2 ),enhanced the immunogenicity of transfected tumor cells.In this experiment, the murine melanoma cell B16-F10oncolysates trans fected by rVV-IL-2 (IL-2VBO) wereused as vaccine. The IL-2VBO or TK-VBO was preparedby incubating B16-F10 cells with rVV-IL-2 or rVV-TK
文摘Direct gene transfer into somatic tissue in vivo is adeveloping technology with potential application forcancer gene therapy. Retrovirus vector, which was aneffective vehicle, still has some disadvantages ingenerating high titer recombinant vectors andmanipulating to mediate in viro gene transfer. In thispaper, recombinant vaccinia virus vector encoding
基金supported by the National Institutes of Health grant U19 A1057229(M.M. D.,X.H.,H.B.G.and S.R.Q.)a National Institutes of Health Pathway to Independence Award K99 AG040149(N.J.)a National Science Foundation graduate fellowship(J.A.W.)
文摘The adaptive immune system produces a large and diverse set of antibodies,each with an individual evolutionary and clonal history.This so called"antibody repertoire"protects each individual against insults such as infection and cancer,and responds to vaccination with B cell proliferation in response to the antigenic stimulation.Hybridomas and antigen-specific FACSbased analysis have given us much insight on how the immune system generates the complex and diverse immune response required to protect the body from the wide variety of potential pathogens.However,these methods have not been sufficient to make global and unbiased characterizations of the clonal structure of the immune system of a particular individual。
文摘Objective The malaria parasites remodel the host erythrocyte structure by exporting parasite proteins that interact with the membrane skeleton proteins of red blood cells(RBCs),facilitating their intracellular survival and pathogenicity.Skeletonbinding protein 1(SBP1)is a conserved exported protein across Plasmodium species.In Plasmodium falciparum,SBP1 has been reported to interact with erythrocyte membrane skeleton proteins 4.1R and spectrin,while its contribution to erythrocyte remodeling and parasite virulence in Plasmodium berghei(Pb)remains unclear.This study aims to determine whether PbSBP1 associates with the host cytoskeletal protein 4.1R and to investigate its role in the remodeling of host RBCs and the pathogenicity of Plasmodium berghei.Methods In Plasmodium berghei,the relationship between PbSBP1 and the erythrocyte cytoskeletal protein 4.1R was examined using co-immunoprecipitation.A Pbsbp1 gene knockout mutant of Plasmodium berghei(Pbsbp1Δ)was generated based on the principle of double crossover homologous recombination.The deformability of erythrocytes infected with Pbsbp1Δparasites was assessed using microfluidic methods.Microchannels with an array of cylindrical pillars were used to detect modifications in infected RBC deformability.The infected RBCs were squashed between the rows and recovered between the columns and the transit velocity(μm/s)of infected RBCs travelling through the microchannel was recorded.The component of the erythrocyte membrane skeleton junctional complex,tropomodulin(TMOD),was fluorescently labeled,and the cytoskeletal network of infected erythrocytes was imaged using super-resolution stochastic optical reconstruction microscopy(STORM)to analyze ultrastructural changes in the cytoskeleton of wild-type(WT)and Pbsbp1Δ-infected erythrocytes.Actin-based junctional complexes were displayed as individual clusters by the labeled TMOD in the STORM images,and the cluster densities and distances between adjacent clusters of infected RBCs were calculated.Additionally,rodent malaria models(BALB/c mice)and experimental cerebral malaria models(C57BL/6 mice)were employed to monitor the growth of Pbsbp1Δand WT parasites during the intraerythrocytic stage and their capacity to induce cerebral malaria in mice.Results PbSBP1 may participate in the remodeling of infected erythrocytes through direct or indirect interaction with the erythrocyte cytoskeletal protein 4.1R.Microfluidic assays revealed that the deformability of erythrocytes infected with Pbsbp1Δparasites was significantly enhanced compared to those infected with WT parasites.STORM imaging further demonstrated that the ultrastructure of the erythrocyte cytoskeleton in Pbsbp1Δ-infected cells was altered relative to that in WTinfected erythrocytes.The distances between nearest neighbors of clusters had a tendency to increase while the cluster densities were decreased in Pbsbp1Δ-infected RBCs compared to WT-infected RBCs.Subsequent phenotypic analysis indicated that the growth rate of Pbsbp1Δparasites during the intraerythrocytic stage was significantly slower than that of WT parasites,and their ability to induce cerebral malaria in mice was also attenuated.These findings suggest that PbSBP1 is involved in the remodeling of the erythrocyte membrane skeleton,likely through its direct or indirect interaction with protein 4.1R,thereby regulating the deformability of infected erythrocytes and influencing the pathogenicity of the blood-stage parasites.Conclusion This study establishes a role for PbSBP1 in host erythrocyte remodeling and parasite virulence,providing new research strategies for the prevention and treatment of malaria.
