Mucosal antibodies are critical for protection against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection by limiting viral replication and transmission.While systemic immunity to SARS-CoV-2 is well-...Mucosal antibodies are critical for protection against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection by limiting viral replication and transmission.While systemic immunity to SARS-CoV-2 is well-characterized,the durability and protective efficacy of mucosal vaccines,especially during the recent JN.1 wave,are less understood.In this study,we conducted a longitudinal assessment of systemic and nasal immune responses in 34 individuals who had received the inhaled Ad5-XBB.1.5 vaccination at days 0,7,14,28,90,and 180 post-vaccination,using neutralization assays,immunoglobulin A/G(IgA/IgG)enzyme-linked immunosorbent assay(ELISA),T-cell staining,and Fc-effector function assays.Our results showed that mucosal vaccination preferentially induces IgA responses in both nasal mucosa and systemic circulation,with nasal IgA showing stronger correlation with neutralizing titers than IgG.However,nasal antibody responses declined significantly by 6 months post-vaccination,with most participants experiencing breakthrough infections during the JN.1 wave.Individuals with high pre-existing anti-Ad5 antibodies exhibited reduced vaccine-induced neutralizing responses.The Ad5-XBB.1.5 mucosal vaccine enhanced antigen-specific CD8^(+)T cell responses with a slight increase in Fc-mediated antibody-dependent cellular phagocytosis(ADCP),but failed to induce detectable CD4^(+)T cell responses.Collectively,our findings elucidate the limited durability of mucosal immunity post-vaccination and highlight the need for improved mucosal vaccine strategies that sustain and optimize nasal IgA responses.展开更多
We have previously reported that AD5-10, a novel agonistic monoclonal antibody against DRS, possessed a strong cytotoxic activity in various tumor cells, via induction of caspase-dependent and -independent signaling p...We have previously reported that AD5-10, a novel agonistic monoclonal antibody against DRS, possessed a strong cytotoxic activity in various tumor cells, via induction of caspase-dependent and -independent signaling pathways. The present study further demonstrates that reactive oxygen species （ROS） were generated in abundance in Jurkat leukemia cells upon AD5-10 stimulation and that ROS accumulation subsequently evoked sustained activation of c-Jun N-terminal kinase （JNK）, loss of mitochondrial membrane potential, and release of endonuclease G （Endo G） from mitochondria into the cytosol. The reducing agent, N-acetylcysteine （NAC）, effectively inhibited the sustained activation of JNK, release of Endo G, and cell death in Jurkat cells treated by AD5-10. Moreover, a dominant-negative form of JNK （but not of p38） enhanced NF-κB activation, suppressed caspase-8 recruitment in death-inducing signaling complexes （DISCs）, and reduced adverse effects on mitochondria, thereby inhibiting AD5-10-induced cell death in Jnrkat leukemia cells. These data provide novel information on the DR5-mediated ceil death-signaling path- way and may shed new light on effective strategies for leukemia and solid tumor therapies.展开更多
We previously demonstrated that gene-modified umbilical cord blood mononuclear cells overexpressing a combination of recombinant neurotrophic factors are a promising therapeutic approach for cell-mediated gene therapy...We previously demonstrated that gene-modified umbilical cord blood mononuclear cells overexpressing a combination of recombinant neurotrophic factors are a promising therapeutic approach for cell-mediated gene therapy for neurodegenerative diseases,neurotrauma,and stroke.In this study,using a mini pig model of spinal cord injury,we proposed for the first time the use of gene-modified leucoconcentrate prepared from peripheral blood in the plastic blood bag for personalized ex vivo gene therapy.Leucoconcentrate obtained from mini pig peripheral blood was transduced with a chimeric adenoviral vector(Ad5/35 F)that carried an enhanced green fluorescent protein(EGFP)reporter gene in the plastic blood bag.The day after blood donation,the mini pigs were subjected to moderate SCI and four hours post-surgery they were intravenously autoinfused with gene-modified leucoconcentrate.A week after gene-modified leucoconcentrate therapy,fluorescent microscopy revealed EGFP-expressing leucocytes in spinal cord at the site of contusion injury.In the spleen the groups of EGFP-positive cells located in the lymphoid follicles were observed.In vitro flow cytometry and fluorescent microscopy studies of the gene-modified leucoconcentrate samples also confirmed the production of EGFP by leucocytes.Thus,the efficacy of leucocytes transduction in the plastic blood bag and their migratory potential suggest their use for temporary production of recombinant biologically active molecules to correct certain pathological conditions.This paper presents a proof-of-concept of simple,safe and effective approach for personalized ex vivo gene therapy based on gene-modified leucoconcentrate autoinfusion.The animal protocols were approved by the Kazan State Medical University Animal Care and Use Committee(approval No.5)on May 27,2014.展开更多
基金supported by grants from the National Key Research and Development Program of China(grant number 2024YFA0920001 to Y.W.)the National Natural Science Foundation of China(grant numbers 82025001,82495200,and 82495203 to J.Z.,92369113 and 82172240 to Y.W.)+5 种基金Guangzhou National Laboratory and State Key Laboratory of Respiratory Disease(grant number GZNL2024B01001 to Y.W.)Selfsupporting Program of Guangzhou National Laboratory(grant number SRPG22-001 to Y.W.)Guangdong Basic and Applied Research Projects(grant number 2023B1515020040 to Y.W.)State Key Laboratory of Respiratory Disease(grant number SKLRD-Z-202411 to L.Z.)Science and Technology Planning Project of Guangzhou City(grant numbers 2023A04J1279 to L.Z.and 2024A03J1230 to J.Z.)the Science and Technology Project of General Administration of Customs,P.R.China(grant number 2023HK065 to L.Z.).
文摘Mucosal antibodies are critical for protection against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection by limiting viral replication and transmission.While systemic immunity to SARS-CoV-2 is well-characterized,the durability and protective efficacy of mucosal vaccines,especially during the recent JN.1 wave,are less understood.In this study,we conducted a longitudinal assessment of systemic and nasal immune responses in 34 individuals who had received the inhaled Ad5-XBB.1.5 vaccination at days 0,7,14,28,90,and 180 post-vaccination,using neutralization assays,immunoglobulin A/G(IgA/IgG)enzyme-linked immunosorbent assay(ELISA),T-cell staining,and Fc-effector function assays.Our results showed that mucosal vaccination preferentially induces IgA responses in both nasal mucosa and systemic circulation,with nasal IgA showing stronger correlation with neutralizing titers than IgG.However,nasal antibody responses declined significantly by 6 months post-vaccination,with most participants experiencing breakthrough infections during the JN.1 wave.Individuals with high pre-existing anti-Ad5 antibodies exhibited reduced vaccine-induced neutralizing responses.The Ad5-XBB.1.5 mucosal vaccine enhanced antigen-specific CD8^(+)T cell responses with a slight increase in Fc-mediated antibody-dependent cellular phagocytosis(ADCP),but failed to induce detectable CD4^(+)T cell responses.Collectively,our findings elucidate the limited durability of mucosal immunity post-vaccination and highlight the need for improved mucosal vaccine strategies that sustain and optimize nasal IgA responses.
基金Acknowledgments We thank Dr Shimin Hu for his generous gifts of constructs. This work was partially supported by National Natural Science Foundation of China （Grant Nos. 30571687 and 30721063） and by State Key Basic Research Program of China （Grant No. 2007CB507404）.
文摘We have previously reported that AD5-10, a novel agonistic monoclonal antibody against DRS, possessed a strong cytotoxic activity in various tumor cells, via induction of caspase-dependent and -independent signaling pathways. The present study further demonstrates that reactive oxygen species （ROS） were generated in abundance in Jurkat leukemia cells upon AD5-10 stimulation and that ROS accumulation subsequently evoked sustained activation of c-Jun N-terminal kinase （JNK）, loss of mitochondrial membrane potential, and release of endonuclease G （Endo G） from mitochondria into the cytosol. The reducing agent, N-acetylcysteine （NAC）, effectively inhibited the sustained activation of JNK, release of Endo G, and cell death in Jurkat cells treated by AD5-10. Moreover, a dominant-negative form of JNK （but not of p38） enhanced NF-κB activation, suppressed caspase-8 recruitment in death-inducing signaling complexes （DISCs）, and reduced adverse effects on mitochondria, thereby inhibiting AD5-10-induced cell death in Jnrkat leukemia cells. These data provide novel information on the DR5-mediated ceil death-signaling path- way and may shed new light on effective strategies for leukemia and solid tumor therapies.
基金the Russian Science Foundation(No.16-15-00010to RRI)the Russian Government Program of Competitive Growth of Kazan Federal University。
文摘We previously demonstrated that gene-modified umbilical cord blood mononuclear cells overexpressing a combination of recombinant neurotrophic factors are a promising therapeutic approach for cell-mediated gene therapy for neurodegenerative diseases,neurotrauma,and stroke.In this study,using a mini pig model of spinal cord injury,we proposed for the first time the use of gene-modified leucoconcentrate prepared from peripheral blood in the plastic blood bag for personalized ex vivo gene therapy.Leucoconcentrate obtained from mini pig peripheral blood was transduced with a chimeric adenoviral vector(Ad5/35 F)that carried an enhanced green fluorescent protein(EGFP)reporter gene in the plastic blood bag.The day after blood donation,the mini pigs were subjected to moderate SCI and four hours post-surgery they were intravenously autoinfused with gene-modified leucoconcentrate.A week after gene-modified leucoconcentrate therapy,fluorescent microscopy revealed EGFP-expressing leucocytes in spinal cord at the site of contusion injury.In the spleen the groups of EGFP-positive cells located in the lymphoid follicles were observed.In vitro flow cytometry and fluorescent microscopy studies of the gene-modified leucoconcentrate samples also confirmed the production of EGFP by leucocytes.Thus,the efficacy of leucocytes transduction in the plastic blood bag and their migratory potential suggest their use for temporary production of recombinant biologically active molecules to correct certain pathological conditions.This paper presents a proof-of-concept of simple,safe and effective approach for personalized ex vivo gene therapy based on gene-modified leucoconcentrate autoinfusion.The animal protocols were approved by the Kazan State Medical University Animal Care and Use Committee(approval No.5)on May 27,2014.
