Background:Goat milk is very similar to human milk in terms of its abundant nutrients and ease of digestion.To derive greater economic benefit,farmers require more female offspring(does);however,the buck-to-doe offspr...Background:Goat milk is very similar to human milk in terms of its abundant nutrients and ease of digestion.To derive greater economic benefit,farmers require more female offspring(does);however,the buck-to-doe offspring sex ratio is approximately 50%.At present,artificial insemination after the separation of X/Y sperm using flow cytometry is the primary means of controlling the sex of livestock offspring.However,flow cytometry has not been successfully utilised for the separation of X/Y sperm aimed at sexing control in dairy goats.Results:In this study,a novel,simple goat sperm sexing technology that activates the toll-like receptor 7/8(TLR7/8),thereby inhibiting X-sperm motility,was investigated.Our results showed that the TLR7/8 coding goat Xchromosome was expressed in approximately 50%of round spermatids in the testis and sperm,as measured from cross-sections of the epididymis and ejaculate,respectively.Importantly,TLR7/8 was located at the tail of the Xsperm.Upon TLR7/8 activation,phosphorylated forms of glycogen synthase kinaseα/β(GSK3α/β)and nuclear factor kappa-B(NF-κB)were detected in the X-sperm,causing reduced mitochondrial activity,ATP levels,and sperm motility.High-motility Y-sperm segregated to the upper layer and the low-motility X-sperm,to the lower layer.Following in vitro fertilisation using the TLR7/8-activated sperm from the lower layer,80.52±6.75%of the embryos were XX females.The TLR7/8-activated sperm were subsequently used for in vivo embryo production via the superovulatory response;nine embryos were collected from the uterus of two does that conceived.Eight of these were XX embryos,and one was an XY embryo.Conclusions:Our study reveals a novel TLR7/8 signalling mechanism that affects X-sperm motility via the GSK3α/β-hexokinase pathway;this technique could be used to facilitate the efficient production of sexed dairy goat embryos.展开更多
The demand for safe vaccines that ensure long-term and broad protection against multiple viral variants has dramatically increased after the emergence of catastrophic infectious diseases such as COVID-19.To ensure lon...The demand for safe vaccines that ensure long-term and broad protection against multiple viral variants has dramatically increased after the emergence of catastrophic infectious diseases such as COVID-19.To ensure long-term and broad protection against heterologous virus variants,antigen-specific polyfunctional T cells should be orchestrated with the activation of follicular helper T(TFH)cells and germinal center(GC)B cells.Herein,we suggest a novel engineered nanoadjuvant(SE(Trojan-TLR7/8a))that enhances the migration of nonexhausted antigen-presenting cells(APCs)into lymph nodes and elicits the activation of TFH cells,the generation of GC B cells,and polyfunctional T cells via multiscale dynamic immunomodulation through squalene nanoemulsion(SE)-mediated macroscopic control of vaccine delivery and Trojan-TLR7/8a-enabled dynamic and sustained activation of APCs at the cellular level.SE(Trojan-TLR7/8a)can be lyophilized,reduce systemic toxicity,and outperform current commercial vaccine adjuvants(Alum or AS03)and mRNA vaccines.SE(Trojan-TLR7/8a)ensures cross-protection against diverse influenza and SARS-CoV-2 variants,providing 100%protection while maintaining a healthy state.SE(Trojan-TLR7/8a)also sustains a potent T-cell response in an aged ferret model of SFTSV infection.SE(Trojan-TLR7/8a)suggested herein provides a novel vaccine design principle for dynamic modulation at the multiscale level and demonstrates long-term and broad protective immunity against emerging pandemic and endemic infectious viruses.展开更多
Toll-like receptors(TLRs)are a family of highly conserved germline-encoded pattern-recognition receptors that are essential for host immune responses.TLR ligands represent a promising class of immunotherapeutics or va...Toll-like receptors(TLRs)are a family of highly conserved germline-encoded pattern-recognition receptors that are essential for host immune responses.TLR ligands represent a promising class of immunotherapeutics or vaccine adjuvants with the potential to generate an effective antitumor immune response.The TLR7/8 agonists have aroused interest because they not only activate antigen-presenting cells but also promote activation of T and natural killer(NK)cells.However,the exact mechanism by which stimulation of these TLRs promotes immune responses remains unclear,and different TLR7/8 agonists have been found to induce different responses.In this study,we demonstrate that both gardiquimod and imiquimod promote the proliferation of murine splenocytes,stimulate the activation of splenic T,NK and natural killer T(NKT)cells,increase the cytolytic activity of splenocytes against B16 and MCA-38 tumor cell lines,and enhance the expression of costimulatory molecules and IL-12 by macrophages and bone marrow-derived dendritic cells(DCs).In a murine model,both agonists improved the antitumor effects of tumor lysate-loaded DCs,resulting in delayed growth of subcutaneous B16 melanoma tumors and suppression of pulmonary metastasis.Further,we found that gardiquimod demonstrated more potent antitumor activity than imiquimod.These results suggest that TLR7/8 agonists may serve as potent innate and adaptive immune response modifiers in tumor therapy.More importantly,they can be used as vaccine adjuvants to potentiate the efficiency of DC-based tumor immunotherapy.展开更多
Due to the heterogeneity of tumors,single phototherapy cannot completely ablate tumors and inhibit tumor metastasis.To overcome these,we formulated targeted and multifunctional polymersomes ABC@ICGIMQ-LHRH(AIRL)that e...Due to the heterogeneity of tumors,single phototherapy cannot completely ablate tumors and inhibit tumor metastasis.To overcome these,we formulated targeted and multifunctional polymersomes ABC@ICGIMQ-LHRH(AIRL)that encapsulated Toll-like receptor(TLR)7/8 agonist imiquimod(IMQ)and photosensitizer indocyanine green(ICG)in the hydrophobic layer as well as bubble-generator NH_(4)HCO_(3) in the hydrophilic cavity to inhibit the growth of primary and distant tumors,and prevent tumor metastasis through synergistic photoimmunotherapy.The AIRL polymersomes exhibited uniform and stable size,and high drug encapsulation efficiency,acid/reduction/laser responsiveness,excellent photothermal conversion efficiency,effective reactive oxygen species generation,high tumor accumulation.AIRL could be effectively internalized by dendritic cells(DCs),achieve lysosome escape and enhance DCs maturation.The synergistic photoimmunotherapy via AIRL polymersomes remarkably promoted the differentiation and activation of T cells,elevated strong systemic immune response to eradicate primary tumors and inhibit the growth of distant tumors.Simultaneously,the endurable immunological memory prevented tumor metastasis,which provided a promising nanoplatform for the combination therapy of cancer.展开更多
Radiotherapy(RT)is a cornerstone of cancer treatment but limited by its dual role in modulating the tumor immune microenvironment:while promoting immunogenic cell death(ICD),RT concurrently upregulates PD-L1 to suppre...Radiotherapy(RT)is a cornerstone of cancer treatment but limited by its dual role in modulating the tumor immune microenvironment:while promoting immunogenic cell death(ICD),RT concurrently upregulates PD-L1 to suppress antitumor immunity.To address this limitation,we developed PEP-PLG-IMDQ,a polymer-peptideimmune agonist nanomedicine that synergizes RT with immunotherapy.This nanoplatform employs a poly(L-glutamic acid)carrier conjugated with a PD-L1-targeting peptide and the TLR7/8 agonist imidazoquinoline(IMDQ),enabling three-pronged action:(1)PD-L1-mediated tumor targeting,(2)TLR7/8-driven dendritic cell activation,and(3)reinforcement of the anti-tumor immune cycle.In CT26 tumor-bearing mice,RT combined with PEP-PLG-IMDQ achieved 98.1%tumor suppression,with 83%long-term survival and complete resistance to tumor rechallenge.Mechanistically,the combination therapy enhanced CD8^(+)T cell infiltration(5.3-fold vs.RT alone)and established durable immune memory.Our work provides a translatable strategy to overcome radioresistance through spatiotemporal immune modulation.展开更多
基金This research was supported by the National Natural Science Foundation of China(31672425)Shaanxi Province Key R&D Program(2018ZDXM-NY-043,2020ZDLNY02–04).
文摘Background:Goat milk is very similar to human milk in terms of its abundant nutrients and ease of digestion.To derive greater economic benefit,farmers require more female offspring(does);however,the buck-to-doe offspring sex ratio is approximately 50%.At present,artificial insemination after the separation of X/Y sperm using flow cytometry is the primary means of controlling the sex of livestock offspring.However,flow cytometry has not been successfully utilised for the separation of X/Y sperm aimed at sexing control in dairy goats.Results:In this study,a novel,simple goat sperm sexing technology that activates the toll-like receptor 7/8(TLR7/8),thereby inhibiting X-sperm motility,was investigated.Our results showed that the TLR7/8 coding goat Xchromosome was expressed in approximately 50%of round spermatids in the testis and sperm,as measured from cross-sections of the epididymis and ejaculate,respectively.Importantly,TLR7/8 was located at the tail of the Xsperm.Upon TLR7/8 activation,phosphorylated forms of glycogen synthase kinaseα/β(GSK3α/β)and nuclear factor kappa-B(NF-κB)were detected in the X-sperm,causing reduced mitochondrial activity,ATP levels,and sperm motility.High-motility Y-sperm segregated to the upper layer and the low-motility X-sperm,to the lower layer.Following in vitro fertilisation using the TLR7/8-activated sperm from the lower layer,80.52±6.75%of the embryos were XX females.The TLR7/8-activated sperm were subsequently used for in vivo embryo production via the superovulatory response;nine embryos were collected from the uterus of two does that conceived.Eight of these were XX embryos,and one was an XY embryo.Conclusions:Our study reveals a novel TLR7/8 signalling mechanism that affects X-sperm motility via the GSK3α/β-hexokinase pathway;this technique could be used to facilitate the efficient production of sexed dairy goat embryos.
基金supported by National Research Foundation(NRF)grants funded by the Korean government(grant numbers RS-2025-00513566 and RS-2023-00218648),Republic of Korea(Prof.Yong Taik Lim)supported by NRF grants funded by Korean government(grant number 2021R1A6A1A03045495),Republic of Korea(Prof.Jong-Soo Lee)supported by the Institute for Basic Science(grant number IBS-R801-D1),Republic of Korea(Director Young Ki Choi).
文摘The demand for safe vaccines that ensure long-term and broad protection against multiple viral variants has dramatically increased after the emergence of catastrophic infectious diseases such as COVID-19.To ensure long-term and broad protection against heterologous virus variants,antigen-specific polyfunctional T cells should be orchestrated with the activation of follicular helper T(TFH)cells and germinal center(GC)B cells.Herein,we suggest a novel engineered nanoadjuvant(SE(Trojan-TLR7/8a))that enhances the migration of nonexhausted antigen-presenting cells(APCs)into lymph nodes and elicits the activation of TFH cells,the generation of GC B cells,and polyfunctional T cells via multiscale dynamic immunomodulation through squalene nanoemulsion(SE)-mediated macroscopic control of vaccine delivery and Trojan-TLR7/8a-enabled dynamic and sustained activation of APCs at the cellular level.SE(Trojan-TLR7/8a)can be lyophilized,reduce systemic toxicity,and outperform current commercial vaccine adjuvants(Alum or AS03)and mRNA vaccines.SE(Trojan-TLR7/8a)ensures cross-protection against diverse influenza and SARS-CoV-2 variants,providing 100%protection while maintaining a healthy state.SE(Trojan-TLR7/8a)also sustains a potent T-cell response in an aged ferret model of SFTSV infection.SE(Trojan-TLR7/8a)suggested herein provides a novel vaccine design principle for dynamic modulation at the multiscale level and demonstrates long-term and broad protective immunity against emerging pandemic and endemic infectious viruses.
基金supported by grants from the Natural Science Foundation of China(no.90713033)the National 973 Basic Research Program of China(no.2007CB815800)the National 115 Key Project for HBV Research(no.2008ZX10002-008).
文摘Toll-like receptors(TLRs)are a family of highly conserved germline-encoded pattern-recognition receptors that are essential for host immune responses.TLR ligands represent a promising class of immunotherapeutics or vaccine adjuvants with the potential to generate an effective antitumor immune response.The TLR7/8 agonists have aroused interest because they not only activate antigen-presenting cells but also promote activation of T and natural killer(NK)cells.However,the exact mechanism by which stimulation of these TLRs promotes immune responses remains unclear,and different TLR7/8 agonists have been found to induce different responses.In this study,we demonstrate that both gardiquimod and imiquimod promote the proliferation of murine splenocytes,stimulate the activation of splenic T,NK and natural killer T(NKT)cells,increase the cytolytic activity of splenocytes against B16 and MCA-38 tumor cell lines,and enhance the expression of costimulatory molecules and IL-12 by macrophages and bone marrow-derived dendritic cells(DCs).In a murine model,both agonists improved the antitumor effects of tumor lysate-loaded DCs,resulting in delayed growth of subcutaneous B16 melanoma tumors and suppression of pulmonary metastasis.Further,we found that gardiquimod demonstrated more potent antitumor activity than imiquimod.These results suggest that TLR7/8 agonists may serve as potent innate and adaptive immune response modifiers in tumor therapy.More importantly,they can be used as vaccine adjuvants to potentiate the efficiency of DC-based tumor immunotherapy.
基金financially supported by National Natural Science Foundation of China(Nos.82172090,82302390 and 82072059)CAMS Innovation Fund for Medical Sciences(Nos.2021-I2M-1-058 and 2022-I2M-1-023)+3 种基金China Postdoctoral Science Foundation(No.2022M720502)Tianjin Municipal Natural Science Foundation(No.22JCQNJC00070)CAMS Union Young Scholars Support Program(No.2022051)Fundamental Research Funds for the Central Universities(No.2019PT320028)。
文摘Due to the heterogeneity of tumors,single phototherapy cannot completely ablate tumors and inhibit tumor metastasis.To overcome these,we formulated targeted and multifunctional polymersomes ABC@ICGIMQ-LHRH(AIRL)that encapsulated Toll-like receptor(TLR)7/8 agonist imiquimod(IMQ)and photosensitizer indocyanine green(ICG)in the hydrophobic layer as well as bubble-generator NH_(4)HCO_(3) in the hydrophilic cavity to inhibit the growth of primary and distant tumors,and prevent tumor metastasis through synergistic photoimmunotherapy.The AIRL polymersomes exhibited uniform and stable size,and high drug encapsulation efficiency,acid/reduction/laser responsiveness,excellent photothermal conversion efficiency,effective reactive oxygen species generation,high tumor accumulation.AIRL could be effectively internalized by dendritic cells(DCs),achieve lysosome escape and enhance DCs maturation.The synergistic photoimmunotherapy via AIRL polymersomes remarkably promoted the differentiation and activation of T cells,elevated strong systemic immune response to eradicate primary tumors and inhibit the growth of distant tumors.Simultaneously,the endurable immunological memory prevented tumor metastasis,which provided a promising nanoplatform for the combination therapy of cancer.
基金supported by National Natural Science Foundation of China(52403211,52025035,U24A20477)the National Key R&D Program of China(2022YFE0110200)+1 种基金Jilin Provincial Department of Education Scientific Research Project(JJKH20250194BS)Jilin Provincial International Cooperation Key Laboratory of Biomedical Polymers(YDZJ202402077CXJD).
文摘Radiotherapy(RT)is a cornerstone of cancer treatment but limited by its dual role in modulating the tumor immune microenvironment:while promoting immunogenic cell death(ICD),RT concurrently upregulates PD-L1 to suppress antitumor immunity.To address this limitation,we developed PEP-PLG-IMDQ,a polymer-peptideimmune agonist nanomedicine that synergizes RT with immunotherapy.This nanoplatform employs a poly(L-glutamic acid)carrier conjugated with a PD-L1-targeting peptide and the TLR7/8 agonist imidazoquinoline(IMDQ),enabling three-pronged action:(1)PD-L1-mediated tumor targeting,(2)TLR7/8-driven dendritic cell activation,and(3)reinforcement of the anti-tumor immune cycle.In CT26 tumor-bearing mice,RT combined with PEP-PLG-IMDQ achieved 98.1%tumor suppression,with 83%long-term survival and complete resistance to tumor rechallenge.Mechanistically,the combination therapy enhanced CD8^(+)T cell infiltration(5.3-fold vs.RT alone)and established durable immune memory.Our work provides a translatable strategy to overcome radioresistance through spatiotemporal immune modulation.
