Objective To develop an efficient expression, purification system of recombinant Escherichia coli heat labile enterotoxin B subunit (rLTB) and study its activity against mucosal immunoadjuvant by nasal immunization ...Objective To develop an efficient expression, purification system of recombinant Escherichia coli heat labile enterotoxin B subunit (rLTB) and study its activity against mucosal immunoadjuvant by nasal immunization Methods A recombinant, pMMB68 LTB was generated by cloning the LTB cDNA fragment into an expression vector (pMMB68) and transformed it into the host strain marine vibrio VSP60 The relevant target protein was identified using SDS polyacrylamide gel electrophoresis (SDS PAGE) and Western blot Sephacryl S 100 gel filtration chromatography was carried out for purification of rLTB in engineering bacteria VSP60 BALB/c mice received hen egg lysozyme (HEL) alone or combined with rLTB by nasal administration After three times immunization, IgG and IgA antibody levels in serum or small intestine wash samples were determined using ELISA Results rLTB protein was highly expressed in VSP60 After gel filtration with Sephacryl S 100, the purity of rLTB reached 98 1%, the yield rate was about 52% After immunization, IgG and IgA antibody responses specific to HEL in system and mucosa of HEL+rLTB groups were significantly increased, compared with the HEL alone group ( P <0 001) Conclusions A set of protocols for large scale rLTB preparation has been established, which is simple, efficient and applicable The rLTB protein we prepared was proved to be a powerful mucocal adjuvant, which could greatly enhance systemic and mucosal immune responses to nasally co administered antigen展开更多
The conventional immunoadjuvants in vaccine have weak effect on stimulating antigen presentation and activating anti-tumor immunity.Unexpectedly,we discovered that non-pathogenic Sendai virus(SeV)could activate antige...The conventional immunoadjuvants in vaccine have weak effect on stimulating antigen presentation and activating anti-tumor immunity.Unexpectedly,we discovered that non-pathogenic Sendai virus(SeV)could activate antigen-presenting cells(APCs)represented by dendritic cells(DCs).Here,we designed an injectable SeV-based hydrogel vaccine(SHV)to execute multi-channel recruitment and stimulation of DCs for boosting the specific immune response against tumors.After the release of the NIR-triggered antigens from tumor cells,dendritic cells around the vaccine efficiently transport the antigens to lymph nodes and present them to T lymphocytes,thereby inducing systemic anti-tumor immune memory.Our findings demonstrated that the SHV with excellent universality,convenience and flexibility has achieved better immune protection effects in inhibiting the occurrence of melanoma and breast cancer.In conclusion,the SHV system might serve as the next generation of personalized anti-tumor vaccines with enhanced features over standard vaccination regimens,and represented an alternative way to suppress tumorigenesis.展开更多
The authors regret that the qPCR primer sequence and Supplemen-tary Fig.S50 in supplementary data are incorrect.After reviewing our published work,we realized that the primer sequence for Bcl2 was written incorrectly ...The authors regret that the qPCR primer sequence and Supplemen-tary Fig.S50 in supplementary data are incorrect.After reviewing our published work,we realized that the primer sequence for Bcl2 was written incorrectly due to copy and paste during the writing process.And the primer sequence for Actin was omitted.In Supplementary Fig.S50,the H&E staining image corresponding to IV group was incorrectly provided.The corrected qPCR primer sequence and Fig.S50 is provided in this corrigendum.The authors are sorry for not checking the supplementary documents carefully before publication。展开更多
Treatment of metastatic cancer remains a great challenge and needs novel approaches.Combining a selective photothermal therapy with an active immunological stimulation,laser immunotherapy(LIT) was developed to induce ...Treatment of metastatic cancer remains a great challenge and needs novel approaches.Combining a selective photothermal therapy with an active immunological stimulation,laser immunotherapy(LIT) was developed to induce systemic immune responses through local intervention.LIT consists of three major components: a near-infrared laser,a light-absorbing agent,and an immunological stimulant.Its effect relies on two major interactions: a selective photothermal interaction and an active immunological stimulation.The selective photothermal interaction can reduce the tumor burden and at the same time release the tumor antigens,which can induce specific antitumor immune response.The expression of heat shock protein and the application of immunoadjuvant further enhance the host immunity.It has been proved in pre-clinical studies that LIT could not only eradicate treated local tumors but also regress and eliminate untreated metastases at distant sites.Moreover,LIT is well tolerated and has shown to have many advantages for cancer treatment compared with other traditional modalities.展开更多
As a non-invasive tumor therapy,photothermal therapy(PTT)has garnered considerable attention for its controllability,non-drug resistance and precise tumor ablation.However,its efficacy is often hindered by limited pho...As a non-invasive tumor therapy,photothermal therapy(PTT)has garnered considerable attention for its controllability,non-drug resistance and precise tumor ablation.However,its efficacy is often hindered by limited photothermal damage resulting from uncontrollable heat transfer distance and weak immune response caused by insufficient antigen presentation,resulting in tumor metastasis.Herein,a chondroitin sulfate-modified Prussian blue-montmorillonite immunoregulator(PM@CS)is developed to enhance both photothermal ablation and immune response.By integrating the tumor cell adhesion of montmorillonite and Golgi-targeting of CS,PM@CS accumulates on the Golgi apparatus of tumor cells,significantly reducing heat transfer distance compared to Prussian blue alone,thereby enhancing photothermal efficacy.Furthermore,the enhanced photothermal damage can effectively interfere with post-translational modification and secretion of metastasis-associated proteins(the expression of GOLPH3 and GOLM1 reduced by 63.4%and 70.3%,respectively).Besides,PM@CS leads to a 3.3-fold increase in dendritic cell maturation(CD80^(+) and CD86^(+) populations)and promotes the proliferation of antigen-specific CD4^(+) and CD8^(+)T cells,which is related to the immune potentiator property of montmorillonite and facilitates antigen presentation.Notably,the voltage-gated calcium channels(CaV)were upregulated and Ca^(2+) inflow was enhanced after PM@CS treatment,ultimately activating calcium signaling cascades.This is conducive to amplifying cascade immunotherapy,thus synergistically inhibiting primary tumor growth and lung metastasis.展开更多
文摘Objective To develop an efficient expression, purification system of recombinant Escherichia coli heat labile enterotoxin B subunit (rLTB) and study its activity against mucosal immunoadjuvant by nasal immunization Methods A recombinant, pMMB68 LTB was generated by cloning the LTB cDNA fragment into an expression vector (pMMB68) and transformed it into the host strain marine vibrio VSP60 The relevant target protein was identified using SDS polyacrylamide gel electrophoresis (SDS PAGE) and Western blot Sephacryl S 100 gel filtration chromatography was carried out for purification of rLTB in engineering bacteria VSP60 BALB/c mice received hen egg lysozyme (HEL) alone or combined with rLTB by nasal administration After three times immunization, IgG and IgA antibody levels in serum or small intestine wash samples were determined using ELISA Results rLTB protein was highly expressed in VSP60 After gel filtration with Sephacryl S 100, the purity of rLTB reached 98 1%, the yield rate was about 52% After immunization, IgG and IgA antibody responses specific to HEL in system and mucosa of HEL+rLTB groups were significantly increased, compared with the HEL alone group ( P <0 001) Conclusions A set of protocols for large scale rLTB preparation has been established, which is simple, efficient and applicable The rLTB protein we prepared was proved to be a powerful mucocal adjuvant, which could greatly enhance systemic and mucosal immune responses to nasally co administered antigen
基金the National Natural Science Foundation of China(32000999,81925020 and 81630051)the Key Project of Tianjin Natural Science Foundation(19JCZDJC34100)Open Project of State Key Laboratory of Oral Diseases(SKLOD2021OF07).
文摘The conventional immunoadjuvants in vaccine have weak effect on stimulating antigen presentation and activating anti-tumor immunity.Unexpectedly,we discovered that non-pathogenic Sendai virus(SeV)could activate antigen-presenting cells(APCs)represented by dendritic cells(DCs).Here,we designed an injectable SeV-based hydrogel vaccine(SHV)to execute multi-channel recruitment and stimulation of DCs for boosting the specific immune response against tumors.After the release of the NIR-triggered antigens from tumor cells,dendritic cells around the vaccine efficiently transport the antigens to lymph nodes and present them to T lymphocytes,thereby inducing systemic anti-tumor immune memory.Our findings demonstrated that the SHV with excellent universality,convenience and flexibility has achieved better immune protection effects in inhibiting the occurrence of melanoma and breast cancer.In conclusion,the SHV system might serve as the next generation of personalized anti-tumor vaccines with enhanced features over standard vaccination regimens,and represented an alternative way to suppress tumorigenesis.
文摘The authors regret that the qPCR primer sequence and Supplemen-tary Fig.S50 in supplementary data are incorrect.After reviewing our published work,we realized that the primer sequence for Bcl2 was written incorrectly due to copy and paste during the writing process.And the primer sequence for Actin was omitted.In Supplementary Fig.S50,the H&E staining image corresponding to IV group was incorrectly provided.The corrected qPCR primer sequence and Fig.S50 is provided in this corrigendum.The authors are sorry for not checking the supplementary documents carefully before publication。
基金supported in part by the US National Institutes of Health(P20 PR016478 from the INBRE Program of the National Center for Research Resources)
文摘Treatment of metastatic cancer remains a great challenge and needs novel approaches.Combining a selective photothermal therapy with an active immunological stimulation,laser immunotherapy(LIT) was developed to induce systemic immune responses through local intervention.LIT consists of three major components: a near-infrared laser,a light-absorbing agent,and an immunological stimulant.Its effect relies on two major interactions: a selective photothermal interaction and an active immunological stimulation.The selective photothermal interaction can reduce the tumor burden and at the same time release the tumor antigens,which can induce specific antitumor immune response.The expression of heat shock protein and the application of immunoadjuvant further enhance the host immunity.It has been proved in pre-clinical studies that LIT could not only eradicate treated local tumors but also regress and eliminate untreated metastases at distant sites.Moreover,LIT is well tolerated and has shown to have many advantages for cancer treatment compared with other traditional modalities.
基金supported by the National Natural Science Foundation of China(52203195)the China Postdoctoral Science Foundation(2023M733303)+2 种基金the 10th Young Elite Scientists Sponsorship Program by CASTthe CUG Scholar Scientific Research Funds at China University of Geosciences(Wuhan)(20222021178)the National Science Fund for Distinguished Young Scholars(51225403)。
文摘As a non-invasive tumor therapy,photothermal therapy(PTT)has garnered considerable attention for its controllability,non-drug resistance and precise tumor ablation.However,its efficacy is often hindered by limited photothermal damage resulting from uncontrollable heat transfer distance and weak immune response caused by insufficient antigen presentation,resulting in tumor metastasis.Herein,a chondroitin sulfate-modified Prussian blue-montmorillonite immunoregulator(PM@CS)is developed to enhance both photothermal ablation and immune response.By integrating the tumor cell adhesion of montmorillonite and Golgi-targeting of CS,PM@CS accumulates on the Golgi apparatus of tumor cells,significantly reducing heat transfer distance compared to Prussian blue alone,thereby enhancing photothermal efficacy.Furthermore,the enhanced photothermal damage can effectively interfere with post-translational modification and secretion of metastasis-associated proteins(the expression of GOLPH3 and GOLM1 reduced by 63.4%and 70.3%,respectively).Besides,PM@CS leads to a 3.3-fold increase in dendritic cell maturation(CD80^(+) and CD86^(+) populations)and promotes the proliferation of antigen-specific CD4^(+) and CD8^(+)T cells,which is related to the immune potentiator property of montmorillonite and facilitates antigen presentation.Notably,the voltage-gated calcium channels(CaV)were upregulated and Ca^(2+) inflow was enhanced after PM@CS treatment,ultimately activating calcium signaling cascades.This is conducive to amplifying cascade immunotherapy,thus synergistically inhibiting primary tumor growth and lung metastasis.
