To treat cancer and inhibit its metastasis to the greatest extent,we proposed to develop an Au(III)agent to induce immunogenic cell death(ICD)and establish long-term immunity.To this end,we optimized a series of Au(II...To treat cancer and inhibit its metastasis to the greatest extent,we proposed to develop an Au(III)agent to induce immunogenic cell death(ICD)and establish long-term immunity.To this end,we optimized a series of Au(III)2-benzoylpyridine thiosemicarbazone complexes to obtain an Au(III)agent(5b)with excellent cytotoxicity to cancer.The results show that 5b effectively inhibits tumor growth and its metastasis in vivo.Interestingly,we revealed a new mechanism of 5b inhibiting tumor growth and metastasis:5b releases ICD-related damage-associated molecular patterns(DAMPs),such as calreticulin(CRT),ATP and high mobility group box 1(HMGB1)by inducing endoplasmic reticulum stress(ERS)and mitochondrial dysfunction,which then stimulated an antitumor CD8^(+)T cell response and Foxp^(3+)T cell depletion,thus establishing long-action antitumor immunity.展开更多
Chemotherapeutic drugs such as doxorubicin(DOx)and oxaliplatin can induce immunogenic cell death(ICD)in tumor cells.Current studies have demonstrated that peptide-based drugs can also induce ICD in tumor cells.Unlike ...Chemotherapeutic drugs such as doxorubicin(DOx)and oxaliplatin can induce immunogenic cell death(ICD)in tumor cells.Current studies have demonstrated that peptide-based drugs can also induce ICD in tumor cells.Unlike small molecule drugs,peptide drugs not only enhance the effectiveness of immunotherapy but also offer the advantage of low biotoxicity[1].展开更多
Ferroptosis in combination with immune therapy emerges as a promising approach for cancer therapy.Herein,dual-responsive metal-polyphenol coordinated nanomedicines were developed for pH/glutathione(GSH)-responsive syn...Ferroptosis in combination with immune therapy emerges as a promising approach for cancer therapy.Herein,dual-responsive metal-polyphenol coordinated nanomedicines were developed for pH/glutathione(GSH)-responsive synergistic ferroptosis and immunotherapy.Our innovative strategy involves the development of a manganese-polyphenol coordinated nanostructure,leveraging the biocompatibility of bovine serum albumin(BSA)as a template to encapsulate the anticancer drug sorafenib.The tumor microenvironment(pH/GSH)prompts the disassembly of MnO_(2)and epigallocatechin gallate(EGCG),thereby releases the anticancer payload.Concurrently,MnO_(2)acts to deplete intracellular GSH,which in turn suppresses glutathione peroxidase activity,leading to an accumulation of lipid peroxides with cell ferroptosis.Additionally,the release of Mn^(2+)ions bolster the cyclic guanosine monophosphlic acid(GMP)-adenosine monophosphlic acid(AMP)synthase-stimulator of interferon gene(cGAS-STING)pathway,which,in conjunction with the immunogenic cell death(ICD)effect induced by tumor cell apoptosis,significantly promotes dendritic cell(DC)maturation and enhances the presentation of tumor antigens.This successively ignites a robust innate and adaptive immune response.Both in vitro and in vivo experiments have demonstrated that the concurrent administration of ferroptosis-inducing and immune-stimulating therapies can significantly inhibit tumor growth.展开更多
Scrub typhus is an acute undifferentiated febrile infectious disease transmitted by a chigger(genus Leptotrombidium)bite carrying Orientia(O.)tsutsugamushi,affecting millions of people annually while more than one bil...Scrub typhus is an acute undifferentiated febrile infectious disease transmitted by a chigger(genus Leptotrombidium)bite carrying Orientia(O.)tsutsugamushi,affecting millions of people annually while more than one billion people are susceptible.Endemic areas are expanding to Africa,Europe,Middle East,and South America which is concerning,as despite best efforts,there is no vaccine to combat the bacteria.There are now three species of Orientia and over 20 strains of O.tsutsugamushi.The past attempts to develop a vaccine have been ineffective as they confer homologous strain-specific immunity.Various immunogenic proteins of O.tsutsugamushi have been identified that interact with the extracellular matrix(fibronectin)or vMLL5 receptor and modify the cytoskeleton of non-phagocytic host cells,which aids in host cell adhesion and invasion.These highly conserved proteins involve type specific antigen 56(TSA56),47 kDa,OmpA,and autotransporter proteins(ScaA,ScaB and ScaC).TSA56 is the most immunogenic and contains four types of hypervariable regions.Out of all autotransporter proteins,ScaA provides the homologous strains specific immunity and when coupled with TSA56 it shows better protective immunity against heterologous strains.The review provides detailed insight into the potential immunogenic proteins of Orientia which can be utilized to develop the vaccine.Furthermore,studies focused on highly antigenic proteins will provide more insight into their roles in developing therapeutics and easy-to-handle rapid diagnostic kits.展开更多
Objectives:Hepatocellular carcinoma(HCC)is among the most frequently occurring malignant tumors of the digestive tract and is associated with an increased mortality rate worldwide.This study aimed to develop and valid...Objectives:Hepatocellular carcinoma(HCC)is among the most frequently occurring malignant tumors of the digestive tract and is associated with an increased mortality rate worldwide.This study aimed to develop and validate a prognostic model based on immunogenic cell death(ICD)-related genes to predict patient survival and guide individualized treatment strategies for HCC.Methods:ICD-related genes were identified from the GeneCards database using a relevance score threshold of A combination of least absolute shrinkage and selection operator(LASSO)>10.regression and multivariate Cox analysis was used to screen prognostic genes and construct a risk score model.Immune cell infiltration was evaluated through single-sample gene set enrichment analysis(ssGSEA)and cell-type identification by estimating relative subsets of RNA transcripts(CIBERSORT)algorithms.Associations between risk groups and the tumor microenvironment(TME),N6-methyladenosine(m6A)regulators,and immune checkpoint expression were analyzed.Drug sensitivity was predicted based on the risk stratification.The reliability of the model was validated in internal cohorts and further confirmed by quantitative reverse transcription polymerase chain reaction(qRT-PCR)and immunohistochemistry(IHC).Results:A six-gene signature(CFHR3,G6PD,IGHM,KPNA2,PON1,and SERPINE1)was identified and used to calculate the risk scores.This study found that high-risk patients exhibited significantly poorer overall survival in both the training and validation datasets.The nomogram integrating the risk score and clinical factors showed strong predictive performance.High-risk patients demonstrated reduced immune cell infiltration,altered expression of immune checkpoints and immunosuppressive factors,and a distinct m6A modification pattern,suggesting a higher likelihood of immune escape.This study also revealed that the risk model effectively predicted sensitivity to multiple anticancer drugs.Conclusion:This study developed a robust ICD-related six-gene prognostic model for HCC that can accurately stratify patient risk,reflect the tumor immune landscape,and provide guidance for immunotherapy and personalized treatment strategies.展开更多
Antibody humanization is critical to reduce immunogenicity and enhance efficacy in the preclinical phase of the development of therapeutic antibodies originated from animal models.Computational suggestions have long b...Antibody humanization is critical to reduce immunogenicity and enhance efficacy in the preclinical phase of the development of therapeutic antibodies originated from animal models.Computational suggestions have long been desired,but available tools focused on immunogenicity calculation of whole antibody sequences and sequence segments,missing the individual residue sites.This study introduces Site-specific Immunogenicity for Therapeutic Antibody(SITA),a novel computational framework that predicts B-cell immunogenicity score for not only the overall antibody,but also individual residues,based on a comprehensive set of amino acid descriptors characterizing physicochemical and spatial features for antibody structures.A transfer-learning-inspired framework was purposely adopted to overcome the scarcity of Antibody-Antibody structural complexes.On an independent testing dataset derived from 13 Antibody-Antibody structural complexes,SITA successfully predicted the epitope sites for Antibody-Antibody structures with a receiver operating characteristic(ROC)-area unver the ROC curve(AUC)of 0.85 and a precision-recall(PR)-AUC of 0.305 at the residue level.Furthermore,the SITA score can significantly distinguish immunogenicity levels of whole human antibodies,therapeutic antibodies and non-human-derived antibodies.More importantly,analysis of an additional 25 thera-peutic antibodies revealed that over 70%of them were detected with decreased immunogenicity after modification compared to their parent variants.Among these,nearly 66%antibodies successfully iden-tified actual modification sites from the top five sites with the highest SITA scores,suggesting the ability of SITA scores for guide the humanization of antibody.Overall,these findings highlight the potential of SITA in optimizing immunogenicity assessments during the process of therapeutic antibody design.展开更多
African swine fever(ASF),caused by African swine fever virus(ASFV),is a highly contagious swine disease that has spread globally.Effective control strategies are not yet available.In this study,we prepared K205R mRNA,...African swine fever(ASF),caused by African swine fever virus(ASFV),is a highly contagious swine disease that has spread globally.Effective control strategies are not yet available.In this study,we prepared K205R mRNA,which was then formulated using Lipid Nanoparticle(LNP).The resulting K205R mRNA-LNP showed a particle size of approximately 86.27 nm and an mRNA encapsulation efficiency of 96.24%.Efficient expression of the K205R protein was confirmed in both HEK293T and PK15 cells.We further evaluated the immunogenicity of K205R mRNA-LNP in mice and pigs.All immunized animals developed significantly higher levels of IgG antibodies against K205R compared to the control group in the first week after the second immunization,with antibody titers reaching up to 105.Challenge experiments showed that K205R mRNA delayed the time of death.Our results suggested the successful implementation of the mRNA platform in the preparation and application of ASFV mRNA.展开更多
Objective:To evaluate the effectiveness of DTwP-HB-Hib vaccination dosing intervals at eight weeks versus four weeks on the immunogenicity of the diphtheria component.Methods:This is a randomized,open-label,parallel,c...Objective:To evaluate the effectiveness of DTwP-HB-Hib vaccination dosing intervals at eight weeks versus four weeks on the immunogenicity of the diphtheria component.Methods:This is a randomized,open-label,parallel,controlled trial on healthy two-month-old infants who had not received DTwP-HB-Hib vaccinations.The infants received three doses of the vaccine either at an eight-week or four-week interval.The anti-diphtheria toxoid IgG antibody levels before and after three doses of the vaccination were measured using ELISA.Results:Eighty infants were enrolled in this study,with 64 fulfilling the study requirements and randomized into two groups.All study participants exhibited uncertain protection against diphtheria antibodies(0.01-0.99 IU/mL)at the beginning of the study with an average of(0.023±0.009)IU/mL and(0.026±0.009)IU/mL in eight weeks and four weeks groups.Following three doses of the vaccination,the antibody level rose to an average of(1.320±0.234)IU/mL in the eight-week group and(1.307±0.186)IU/mL in the four-week group,with no statistically significant difference in the antibody levels observed(P=0.814).Conclusions:DTwP-HB-Hib vaccinations can be administered at an eight-week or four-week interval,as they do not significantly affect the level of anti-diphtheria IgG antibodies.展开更多
As PEGylated liposomes have witnessed remarkable advancements in drug delivery,their immunogenicity has emerged as a notable challenge.In this study,we discovered that a simple pre-injection of folic acid(FA)effective...As PEGylated liposomes have witnessed remarkable advancements in drug delivery,their immunogenicity has emerged as a notable challenge.In this study,we discovered that a simple pre-injection of folic acid(FA)effectively mitigated the immunogenicity of PEGylated liposomes and enhanced their in vivo performance by tolerating splenic marginal zone B cells.FA specifically inhibited the internalization of PEGylated liposomes by splenic marginal zone B cells,thereby reducing splenic lymphocyte proliferation and specific IgM secretion.This modulation alleviated Ig M-mediated accelerated blood clearance and adverse accumulation of the PEGylated liposomes in the skin.These findings provide new insights into the immunomodulatory effects of FA and promising avenues to enhance the efficacy and safety of PEGylated liposomal nanomedicines.展开更多
[ Objective ] The aim of the study was to construct associated DNA vaccine of PRRS （Porcine reproductive and respiratory syndrome） and PCV-2 （Porcine circovirus type 2） disease and study its immunogenicity. [ Meth...[ Objective ] The aim of the study was to construct associated DNA vaccine of PRRS （Porcine reproductive and respiratory syndrome） and PCV-2 （Porcine circovirus type 2） disease and study its immunogenicity. [ Method] In_ this study, the ORF5 gene of PRRSV isolated in Liaoning was cloned into plRES-neo expression vector, and the neo gene of plRES-neo expression vector was substituted by the ORF2 gene of the PCV-2 Mongolia strain to construct the recombinant expression vector. The expression in BHK cells was detected through Western blot and IFA. Then the ELISA antibody level and the number of spleen T lymphocytes were detected after Balb/c mice were immunized with this DNA vaccine. E Result] The recombinant plasmid plRES-ORF2-ORF5 was constructed successfully and could express the target proteins in BHK cells, as indicated by Western blot and IFA. There was no significant difference in ELISA antibody between plRES-ORF2-ORF5 immunized group and inactived vaccine immunized groups, while the number of spleen T lymphocytes induced by DNA vaccine was higher than that induced by inactived vaccine. [ Conclusion] The recombinant plasmid plRES-ORF2-ORF5 should induce good humoral immune response and cellular immune response in mice, providing the conditions for better prevention and control of PRRS and PCV-2 disease.展开更多
Immunotherapy has revolutionized cancer treatment and substantially improved patient outcomes with respect to multiple types of tumors.However,most patients cannot benefit from such therapies,mainly due to the intrins...Immunotherapy has revolutionized cancer treatment and substantially improved patient outcomes with respect to multiple types of tumors.However,most patients cannot benefit from such therapies,mainly due to the intrinsic low immunogenicity of cancer cells(CCs)that allows them to escape recognition by immune cells of the body.Immunogenic cell death(ICD),which is a form of regulated cell death,engages in a complex dialogue between dying CCs and immune cells in the tumor microenvironment(TME),ultimately evoking the damage-associated molecular pattern(DAMP)signals to activate tumor-specific immunity.The ICD inducers mediate the death of CCs and improve both antigenicity and adjuvanticity.At the same time,they reprogram TME with a“cold-warmhot”immune status,ultimately amplifying and sustaining dendritic cell-and T cell-dependent innate sensing as well as the antitumor immune responses.In this review,we discuss how to stimulate ICD based upon the biological properties of CCs that have evolved under diverse stress conditions.Additionally,we highlight how this dynamic interaction contributes to priming tumor immunogenicity,thereby boosting anticancer immune responses.We believe that a deep understanding of these ICD processes will provide a framework for evaluating its vital role in cancer immunotherapy.展开更多
Tumor vaccines are a promising avenue in cancer immunotherapy.Despite the progress in targeting specific immune epitopes,tumor cells lacking these epitopes can evade the treatment.Here,we aimed to construct an efficie...Tumor vaccines are a promising avenue in cancer immunotherapy.Despite the progress in targeting specific immune epitopes,tumor cells lacking these epitopes can evade the treatment.Here,we aimed to construct an efficient in situ tumor vaccine called Vac-SM,utilizing shikonin(SKN)to induce immunogenic cell death(ICD)and Mycobacterium smegmatis as an immune adjuvant to enhance in situ tumor vaccine efficacy.SKN showed a dose-dependent and time-dependent cytotoxic effect on the tumor cell line and induced ICD in tumor cells as evidenced by the CCK-8 assay and the detection of the expression of relevant indicators,respectively.Compared with the control group,the in situ Vac-SM injection in mouse subcutaneous metastatic tumors significantly inhibited tumor growth and distant tumor metastasis,while also improving survival rates.Mycobacterium smegmatis effectively induced maturation and activation of bone marrow-derived dendritic cells(DCs),and in vivo tumor-draining lymph nodes showed an increased maturation of DCs and a higher proportion of effector memory T-cell subsets with the Vac-SM treatment,based on flow cytometry analysis results.Collectively,the Vac-SM vaccine effectively induces ICD,improves antigen presentation by DCs,activates a specific systemic antitumor T-cell immune response,exhibits a favorable safety profile,and holds the promise for clinical translation for local tumor immunotherapy.展开更多
AIM: The incorporation of hepatitis B virus (HBV) preS1 region into epitope-based vaccines against HBV has been accepted widely, but the incorporate site and size of preS1 sequence is controversial. Therefore our purp...AIM: The incorporation of hepatitis B virus (HBV) preS1 region into epitope-based vaccines against HBV has been accepted widely, but the incorporate site and size of preS1 sequence is controversial. Therefore our purpose was to further investigate its immunogenic domains for the epitopebased hepatitis B vaccine design.METHODS: Eight GST fusion proteins containing overlapping preS1 fragments in preS1 (21-119) region were expressed in E.coli. Using these purified fusion proteins, the immunogenic domains in preS1 region were identified in detail in mice and humans by Western blot analysis and ELISA.RESULTS: The results in mice showed that the immunogenic domains mainly existed in preS1 (21-59) and preS1 (95-109). Similarly, these fragments had strong immunogenicity in humans; whereas the other parts except for preS1 (60-70) also had some immunogenicity.More importantly, a major immunogenic domain, preS1 (34-59), which has much stronger immunogenicity, was identified. Additionally, the antibodies against some preS1 fragments, especially preS1 (34-59), were speculated to be virus-neutralizing.CONCLUSION: Eight GST fusion proteins containing overlapping preS1 fragments were prepared successfully. They were used for the study on the immunogenic domains in preS1 (21-119) region. The preS1 (34-59) fragments were the major immunogenic domains in the preS1 region, and the antibodies against these fragments were speculated to be virus-neutralizing. Therefore, the incorporation of preS1 (34-59) fragments into epitopebased HBV vaccines may be efficient for enhancement of immune response. Additionally, the results also imply that there are more complex immune responses to preS1 region and more abundant immunogenic domains in humans.展开更多
Stem cell therapy is a promising strategy for the treatment of traumatic brain injury(TBI). However, animal experiments are needed to evaluate safety;in particular, to examine the immunogenicity and tumorigenicity of ...Stem cell therapy is a promising strategy for the treatment of traumatic brain injury(TBI). However, animal experiments are needed to evaluate safety;in particular, to examine the immunogenicity and tumorigenicity of human umbilical cord mesenchymal stem cells(hu MSCs) before clinical application. In this study, hu MSCs were harvested from human amniotic membrane and umbilical cord vascular tissue. A rat model of TBI was established using the controlled cortical impact method. Starting from the third day after injury, the rats were injected with 10 μL of 5 × 10^(6)/m L hu MSCs by cerebral stereotaxis or with 500 μL of 1 × 10^(6)/m L hu MSCs via the tail vein for 3 successive days. hu MSC transplantation decreased the serum levels of proinflammatory cytokines in rats with TBI and increased the serum levels of anti-inflammatory cytokines, thereby exhibiting good immunoregulatory function. The transplanted hu MSCs were distributed in the liver, lung and brain injury sites. No abnormal proliferation or tumorigenesis was found in these organs up to 12 months after transplantation. The transplanted hu MSCs negligibly proliferated in vivo, and apoptosis was gradually observed at later stages. These findings suggest that hu MSC transplantation for the treatment of traumatic brain injury displays good safety. In addition, hu MSCs exhibit good immunoregulatory function, which can help prevent and reduce secondary brain injury caused by the rapid release of inflammatory factors after TBI. This study was approved by the Ethics Committee of Wuhan General Hospital of PLA(approval No. 20160054) on November 1, 2016.展开更多
AIM： To construct a prokaryotic expression vector carrying Campylobacterjejuni peblA gene and express it in Escherichia coli. Immunoreactivity and antigenicity of rPEB1 were evaluated. The ability of rPEB1 to induce ...AIM： To construct a prokaryotic expression vector carrying Campylobacterjejuni peblA gene and express it in Escherichia coli. Immunoreactivity and antigenicity of rPEB1 were evaluated. The ability of rPEB1 to induce antibody responses and protective efficacy was identified. METHODS： peblA gene was amplified by PCR, target gene and prokaryotic expression ptasmid pET28a （＋） was digested with BamHI and XhoI, respectively. DNA was ligated with T4 DNA ligase to construct recombinant plasmid pET28a（＋）-peblA. The rPEB1 was expressed in E. coli BL21 （DE3） and identified by SDS-PAGE. BALB/c mice were immunized with rPEBI. ELISA was used to detect the specific antibody titer and MTT method was used to measure the stimulation index of spleen lymphocyte transformation. RESULTS： The recombinant plasmid pET28a （＋）-peblA was correctly constructed. The expression output of PEB1 protein in pET28a （＋）-peblA system was approximately 33% of total proteins in E. coil The specific IgG antibody was detected in serum of BALB/c mice immunized with rPEB1 protein. Effective immunological protection with a lower sickness incidence and mortality was seen in the mice suffering from massive C. jejuni infection. CONCLUSION： rPEB1 protein is a valuable candidate for C. jejuni subunit vaccine.展开更多
Tumor immunotherapy,especially immune checkpoint blockade(ICB),has revolutionized the cancer field.However,the limited response of tumors to immunotherapy is a major obstacle.Tumor immunogenic cell death(ICD)is a deat...Tumor immunotherapy,especially immune checkpoint blockade(ICB),has revolutionized the cancer field.However,the limited response of tumors to immunotherapy is a major obstacle.Tumor immunogenic cell death(ICD)is a death mode of tumor cells that can promote tumor immunity.ICD can induce strong an-titumor immune responses through the ectopic exposure of calreticulin on the plasma membrane surface and the release of the non-histone nuclear protein high-mobility group box 1(HMGB1),ATP,and in-terferon(IFN),thus activating an adaptive immune response against dead cell-associated antigens and enhancing the therapeutic effect of tumor immunotherapy.Chemotherapy,radiotherapy,photothermal therapy,magneto-thermodynamics therapy,nanopulse stimulation,and oncolytic virus therapy can all induce a strong antitumor immune response by ICD.In addition,the application of nanotechnology can precisely target drug delivery and improve the efficacy of immunotherapy.Here we introduce the basic concepts and molecular mechanisms underlying the induction of ICD.Then,we summarize and discuss the progress in the application of nanotechnology in immunotherapy to promote ICD.Finally,we attempt to define the challenges and future directions in this area to extend the benefits of ICD to a broader patient population.展开更多
To purify the protein encoding the small capsid protein （SCP） of KSHV and analyze its immunogenicity, the carboxyl terminus of orf65 of Kaposi＇s sarcoma associated-herpesvirus （KSHV） was expressed in a prokaryoti...To purify the protein encoding the small capsid protein （SCP） of KSHV and analyze its immunogenicity, the carboxyl terminus of orf65 of Kaposi＇s sarcoma associated-herpesvirus （KSHV） was expressed in a prokaryotic expression system. The expression of recombinant E.coli containing pQE-80L-orf65 was induced by isopropyl-13-D-thiogalactopyranoside （IPTG） and the fusion protein was purified by chromatography. The expressed protein and its purified product were identified by sodium dodecyl sulfate- polyacrylamide gel electrophoresis （SDS-PAGE） and showed that 9 kDa was the expected size of the purified orf65 protein. The antiserum was produced in rabbit which was immunized by purified orf65 protein. An ELISA assay was established to analyze the immunogenicity of the purified orf65 protein. The ELISA analysis demonstrated that orf65 protein has strong immune activity, and the immune activity of polyclonal antibody against orf65 was more than 4 fold higher than that in the serum of the non-immunized rabbit. These results demonstrate that purified orf65 protein has very strong immunogenicity and can be used in screening KSHV infection in the general population using ELISA.展开更多
Enterovirus （EV71） can cause severe neurological diseases, but the underlying pathogenesis remains unclear. The capsid protein, viral protein 1 （VP1）, plays a critical role in the pathogenicity of EVT1. High level...Enterovirus （EV71） can cause severe neurological diseases, but the underlying pathogenesis remains unclear. The capsid protein, viral protein 1 （VP1）, plays a critical role in the pathogenicity of EVT1. High level expression and secretion ofVP 1 protein are necessary for structure, function and immunogenicity in its natural conformation. In our previous studies, 5 codon-optimized VP 1 DNA vaccines, including wt-VP 1, tPA-VP 1, VP l-d, VP 1-hFc and VP 1 - mFc, were constructed and analyzed. They expressed VP1 protein, but the levels of secretion and immunogenicity of these VP1 constructs were significantly different （P〈0.05）. In this study, we further investigated the protein lev- els of these constructs and determined that all of these constructs expressed VP1 protein. The secretion level was increased by including a tPA leader sequence, which was further increased by fusing human IgG Fc （hFc） to VP1. VP 1-hFc demonstrated the most potent immunogenicity in mice. Furthermore, hFc domain could be used to purify VPI-hFc protein for additional studies.展开更多
Photodynamic therapy(PDT)not only destroys tumor cells directly but also induced anti-tumor immune response through damage-associated molecular patterns(DAMPs).It is reported that anti-tumor response was associated wi...Photodynamic therapy(PDT)not only destroys tumor cells directly but also induced anti-tumor immune response through damage-associated molecular patterns(DAMPs).It is reported that anti-tumor response was associated with light dose and photosensitizer used in PDT.In this study,4T1 tumor cells were implanted on both the right and left flanks of mice.Only the right tumor was treated by HpD-PDT,while the left tumor was not irradiated.The anti-tumor immune response induced by HpD-PDT was investigated.The expression of DAMPs and costimulatory molecules induced by HpD-PDT were tested by immuno fluorescence and flow cytometry in vivo.Different light doses of PDT were designed to treat 4T1 cells.The killing effect was assessed by CCK-8 kit and apoptosis kit.The expression of DAMPs on 4T1 cells after HpDPDT were evaluated by flow cytometry,western blot and ATP kit.This study showed that CD4^(+)T,CD8^(+)T and the production of IFN-γwere increased significantly on day 10 in righttumor after PDT treatment compared with control group.HpD-PDT enhanced the expression of calreticulin(CRT)on tumor tissue.Importantly,co-stimulatory molecular OX-40 and 4-1BB were elevated on CD8^(+)T cells.In vitro,immunogenic death of 4T1 cells was induced after PDT.Besides,the expression of DAMPs increased with the increasing of energy density.This study indicates that anti-tumor immune effect was induced by HpD-PDT.The knowledge of the involvement of CRT,ATP and co-stimulatory molecules uncovers important mechanistic insight into the anti-tumor immunogenicity.It was the first time that co-stimulatory molecules were investigated and found to elevate after PDT.展开更多
Grass carp reovirus (GCRV) is a tentative member of the Aquareovirus genus in the family Reoviridae. The mature virion comprises 11 dsRNA genomes enclosed by two concentric icosahedral proteins shells that is comprise...Grass carp reovirus (GCRV) is a tentative member of the Aquareovirus genus in the family Reoviridae. The mature virion comprises 11 dsRNA genomes enclosed by two concentric icosahedral proteins shells that is comprised of five core proteins and two outer capsid proteins. The genome sequence and 3D structure demonstrate there is a higher level of sequence homology in structural proteins between GCRV and mammalian orthoreoviruses (MRV) compared to other members of the family. To understand the pathogenesis of GCRV infection, the outer capsid protein VP5, a homology of the μ1 protein of MRV, was expressed in E.coli. It was found that the recombinant VP5 was highly expressed, and the expressed His-tag fusion protein was involved in the formation of the inclusion body. Additionally, specific anti-VP5 serum was prepared from purified protein and western blot demonstrated that the expressed protein was able to bind immunologically to rabbit anti GCRV particle serum and the immunogenicity was determined by ELISA assay. Additional experiments in investigating the functional properties of VP5 will further elucidate the role of the GCRV outer capsid protein VP5 during entry into host cells, and its interaction among viral proteins and host cells during the infection process.展开更多
基金supported by the Natural Science Foundation of Guangxi(No.2022GXNSFGA035003)the National Natural Science Foundation of China(No.22077021).
文摘To treat cancer and inhibit its metastasis to the greatest extent,we proposed to develop an Au(III)agent to induce immunogenic cell death(ICD)and establish long-term immunity.To this end,we optimized a series of Au(III)2-benzoylpyridine thiosemicarbazone complexes to obtain an Au(III)agent(5b)with excellent cytotoxicity to cancer.The results show that 5b effectively inhibits tumor growth and its metastasis in vivo.Interestingly,we revealed a new mechanism of 5b inhibiting tumor growth and metastasis:5b releases ICD-related damage-associated molecular patterns(DAMPs),such as calreticulin(CRT),ATP and high mobility group box 1(HMGB1)by inducing endoplasmic reticulum stress(ERS)and mitochondrial dysfunction,which then stimulated an antitumor CD8^(+)T cell response and Foxp^(3+)T cell depletion,thus establishing long-action antitumor immunity.
基金supported by the National Natural Science Foundation of China(Grant Nos.:82073975,81673563,81102762,82204715,and 82304743).
文摘Chemotherapeutic drugs such as doxorubicin(DOx)and oxaliplatin can induce immunogenic cell death(ICD)in tumor cells.Current studies have demonstrated that peptide-based drugs can also induce ICD in tumor cells.Unlike small molecule drugs,peptide drugs not only enhance the effectiveness of immunotherapy but also offer the advantage of low biotoxicity[1].
基金supported by the National Key R&D Program of China(No.2022YFC2304205)National Natural Science Foundation of China(Nos.51903062,52273128)the Plan on Enhancing Scientific Research in GMU(No.02-410-2405033).
文摘Ferroptosis in combination with immune therapy emerges as a promising approach for cancer therapy.Herein,dual-responsive metal-polyphenol coordinated nanomedicines were developed for pH/glutathione(GSH)-responsive synergistic ferroptosis and immunotherapy.Our innovative strategy involves the development of a manganese-polyphenol coordinated nanostructure,leveraging the biocompatibility of bovine serum albumin(BSA)as a template to encapsulate the anticancer drug sorafenib.The tumor microenvironment(pH/GSH)prompts the disassembly of MnO_(2)and epigallocatechin gallate(EGCG),thereby releases the anticancer payload.Concurrently,MnO_(2)acts to deplete intracellular GSH,which in turn suppresses glutathione peroxidase activity,leading to an accumulation of lipid peroxides with cell ferroptosis.Additionally,the release of Mn^(2+)ions bolster the cyclic guanosine monophosphlic acid(GMP)-adenosine monophosphlic acid(AMP)synthase-stimulator of interferon gene(cGAS-STING)pathway,which,in conjunction with the immunogenic cell death(ICD)effect induced by tumor cell apoptosis,significantly promotes dendritic cell(DC)maturation and enhances the presentation of tumor antigens.This successively ignites a robust innate and adaptive immune response.Both in vitro and in vivo experiments have demonstrated that the concurrent administration of ferroptosis-inducing and immune-stimulating therapies can significantly inhibit tumor growth.
基金funded by Department of Health Research,Government of India,New Delhi,India(grant number:YSS/2020/000116/PRCYSS).
文摘Scrub typhus is an acute undifferentiated febrile infectious disease transmitted by a chigger(genus Leptotrombidium)bite carrying Orientia(O.)tsutsugamushi,affecting millions of people annually while more than one billion people are susceptible.Endemic areas are expanding to Africa,Europe,Middle East,and South America which is concerning,as despite best efforts,there is no vaccine to combat the bacteria.There are now three species of Orientia and over 20 strains of O.tsutsugamushi.The past attempts to develop a vaccine have been ineffective as they confer homologous strain-specific immunity.Various immunogenic proteins of O.tsutsugamushi have been identified that interact with the extracellular matrix(fibronectin)or vMLL5 receptor and modify the cytoskeleton of non-phagocytic host cells,which aids in host cell adhesion and invasion.These highly conserved proteins involve type specific antigen 56(TSA56),47 kDa,OmpA,and autotransporter proteins(ScaA,ScaB and ScaC).TSA56 is the most immunogenic and contains four types of hypervariable regions.Out of all autotransporter proteins,ScaA provides the homologous strains specific immunity and when coupled with TSA56 it shows better protective immunity against heterologous strains.The review provides detailed insight into the potential immunogenic proteins of Orientia which can be utilized to develop the vaccine.Furthermore,studies focused on highly antigenic proteins will provide more insight into their roles in developing therapeutics and easy-to-handle rapid diagnostic kits.
基金supported by 1.3.5 project for disciplines of excellence,West China Hospital,Sichuan University(ZYJC21046)1.3.5 project for disciplines of excellence-Clinical Research Incubation Project,West China Hospital,Sichuan University(2021HXFH001)+4 种基金Natural Science Foundation of Sichuan Province(2022NSFSC0806)National Natural Science Foundation of China for Young Scientists Fund(82203782)Sichuan University-Zigong School-Local Cooperation Project(2021CDZG-23)Sichuan University-Sui Lin School-Local Cooperation Project(2022CDSN-18)China Telecom Sichuan Company Biliary tract Tumor Big Data Platform and Application Phase I R&D Project(312230752).
文摘Objectives:Hepatocellular carcinoma(HCC)is among the most frequently occurring malignant tumors of the digestive tract and is associated with an increased mortality rate worldwide.This study aimed to develop and validate a prognostic model based on immunogenic cell death(ICD)-related genes to predict patient survival and guide individualized treatment strategies for HCC.Methods:ICD-related genes were identified from the GeneCards database using a relevance score threshold of A combination of least absolute shrinkage and selection operator(LASSO)>10.regression and multivariate Cox analysis was used to screen prognostic genes and construct a risk score model.Immune cell infiltration was evaluated through single-sample gene set enrichment analysis(ssGSEA)and cell-type identification by estimating relative subsets of RNA transcripts(CIBERSORT)algorithms.Associations between risk groups and the tumor microenvironment(TME),N6-methyladenosine(m6A)regulators,and immune checkpoint expression were analyzed.Drug sensitivity was predicted based on the risk stratification.The reliability of the model was validated in internal cohorts and further confirmed by quantitative reverse transcription polymerase chain reaction(qRT-PCR)and immunohistochemistry(IHC).Results:A six-gene signature(CFHR3,G6PD,IGHM,KPNA2,PON1,and SERPINE1)was identified and used to calculate the risk scores.This study found that high-risk patients exhibited significantly poorer overall survival in both the training and validation datasets.The nomogram integrating the risk score and clinical factors showed strong predictive performance.High-risk patients demonstrated reduced immune cell infiltration,altered expression of immune checkpoints and immunosuppressive factors,and a distinct m6A modification pattern,suggesting a higher likelihood of immune escape.This study also revealed that the risk model effectively predicted sensitivity to multiple anticancer drugs.Conclusion:This study developed a robust ICD-related six-gene prognostic model for HCC that can accurately stratify patient risk,reflect the tumor immune landscape,and provide guidance for immunotherapy and personalized treatment strategies.
基金supported by funding from the National Key R&D Program of China(Grant Nos.:2023YFC3404000 and 2019YFA0905900)the National Natural Science Foundation of China(Grant Nos.:32370697 and 32070657)AI for the Science project of Fudan University,China(Project No.:XM06231724)。
文摘Antibody humanization is critical to reduce immunogenicity and enhance efficacy in the preclinical phase of the development of therapeutic antibodies originated from animal models.Computational suggestions have long been desired,but available tools focused on immunogenicity calculation of whole antibody sequences and sequence segments,missing the individual residue sites.This study introduces Site-specific Immunogenicity for Therapeutic Antibody(SITA),a novel computational framework that predicts B-cell immunogenicity score for not only the overall antibody,but also individual residues,based on a comprehensive set of amino acid descriptors characterizing physicochemical and spatial features for antibody structures.A transfer-learning-inspired framework was purposely adopted to overcome the scarcity of Antibody-Antibody structural complexes.On an independent testing dataset derived from 13 Antibody-Antibody structural complexes,SITA successfully predicted the epitope sites for Antibody-Antibody structures with a receiver operating characteristic(ROC)-area unver the ROC curve(AUC)of 0.85 and a precision-recall(PR)-AUC of 0.305 at the residue level.Furthermore,the SITA score can significantly distinguish immunogenicity levels of whole human antibodies,therapeutic antibodies and non-human-derived antibodies.More importantly,analysis of an additional 25 thera-peutic antibodies revealed that over 70%of them were detected with decreased immunogenicity after modification compared to their parent variants.Among these,nearly 66%antibodies successfully iden-tified actual modification sites from the top five sites with the highest SITA scores,suggesting the ability of SITA scores for guide the humanization of antibody.Overall,these findings highlight the potential of SITA in optimizing immunogenicity assessments during the process of therapeutic antibody design.
基金funded by the National Key Research and Development Program of China(2022YFD1800500,2021YFD1801401,2023YFD1802600)the Central Public-interest Scientific Institution Basal Research Fund,China(Y2022PT11)the Shanghai Sailing Program,China(23YF1457400).
文摘African swine fever(ASF),caused by African swine fever virus(ASFV),is a highly contagious swine disease that has spread globally.Effective control strategies are not yet available.In this study,we prepared K205R mRNA,which was then formulated using Lipid Nanoparticle(LNP).The resulting K205R mRNA-LNP showed a particle size of approximately 86.27 nm and an mRNA encapsulation efficiency of 96.24%.Efficient expression of the K205R protein was confirmed in both HEK293T and PK15 cells.We further evaluated the immunogenicity of K205R mRNA-LNP in mice and pigs.All immunized animals developed significantly higher levels of IgG antibodies against K205R compared to the control group in the first week after the second immunization,with antibody titers reaching up to 105.Challenge experiments showed that K205R mRNA delayed the time of death.Our results suggested the successful implementation of the mRNA platform in the preparation and application of ASFV mRNA.
文摘Objective:To evaluate the effectiveness of DTwP-HB-Hib vaccination dosing intervals at eight weeks versus four weeks on the immunogenicity of the diphtheria component.Methods:This is a randomized,open-label,parallel,controlled trial on healthy two-month-old infants who had not received DTwP-HB-Hib vaccinations.The infants received three doses of the vaccine either at an eight-week or four-week interval.The anti-diphtheria toxoid IgG antibody levels before and after three doses of the vaccination were measured using ELISA.Results:Eighty infants were enrolled in this study,with 64 fulfilling the study requirements and randomized into two groups.All study participants exhibited uncertain protection against diphtheria antibodies(0.01-0.99 IU/mL)at the beginning of the study with an average of(0.023±0.009)IU/mL and(0.026±0.009)IU/mL in eight weeks and four weeks groups.Following three doses of the vaccination,the antibody level rose to an average of(1.320±0.234)IU/mL in the eight-week group and(1.307±0.186)IU/mL in the four-week group,with no statistically significant difference in the antibody levels observed(P=0.814).Conclusions:DTwP-HB-Hib vaccinations can be administered at an eight-week or four-week interval,as they do not significantly affect the level of anti-diphtheria IgG antibodies.
基金supported by the National Natural Science Foundation of China(Nos.82373817 and 82003659)Shanghai Natural Science Foundation(No.23ZR1477500)Pudong Health Bureau of Shanghai(No.YC-2023-0401)。
文摘As PEGylated liposomes have witnessed remarkable advancements in drug delivery,their immunogenicity has emerged as a notable challenge.In this study,we discovered that a simple pre-injection of folic acid(FA)effectively mitigated the immunogenicity of PEGylated liposomes and enhanced their in vivo performance by tolerating splenic marginal zone B cells.FA specifically inhibited the internalization of PEGylated liposomes by splenic marginal zone B cells,thereby reducing splenic lymphocyte proliferation and specific IgM secretion.This modulation alleviated Ig M-mediated accelerated blood clearance and adverse accumulation of the PEGylated liposomes in the skin.These findings provide new insights into the immunomodulatory effects of FA and promising avenues to enhance the efficacy and safety of PEGylated liposomal nanomedicines.
文摘[ Objective ] The aim of the study was to construct associated DNA vaccine of PRRS （Porcine reproductive and respiratory syndrome） and PCV-2 （Porcine circovirus type 2） disease and study its immunogenicity. [ Method] In_ this study, the ORF5 gene of PRRSV isolated in Liaoning was cloned into plRES-neo expression vector, and the neo gene of plRES-neo expression vector was substituted by the ORF2 gene of the PCV-2 Mongolia strain to construct the recombinant expression vector. The expression in BHK cells was detected through Western blot and IFA. Then the ELISA antibody level and the number of spleen T lymphocytes were detected after Balb/c mice were immunized with this DNA vaccine. E Result] The recombinant plasmid plRES-ORF2-ORF5 was constructed successfully and could express the target proteins in BHK cells, as indicated by Western blot and IFA. There was no significant difference in ELISA antibody between plRES-ORF2-ORF5 immunized group and inactived vaccine immunized groups, while the number of spleen T lymphocytes induced by DNA vaccine was higher than that induced by inactived vaccine. [ Conclusion] The recombinant plasmid plRES-ORF2-ORF5 should induce good humoral immune response and cellular immune response in mice, providing the conditions for better prevention and control of PRRS and PCV-2 disease.
基金supported by the National Natural Science Foundation of China (No. 31971378, 81830002, 31870873 and 31991171)
文摘Immunotherapy has revolutionized cancer treatment and substantially improved patient outcomes with respect to multiple types of tumors.However,most patients cannot benefit from such therapies,mainly due to the intrinsic low immunogenicity of cancer cells(CCs)that allows them to escape recognition by immune cells of the body.Immunogenic cell death(ICD),which is a form of regulated cell death,engages in a complex dialogue between dying CCs and immune cells in the tumor microenvironment(TME),ultimately evoking the damage-associated molecular pattern(DAMP)signals to activate tumor-specific immunity.The ICD inducers mediate the death of CCs and improve both antigenicity and adjuvanticity.At the same time,they reprogram TME with a“cold-warmhot”immune status,ultimately amplifying and sustaining dendritic cell-and T cell-dependent innate sensing as well as the antitumor immune responses.In this review,we discuss how to stimulate ICD based upon the biological properties of CCs that have evolved under diverse stress conditions.Additionally,we highlight how this dynamic interaction contributes to priming tumor immunogenicity,thereby boosting anticancer immune responses.We believe that a deep understanding of these ICD processes will provide a framework for evaluating its vital role in cancer immunotherapy.
基金supported by grants from the Natural Science Foundation of Huai'an Science and Technology Bureau(Grant No.HAB202312)the Science and Technology Development Fund of the Affiliated Hospital of Xuzhou Medical University(Grant No.XYFY2021018).
文摘Tumor vaccines are a promising avenue in cancer immunotherapy.Despite the progress in targeting specific immune epitopes,tumor cells lacking these epitopes can evade the treatment.Here,we aimed to construct an efficient in situ tumor vaccine called Vac-SM,utilizing shikonin(SKN)to induce immunogenic cell death(ICD)and Mycobacterium smegmatis as an immune adjuvant to enhance in situ tumor vaccine efficacy.SKN showed a dose-dependent and time-dependent cytotoxic effect on the tumor cell line and induced ICD in tumor cells as evidenced by the CCK-8 assay and the detection of the expression of relevant indicators,respectively.Compared with the control group,the in situ Vac-SM injection in mouse subcutaneous metastatic tumors significantly inhibited tumor growth and distant tumor metastasis,while also improving survival rates.Mycobacterium smegmatis effectively induced maturation and activation of bone marrow-derived dendritic cells(DCs),and in vivo tumor-draining lymph nodes showed an increased maturation of DCs and a higher proportion of effector memory T-cell subsets with the Vac-SM treatment,based on flow cytometry analysis results.Collectively,the Vac-SM vaccine effectively induces ICD,improves antigen presentation by DCs,activates a specific systemic antitumor T-cell immune response,exhibits a favorable safety profile,and holds the promise for clinical translation for local tumor immunotherapy.
文摘AIM: The incorporation of hepatitis B virus (HBV) preS1 region into epitope-based vaccines against HBV has been accepted widely, but the incorporate site and size of preS1 sequence is controversial. Therefore our purpose was to further investigate its immunogenic domains for the epitopebased hepatitis B vaccine design.METHODS: Eight GST fusion proteins containing overlapping preS1 fragments in preS1 (21-119) region were expressed in E.coli. Using these purified fusion proteins, the immunogenic domains in preS1 region were identified in detail in mice and humans by Western blot analysis and ELISA.RESULTS: The results in mice showed that the immunogenic domains mainly existed in preS1 (21-59) and preS1 (95-109). Similarly, these fragments had strong immunogenicity in humans; whereas the other parts except for preS1 (60-70) also had some immunogenicity.More importantly, a major immunogenic domain, preS1 (34-59), which has much stronger immunogenicity, was identified. Additionally, the antibodies against some preS1 fragments, especially preS1 (34-59), were speculated to be virus-neutralizing.CONCLUSION: Eight GST fusion proteins containing overlapping preS1 fragments were prepared successfully. They were used for the study on the immunogenic domains in preS1 (21-119) region. The preS1 (34-59) fragments were the major immunogenic domains in the preS1 region, and the antibodies against these fragments were speculated to be virus-neutralizing. Therefore, the incorporation of preS1 (34-59) fragments into epitopebased HBV vaccines may be efficient for enhancement of immune response. Additionally, the results also imply that there are more complex immune responses to preS1 region and more abundant immunogenic domains in humans.
基金supported by the General Project of Hubei Health Committee of China,No.WJ2019M263(to GW)。
文摘Stem cell therapy is a promising strategy for the treatment of traumatic brain injury(TBI). However, animal experiments are needed to evaluate safety;in particular, to examine the immunogenicity and tumorigenicity of human umbilical cord mesenchymal stem cells(hu MSCs) before clinical application. In this study, hu MSCs were harvested from human amniotic membrane and umbilical cord vascular tissue. A rat model of TBI was established using the controlled cortical impact method. Starting from the third day after injury, the rats were injected with 10 μL of 5 × 10^(6)/m L hu MSCs by cerebral stereotaxis or with 500 μL of 1 × 10^(6)/m L hu MSCs via the tail vein for 3 successive days. hu MSC transplantation decreased the serum levels of proinflammatory cytokines in rats with TBI and increased the serum levels of anti-inflammatory cytokines, thereby exhibiting good immunoregulatory function. The transplanted hu MSCs were distributed in the liver, lung and brain injury sites. No abnormal proliferation or tumorigenesis was found in these organs up to 12 months after transplantation. The transplanted hu MSCs negligibly proliferated in vivo, and apoptosis was gradually observed at later stages. These findings suggest that hu MSC transplantation for the treatment of traumatic brain injury displays good safety. In addition, hu MSCs exhibit good immunoregulatory function, which can help prevent and reduce secondary brain injury caused by the rapid release of inflammatory factors after TBI. This study was approved by the Ethics Committee of Wuhan General Hospital of PLA(approval No. 20160054) on November 1, 2016.
基金Grants from the Governor Foundation for Excellent Talents of Guizhou Province,No.200607
文摘AIM： To construct a prokaryotic expression vector carrying Campylobacterjejuni peblA gene and express it in Escherichia coli. Immunoreactivity and antigenicity of rPEB1 were evaluated. The ability of rPEB1 to induce antibody responses and protective efficacy was identified. METHODS： peblA gene was amplified by PCR, target gene and prokaryotic expression ptasmid pET28a （＋） was digested with BamHI and XhoI, respectively. DNA was ligated with T4 DNA ligase to construct recombinant plasmid pET28a（＋）-peblA. The rPEB1 was expressed in E. coli BL21 （DE3） and identified by SDS-PAGE. BALB/c mice were immunized with rPEBI. ELISA was used to detect the specific antibody titer and MTT method was used to measure the stimulation index of spleen lymphocyte transformation. RESULTS： The recombinant plasmid pET28a （＋）-peblA was correctly constructed. The expression output of PEB1 protein in pET28a （＋）-peblA system was approximately 33% of total proteins in E. coil The specific IgG antibody was detected in serum of BALB/c mice immunized with rPEB1 protein. Effective immunological protection with a lower sickness incidence and mortality was seen in the mice suffering from massive C. jejuni infection. CONCLUSION： rPEB1 protein is a valuable candidate for C. jejuni subunit vaccine.
基金financially supported by National Natural Science Foundation of China[Nos.82072996(Z.Sun.),81874131(Z.Sun),51703187(Z.Xu)]National Key Research and Development Program(No.2017YFSF090107)+2 种基金the Chongqing Talent Plan for Young Top Notch Talents(No.CQYC202005029)Hubei Province Natural Science Funds for Distinguished Young Scholar[No.2017CFA062(Z.Sun)]Innovative Research Team of High-level Local Universities in Shanghai[No.ZLCX20180500(Z.Sun)].
文摘Tumor immunotherapy,especially immune checkpoint blockade(ICB),has revolutionized the cancer field.However,the limited response of tumors to immunotherapy is a major obstacle.Tumor immunogenic cell death(ICD)is a death mode of tumor cells that can promote tumor immunity.ICD can induce strong an-titumor immune responses through the ectopic exposure of calreticulin on the plasma membrane surface and the release of the non-histone nuclear protein high-mobility group box 1(HMGB1),ATP,and in-terferon(IFN),thus activating an adaptive immune response against dead cell-associated antigens and enhancing the therapeutic effect of tumor immunotherapy.Chemotherapy,radiotherapy,photothermal therapy,magneto-thermodynamics therapy,nanopulse stimulation,and oncolytic virus therapy can all induce a strong antitumor immune response by ICD.In addition,the application of nanotechnology can precisely target drug delivery and improve the efficacy of immunotherapy.Here we introduce the basic concepts and molecular mechanisms underlying the induction of ICD.Then,we summarize and discuss the progress in the application of nanotechnology in immunotherapy to promote ICD.Finally,we attempt to define the challenges and future directions in this area to extend the benefits of ICD to a broader patient population.
基金Knowledge Innovation Program of the Chinese Academy of Sciences(0702121Y-J1)
文摘To purify the protein encoding the small capsid protein （SCP） of KSHV and analyze its immunogenicity, the carboxyl terminus of orf65 of Kaposi＇s sarcoma associated-herpesvirus （KSHV） was expressed in a prokaryotic expression system. The expression of recombinant E.coli containing pQE-80L-orf65 was induced by isopropyl-13-D-thiogalactopyranoside （IPTG） and the fusion protein was purified by chromatography. The expressed protein and its purified product were identified by sodium dodecyl sulfate- polyacrylamide gel electrophoresis （SDS-PAGE） and showed that 9 kDa was the expected size of the purified orf65 protein. The antiserum was produced in rabbit which was immunized by purified orf65 protein. An ELISA assay was established to analyze the immunogenicity of the purified orf65 protein. The ELISA analysis demonstrated that orf65 protein has strong immune activity, and the immune activity of polyclonal antibody against orf65 was more than 4 fold higher than that in the serum of the non-immunized rabbit. These results demonstrate that purified orf65 protein has very strong immunogenicity and can be used in screening KSHV infection in the general population using ELISA.
基金supported by the National Natural Science Foundation of China(Grant No.81000725 and 31470889)the Priority Academic Program of Basic Medical Science of Nanjing Medical University(Grant No.JX10131801060)
文摘Enterovirus （EV71） can cause severe neurological diseases, but the underlying pathogenesis remains unclear. The capsid protein, viral protein 1 （VP1）, plays a critical role in the pathogenicity of EVT1. High level expression and secretion ofVP 1 protein are necessary for structure, function and immunogenicity in its natural conformation. In our previous studies, 5 codon-optimized VP 1 DNA vaccines, including wt-VP 1, tPA-VP 1, VP l-d, VP 1-hFc and VP 1 - mFc, were constructed and analyzed. They expressed VP1 protein, but the levels of secretion and immunogenicity of these VP1 constructs were significantly different （P〈0.05）. In this study, we further investigated the protein lev- els of these constructs and determined that all of these constructs expressed VP1 protein. The secretion level was increased by including a tPA leader sequence, which was further increased by fusing human IgG Fc （hFc） to VP1. VP 1-hFc demonstrated the most potent immunogenicity in mice. Furthermore, hFc domain could be used to purify VPI-hFc protein for additional studies.
基金This work was supported by the National Key Research and Development Program of China[2018YFB0407200]National Natural Science Foundation of China[61975239]Medical and Health Technology Innovation Project of the Chinese Academy of Medical Sciences[2019-I2M-5-061].
文摘Photodynamic therapy(PDT)not only destroys tumor cells directly but also induced anti-tumor immune response through damage-associated molecular patterns(DAMPs).It is reported that anti-tumor response was associated with light dose and photosensitizer used in PDT.In this study,4T1 tumor cells were implanted on both the right and left flanks of mice.Only the right tumor was treated by HpD-PDT,while the left tumor was not irradiated.The anti-tumor immune response induced by HpD-PDT was investigated.The expression of DAMPs and costimulatory molecules induced by HpD-PDT were tested by immuno fluorescence and flow cytometry in vivo.Different light doses of PDT were designed to treat 4T1 cells.The killing effect was assessed by CCK-8 kit and apoptosis kit.The expression of DAMPs on 4T1 cells after HpDPDT were evaluated by flow cytometry,western blot and ATP kit.This study showed that CD4^(+)T,CD8^(+)T and the production of IFN-γwere increased significantly on day 10 in righttumor after PDT treatment compared with control group.HpD-PDT enhanced the expression of calreticulin(CRT)on tumor tissue.Importantly,co-stimulatory molecular OX-40 and 4-1BB were elevated on CD8^(+)T cells.In vitro,immunogenic death of 4T1 cells was induced after PDT.Besides,the expression of DAMPs increased with the increasing of energy density.This study indicates that anti-tumor immune effect was induced by HpD-PDT.The knowledge of the involvement of CRT,ATP and co-stimulatory molecules uncovers important mechanistic insight into the anti-tumor immunogenicity.It was the first time that co-stimulatory molecules were investigated and found to elevate after PDT.
基金National Basic Research Program ofChina (973 Program) (2009CB118701)National NaturalScientific Foundation of China (30671615, 30871940)+1 种基金Innovation Project of the Chinese Academy of Sciences(KSCX2-YW-N-021)Science and Technology Foundation of Zhejiang Province (2007C22052)
文摘Grass carp reovirus (GCRV) is a tentative member of the Aquareovirus genus in the family Reoviridae. The mature virion comprises 11 dsRNA genomes enclosed by two concentric icosahedral proteins shells that is comprised of five core proteins and two outer capsid proteins. The genome sequence and 3D structure demonstrate there is a higher level of sequence homology in structural proteins between GCRV and mammalian orthoreoviruses (MRV) compared to other members of the family. To understand the pathogenesis of GCRV infection, the outer capsid protein VP5, a homology of the μ1 protein of MRV, was expressed in E.coli. It was found that the recombinant VP5 was highly expressed, and the expressed His-tag fusion protein was involved in the formation of the inclusion body. Additionally, specific anti-VP5 serum was prepared from purified protein and western blot demonstrated that the expressed protein was able to bind immunologically to rabbit anti GCRV particle serum and the immunogenicity was determined by ELISA assay. Additional experiments in investigating the functional properties of VP5 will further elucidate the role of the GCRV outer capsid protein VP5 during entry into host cells, and its interaction among viral proteins and host cells during the infection process.
