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
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
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
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
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
AIEgens can serve as an effective platform for the construction of photosensitizer-based immunogenic cell death(ICD)inducers.To date,several mitochondria or endoplasmic reticulum(ER)-targeted aggregationinduced emissi...AIEgens can serve as an effective platform for the construction of photosensitizer-based immunogenic cell death(ICD)inducers.To date,several mitochondria or endoplasmic reticulum(ER)-targeted aggregationinduced emission(AIE)molecules have been developed and have evoked massive ICD in cells.However,due to the complex physicochemical environment in cells,these small AIE molecules cannot maintain a stable aggregate state,which not only affects the fluorescence intensity of the photosensitizer but also decreases the generation of reactive oxygen species(ROS),and thus reducing the effect of the photosensitizer to elicit ICD.AIEgen-based nanomicelles,which maintain a stable micellar structure,can prevent defects of AIE molecules in photodynamic therapy(PDT)applications.Therefore,in this study,a mitochondria-targeted AIE nanophotosensitizer was synthesized and used as a highly potent ICD inducer for vaccine preparation and tumor prevention.展开更多
Objective The present study aimed to evaluate the immunogenicity of BA.2 variant receptor binding domain(RBD)recombinant protein formulated with CpG 1826 plus alum dual adjuvant.Methods The BA.2 variant RBD(residues 3...Objective The present study aimed to evaluate the immunogenicity of BA.2 variant receptor binding domain(RBD)recombinant protein formulated with CpG 1826 plus alum dual adjuvant.Methods The BA.2 variant RBD(residues 308-548)fusing TT-P2 epitope was obtained from prokaryotic expression system,purification technology and dialysis renaturation,which was designated as Sot protein.The soluble Sot protein formulated with CpG 1826 plus alum dual adjuvant was designated as Sot/CA subunit vaccine and then the BALB/c mice were intramuscularly administrated with two doses of the Sot/CA subunit vaccine at 14-day interval(day 0 and 14).On day 28,the number of effector T lymphocytes secreting IFN-γand IL-4 in mice spleen were determined by enzyme-linked immunospot(ELISpot)assay.The serum IgG,IgG1 and IgG2a antibodies were examined by enzyme-linked immunosorbent assay(ELISA).In addition,the level of neutralizing antibodies(NAbs)induced by Sot/CA subunit vaccine was also evaluated by the microneutralization assay.Results The high-purity soluble Sot protein with antigenicity was successfully obtained by the prokaryotic expression,protein purification and dialysis renaturation.The Sot/CA subunit vaccine induced a high level of IgG antibodies and NAbs,which were of cross-neutralizing activity against SARS-CoV-2 BA.2 and XBB.1.5 variants.Meanwhile,Sot/CA subunit vaccine also induced a high level of effector T lymphocytes secreting IFN-γ(635.00±17.62)and IL-4(279.20±13.10),respectively.Combined with a decreased IgG1/IgG2a ratio in the serum,which indicating Sot/CA subunit vaccine induced a Th1-type predominant immune response.Conclusion The Sot protein formulated with CpG 1826 plus alum dual adjuvant showed that the excellent cellular and humoral immunogenicity,which provided a scientific basis for the development of BA.2 variant subunit vaccines and references for the adjuvant application of subunit vaccines.展开更多
Food allergens are mainly naturally-occurring proteins with immunoglobulin E(IgE)-binding epitopes.Understanding the structural and immunogenic characteristics of allergenic proteins is essential in assessing whether ...Food allergens are mainly naturally-occurring proteins with immunoglobulin E(IgE)-binding epitopes.Understanding the structural and immunogenic characteristics of allergenic proteins is essential in assessing whether and how food processing techniques reduce allergenicity.We here discuss the impacts of food processing technologies on the modification of physicochemical,structural,and immunogenic properties of allergenic proteins.Detection techniques for characterizing changes in these properties of food allergens are summarized.Food processing helps to reduce allergenicity by aggregating or denaturing proteins,which masks,modifies,or destroys antigenic epitopes,whereas,it cannot eliminate allergenicity completely,and sometimes even improves allergenicity by exposing new epitopes.Moreover,most food processing techniques have been tested on purified food allergens rather than food products due to potential interference of other food components.We provide guidance for further development of processing operations that can decrease the allergenicity of allergenic food proteins without negatively impacting the nutritional profile.展开更多
The modulated electro-hyperthermia (mEHT) method is a unique approach that utilizes all the essential apoptotic pathways through an external radiofrequency (RF) signal. The high-frequency RF is amplitude-modulated and...The modulated electro-hyperthermia (mEHT) method is a unique approach that utilizes all the essential apoptotic pathways through an external radiofrequency (RF) signal. The high-frequency RF is amplitude-modulated and coupled capacitively to the target. The provided energy triggers the death receptors and FAS-FADD complexes in the malignant cells. Multi-pathway apoptosis produces immunogenic cell death (ICD). This ICD provides intracellular information about cancer cells by releasing damage-associated molecular patterns (DAMP), including membrane expression of calreticulin (CRT) and extracellular ATP, HMGB1, and HSP70, executing tumor-specific antigen presentation. The antigen-presenting cells (APCs) play a crucial role in reestablishing immune surveillance and hampering the tumor cells’ ability to hide, thereby evading immune attacks. The matured DCs (generally APCs) produce tumor-specific killer and helper T-cells, which have the potential to be active in distant metastases from the treated location. This unique mechanism of action underscores its potential in cancer treatment and extends the local mEHT treatment to the whole body anticancer therapy with an abscopal effect.展开更多
Background:Colon adenocarcinoma(COAD)is a gastrointestinal malignancy with a high mortality rate.Studies have confirmed the role of immunogenic cell death(ICD)in different cancer types.However,there is a lack of resea...Background:Colon adenocarcinoma(COAD)is a gastrointestinal malignancy with a high mortality rate.Studies have confirmed the role of immunogenic cell death(ICD)in different cancer types.However,there is a lack of research on ICD-related genes(ICD-RGs)in COAD.This study aimed to examine the impact of ICD-RGs on COAD and their interaction with the immune microenvironment.Methods:Using data from The Cancer Genome Atlas and Gene Expression Omnibus databases,we identified 107 ICD-RGs in COAD.Using a one-way Cox regression analysis,we examined the relationship between these ICD-RGs and overall survival in COAD.Results:Following the regression analyses,we identified 14 overall survival-related genes.Furthermore,we examined the predictive impact of the ICD-RGs using the least absolute shrinkage and selection operator regression analysis and developed a nine-genes prognostic model.The Cancer Genome Atlas and Gene Expression Omnibus datasets were used for training and validation.Kaplan-Meier analysis was used to confirm that the high-risk group had a lower survival rate than the low-risk group.Finally,following a multifactorial analysis,we created a prognostic nomogram that integrated clinical data and risk scores.Conclusions:The nine-genes model exhibits robust stability and can provide valuable insights for guiding the development of tumor immunotherapy strategies and personalized drug selection for patients with COAD.展开更多
Head and neck cancer(HNC)is a prevalent malignant tumor that is aggressive and frequently resistant to treatment.Immunotherapy,a new approach for treating head and neck tumors,is primarily used in patients with recurr...Head and neck cancer(HNC)is a prevalent malignant tumor that is aggressive and frequently resistant to treatment.Immunotherapy,a new approach for treating head and neck tumors,is primarily used in patients with recurrent or metastatic cancer;however,some patients do not respond well to this treatment or experience significant side effects.Further research is needed to identify better treatment options for head and neck tumors.Dendritic cells(DCs)are pivotal components of the immune system and play a crucial role in initiating and regulating immune responses by presenting antigens to other immune cells.Nonetheless,DCs are frequently suppressed in the tumor immune microenvironment.Immunogenic cell death(ICD)is a form of regulated cell death capable of activating DCs by releasing damage-associated molecular patterns(DAMPs),thus potentially enhancing therapeutic efficacy against HNC.This review summarizes the evidence for the involvement of DCs in the anti-tumor immune response against HNC and their status within the tumor microenvironment.We also outline how DAMPs in ICD can activate DCs and discuss the prospects of ICD-based DC vaccine therapy,with the aim of providing new insights into immunotherapy of patients with HNC.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
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
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 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 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.
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.
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.
基金supported by the National Natural Science Foundation of China(Nos.52173137 and 51873163).
文摘AIEgens can serve as an effective platform for the construction of photosensitizer-based immunogenic cell death(ICD)inducers.To date,several mitochondria or endoplasmic reticulum(ER)-targeted aggregationinduced emission(AIE)molecules have been developed and have evoked massive ICD in cells.However,due to the complex physicochemical environment in cells,these small AIE molecules cannot maintain a stable aggregate state,which not only affects the fluorescence intensity of the photosensitizer but also decreases the generation of reactive oxygen species(ROS),and thus reducing the effect of the photosensitizer to elicit ICD.AIEgen-based nanomicelles,which maintain a stable micellar structure,can prevent defects of AIE molecules in photodynamic therapy(PDT)applications.Therefore,in this study,a mitochondria-targeted AIE nanophotosensitizer was synthesized and used as a highly potent ICD inducer for vaccine preparation and tumor prevention.
基金funded by the National Key R&D Program of China(2023YFC2605302).
文摘Objective The present study aimed to evaluate the immunogenicity of BA.2 variant receptor binding domain(RBD)recombinant protein formulated with CpG 1826 plus alum dual adjuvant.Methods The BA.2 variant RBD(residues 308-548)fusing TT-P2 epitope was obtained from prokaryotic expression system,purification technology and dialysis renaturation,which was designated as Sot protein.The soluble Sot protein formulated with CpG 1826 plus alum dual adjuvant was designated as Sot/CA subunit vaccine and then the BALB/c mice were intramuscularly administrated with two doses of the Sot/CA subunit vaccine at 14-day interval(day 0 and 14).On day 28,the number of effector T lymphocytes secreting IFN-γand IL-4 in mice spleen were determined by enzyme-linked immunospot(ELISpot)assay.The serum IgG,IgG1 and IgG2a antibodies were examined by enzyme-linked immunosorbent assay(ELISA).In addition,the level of neutralizing antibodies(NAbs)induced by Sot/CA subunit vaccine was also evaluated by the microneutralization assay.Results The high-purity soluble Sot protein with antigenicity was successfully obtained by the prokaryotic expression,protein purification and dialysis renaturation.The Sot/CA subunit vaccine induced a high level of IgG antibodies and NAbs,which were of cross-neutralizing activity against SARS-CoV-2 BA.2 and XBB.1.5 variants.Meanwhile,Sot/CA subunit vaccine also induced a high level of effector T lymphocytes secreting IFN-γ(635.00±17.62)and IL-4(279.20±13.10),respectively.Combined with a decreased IgG1/IgG2a ratio in the serum,which indicating Sot/CA subunit vaccine induced a Th1-type predominant immune response.Conclusion The Sot protein formulated with CpG 1826 plus alum dual adjuvant showed that the excellent cellular and humoral immunogenicity,which provided a scientific basis for the development of BA.2 variant subunit vaccines and references for the adjuvant application of subunit vaccines.
基金supported by the National Natural Science Foundation of China (32102605)the Agricultural Science and Technology Innovation Program under Grant (CAAS-ASTIP-2020IAR)the Earmarked Fund for CARS (CARS-44)。
文摘Food allergens are mainly naturally-occurring proteins with immunoglobulin E(IgE)-binding epitopes.Understanding the structural and immunogenic characteristics of allergenic proteins is essential in assessing whether and how food processing techniques reduce allergenicity.We here discuss the impacts of food processing technologies on the modification of physicochemical,structural,and immunogenic properties of allergenic proteins.Detection techniques for characterizing changes in these properties of food allergens are summarized.Food processing helps to reduce allergenicity by aggregating or denaturing proteins,which masks,modifies,or destroys antigenic epitopes,whereas,it cannot eliminate allergenicity completely,and sometimes even improves allergenicity by exposing new epitopes.Moreover,most food processing techniques have been tested on purified food allergens rather than food products due to potential interference of other food components.We provide guidance for further development of processing operations that can decrease the allergenicity of allergenic food proteins without negatively impacting the nutritional profile.
文摘The modulated electro-hyperthermia (mEHT) method is a unique approach that utilizes all the essential apoptotic pathways through an external radiofrequency (RF) signal. The high-frequency RF is amplitude-modulated and coupled capacitively to the target. The provided energy triggers the death receptors and FAS-FADD complexes in the malignant cells. Multi-pathway apoptosis produces immunogenic cell death (ICD). This ICD provides intracellular information about cancer cells by releasing damage-associated molecular patterns (DAMP), including membrane expression of calreticulin (CRT) and extracellular ATP, HMGB1, and HSP70, executing tumor-specific antigen presentation. The antigen-presenting cells (APCs) play a crucial role in reestablishing immune surveillance and hampering the tumor cells’ ability to hide, thereby evading immune attacks. The matured DCs (generally APCs) produce tumor-specific killer and helper T-cells, which have the potential to be active in distant metastases from the treated location. This unique mechanism of action underscores its potential in cancer treatment and extends the local mEHT treatment to the whole body anticancer therapy with an abscopal effect.
文摘Background:Colon adenocarcinoma(COAD)is a gastrointestinal malignancy with a high mortality rate.Studies have confirmed the role of immunogenic cell death(ICD)in different cancer types.However,there is a lack of research on ICD-related genes(ICD-RGs)in COAD.This study aimed to examine the impact of ICD-RGs on COAD and their interaction with the immune microenvironment.Methods:Using data from The Cancer Genome Atlas and Gene Expression Omnibus databases,we identified 107 ICD-RGs in COAD.Using a one-way Cox regression analysis,we examined the relationship between these ICD-RGs and overall survival in COAD.Results:Following the regression analyses,we identified 14 overall survival-related genes.Furthermore,we examined the predictive impact of the ICD-RGs using the least absolute shrinkage and selection operator regression analysis and developed a nine-genes prognostic model.The Cancer Genome Atlas and Gene Expression Omnibus datasets were used for training and validation.Kaplan-Meier analysis was used to confirm that the high-risk group had a lower survival rate than the low-risk group.Finally,following a multifactorial analysis,we created a prognostic nomogram that integrated clinical data and risk scores.Conclusions:The nine-genes model exhibits robust stability and can provide valuable insights for guiding the development of tumor immunotherapy strategies and personalized drug selection for patients with COAD.
基金supported by the Provincial Science and Technology Plan Project of Sichuan Provincial Department of Science and Technology(2022SNZY001)the Science and Technology Project of Sichuan Provincial Health Commission(Appropriate Technology Base)(2022JDXM021)the Science and Technology Research Project of Sichuan Administration of Traditional Chinese Medicine(2024MS643).
文摘Head and neck cancer(HNC)is a prevalent malignant tumor that is aggressive and frequently resistant to treatment.Immunotherapy,a new approach for treating head and neck tumors,is primarily used in patients with recurrent or metastatic cancer;however,some patients do not respond well to this treatment or experience significant side effects.Further research is needed to identify better treatment options for head and neck tumors.Dendritic cells(DCs)are pivotal components of the immune system and play a crucial role in initiating and regulating immune responses by presenting antigens to other immune cells.Nonetheless,DCs are frequently suppressed in the tumor immune microenvironment.Immunogenic cell death(ICD)is a form of regulated cell death capable of activating DCs by releasing damage-associated molecular patterns(DAMPs),thus potentially enhancing therapeutic efficacy against HNC.This review summarizes the evidence for the involvement of DCs in the anti-tumor immune response against HNC and their status within the tumor microenvironment.We also outline how DAMPs in ICD can activate DCs and discuss the prospects of ICD-based DC vaccine therapy,with the aim of providing new insights into immunotherapy of patients with HNC.
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.
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.
