Objective To eliminate the side effects of aluminum adjuvant and His-tag,we constructed chimeric VLPs displaying the epitope of EV71(SP70) without His-tagged.Then evaluating whether the VLPs could efficiently evoke ...Objective To eliminate the side effects of aluminum adjuvant and His-tag,we constructed chimeric VLPs displaying the epitope of EV71(SP70) without His-tagged.Then evaluating whether the VLPs could efficiently evoke not only humoral but also cellular immune responses against EV71 without adjuvant.Methods The fusion protein was constructed by inserting SP70 into the MIR of truncated HBc Ag sequence,expressed in E.Coli,and purified through ion exchange chromatography and density gradient centrifugation.Mice were immunized with the VLPs and sera were collected afterwards.The specific antibody titers,Ig G subtypes and neutralizing efficacy were detected by ELISA,neutralization assay,and EV71 lethal challenge.IFN-γ and IL-4 secreted by splenocytes were tested by ELISPOT assay.Results HBc-SP70 proteins can self-assemble into empty VLPs.After immunization with HBc-SP70 VLPs,the detectable anti-EV71 antibodies were effective in neutralizing EV71 and protected newborn mice from EV71 lethal challenge.There was no significant difference for the immune efficacy whether the aluminum adjuvant was added or not.The specific Ig G subtypes were mainly IgG1 and IgG2 b and splenocytes from the mice immunized produced high levels of IFN-γ and IL-4.Conclusion The fusion proteins without His-tagged was expressed and purified as soluble chimeric HBc-SP70 VLPs without renaturation.In the absence of adjuvant,they were efficient to elicit high levels of Th1/Th2 mixed immune response as well as assisted by aluminum adjuvant.Furthermore,the chimeric VLPs have potential to prevent HBV and EV71 infection simultaneously.展开更多
Objective To construct eukaryotic expression vector of HPV18 L1-E6, E7 chimeric gene and examine the humoral and cellular immune responses induced by this DNA vaccines in mice. Methods The C-terminal of major capsid p...Objective To construct eukaryotic expression vector of HPV18 L1-E6, E7 chimeric gene and examine the humoral and cellular immune responses induced by this DNA vaccines in mice. Methods The C-terminal of major capsid protein L1 gene and mutant zinc finger domains of early E6/7 oncogenes in HPV18 were integrated and inserted into eukaryotic expression vector pVAX1 to generate vaccines pVAX1-L1E6Mxx, E7Mxx. CHO cells were transiently transfected with the individual construct. Target protein expressions in the lysate of the transfected cells were measured by ELISA and immunocytochemistry. After BALB/c mice were vaccinated with various recombinant plasmids(pVAX1-L1-E6M3 or pVAX1-L1-E7M3) and immunie adjuvants (pLXHDmB7-2 or LTB) through different administration routes (intramuscular or intranasal) , the great cellular immune responses were produced as revealed by delayed-type hypersensitivity (DTH) and lymphocyte proliferation, and the expression of IL-4 and IFN-γ cells in CD4 + and CD8 + subpopulations. Results The highly efficient expression of pVAX1-L1E6Mxx, E7Mxx vector in host eukaryotic cells were demonstrated both by ELISA and immunocytochemistry. The level of specific serum IgG against HPV in experiment groups mice was much higher than that of control group, and intranuscular immunization group had the highest antibody level. Intramuscular immunization groups were superior to intranasal immunization groups in DTH response, splenocyte proliferation and CD8+ IFN-γ + cells number, but CD4 + IL4 + cell number was higher in intranasal immunization groups. The immunization groups using pLXHDmB7-2 as adjuvant were superior to other groups in immunoresponse. Conclusion These DNA vaccines produce remarkable cellular and humoral immune responses in the mouse and may provide as prophylatic and therapeutic candidates for HPV induced cancer treatment.展开更多
With the increasing immunological studies on camels due to the advantage of their single-chain antibodies for humanizations,it is demanding to develop an easy-to-handle evaluation method of their humoral immune respon...With the increasing immunological studies on camels due to the advantage of their single-chain antibodies for humanizations,it is demanding to develop an easy-to-handle evaluation method of their humoral immune response before proceeding with immunization of foreign antigens that may be toxic to camels.In this study,we quantitatively determined the expression levels of T-helper 2(Th2) cytokines in peripheral blood lymphocytes obtained from Bactrian camels by real-time PCR.The recorded kinetic profiles resulting from the immunization of ovalbumin(OVA) indicated that after immunization,Th2 cytokines including interleukin(IL) families such as IL-4,IL-10,and IL-13 in the camels were up-regulated by a factor of 1.78,3.15,and 1.22,respectively,which was validated by traditional enzyme-linked immunosorbent assay(ELISA) methods.Unlike ELISA which requires specific enzyme-labeled antibodies,this established method based on the minimal amount of blood samples holds an advantage in the preliminary evaluation of camel humoral immune response with desirable precision,which is meaningful for biomedical explorations of camel-derived antibodies.展开更多
[ Objective] The paper was to study the immune enhancement of ATRA on Newcastle Disease (ND) vaccine. [ Method ] The 1-day-old AA broilers were treated with ATRA at the doses of I and 5 p.mol/kg, respectively. At 7 ...[ Objective] The paper was to study the immune enhancement of ATRA on Newcastle Disease (ND) vaccine. [ Method ] The 1-day-old AA broilers were treated with ATRA at the doses of I and 5 p.mol/kg, respectively. At 7 and 28 days of age, broilers in drug control group, low dose group and high dose group were immunized with ND vaccine by intranasal and eye immunization approach. At 7, 14, 21, 28, 35, 42 and 49 clays of age, seven chickens were randomly se- lected from each group and weighed. The thymus, spleen, bursa of fabrieius and serum were collected for calculating immune organ index of thymus, spleen and bursa of fabricius. The ND specific antibody titers in serum were determined with HI test. [Result] ATRA promoted the growth of thymus, spleen and bursa of fabricius, and improved the immune organ index and ND specific antibody titers of chicks. [ Conclusion] ATRA enhanced the humoral immune response of chicks, and ATRT at the dose of 5 μmol/kg presented more prominent immune enhancement effect on ND vaccine.展开更多
[Objectives]The paper was to study the effects of combined application of nonreplicating Toxoplasma uracil auxotrophs(NRTUAs)and Agaricus blazei Murill polysaccharide(ABP)on growth and humoral immunity of chicks.[Meth...[Objectives]The paper was to study the effects of combined application of nonreplicating Toxoplasma uracil auxotrophs(NRTUAs)and Agaricus blazei Murill polysaccharide(ABP)on growth and humoral immunity of chicks.[Methods]A total of 120 one-day old female Hyline brown laying hens were randomly divided into 4 groups,30 hens for each group.The chicks in group 1 were subcutaneously injected with NRTUAs and fed on the diet containing with ABP;the chicks in group 2 were subcutaneously injected with NRTUAs;the chicks in group 3 were subcutaneously injected with equal volume of PBS,and fed on the diet containing with ABP;the chicks in group 4 were subcutaneously injected with equal volume of PBS.The body weight of chicks in each group was counted at the 21^(st),42^(nd),84^(th)and 112^(th)week.During this period,blood samples were collected from chicks in each group at 0,7,14,21,28 and 35 d post immunization against Newcastle disease(ND),and serum was separated to detect the antibody titer of ND.[Results]The combined application of NRTUAs and ABP had no effect on growth of chicks,but promoted the humoral immune response of chicks,significantly improved the ND antibody level of chicks,and could maintain high levels of antibodies in the body for a long time.[Conclusions]The study lays a theoretical foundation for further developing the clinical application of NRTUAs and ABP.展开更多
Background:B cells represent a crucial component of adaptive immunity that ensures long-term protection from infection by generating pathogen-specific immunoglobulins.Exercise alters B cell counts and immunoglobulin l...Background:B cells represent a crucial component of adaptive immunity that ensures long-term protection from infection by generating pathogen-specific immunoglobulins.Exercise alters B cell counts and immunoglobulin levels,but evidence-based conclusions on potential benefits for adaptive immunity are lacking.This systematic review assessed current literatures on the impact of acute exercise and exercise training on B cells,immunoglobulins,and markers of secretory immunity in human biofluids.Methods:According to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses(PRISMA)guidelines,MEDLINE,Web of Science,and Embase were searched on March 8,2023.Non-randomized controlled trials and crossover trials investigating the impact of acute exercise or exercise training on B cell counts and proportions,immunoglobulin levels,salivary flow rate,or secretory immunoglobulin A secretion rate were included.Quality and reporting of exercise training studies were assessed using the Tool for the Assessment of Study Quality and reporting in Exercise.Study characteristics,outcome measures,and statistically significant changes were summarized tabularly.Results:Of the 67 eligible studies,22 applied acute exercise and 45 applied exercise training.All included outcomes revealed significant alterations over time in acute exercise and exercise training context,but only a few investigations showed significant differences compared to control conditions.Secretory and plasma immunoglobulin A levels were most consistently increased in response to exercise training.Conclusion:B cell-related outcomes are altered by acute exercise and exercise training,but evidence-based conclusions cannot be drawn with high confidence due to the large heterogeneity in populations and exercise modalities.Well-designed trials with large sample sizes are needed to clarify how exercise shapes B cell-related immunity.展开更多
Objective To evaluate the effects of ethyl-acetate fraction (EAF) of extracts from Tetrastigma hemsleyanum Diels et. Gilg (TDG) on immune functions of ICR mice. Methods ICR mice were exposed to different doses of ...Objective To evaluate the effects of ethyl-acetate fraction (EAF) of extracts from Tetrastigma hemsleyanum Diels et. Gilg (TDG) on immune functions of ICR mice. Methods ICR mice were exposed to different doses of EAF for 15 or 30 days and then their immune functions were analyzed, including ConA-induced splenic lymphocyte transformation, SRBC- induced delayed type hypersensitivity response, serum hemolysin analysis, antibody-producing cells, peritoneal macrophage phagocytized chicken red blood cells, natural killer cell activity, and serum level of cytoldnes. Results EAF of extracts from TDG at different doses had various effects on immune functions of ICR mice. As compared with the controls, it increased the mouse spleen lymphocyte transformation induced by ConA, the left-hind voix pedis thickness and the number of plague forming cells (PFCs) at the dose of 1.82 mg/mL, 5.48 mg/mL, and 9.12 mg/mL, respectively; increased the ink clearance ability at the dose of 0.91 mg/mL, 1.82 mg/mL, 5.48 mg/mL, and 9.12 mg/mL, respectively; increased the phagocytosis index of mononuclear-macrophages and production of serum interferon-gamma (IFN-?) at the dose of 5.48 mg/mL; and could promote the production of serum tumor necrosis factor-alpha (TNF-α) at the dose of 9.12 mg/mL. Conclusion EAF of extracts from TDG can regulate mouse immune functions in vivo.展开更多
It is well known that IL4 plays important roles in the induction of humoral immunity. In the present study, the humoral antitumor responses induced by IL4 genemodified tumor vaccine were investigated. The mice were va...It is well known that IL4 plays important roles in the induction of humoral immunity. In the present study, the humoral antitumor responses induced by IL4 genemodified tumor vaccine were investigated. The mice were vaccinated with the IL4 genemodified B16 melanoma cells. Then the proliferative capacity of the splenic lymphocytes, the levels of the antibodies in the murine serum against wildtype B16 cells and the cytotoxicity of the serum to wildtype melanoma cells were detected. Our data showed that the LPSinduced proliferation of the splenic lymphocytes from the mice vaccinated with the IL4 genemodified tumor vaccine increased more significantly than that from mice vaccinated with wildtype tumor vaccine. The cytotoxicity of the serum to wildtype melanoma cells also increased markedly when detected. It was also observed that the number of pulmonary metastases decreased more obviously when the mice were intravenously injected with the mixture of wildtype B16 cells and the serum from the mice vaccinated with the IL4 genemodified B16 cells. Our data demonstrated that humoral immunity might contribute to the antitumor effect of IL4 gene therapy.展开更多
Objective To study the humoral immunity status and distribution pattern of lymphocyte subgroups of peripheral blood mononuclear cell (PBMC) in patients with Kashin-Beck Disease (KBD), and their relationship with eryt...Objective To study the humoral immunity status and distribution pattern of lymphocyte subgroups of peripheral blood mononuclear cell (PBMC) in patients with Kashin-Beck Disease (KBD), and their relationship with erythrocyte selenium. Methods 23 X-ray diagnosed patients, 22 age- and sex- matched healthy children in KBD affected area (KAA), and 25 in KBD non-affected area (KNAA) were randomly selected. Immunohistochemistry with monoclonal antibodies anti-CD4, anti-CD8, anti-CD20 was conducted to analyze the lymphocyte subsets. Serum IgM, IgA, IgG, Complement C3 and C4 were assayed using rate nephelometry (Array 360 System, USA). The contents of erythrocyte selenium was determined by 2,3-diaminonaphthalene fluorescence assay. Results CD4+ and CD8+ cells percentage in PBMCs and serum IgA were significantly lower in KAA than those in KNAA(P< 0.05). CD20+ percentage in KAA displayed a decreasing trend compared to KNAA, although not statistically significantly. No statistical differences were found in CD4/CD8 ratio, serum IgG, IgM, C3 and C4 levels. Erythrocyte selenium level in KAA still showed a pronounced decrease compared to that in KNAA. Correlation analysis showed that erythrocyte selenium contents had a strong association with the CD4 cell percentage (r= 0.625, P< 0.05), as well as serum IgA (r= 0.462, P< 0.05). In addition, a moderate correlation between the serum IgA and CD4+ percentage (r= 0.130, P> 0.05) was found. Conclusion These results suggested that children in KAA had a comparably low cellular immunity level and their humoral immunity status was also in a state of moderate immune suppression. Of this immune disorder in Kashin-Beck disease patients, selenium deficiency probably played a critical role via affecting the distribution pattern of peripheral blood lymphocyte. Selenium-deficiency and immune impairment maybe both have something to do with the cause-effect chain of KBD.展开更多
[ Objective ] The paper was to screen the active ingredients of traditional Chinese medicine ( TCM), which significantly enhanced the immune effect of porcine circovirus type 2 (PCV2) vaccine. [Method]Sixty-six 14...[ Objective ] The paper was to screen the active ingredients of traditional Chinese medicine ( TCM), which significantly enhanced the immune effect of porcine circovirus type 2 (PCV2) vaccine. [Method]Sixty-six 14-day-old piglets were randomly divided into 11 groups, including immune control group, blank control group and nine drug groups. Piglets that were reared for 21 d were intramuscularly injected with 2 mL of inactivated vaccine in cervical region expect for blank control group. Piglets in nine drug groups were administrated with 5 mL of matrine, scutellarin and astragalus polysaecharide (ASP) at three doses ( high dose 12 mg, medium dose 8 nag, low dose 4 mg) three days before and after immunization; piglets in blank control group and irmnune control group were administrated with equal volume of normal saline. Five piglets were randomly selected from each group to collect venous blood at 7, 14, 21 and 28 d post immunization, and the PCV2 antibody level, the concentration of specific immunoglobulin [gG, the changes in peripheral blood lymphocyte subsets, and the content of serum IL-2 and IFN-γ were determined. [ Result] The immunological effects of active ingredients of three TCM on PCV2 vaccine all enhanced at different degree. The effects of scutellarin middle dose group was the best. [ Conclusion ] Scutellarin could be used as a candidate drug for PCV2 vaccine immunoenhancer.展开更多
Salmonella is a ubiquitous pathogen which, in addition to causing poultry diseases, has a growing zoonotic impact. It has demanded the implementation of diverse control strategies, in which vaccines play a major role....Salmonella is a ubiquitous pathogen which, in addition to causing poultry diseases, has a growing zoonotic impact. It has demanded the implementation of diverse control strategies, in which vaccines play a major role. The understanding of the immune pathways elicited by the different vaccines is important, contributing for the establishment of strong immune correlates of protection, for instance. With the purpose of determining the dynamics of the humoral and cellular immune responses to vaccination, broiler breeders (Cobb Slow) were immunized with live or inactivated vaccines against Salmonella Enteritidis. Lymphocyte and macrophage subsets were analyzed in the peripheral blood by flow cytometry and antigen-specific circulating IgY and mucosal IgA were quantified. The markers analyzed by flow cytometry were CD8/CD28, CD4/TCRVβ1, Kul/ MHC II and Bu-1. Both live and inactivated vaccines induced an increase in the proportion of circulating monocytes (Kul+MHCII+) in some time points compared to non-vaccinated controls. However, whereas the live vaccine leads to an increase in CD8-CD28+ and Bu-1+ lymphocytescompared to the control group, the inactivated vaccine prompteda reduction in the percentage of severalleucocyte subsets (Kul-MHCII+, Bu-1+, CD8+CD28+, CD8-CD28+, CD4+TCRVβ1-, CD4+TCRVβ1+, CD4-TCRVβ1+) after the boost dose. Both vaccines induced specific serum IgY and mucosal IgA production;however, the inactivated vaccine stimulated higher titers in a shorter period. These results contribute to the understanding of mechanisms of action of live and inactivated Salmonella vaccines in chickens.展开更多
Humanized immune system(HIS)mice have been developed and used as a small surrogate model to study human immune function under normal or disease conditions.Although variations are found between models,most HIS mice sho...Humanized immune system(HIS)mice have been developed and used as a small surrogate model to study human immune function under normal or disease conditions.Although variations are found between models,most HIS mice show robust human T cell responses.However,there has been unsuccessful in constructing HIS mice that produce high-affinity human antibodies,primarily due to defects in terminal B cell differentiation,antibody affinity maturation,and development of primary follicles and germinal centers.In this review,we elaborate on the current knowledge about and previous attempts to improve human B cell development in HIS mice,and propose a potential strategy for constructing HIS mice with improved humoral immunity by transplantation of human follicular dendritic cells(FDCs)to facilitate the development of secondary follicles.展开更多
Background We have previously developed and characterized a monoclonal anti-idiotype antibody, designated 6B 11, which mimics an ovarian carcinoma associated antigen OC166-9 and whose corresponding monoclonal antibody...Background We have previously developed and characterized a monoclonal anti-idiotype antibody, designated 6B 11, which mimics an ovarian carcinoma associated antigen OC166-9 and whose corresponding monoclonal antibody is COC166-9 (Abl). In this study, we evaluate the humoral immune responses induced by the fusion protein 6B11 single-chain variable fragment (scFv)/human granulocyte macrophage colony-stimulating factor (hGM-CSF) and 6B 1 lscFv in BALB/c mice. Methods The fusion protein 6B 11 scFv/hGM-CSF was constructed by fusing a recombinant single-chain variable fragment of 6B11scFv to GM-CSE BALB/c mice were administrated by 6B11scFv/hGM-CSF and 6B11scFv, respectively. Results The fusion protein 6B11scFv/hGM-CSF retained binding to the anti-mouse F(ab)2' and was also biologically active as measured by proliferation of human GM-CSF dependent cell TF1 in vitro. After immunization with the 6B11scFv/hGM-CSF and 6BllScFv, BALB/c mice showed significantly enhanced Ab3 antibody responses to 6B11 scFv/hGM-CSF compared with the 6B11 scFv alone. The level of Ab3 was the highest after the first week and maintained for five weeks after the last immunization. Another booster was given when the Ab3 titer descended, and it would reach to the high level in a week. Conclusion The fusion protein 6B11scFv/hGM-CSF can induce humoral immunity against ovarian carcinoma in vivo. We also provide the theoretical foundation for the application of the fusion protein 6B11 scFv/hGM-CSF for active immunotherapy of ovarian cancer.展开更多
Colorectal cancer(CRC)is the second leading cause of cancer-related death worldwide.The five-year survival rate of CRC patients depends on the stage at diagnosis,being higher than 80%when CRC is diagnosed in the early...Colorectal cancer(CRC)is the second leading cause of cancer-related death worldwide.The five-year survival rate of CRC patients depends on the stage at diagnosis,being higher than 80%when CRC is diagnosed in the early stages but lower than 10%when CRC is diagnosed in advanced stages.Autoantibodies against specific CRC autoantigens(tumor-associated antigens(TAAs))in the sera of patients have been widely demonstrated to aid in early diagnosis.Thus,we herein aim to identify autoantigens target of autoantibodies specific to CRC that possess a significant ability to discriminate between CRC patients and healthy individuals by means of liquid biopsy.To that end,we examined the protein content of the exosomes released by five CRC cell lines and tissue samples from CRC patients by means of immunoprecipitation coupled with mass spectrometry analysis.A total of 103 proteins were identified as potential autoantigens specific to CRC.After bioinformatics and meta-analysis,we selected 15 proteins that are more likely to be actual CRC autoantigens in order to evaluate their role in CRC prognosis by Western blot(WB)and immunohistochemistry(IHC).We found dysregulation at the protein level for 11 of these proteins in both tissue and plasma exosome samples from patients,along with an association of nine of these proteins with CRC prognosis.After validation,all but one showed a statistically significant high diagnostic ability to distinguish CRC patients and individuals with premalignant lesions from healthy individuals,either by luminescence Halotag-based beads,or by a multiplexed biosensing platform involving the use of magnetic microcarriers as solid support modified with covalently immobilized Halotag fusion proteins constructed for CRC detection.Taken together,our results highlight the usefulness of the approach defined here to identify the TAAs specific to chronic diseases;they also demonstrate that the measurement of autoantibody levels in plasma against the TAAs identified here could be integrated into a point-of-care(POC)device for CRC detection with high diagnostic ability.展开更多
BACKGROUND Helicobacter pylori(H.pylori)has been found to be associated with extragastrointestinal diseases,possibly including adverse food reactions(such as food allergy or intolerance).However,there are few studies ...BACKGROUND Helicobacter pylori(H.pylori)has been found to be associated with extragastrointestinal diseases,possibly including adverse food reactions(such as food allergy or intolerance).However,there are few studies on H.pylori and food allergy or intolerance,and the results are inconsistent.Food-specific immunoglobulin(Ig)G has been revealed to be associated with food allergy or intolerance and can be used as a marker to explore the correlation between H.pylori infection and food allergy or intolerance.AIM To explore the relationship between H.pylori infection and food-specific IgG METHODS We retrospectively analyzed the physical examination data of 21822 subjects from February 2014 to December 2018 in this study.H.pylori infection was detected using the 13C urea breath test.Food-specific IgG of eggs,milk and wheat in serum was assessed.Subjects were grouped according to H.pylori positivity,and the positive rates of three kinds of food-specific IgG were compared between the two groups.Multivariable logistic regression analysis was performed to elucidate the association between H.pylori infection and food-specific IgG.RESULTS The total infection rate of H.pylori was 39.3%,and the total food-specific IgGpositive rates of eggs,milk and wheat were 25.2%,9.0%and 4.9%,respectively.The infection rate of H.pylori was higher in males than in females,while the positive rates of food-specific IgG were lower in males than in females.The positive rates of food-specific IgG decreased with age in both males and females.In the H.pylori-positive groups,the positive rates of food-specific IgG of eggs,milk and wheat were all lower than those in the H.pylori-negative groups.Multivariate logistic regression analysis revealed that H.pylori infection was negatively correlated with the food-specific IgG-positive rates of eggs,milk and wheat(odds ratio value of eggs 0.844-0.873,milk 0.741-0.751 and wheat 0.755-0.788,in different models).CONCLUSION H.pylori infection was found to be negatively associated with the food-specific IgG of eggs,milk and wheat in Southwest China.展开更多
To explore the primary humoral and cellular immunological mechanism of the combined hepatitis A-measles-varicella vaccine, the mice were inoculated with hepatitis A-measles-varicella vaccine by intraperitoneally and t...To explore the primary humoral and cellular immunological mechanism of the combined hepatitis A-measles-varicella vaccine, the mice were inoculated with hepatitis A-measles-varicella vaccine by intraperitoneally and two weeks later, blood was collected to observe the mice's immunological status. Antibody level was measured to appraise the humoral immunity. At the same time, T lymphocyte surface marker, NK cell activity, LAK cell activity, delayed type hypersensitivity of skin, Mφ phagocytic function, mRNA level of cytokine IL-2 and IFN-γ plus lymphocyte transformation test were used to analyze the cellular immunity. The humoral immunity results show that the combined hepatitis A-measles-varicella vaccine produce the same antibody level as their corresponding univalent vaccine, and maintained fine immunogenicity and security. The result of cellular immunity shows that the combined vaccine could activate physical immunocyte, increase the regulative ability of cytokine, enhance the physical immune function and immune defense ability. The present research proved the security and better humoral and cellular immunity of combined hepatitis A-measles-varicella vaccine from the immunological point of view, which laid good foundation for further study and development.展开更多
C-type lectins(CTLs)are a superfamily of Ca^(2+)-dependent carbohydrate-recognition proteins,and an important pattern recognition receptor(PRR)in insect innate immunity which can mediate humoral and cellular immunity ...C-type lectins(CTLs)are a superfamily of Ca^(2+)-dependent carbohydrate-recognition proteins,and an important pattern recognition receptor(PRR)in insect innate immunity which can mediate humoral and cellular immunity in insects.In this study,we report a novel dual carbohydrate-recognition domain(CRD)CTL from Plutella xylostella which we designate PxIML.PxIML is a protein with a 969 bp open reading frame(ORF)encoding 322 amino acids,containing a signal peptide and a dual-CRD with EPN(Glu_(124)-Pro_(125)-Asn_(126))and QPD(Gln_(274)-Pro_(275)-Asp_(276))motifs.The expression of PxIML mRNA in the fat body was significantly higher than in hemocytes and midgut.The relative expression levels of PxIML in the whole insect and the fat body were significantly inhibited after infection with Bacillus thuringiensis 8010(Bt8010)at 18 h,while they were significantly upregulated after infection with Serratia marcescens IAE6 or Pichia pastoris.The recombinant PxIML(rPxIML)protein could bind to the tested pathogen-associated molecular patterns(PAMPs),and the bacteria of Enterobacter sp.IAE5,S.marcescens IAE6,Staphylococcus aureus,Escherichia coli BL21,and Bt8010 in a Ca^(2+)-dependent manner,however,it showed limited binding to the fungus,P.pastoris.The rPxIML exhibited strong activity in the presence of Ca^(2+) to agglutinate Bt8010,Enterobacter sp.IAE5 and S.aureus,but it only weakly agglutinated with E.coli BL21,and could not agglutinate with S.marcescens IAE6 or P.pastoris.Furthermore,the rPxIML could bind to hemocytes,promote the adsorption of hemocytes to beads,and enhance the phenoloxidase(PO)activity and melanization of P.xylostella.Our results suggest that PxIML plays an important role in pathogen recognition and in mediating subsequent humoral and cellular immunity of P.xylostella.展开更多
[ Objective] To study the effects of different oligosaccharides on immunity and production performance of piglets.[ Method] A total of 75 Yorkshire x Rongchang piglets were assigned into five groups, 15 in each group....[ Objective] To study the effects of different oligosaccharides on immunity and production performance of piglets.[ Method] A total of 75 Yorkshire x Rongchang piglets were assigned into five groups, 15 in each group. Isomalto-oligosaccharide ( IMO), fructo-oligosaccharide ( FOS), mannan-oligosacchadde (MOS), and mixed oligosacchadde were supplemented to basal diet (7.5 g/kg), respectively. The control group was also set. The experimental period lasted for 53 d. The clinic symptoms and diarrhea were observed. All piglets were weighed once a week. Feed intake was also recorded. Blood was collected via precaval vein for determination of immunology indexes at the age of 30 and 60 d, respectively. [ Result] The IMO and FOS enhanced cellular immunity of 30-day-old piglets significantly (P 〈 0.05), and the IMO also enhanced their humoral immunity. The MOS enhanced cellular immunity and humoral immunity of 60-day-old piglets significantly (P 〈 0.05). It also improved production performance largest, and its diarrhea rate was the lowest. [ Conclusion] Different oligosaccharides have various effects on immunity and production performance of oiolets.展开更多
Activating humoral and cellular immunity in lymph nodes(LNs)of nanoparticle-based vaccines is critical to controlling tumors.However,how the physical properties of nanovaccine carriers orchestrate antigen capture,lymp...Activating humoral and cellular immunity in lymph nodes(LNs)of nanoparticle-based vaccines is critical to controlling tumors.However,how the physical properties of nanovaccine carriers orchestrate antigen capture,lymphatic delivery,antigen presentation and immune response in LNs is largely unclear.Here,we manufactured gold nanoparticles(AuNPs)with the same size but different shapes(cages,rods,and stars),and loaded tumor antigen as nanovaccines to explore their disparate characters on above four areas.Results revealed that star-shaped AuNPs captured and retained more repetitive antigen epitopes.On lymphatic delivery,both rods and star-shaped nanovaccines mainly drain into the LN follicles region while cage-shaped showed stronger paracortex retention.A surprising finding is that the star-shaped nanovaccines elicited potent humoral immunity,which is mediated by CD4^(+)T helper cell and follicle B cell cooperation significantly preventing tumor growth in the prophylactic study.Interestingly,cage-shaped nanovaccines preferentially presented peptide-MHC I complexes to evoke robust CD8^(+)T cell immunity and showed the strongest therapeutic efficacy when combined with the PD-1 checkpoint inhibitor in established tumor study.These results highlight the importance of nanoparticle shape on antigen delivery and presentation for immune response in LNs,and our findings support the notion that different design strategies are required for prophylactic and therapeutic vaccines.展开更多
Three-month-old Alzheimer's disease model transgenic mice were immunized with Aβ1-42, Plp-Adenovirus [Ad]-X-CMV-(Aβ3-10)lo-CpG [AdCpG-(Aβ3-10)1] or AdCpG virus fluid via na- sal mucosal inhalation, respectivel...Three-month-old Alzheimer's disease model transgenic mice were immunized with Aβ1-42, Plp-Adenovirus [Ad]-X-CMV-(Aβ3-10)lo-CpG [AdCpG-(Aβ3-10)1] or AdCpG virus fluid via na- sal mucosal inhalation, respectively. ELISA analysis of serum showed Aβ42 antibody titers were significantly increased in mice immunized with Aβ1-42 and AdCpG-(Aβ3-10)10. Concanavalin A and AdCpG-(Aβ3-10)10 stimulation significantly increased the number of proliferating spleen cells cultured from AdCpG(Aβ3-10)Io and Aβ42 groups compared with the control group. In the AdCp- G(Aβ3-10)10 group, levels of interleukin (IL)-4 and IL-10 were increased, while those of IL-2 and interferon-y were decreased. In the A[342 group, levels of IL-4, IL-10, IL-2 and interferon-y were all increased. Experimental findings indicate that AdCpG-(Aβ3-10)10 vaccine can produce strong T helper 2 (Th2) humoral immune responses in addition to the production of Aβ42 antibody. The cellular immunologic response was weak and avoided Aβ1-42-mediated cytotoxicity.展开更多
基金supported by the National Science-technology Support Plan Projects 'The development of EV71 genetic engineering vaccine'[2008BAI69B02]
文摘Objective To eliminate the side effects of aluminum adjuvant and His-tag,we constructed chimeric VLPs displaying the epitope of EV71(SP70) without His-tagged.Then evaluating whether the VLPs could efficiently evoke not only humoral but also cellular immune responses against EV71 without adjuvant.Methods The fusion protein was constructed by inserting SP70 into the MIR of truncated HBc Ag sequence,expressed in E.Coli,and purified through ion exchange chromatography and density gradient centrifugation.Mice were immunized with the VLPs and sera were collected afterwards.The specific antibody titers,Ig G subtypes and neutralizing efficacy were detected by ELISA,neutralization assay,and EV71 lethal challenge.IFN-γ and IL-4 secreted by splenocytes were tested by ELISPOT assay.Results HBc-SP70 proteins can self-assemble into empty VLPs.After immunization with HBc-SP70 VLPs,the detectable anti-EV71 antibodies were effective in neutralizing EV71 and protected newborn mice from EV71 lethal challenge.There was no significant difference for the immune efficacy whether the aluminum adjuvant was added or not.The specific Ig G subtypes were mainly IgG1 and IgG2 b and splenocytes from the mice immunized produced high levels of IFN-γ and IL-4.Conclusion The fusion proteins without His-tagged was expressed and purified as soluble chimeric HBc-SP70 VLPs without renaturation.In the absence of adjuvant,they were efficient to elicit high levels of Th1/Th2 mixed immune response as well as assisted by aluminum adjuvant.Furthermore,the chimeric VLPs have potential to prevent HBV and EV71 infection simultaneously.
文摘Objective To construct eukaryotic expression vector of HPV18 L1-E6, E7 chimeric gene and examine the humoral and cellular immune responses induced by this DNA vaccines in mice. Methods The C-terminal of major capsid protein L1 gene and mutant zinc finger domains of early E6/7 oncogenes in HPV18 were integrated and inserted into eukaryotic expression vector pVAX1 to generate vaccines pVAX1-L1E6Mxx, E7Mxx. CHO cells were transiently transfected with the individual construct. Target protein expressions in the lysate of the transfected cells were measured by ELISA and immunocytochemistry. After BALB/c mice were vaccinated with various recombinant plasmids(pVAX1-L1-E6M3 or pVAX1-L1-E7M3) and immunie adjuvants (pLXHDmB7-2 or LTB) through different administration routes (intramuscular or intranasal) , the great cellular immune responses were produced as revealed by delayed-type hypersensitivity (DTH) and lymphocyte proliferation, and the expression of IL-4 and IFN-γ cells in CD4 + and CD8 + subpopulations. Results The highly efficient expression of pVAX1-L1E6Mxx, E7Mxx vector in host eukaryotic cells were demonstrated both by ELISA and immunocytochemistry. The level of specific serum IgG against HPV in experiment groups mice was much higher than that of control group, and intranuscular immunization group had the highest antibody level. Intramuscular immunization groups were superior to intranasal immunization groups in DTH response, splenocyte proliferation and CD8+ IFN-γ + cells number, but CD4 + IL4 + cell number was higher in intranasal immunization groups. The immunization groups using pLXHDmB7-2 as adjuvant were superior to other groups in immunoresponse. Conclusion These DNA vaccines produce remarkable cellular and humoral immune responses in the mouse and may provide as prophylatic and therapeutic candidates for HPV induced cancer treatment.
基金supported by the National Natural Science Foundation of China(U1703118)Natural Science Foundation of Jiangsu Province(No.BK20181364)+6 种基金Natural Science Foundation of Jiangsu Higher Education Institutions of China(No.19KJA310003)Scientific Research Foundation of Jiangsu health and Health Committee(No.H2018087)a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD),Jiangsu Shuangchuang ProgramOpen Funds of the State Key Laboratory for Chemo/Biosensing and Chemometrics(2016015)Open project of the National Laboratory of Biomacromolecules(2017kf05)the cooperative project between Southeast University and Nanjing Medical University(2018DN0004)Jiangsu Specially-Appointed Professor project,China。
文摘With the increasing immunological studies on camels due to the advantage of their single-chain antibodies for humanizations,it is demanding to develop an easy-to-handle evaluation method of their humoral immune response before proceeding with immunization of foreign antigens that may be toxic to camels.In this study,we quantitatively determined the expression levels of T-helper 2(Th2) cytokines in peripheral blood lymphocytes obtained from Bactrian camels by real-time PCR.The recorded kinetic profiles resulting from the immunization of ovalbumin(OVA) indicated that after immunization,Th2 cytokines including interleukin(IL) families such as IL-4,IL-10,and IL-13 in the camels were up-regulated by a factor of 1.78,3.15,and 1.22,respectively,which was validated by traditional enzyme-linked immunosorbent assay(ELISA) methods.Unlike ELISA which requires specific enzyme-labeled antibodies,this established method based on the minimal amount of blood samples holds an advantage in the preliminary evaluation of camel humoral immune response with desirable precision,which is meaningful for biomedical explorations of camel-derived antibodies.
基金Supported by Science and Technology Development Fund of Tianjin Agricultural University(2013N09)"Veterinary Biological Technology"Innovation Team Project of Colleges and Universities in Tianjin(TD12-5019)+1 种基金Innovative Team Training Program Fund of Colleges and Universities in Tianjin(TNTD2015015)Development Program Leading Education Reform and Innovation of College Teachers at Tianjin Agricultural University(20171003)
文摘[ Objective] The paper was to study the immune enhancement of ATRA on Newcastle Disease (ND) vaccine. [ Method ] The 1-day-old AA broilers were treated with ATRA at the doses of I and 5 p.mol/kg, respectively. At 7 and 28 days of age, broilers in drug control group, low dose group and high dose group were immunized with ND vaccine by intranasal and eye immunization approach. At 7, 14, 21, 28, 35, 42 and 49 clays of age, seven chickens were randomly se- lected from each group and weighed. The thymus, spleen, bursa of fabrieius and serum were collected for calculating immune organ index of thymus, spleen and bursa of fabricius. The ND specific antibody titers in serum were determined with HI test. [Result] ATRA promoted the growth of thymus, spleen and bursa of fabricius, and improved the immune organ index and ND specific antibody titers of chicks. [ Conclusion] ATRA enhanced the humoral immune response of chicks, and ATRT at the dose of 5 μmol/kg presented more prominent immune enhancement effect on ND vaccine.
基金Supported by National Natural Science Foundation of China(31902277)。
文摘[Objectives]The paper was to study the effects of combined application of nonreplicating Toxoplasma uracil auxotrophs(NRTUAs)and Agaricus blazei Murill polysaccharide(ABP)on growth and humoral immunity of chicks.[Methods]A total of 120 one-day old female Hyline brown laying hens were randomly divided into 4 groups,30 hens for each group.The chicks in group 1 were subcutaneously injected with NRTUAs and fed on the diet containing with ABP;the chicks in group 2 were subcutaneously injected with NRTUAs;the chicks in group 3 were subcutaneously injected with equal volume of PBS,and fed on the diet containing with ABP;the chicks in group 4 were subcutaneously injected with equal volume of PBS.The body weight of chicks in each group was counted at the 21^(st),42^(nd),84^(th)and 112^(th)week.During this period,blood samples were collected from chicks in each group at 0,7,14,21,28 and 35 d post immunization against Newcastle disease(ND),and serum was separated to detect the antibody titer of ND.[Results]The combined application of NRTUAs and ABP had no effect on growth of chicks,but promoted the humoral immune response of chicks,significantly improved the ND antibody level of chicks,and could maintain high levels of antibodies in the body for a long time.[Conclusions]The study lays a theoretical foundation for further developing the clinical application of NRTUAs and ABP.
文摘Background:B cells represent a crucial component of adaptive immunity that ensures long-term protection from infection by generating pathogen-specific immunoglobulins.Exercise alters B cell counts and immunoglobulin levels,but evidence-based conclusions on potential benefits for adaptive immunity are lacking.This systematic review assessed current literatures on the impact of acute exercise and exercise training on B cells,immunoglobulins,and markers of secretory immunity in human biofluids.Methods:According to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses(PRISMA)guidelines,MEDLINE,Web of Science,and Embase were searched on March 8,2023.Non-randomized controlled trials and crossover trials investigating the impact of acute exercise or exercise training on B cell counts and proportions,immunoglobulin levels,salivary flow rate,or secretory immunoglobulin A secretion rate were included.Quality and reporting of exercise training studies were assessed using the Tool for the Assessment of Study Quality and reporting in Exercise.Study characteristics,outcome measures,and statistically significant changes were summarized tabularly.Results:Of the 67 eligible studies,22 applied acute exercise and 45 applied exercise training.All included outcomes revealed significant alterations over time in acute exercise and exercise training context,but only a few investigations showed significant differences compared to control conditions.Secretory and plasma immunoglobulin A levels were most consistently increased in response to exercise training.Conclusion:B cell-related outcomes are altered by acute exercise and exercise training,but evidence-based conclusions cannot be drawn with high confidence due to the large heterogeneity in populations and exercise modalities.Well-designed trials with large sample sizes are needed to clarify how exercise shapes B cell-related immunity.
基金the Scientific Research Foundation of Health Bureau of Zhejiang Province (No. 2006c151)
文摘Objective To evaluate the effects of ethyl-acetate fraction (EAF) of extracts from Tetrastigma hemsleyanum Diels et. Gilg (TDG) on immune functions of ICR mice. Methods ICR mice were exposed to different doses of EAF for 15 or 30 days and then their immune functions were analyzed, including ConA-induced splenic lymphocyte transformation, SRBC- induced delayed type hypersensitivity response, serum hemolysin analysis, antibody-producing cells, peritoneal macrophage phagocytized chicken red blood cells, natural killer cell activity, and serum level of cytoldnes. Results EAF of extracts from TDG at different doses had various effects on immune functions of ICR mice. As compared with the controls, it increased the mouse spleen lymphocyte transformation induced by ConA, the left-hind voix pedis thickness and the number of plague forming cells (PFCs) at the dose of 1.82 mg/mL, 5.48 mg/mL, and 9.12 mg/mL, respectively; increased the ink clearance ability at the dose of 0.91 mg/mL, 1.82 mg/mL, 5.48 mg/mL, and 9.12 mg/mL, respectively; increased the phagocytosis index of mononuclear-macrophages and production of serum interferon-gamma (IFN-?) at the dose of 5.48 mg/mL; and could promote the production of serum tumor necrosis factor-alpha (TNF-α) at the dose of 9.12 mg/mL. Conclusion EAF of extracts from TDG can regulate mouse immune functions in vivo.
文摘It is well known that IL4 plays important roles in the induction of humoral immunity. In the present study, the humoral antitumor responses induced by IL4 genemodified tumor vaccine were investigated. The mice were vaccinated with the IL4 genemodified B16 melanoma cells. Then the proliferative capacity of the splenic lymphocytes, the levels of the antibodies in the murine serum against wildtype B16 cells and the cytotoxicity of the serum to wildtype melanoma cells were detected. Our data showed that the LPSinduced proliferation of the splenic lymphocytes from the mice vaccinated with the IL4 genemodified tumor vaccine increased more significantly than that from mice vaccinated with wildtype tumor vaccine. The cytotoxicity of the serum to wildtype melanoma cells also increased markedly when detected. It was also observed that the number of pulmonary metastases decreased more obviously when the mice were intravenously injected with the mixture of wildtype B16 cells and the serum from the mice vaccinated with the IL4 genemodified B16 cells. Our data demonstrated that humoral immunity might contribute to the antitumor effect of IL4 gene therapy.
基金ThisresearchwassupportedbytheNationalNaturalScienceFoundationofChina(No.39770667),theScience&TechnologyDevelopmentFoundationofShaanxiProvince(No.2001SM82)and china and belgium cooperated study
文摘Objective To study the humoral immunity status and distribution pattern of lymphocyte subgroups of peripheral blood mononuclear cell (PBMC) in patients with Kashin-Beck Disease (KBD), and their relationship with erythrocyte selenium. Methods 23 X-ray diagnosed patients, 22 age- and sex- matched healthy children in KBD affected area (KAA), and 25 in KBD non-affected area (KNAA) were randomly selected. Immunohistochemistry with monoclonal antibodies anti-CD4, anti-CD8, anti-CD20 was conducted to analyze the lymphocyte subsets. Serum IgM, IgA, IgG, Complement C3 and C4 were assayed using rate nephelometry (Array 360 System, USA). The contents of erythrocyte selenium was determined by 2,3-diaminonaphthalene fluorescence assay. Results CD4+ and CD8+ cells percentage in PBMCs and serum IgA were significantly lower in KAA than those in KNAA(P< 0.05). CD20+ percentage in KAA displayed a decreasing trend compared to KNAA, although not statistically significantly. No statistical differences were found in CD4/CD8 ratio, serum IgG, IgM, C3 and C4 levels. Erythrocyte selenium level in KAA still showed a pronounced decrease compared to that in KNAA. Correlation analysis showed that erythrocyte selenium contents had a strong association with the CD4 cell percentage (r= 0.625, P< 0.05), as well as serum IgA (r= 0.462, P< 0.05). In addition, a moderate correlation between the serum IgA and CD4+ percentage (r= 0.130, P> 0.05) was found. Conclusion These results suggested that children in KAA had a comparably low cellular immunity level and their humoral immunity status was also in a state of moderate immune suppression. Of this immune disorder in Kashin-Beck disease patients, selenium deficiency probably played a critical role via affecting the distribution pattern of peripheral blood lymphocyte. Selenium-deficiency and immune impairment maybe both have something to do with the cause-effect chain of KBD.
基金Supported by the 333 Project of Jiangsu Province(BRA2016102)Natural Science Foundation of Tibet(ZJ2013018)Phoenix Talent Engineering of Jiangsu Agri-animal Husbandry Vocational College
文摘[ Objective ] The paper was to screen the active ingredients of traditional Chinese medicine ( TCM), which significantly enhanced the immune effect of porcine circovirus type 2 (PCV2) vaccine. [Method]Sixty-six 14-day-old piglets were randomly divided into 11 groups, including immune control group, blank control group and nine drug groups. Piglets that were reared for 21 d were intramuscularly injected with 2 mL of inactivated vaccine in cervical region expect for blank control group. Piglets in nine drug groups were administrated with 5 mL of matrine, scutellarin and astragalus polysaecharide (ASP) at three doses ( high dose 12 mg, medium dose 8 nag, low dose 4 mg) three days before and after immunization; piglets in blank control group and irmnune control group were administrated with equal volume of normal saline. Five piglets were randomly selected from each group to collect venous blood at 7, 14, 21 and 28 d post immunization, and the PCV2 antibody level, the concentration of specific immunoglobulin [gG, the changes in peripheral blood lymphocyte subsets, and the content of serum IL-2 and IFN-γ were determined. [ Result] The immunological effects of active ingredients of three TCM on PCV2 vaccine all enhanced at different degree. The effects of scutellarin middle dose group was the best. [ Conclusion ] Scutellarin could be used as a candidate drug for PCV2 vaccine immunoenhancer.
文摘Salmonella is a ubiquitous pathogen which, in addition to causing poultry diseases, has a growing zoonotic impact. It has demanded the implementation of diverse control strategies, in which vaccines play a major role. The understanding of the immune pathways elicited by the different vaccines is important, contributing for the establishment of strong immune correlates of protection, for instance. With the purpose of determining the dynamics of the humoral and cellular immune responses to vaccination, broiler breeders (Cobb Slow) were immunized with live or inactivated vaccines against Salmonella Enteritidis. Lymphocyte and macrophage subsets were analyzed in the peripheral blood by flow cytometry and antigen-specific circulating IgY and mucosal IgA were quantified. The markers analyzed by flow cytometry were CD8/CD28, CD4/TCRVβ1, Kul/ MHC II and Bu-1. Both live and inactivated vaccines induced an increase in the proportion of circulating monocytes (Kul+MHCII+) in some time points compared to non-vaccinated controls. However, whereas the live vaccine leads to an increase in CD8-CD28+ and Bu-1+ lymphocytescompared to the control group, the inactivated vaccine prompteda reduction in the percentage of severalleucocyte subsets (Kul-MHCII+, Bu-1+, CD8+CD28+, CD8-CD28+, CD4+TCRVβ1-, CD4+TCRVβ1+, CD4-TCRVβ1+) after the boost dose. Both vaccines induced specific serum IgY and mucosal IgA production;however, the inactivated vaccine stimulated higher titers in a shorter period. These results contribute to the understanding of mechanisms of action of live and inactivated Salmonella vaccines in chickens.
基金supported by the National Key Research and Development Program of China(2021YFA1100700)the National Natural Science Foundation of China(81941008)+3 种基金the Natural Science Foundation of Jilin Province,China(20200201191JC)the Department of Human Resource and Social Security of Jilin Province(2022DJ02)the Science Development of Jilin Province,China(20230505029ZP)the Bethune Medical Department of Jilin University(2022JBGS01)。
文摘Humanized immune system(HIS)mice have been developed and used as a small surrogate model to study human immune function under normal or disease conditions.Although variations are found between models,most HIS mice show robust human T cell responses.However,there has been unsuccessful in constructing HIS mice that produce high-affinity human antibodies,primarily due to defects in terminal B cell differentiation,antibody affinity maturation,and development of primary follicles and germinal centers.In this review,we elaborate on the current knowledge about and previous attempts to improve human B cell development in HIS mice,and propose a potential strategy for constructing HIS mice with improved humoral immunity by transplantation of human follicular dendritic cells(FDCs)to facilitate the development of secondary follicles.
基金This study was supported by a grant from the National Natural Science Foundation of China (No. 30471959 and 30571940).
文摘Background We have previously developed and characterized a monoclonal anti-idiotype antibody, designated 6B 11, which mimics an ovarian carcinoma associated antigen OC166-9 and whose corresponding monoclonal antibody is COC166-9 (Abl). In this study, we evaluate the humoral immune responses induced by the fusion protein 6B11 single-chain variable fragment (scFv)/human granulocyte macrophage colony-stimulating factor (hGM-CSF) and 6B 1 lscFv in BALB/c mice. Methods The fusion protein 6B 11 scFv/hGM-CSF was constructed by fusing a recombinant single-chain variable fragment of 6B11scFv to GM-CSE BALB/c mice were administrated by 6B11scFv/hGM-CSF and 6B11scFv, respectively. Results The fusion protein 6B11scFv/hGM-CSF retained binding to the anti-mouse F(ab)2' and was also biologically active as measured by proliferation of human GM-CSF dependent cell TF1 in vitro. After immunization with the 6B11scFv/hGM-CSF and 6BllScFv, BALB/c mice showed significantly enhanced Ab3 antibody responses to 6B11 scFv/hGM-CSF compared with the 6B11 scFv alone. The level of Ab3 was the highest after the first week and maintained for five weeks after the last immunization. Another booster was given when the Ab3 titer descended, and it would reach to the high level in a week. Conclusion The fusion protein 6B11scFv/hGM-CSF can induce humoral immunity against ovarian carcinoma in vivo. We also provide the theoretical foundation for the application of the fusion protein 6B11 scFv/hGM-CSF for active immunotherapy of ovarian cancer.
基金This work was supported by the financial support of the PI17CIII/00045 and PI20CIII/00019 grants from the AES-ISCIII program to R.B.The financial support of the PID2019-103899RB-I00(Ministerio de Ciencia e Innovación)Research Project and the TRANSNANOAVANSENS-CM Program from the Comunidad de Madrid(S2018/NMT-4349)to S.C.are gratefully acknowledged.+3 种基金G.D.acknowledges the financial support of PI15/00246 grant of the FIS and Cátedra UAM-Roche en Medicina de InnovaciónThe FPU predoctoral contract to A.M.-C.is supported by the Spanish Ministerio de Educación,Cultura y Deporte.G.S.-F.is recipient of a predoctoral contract(1193818 N)supported by The Flanders Research Foundation(FWO).M.A.-N.was supported by a contract of the Programa Operativo de Empleo Juvenily la Iniciativa de Empleo Juvenil(YEI)with the participation of the Consejería de Educación,Juventud y Deporte de la Comunidad de Madrid y del Fondo Social EuropeoThe predoctoral contract from the Spanish Ministerio de Economía y Competitividad(BES-2016-076606,E.P.)Talento-Contract from Comunidad de Madrid(2019-T2/IND-15965,R.M.T.-R.)are also gratefully acknowledged.
文摘Colorectal cancer(CRC)is the second leading cause of cancer-related death worldwide.The five-year survival rate of CRC patients depends on the stage at diagnosis,being higher than 80%when CRC is diagnosed in the early stages but lower than 10%when CRC is diagnosed in advanced stages.Autoantibodies against specific CRC autoantigens(tumor-associated antigens(TAAs))in the sera of patients have been widely demonstrated to aid in early diagnosis.Thus,we herein aim to identify autoantigens target of autoantibodies specific to CRC that possess a significant ability to discriminate between CRC patients and healthy individuals by means of liquid biopsy.To that end,we examined the protein content of the exosomes released by five CRC cell lines and tissue samples from CRC patients by means of immunoprecipitation coupled with mass spectrometry analysis.A total of 103 proteins were identified as potential autoantigens specific to CRC.After bioinformatics and meta-analysis,we selected 15 proteins that are more likely to be actual CRC autoantigens in order to evaluate their role in CRC prognosis by Western blot(WB)and immunohistochemistry(IHC).We found dysregulation at the protein level for 11 of these proteins in both tissue and plasma exosome samples from patients,along with an association of nine of these proteins with CRC prognosis.After validation,all but one showed a statistically significant high diagnostic ability to distinguish CRC patients and individuals with premalignant lesions from healthy individuals,either by luminescence Halotag-based beads,or by a multiplexed biosensing platform involving the use of magnetic microcarriers as solid support modified with covalently immobilized Halotag fusion proteins constructed for CRC detection.Taken together,our results highlight the usefulness of the approach defined here to identify the TAAs specific to chronic diseases;they also demonstrate that the measurement of autoantibody levels in plasma against the TAAs identified here could be integrated into a point-of-care(POC)device for CRC detection with high diagnostic ability.
基金by Key Research and Development Projects of the Ministry of Science and Technology,China,No.2017YFC0113901.
文摘BACKGROUND Helicobacter pylori(H.pylori)has been found to be associated with extragastrointestinal diseases,possibly including adverse food reactions(such as food allergy or intolerance).However,there are few studies on H.pylori and food allergy or intolerance,and the results are inconsistent.Food-specific immunoglobulin(Ig)G has been revealed to be associated with food allergy or intolerance and can be used as a marker to explore the correlation between H.pylori infection and food allergy or intolerance.AIM To explore the relationship between H.pylori infection and food-specific IgG METHODS We retrospectively analyzed the physical examination data of 21822 subjects from February 2014 to December 2018 in this study.H.pylori infection was detected using the 13C urea breath test.Food-specific IgG of eggs,milk and wheat in serum was assessed.Subjects were grouped according to H.pylori positivity,and the positive rates of three kinds of food-specific IgG were compared between the two groups.Multivariable logistic regression analysis was performed to elucidate the association between H.pylori infection and food-specific IgG.RESULTS The total infection rate of H.pylori was 39.3%,and the total food-specific IgGpositive rates of eggs,milk and wheat were 25.2%,9.0%and 4.9%,respectively.The infection rate of H.pylori was higher in males than in females,while the positive rates of food-specific IgG were lower in males than in females.The positive rates of food-specific IgG decreased with age in both males and females.In the H.pylori-positive groups,the positive rates of food-specific IgG of eggs,milk and wheat were all lower than those in the H.pylori-negative groups.Multivariate logistic regression analysis revealed that H.pylori infection was negatively correlated with the food-specific IgG-positive rates of eggs,milk and wheat(odds ratio value of eggs 0.844-0.873,milk 0.741-0.751 and wheat 0.755-0.788,in different models).CONCLUSION H.pylori infection was found to be negatively associated with the food-specific IgG of eggs,milk and wheat in Southwest China.
基金Supported by the WU Jie-ping Medical Foundation of Ministry of Health, China(No320675007127)
文摘To explore the primary humoral and cellular immunological mechanism of the combined hepatitis A-measles-varicella vaccine, the mice were inoculated with hepatitis A-measles-varicella vaccine by intraperitoneally and two weeks later, blood was collected to observe the mice's immunological status. Antibody level was measured to appraise the humoral immunity. At the same time, T lymphocyte surface marker, NK cell activity, LAK cell activity, delayed type hypersensitivity of skin, Mφ phagocytic function, mRNA level of cytokine IL-2 and IFN-γ plus lymphocyte transformation test were used to analyze the cellular immunity. The humoral immunity results show that the combined hepatitis A-measles-varicella vaccine produce the same antibody level as their corresponding univalent vaccine, and maintained fine immunogenicity and security. The result of cellular immunity shows that the combined vaccine could activate physical immunocyte, increase the regulative ability of cytokine, enhance the physical immune function and immune defense ability. The present research proved the security and better humoral and cellular immunity of combined hepatitis A-measles-varicella vaccine from the immunological point of view, which laid good foundation for further study and development.
基金the project of the National Key R&D Program of China(2017YFE0122000)the National Natural Science Foundation of China(31871968)the Natural Science Foundation of Fujian Province,China(2018J01614).
文摘C-type lectins(CTLs)are a superfamily of Ca^(2+)-dependent carbohydrate-recognition proteins,and an important pattern recognition receptor(PRR)in insect innate immunity which can mediate humoral and cellular immunity in insects.In this study,we report a novel dual carbohydrate-recognition domain(CRD)CTL from Plutella xylostella which we designate PxIML.PxIML is a protein with a 969 bp open reading frame(ORF)encoding 322 amino acids,containing a signal peptide and a dual-CRD with EPN(Glu_(124)-Pro_(125)-Asn_(126))and QPD(Gln_(274)-Pro_(275)-Asp_(276))motifs.The expression of PxIML mRNA in the fat body was significantly higher than in hemocytes and midgut.The relative expression levels of PxIML in the whole insect and the fat body were significantly inhibited after infection with Bacillus thuringiensis 8010(Bt8010)at 18 h,while they were significantly upregulated after infection with Serratia marcescens IAE6 or Pichia pastoris.The recombinant PxIML(rPxIML)protein could bind to the tested pathogen-associated molecular patterns(PAMPs),and the bacteria of Enterobacter sp.IAE5,S.marcescens IAE6,Staphylococcus aureus,Escherichia coli BL21,and Bt8010 in a Ca^(2+)-dependent manner,however,it showed limited binding to the fungus,P.pastoris.The rPxIML exhibited strong activity in the presence of Ca^(2+) to agglutinate Bt8010,Enterobacter sp.IAE5 and S.aureus,but it only weakly agglutinated with E.coli BL21,and could not agglutinate with S.marcescens IAE6 or P.pastoris.Furthermore,the rPxIML could bind to hemocytes,promote the adsorption of hemocytes to beads,and enhance the phenoloxidase(PO)activity and melanization of P.xylostella.Our results suggest that PxIML plays an important role in pathogen recognition and in mediating subsequent humoral and cellular immunity of P.xylostella.
基金funded by the Technology Programme Project of Zhongshan City (China) (20073A159)the Open Project of the Key Laboratory of Animal Nutrition and Feed of Yunnan Province
文摘[ Objective] To study the effects of different oligosaccharides on immunity and production performance of piglets.[ Method] A total of 75 Yorkshire x Rongchang piglets were assigned into five groups, 15 in each group. Isomalto-oligosaccharide ( IMO), fructo-oligosaccharide ( FOS), mannan-oligosacchadde (MOS), and mixed oligosacchadde were supplemented to basal diet (7.5 g/kg), respectively. The control group was also set. The experimental period lasted for 53 d. The clinic symptoms and diarrhea were observed. All piglets were weighed once a week. Feed intake was also recorded. Blood was collected via precaval vein for determination of immunology indexes at the age of 30 and 60 d, respectively. [ Result] The IMO and FOS enhanced cellular immunity of 30-day-old piglets significantly (P 〈 0.05), and the IMO also enhanced their humoral immunity. The MOS enhanced cellular immunity and humoral immunity of 60-day-old piglets significantly (P 〈 0.05). It also improved production performance largest, and its diarrhea rate was the lowest. [ Conclusion] Different oligosaccharides have various effects on immunity and production performance of oiolets.
基金supported by the National Natural Science Foundation of China(Nos.81901878,U1804183,and 82202318)Key Scientific Research Project(Education Department of Henan Province,China,20HASTIT049)China Postdoctoral Science Foundation(2019M662553,2020T130611,and 2022TQ0310)。
文摘Activating humoral and cellular immunity in lymph nodes(LNs)of nanoparticle-based vaccines is critical to controlling tumors.However,how the physical properties of nanovaccine carriers orchestrate antigen capture,lymphatic delivery,antigen presentation and immune response in LNs is largely unclear.Here,we manufactured gold nanoparticles(AuNPs)with the same size but different shapes(cages,rods,and stars),and loaded tumor antigen as nanovaccines to explore their disparate characters on above four areas.Results revealed that star-shaped AuNPs captured and retained more repetitive antigen epitopes.On lymphatic delivery,both rods and star-shaped nanovaccines mainly drain into the LN follicles region while cage-shaped showed stronger paracortex retention.A surprising finding is that the star-shaped nanovaccines elicited potent humoral immunity,which is mediated by CD4^(+)T helper cell and follicle B cell cooperation significantly preventing tumor growth in the prophylactic study.Interestingly,cage-shaped nanovaccines preferentially presented peptide-MHC I complexes to evoke robust CD8^(+)T cell immunity and showed the strongest therapeutic efficacy when combined with the PD-1 checkpoint inhibitor in established tumor study.These results highlight the importance of nanoparticle shape on antigen delivery and presentation for immune response in LNs,and our findings support the notion that different design strategies are required for prophylactic and therapeutic vaccines.
基金supported by the National Natural Science Foundation of China,No.30471927
文摘Three-month-old Alzheimer's disease model transgenic mice were immunized with Aβ1-42, Plp-Adenovirus [Ad]-X-CMV-(Aβ3-10)lo-CpG [AdCpG-(Aβ3-10)1] or AdCpG virus fluid via na- sal mucosal inhalation, respectively. ELISA analysis of serum showed Aβ42 antibody titers were significantly increased in mice immunized with Aβ1-42 and AdCpG-(Aβ3-10)10. Concanavalin A and AdCpG-(Aβ3-10)10 stimulation significantly increased the number of proliferating spleen cells cultured from AdCpG(Aβ3-10)Io and Aβ42 groups compared with the control group. In the AdCp- G(Aβ3-10)10 group, levels of interleukin (IL)-4 and IL-10 were increased, while those of IL-2 and interferon-y were decreased. In the A[342 group, levels of IL-4, IL-10, IL-2 and interferon-y were all increased. Experimental findings indicate that AdCpG-(Aβ3-10)10 vaccine can produce strong T helper 2 (Th2) humoral immune responses in addition to the production of Aβ42 antibody. The cellular immunologic response was weak and avoided Aβ1-42-mediated cytotoxicity.