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1.
PeerJ ; 12: e17143, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38618563

RESUMO

Phage display technology has become an important research tool in biological research, fundamentally changing the traditional monoclonal antibody preparation process, and has been widely used in the establishment of antigen-antibody libraries, drug design, vaccine research, pathogen detection, gene therapy, antigenic epitope research, and cellular signal transduction research.The phage display is a powerful platform for technology development. Using phage display technology, single chain fragment variable (scFv) can be screened, replacing the disadvantage of the large size of traditional antibodies. Phage display single chain antibody libraries have significant biological implications. Here we describe the types of antibodies, including chimeric antibodies, bispecific antibodies, and scFvs. In addition, we describe the phage display system, phage display single chain antibody libraries, screening of specific antibodies by phage libraries and the application of phage libraries.


Assuntos
Anticorpos Biespecíficos , Bacteriófagos , Anticorpos de Cadeia Única , Anticorpos de Cadeia Única/genética , Anticorpos Monoclonais , Bacteriófagos/genética , Tecnologia
2.
Curr Drug Deliv ; 2023 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-37818569

RESUMO

Immunomodulatory mechanisms are indispensable and key factors in maintaining the balance of the environment in humans. When the immune function of the immune system is impaired, autoimmune diseases occur. Excessive body fatigue, natural aging of the human body, malnutrition, genetic factors and other reasons cause low immune function, due to which the body is prone to being infected by bacteria or cancer. Clinically, the existing therapeutic drugs still have problems such as high toxicity, long treatment cycle, drug resistance and high price, so we still need to explore and develop a high efficiency and low toxicity drug. Poly(lactic-co-glycolic acid) (PLGA) refers to a non-toxic polymer compound that exhibits excellent biocompatibility. Traditional Chinese medicine (TCM) monomers come from natural plants, and have the characteristics of high efficiency and low toxicity. Applying PLGA to TCM monomers can make up for the defects of traditional dosage forms, improve bioavailability, reduce the frequency and dosage of drug use, and reduce toxicity and side effects, thus having the characteristics of sustained release and targeting. Accordingly, PLGA nanoparticles loaded with TCM monomers have been the focus of development. The previous research on drug loading advantages, preparation methods, and immune regulation of TCM PLGA nanoparticles is summarized in the following sections.

3.
PeerJ ; 11: e15790, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37605748

RESUMO

Gene knockout is a widely used method in biology for investigating gene function. Several technologies are available for gene knockout, including zinc-finger nuclease technology (ZFN), suicide plasmid vector systems, transcription activator-like effector protein nuclease technology (TALEN), Red homologous recombination technology, CRISPR/Cas, and others. Of these, Red homologous recombination technology, CRISPR/Cas9 technology, and suicide plasmid vector systems have been the most extensively used for knocking out bacterial drug resistance genes. These three technologies have been shown to yield significant results in researching bacterial gene functions in numerous studies. This study provides an overview of current gene knockout methods that are effective for genetic drug resistance testing in bacteria. The study aims to serve as a reference for selecting appropriate techniques.


Assuntos
Traumatismos Craniocerebrais , Farmacorresistência Bacteriana/genética , Técnicas de Inativação de Genes , Genes Bacterianos , Tecnologia , Nucleases dos Efetores Semelhantes a Ativadores de Transcrição , Nucleases de Dedos de Zinco , Animais
4.
Biomedicines ; 11(6)2023 Jun 09.
Artigo em Inglês | MEDLINE | ID: mdl-37371768

RESUMO

Dendritic cells are the only antigen-presenting cells capable of activating naive T cells in humans and mammals and are the most effective antigen-presenting cells. With deepening research, it has been found that dendritic cells have many subsets, and the surface receptors of each subset are different. Specific receptors targeting different subsets of DCs will cause different immune responses. At present, DC-targeted research plays an important role in the treatment and prevention of dozens of related diseases in the clinic. This article focuses on the current status of DC surface receptors and targeted applications.

5.
Clin Implant Dent Relat Res ; 25(3): 458-472, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37042239

RESUMO

BACKGROUND: We have modified the internal sinus elevation by combining it with the sinus mucoperiosteum stripping procedure, which further increases the indications for the internal lift. Similar long-term clinical follow-up studies and three-dimensional finite element analyses are rare. OBJECTIVE: This study aimed to investigate the feasibility of the modified internal sinus floor elevation method in patients with low residual bone height using a three-dimensional (3D) finite element model and report on the long-term outcomes. MATERIALS AND METHODS: Overall, 99 implants were placed in 86 patients. All patients were followed-up for 3-24 months. The modified internal sinus floor elevation was dynamically simulated using a 3D finite element model, and the stress of the sinus membrane was measured. RESULTS: In trial group A (modified internal sinus floor elevation group), 57 implants were placed in 52 patients. The sinus floor height was lifted by 6.5 mm (95%confidence interval (CI): 6.2-6.8). The perforation rate was 8.8%, and the implant survival rate was 96.5%. In control group B (external sinus floor elevation group), 42 implants were placed in 34 patients. The sinus floor height was lifted by 8.8 mm (95%CI: 8.4-9.3). The perforation rate was 14.3%, and the implant survival rate was 100%. In trial group A, compared with the control group B, perforation decreased by 5.5% (odds ratio = 0.50 and 95%CI: 0.14-1.78; p = 0.282), and the sinus floor lift height was 2.3 mm lower (95%CI, 1.8-2.9; p < 0.001). The finite element analysis showed that the peak stress of the sinus membrane increased with an increase in height elevation and degree of membrane separation. CONCLUSION: Our findings indicate the positive clinical outcomes in patients with low RBH associated with the modified internal sinus elevation procedure.


Assuntos
Implantes Dentários , Levantamento do Assoalho do Seio Maxilar , Humanos , Implantação Dentária Endóssea/métodos , Maxila/cirurgia , Seio Maxilar/cirurgia , Estudos Retrospectivos , Levantamento do Assoalho do Seio Maxilar/métodos , Resultado do Tratamento
6.
Iran J Immunol ; 20(1): 57-66, 2023 03 14.
Artigo em Inglês | MEDLINE | ID: mdl-36932950

RESUMO

Background: Staphylococcus aureus is an opportunistic pathogen responsible for various infections with diverse clinical presentation and severity. The α-hemolysin is a major virulence factor in the pathogenesis of S. aureus infections. Objective: To produce a chimeric fusion protein for hemolytic detection of the S. aureus isolates and as a component of a multi-antigen vaccine. Methods: The fused strategy employed a flexible linker to incorporate the possible B cell and T cell determinants into one chimera (HlaD). The humoral and cellular response to the HlaD in mice was assessed to reveal a non-significant difference compared with the full-length α-hemolysin mutant (Hla H35L). Results: The results of the protective effect, the mimetic lung cell injury, and bacterial clearness demonstrated that the mice vaccinated with the HlaD alleviated the severity of the infection of the S. aureus, and the HlaD could similarly function with Hla H35L. Conclusion: The chimeric fusion (HlaD) provided a diagnostic antigen for hemolysis of the S. aureus strains and a potential vaccine component.


Assuntos
Infecções Estafilocócicas , Staphylococcus aureus , Animais , Camundongos , Staphylococcus aureus/metabolismo , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/metabolismo , Pulmão/metabolismo , Fatores de Virulência/metabolismo
7.
Chem Biol Drug Des ; 101(1): 131-137, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36198093

RESUMO

Gossypin is a flavonoid compound prepared from chinese medicine Hibiscus, which not only has significant pharmacological activities in antioxidant, anti-inflammatory, neuroprotective, anti-cancer, anti-tumor, and anti-diabetic aspects, but also has the advantages of small side effects and easy preparation because it is extracted from traditional chinese medicine, so it has received widespread attention from scholars and researchers. This paper reviews the pharmacological effects and mechanisms of gossypin in recent years, and hopes to provide a theoretical basis for its clinical application.


Assuntos
Anti-Inflamatórios , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico
8.
J Microbiol Biotechnol ; 32(12): 1506-1514, 2022 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-36377198

RESUMO

Allergic contact dermatitis (ACD) is an allergen-specific T-cell-mediated inflammatory response, albeit with unclear pathogenesis. Exosomes are nanoscale extracellular vesicles secreted by several cell types and widely distributed in various biological fluids. Exosomes affect the occurrence and development of ACD through immunoregulation among other ways. Nevertheless, the role of exosomes in ACD warrants further clarification. This review examines the progress of research into exosomes and their involvement in the pathogenesis, diagnosis, and treatment of ACD and provides ideas for exploring new diagnostic and treatment methods for this disease.


Assuntos
Dermatite Alérgica de Contato , Exossomos , Vesículas Extracelulares , Humanos , Exossomos/patologia , Dermatite Alérgica de Contato/diagnóstico , Dermatite Alérgica de Contato/patologia , Alérgenos , Linfócitos T
9.
Comput Math Methods Med ; 2022: 9222541, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35437448

RESUMO

Background: Allergic contact dermatitis (ACD) is a form of chronic cutaneous inflammatory disease of immunological origin that has adverse impacts on patient quality of life, underscoring the need for the development of safe and effective therapeutic agents to treat affected individuals. Fisetin is a Chinese herbal preparation that reportedly exhibits antitumor, antioxidant, antimicrobial, anticoagulatory, and antimalarial activity. In the current report, the immunomodulatory activity of fisetin was appraised by assessing its impact on balance between regulatory T (Treg) and Th17 cells in an ACD model. Methods: BALB/c mice (n = 60) were randomized into control, ACD model, CTX positive control (20 mg/kg), and fisetin treatment groups (three dose levels: 2, 4, or 8 mg/kg). ACD induction was achieved by sensitizing mice on the shaved ventral abdomen via the application of 5% DNFB (50 µL) on days 1 and 2, followed by rechallenge in the right ear with 5% DNFB (20 µL) on day 5. Beginning on day 1, immunized mice were intraperitoneally injected with the appropriate fisetin dose (in saline) once per day for 7 days. On day 7, ear swelling, transcription factor expression, Th17/Treg cell populations, and cytokine production were assessed in vivo. Results: Fisetin treatment significantly suppressed ear swelling and associated inflammatory cell infiltration, besides reducing the production of Th17 cytokines (IL-17, TNF-α, and IL-6) and the expression of the Th17 lineage transcription factor RORγt while simultaneously enhancing Treg-specific cytokine production (TGF-ß and IL-10) and the expression of the Treg lineage transcription factor Foxp3, thereby restoring the Th17/Treg cell in ACD mice. Conclusions: These data indicate that fisetin exhibits immunomodulatory activity and can alter the Th17/Treg cell balance, highlighting its potential value as a treatment drug for ACD.


Assuntos
Dermatite Alérgica de Contato , Linfócitos T Reguladores , Animais , Citocinas , Dinitrofluorbenzeno/farmacologia , Flavonóis , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Qualidade de Vida , Células Th17 , Fatores de Transcrição
10.
J Microbiol Biotechnol ; 31(5): 667-675, 2021 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-33879639

RESUMO

Streptococcus agalactiae is an important bacterial pathogen and causative agent of diseases including neonatal sepsis and meningitis, as well as infections in healthy adults and pregnant women. Although antibiotic treatments effectively relieve symptoms, the emergence and transmission of multidrug-resistant strains indicate the need for an effective immunotherapy. Effector T helper (Th) 17 cells are a relatively newly discovered subpopulation of helper CD4+ T lymphocytes, and which, by expressing interleukin (IL)-17A, play crucial roles in host defenses against a variety of pathogens, including bacteria and viruses. However, whether S. agalactiae infection can induce the differentiation of CD4+ T cells into Th17 cells, and whether IL-17A can play an effective role against S. agalactiae infections, are still unclear. In this study, we analyzed the responses of CD4+ T cells and their defensive effects after S. agalactiae infection. The results showed that S. agalactiae infection induces not only the formation of Th1 cells expressing interferon (IFN)-γ, but also the differentiation of mouse splenic CD4+ T cells into Th17 cells, which highly express IL-17A. In addition, the bacterial load of S. agalactiae was significantly increased and decreased in organs as determined by antibody neutralization and IL-17A addition experiments, respectively. The results confirmed that IL-17A is required by the host to defend against S. agalactiae and that it plays an important role in effectively eliminating S. agalactiae. Our findings therefore prompt us to adopt effective methods to regulate the expression of IL-17A as a potent strategy for the prevention and treatment of S. agalactiae infection.


Assuntos
Interleucina-17/imunologia , Infecções Estreptocócicas/imunologia , Streptococcus agalactiae/fisiologia , Células Th17/imunologia , Animais , Carga Bacteriana/efeitos dos fármacos , Carga Bacteriana/imunologia , Linfócitos T CD4-Positivos/imunologia , Diferenciação Celular , Proliferação de Células , Citocinas/imunologia , Feminino , Interleucina-17/administração & dosagem , Interleucina-17/antagonistas & inibidores , Camundongos , Baço/imunologia , Baço/microbiologia , Infecções Estreptocócicas/microbiologia , Células Th1/imunologia
11.
Res Vet Sci ; 136: 422-429, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33812285

RESUMO

Glyceraldehyde-3-phosphate dehydrogenase C (GapC) of Streptococcus dysgalactiae (S. dysgalactiae) is a highly conserved surface protein that can induce a protective immune response against S. dysgalactiae infection. To investigate the immune response and protective efficacy induced by epitope-vaccines against S. dysgalactiae infection, we constructed epitope-vaccines GTB1, GB1B2, and GTB1B2 using a T cell epitope (GapC63-77, abbreviated as GT) and two B cell epitopes (GapC30-36, abbreviated as GB1, and GapC97-103, abbreviated as GB2), which were identified in GapC1-150 of S. dysgalactiae in tandem by a GSGSGS linker. BALB/c mice were immunized via an intramuscular injection with the epitope vaccines. The levels of the cytokines, IFN-γ, IL-4, and IL-17, secreted by splenic lymphocytes and the antibody levels in the sera of the immunized mice were detected by ELISA. The immunized mice were subsequently challenged with S. dysgalactiae, and the bacterial colonization in the immunized-mouse organs was examined using the plate counting method. The results showed that the level of the cytokines induced by GTB1B2 was lower than that induced by GapC1-150, but higher than that induced by other epitope vaccines. The level of IgG induced by GTB1B2 was lower than that induced by GapC1-150, but higher than the levels induced by other epitope vaccines. The bacterial colonization numbers in the organs of the mice immunized with GTB1B2 were higher those of the mice immunized with GapC1-150, but significantly lower than those from the mice immunized with other epitope-vaccines. Our results demonstrated that the T cell and B cell epitopes in the epitope-vaccines worked synergistically against bacterial challenge. The multi-epitope vaccine, GTB1B2, could induce stronger cellular and humoral immune responses, and provide a better protective effect against S. dysgalactiae infection.


Assuntos
Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Imunogenicidade da Vacina , Vacinas Estreptocócicas/imunologia , Streptococcus/imunologia , Animais , Citocinas/imunologia , Ensaio de Imunoadsorção Enzimática/veterinária , Epitopos de Linfócito B/imunologia , Epitopos de Linfócito T/imunologia , Feminino , Camundongos , Camundongos Endogâmicos BALB C
12.
Immunopharmacol Immunotoxicol ; 43(2): 171-175, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33491523

RESUMO

INTRODUCTION: Most of the immunosuppressive drugs are used for the treatment of autoimmune disease, allergic diseases, and transplant rejection, but toxicity is the major obstacle for the potent drugs in the wide use of these immunosuppressive drugs. Daphnetin, a Chinese herbal product, has been reported that daphnetin possesses antimicrobial, anticoagulation, antimalarial, anticancer, and antioxidant activity. In a previous study, we found that daphnetin exhibited a potential immunosuppressive effect on LPS-induced B lymphocyte cells in vitro, therefore, in this research, we investigated the immunosuppressive effects of daphnetin in BALB/c mice use OVA as a prototype antigen. METHODS: Sixty BALB/c mice were divided into six groups. The emulsion (100 µL containing 100 µg OVA) was injected subcutaneously with OVA + CFA into the shaved backs of the BALB/c mice on day 1, and a boosting injection was administered in OVA + IFA 2 weeks later. Beginning on the day of immunization, the immunized mice were administered intraperitoneally with daphnetin at a dose of 5, 10, and 20 mg/kg in saline solution for 28 consecutive days. We measured the effect of daphnetin on OVA-specific antibody, cytokine production, and Splenocyte proliferation in vivo. RESULTS: The results revealed that daphnetin significantly suppressed serum immunoglobulin G levels (IgG), and the OVA-specific IgG subclasses IgG1 and IgG2b, daphnetin was also significantly decreased the Th1 and Th2 cytokine productions, inhibited the splenocytes proliferation rate in vivo. CONCLUSIONS: It proved that daphnetin could suppress humoral response activity on OVA-sensitized mice, suggesting a potential role on daphnetin as a new immunosuppressive drug.


Assuntos
Imunidade Humoral/efeitos dos fármacos , Imunidade Humoral/imunologia , Imunossupressores/farmacologia , Ovalbumina/toxicidade , Umbeliferonas/farmacologia , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Injeções Subcutâneas , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/administração & dosagem , Baço/efeitos dos fármacos , Baço/imunologia
13.
Microb Pathog ; 144: 104167, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32222538

RESUMO

Glyceraldehyde-3-phosphate dehydrogenase-C (GapC) is a highly conserved surface protein of Staphylococcus aureus, with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity, which represents an excellent vaccine candidate antigen. It can induce protective immune responses to S. aureus infections. However, CD4+ T cell epitopes of GapC that induce CD4+ T cell immune responses are currently unclear. In this study, we used bioinformatics prediction algorithms to predict CD4+ T cell epitopes of GapC. Ten peptides were synthesized to investigate the candidate epitopes. Our results showed that the peptides, G4 (GapC 104-123) and G10 (GapC 314-333) were able to induce proliferation of CD4+ T cells and secrete high levels of interferon (IFN)-γ, respectively. In addition, they significantly reduced bacterial loads in tissue and induced immunoprotective effects. It is suggested that G4 and G10 are Th1-type epitopes of S. aureus GapC. This study provides the potential development of the design of epitope-based vaccine against S. aureus.


Assuntos
Anticorpos Antibacterianos/imunologia , Linfócitos T CD4-Positivos/imunologia , Epitopos de Linfócito T/imunologia , Gliceraldeído-3-Fosfato Desidrogenases/imunologia , Staphylococcus aureus/imunologia , Algoritmos , Animais , Carga Bacteriana/imunologia , Vacinas Bacterianas/imunologia , Proliferação de Células/fisiologia , Biologia Computacional , Feminino , Gliceraldeído-3-Fosfato Desidrogenases/genética , Interferon gama/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Infecções Estafilocócicas/imunologia , Infecções Estafilocócicas/prevenção & controle , Staphylococcus aureus/genética , Staphylococcus aureus/metabolismo
14.
Sci Rep ; 8(1): 3580, 2018 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-29483570

RESUMO

Staphylococcus aureus can cause different types of diseases from mild skin infections to life-threatening sepsis worldwide. Owing to the emergence and transmission of multidrug-resistant strains, developing an impactful immunotherapy especially vaccine control approach against S. aureus infections is increasingly encouraged and supported. S. aureus manganese transport protein C (MntC), which is a highly-conserved cell surface protein, can elicit protective immunity against S. aureus and Staphylococcus epidermidis. In this study, we evaluated the humoral immune response and CD4+ T cell-mediated immune responses in a mouse peritonitis model. The results showed that MntC-specific antibodies conferred an essential protection for mice to reduce invasion of S. aureus, which was corroborated via the opsonophagocytic killing assay and passive immunization experiment in mice, and moreover MntC-induced Th17 played a remarkable part in preventing S. aureus infection since the MntC-induced protective immunity decreased after neutralization of IL-17 by antibody in vivo and the Th17 adoptive transferred-mice could partly resist S. aureus challenge. In conclusion, we considered that the MntC-specific antibodies and MntC-specific Th17 cells play cooperative roles in the prevention of S. aureus infection.


Assuntos
Proteínas de Bactérias/imunologia , Peritonite/imunologia , Peritonite/microbiologia , Infecções Estafilocócicas/imunologia , Vacinas Antiestafilocócicas/imunologia , Staphylococcus aureus/imunologia , Células Th17/imunologia , Animais , Anticorpos Antibacterianos , Proliferação de Células/efeitos dos fármacos , Modelos Animais de Doenças , Feminino , Imunidade Celular , Imunidade Humoral , Imunização Passiva/psicologia , Interferon gama/metabolismo , Interleucina-17/metabolismo , Manganês/metabolismo , Camundongos , Camundongos Endogâmicos BALB C
15.
Microb Pathog ; 91: 46-53, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26657721

RESUMO

The GapC protein is highly conserved surface dehydrogenase among Streptococcus dysgalactiae (S. dysgalactiae) and is shown to be involved in bacterial virulence. Immunization of GapC protein can induce specific CD4(+) T-cell immune responses and protect against S. dysgalactiae infection. However, there are no studies to identify immunodominant CD4(+) T-cell epitopes on GapC protein. In this study, in silico MHC affinity measurement method was firstly used to predict potential CD4(+) T-cell epitopes on GapC protein. Six predictive 15-mer peptides were synthesized and two novel GapC CD4(+) T-cell epitopes, GapC63-77 and GapC96-110, were for the first time identified using CD4(+) T-cells obtained from GapC-immunized BALB/c (H-2(d)) and C57BL/6 (H-2(b)) mice spleen based on cell proliferation and cytokines response. The results showed that peptides containing 63-77 and 96-110 induced significant antigen-specific CD4(+) T-cells proliferation response in vivo. At the same time, high levels of IFN-γ and IL-17A, as well as moderate levels of IL-10 and IL-4 were detected in CD4(+) T-cells isolated from both GapC and peptide-immunized mice in vivo, suggesting that GapC63-77 and GapC96-110 preferentially elicited polarized Th1/Th17-type responses. The characterization of GapC CD4(+) T-cell epitopes not only helps us understand its protective immunity, but also contributes to design effective T-cell epitope-based vaccine against S. dysgalactiae infection.


Assuntos
Antígenos de Bactérias/química , Antígenos de Bactérias/imunologia , Proteínas de Bactérias/química , Proteínas de Bactérias/imunologia , Linfócitos T CD4-Positivos/imunologia , Epitopos de Linfócito T/química , Epitopos de Linfócito T/imunologia , Infecções Estreptocócicas/microbiologia , Streptococcus/imunologia , Animais , Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Mapeamento de Epitopos , Epitopos de Linfócito T/genética , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Infecções Estreptocócicas/genética , Infecções Estreptocócicas/imunologia , Streptococcus/genética
16.
Microb Pathog ; 89: 108-13, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26423555

RESUMO

Iron-regulated surface determinant B (IsdB) of Staphylococcus aureus (S. aureus) is a highly conserved surface protein that can induce protective CD4(+) T-cell immune response. A pivotal role of CD4(+) T-cells in effective immunity against S. aureus infection has been proved, but CD4(+) T-cell epitopes on the S. aureus IsdB have not been well identified. In this study, MHC binding assay was firstly used to predict CD4(+) T-cell epitopes on S. aureus IsdB protein, and six peptides were synthesized to validate the probable epitopes. Two novel IsdB CD4(+) T-cell epitopes, P1 (residues 159-178) and P4 (residues 287-306), were for the first time identified using CD4(+) T-cells obtained from IsdB-immunized C57BL/6 (H-2(b)) and BALB/c (H-2(d)) mice spleen based on cell proliferation and cytokines response. The results showed that P1 and P4 emulsified in Freund's adjuvant (FA) induced much higher cell proliferation compared with PBS emulsified in FA. CD4(+) T-cells stimulated with peptides P1 and P4 secreted significantly higher levels of IFN-γ and IL-17A. However, the level of the cytokine IL-4 almost remained unchanged, suggesting that P1 and P4 preferentially elicited polarized Th1-type responses. In addition, BALB/c mice just respond to P4 not P1, while C57BL/6 mice respond to P1 not P4, implying that epitope P1 and P4 were determined as H-2(b) and H-2(d) restricted epitope, respectively. Taken together, our data may provide an explanation of the IsdB-induced protection against S. aureus and highlight the possibility of developing the epitope-based vaccine against the S. aureus.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Proteínas de Transporte de Cátions/imunologia , Mapeamento de Epitopos , Epitopos de Linfócito T/imunologia , Staphylococcus aureus/imunologia , Animais , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL
17.
Microbiol Immunol ; 59(9): 533-44, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26249788

RESUMO

Pseudomonas aeruginosa poses a major threat to human health and to the mink industry. Thus, development of vaccines that elicit robust humoral and cellular immunity against P. aeruginosa is greatly needed. In this study, a recombinant attenuated Salmonella vaccine (RASV) that expresses the outer membrane proteins fusion OprF190-342 -OprI21-83 (F1I2) from P. aeruginosa was constructed and the potency of this vaccine candidate assessed by measuring F1I2-specific humoral immune responses upon vaccination through s.c. or oral routes. S.C. administration achieved higher serum IgG titers and IgA titers in the intestine and induced stronger F1I2-specific IgG and IgA titers in lung homogenate than did oral administration, which resulted in low IgG titers and no local IgA production. High titers of IFN-γ, IL-4, and T-lymphocyte subsets induced a mixed Th1/Th2 response in mice immunized s.c., indicating elicitation of cellular immunity. Importantly, when immunized mice were challenged with P. aeruginosa by the intranasal route 30 days after the initial immunization, s.c. vaccination achieved 77.78% protection, in contrast to 41.18% via oral administration and 66.67% via Escherichia coli-expressed F1I2 (His-F1I2) vaccination. These results indicate that s.c. vaccination provides a better protective response against P. aeruginosa infection than do oral administration and the His-F1I2 vaccine.


Assuntos
Proteínas de Bactérias/imunologia , Portadores de Fármacos , Lipoproteínas/imunologia , Infecções por Pseudomonas/prevenção & controle , Vacinas contra Pseudomonas/imunologia , Pseudomonas aeruginosa/imunologia , Salmonella typhimurium/genética , Administração Oral , Animais , Anticorpos Antibacterianos/análise , Anticorpos Antibacterianos/sangue , Proteínas de Bactérias/genética , Modelos Animais de Doenças , Escherichia coli , Feminino , Imunoglobulina A/análise , Imunoglobulina G/sangue , Injeções Subcutâneas , Interferon gama/metabolismo , Interleucina-4/metabolismo , Mucosa Intestinal/imunologia , Lipoproteínas/genética , Pulmão/imunologia , Camundongos Endogâmicos BALB C , Vison , Vacinas contra Pseudomonas/administração & dosagem , Vacinas contra Pseudomonas/genética , Pseudomonas aeruginosa/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Vacinas contra Salmonella/administração & dosagem , Vacinas contra Salmonella/genética , Salmonella typhimurium/crescimento & desenvolvimento , Soro/imunologia , Análise de Sobrevida , Subpopulações de Linfócitos T/imunologia , Resultado do Tratamento , Vacinas Atenuadas/administração & dosagem , Vacinas Atenuadas/genética , Vacinas Atenuadas/imunologia
18.
PLoS One ; 10(6): e0131221, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26121648

RESUMO

The GapC of Streptococcus dysgalactiae (S. dysgalactiae) is a highly conserved surface protein that can induce protective humoral immune response in animals. However, B-cell epitopes on the S. dysgalactiae GapC have not been well identified. In this study, a monoclonal antibody (mAb5B7) against the GapC1-150 protein was prepared. After passive transfer, mAb5B7 could partially protect mice against S. dysgalactiae infection. Eleven positive phage clones recognized by mAb5B7 were identified by screening phage-displayed random 12-peptide library, most of which matched the consensus motif DTTQGRFD. The motif sequence exactly matches amino acids 48-55 of the S. dysgalactiae GapC protein. In addition, the motif 48DTTQGRFD55 shows high homology among various streptococcus species. Site-directed mutagenic analysis further confirmed that residues D48, T50, Q51, G52 and F54 formed the core motif of 48DTTQGRFD55. This motif was the minimal determinant of the B-cell epitope recognized by the mAb5B7. As expected, epitope-peptide evoked protective immune response against S. dysgalactiae infection in immunized mice. Taken together, this identified conserved B-cell epitope within S. dysgalactiae GapC could provide very valuable insights for vaccine design against S. dysgalactiae infection.


Assuntos
Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Sequência Conservada , Epitopos de Linfócito B/química , Epitopos de Linfócito B/imunologia , Biblioteca de Peptídeos , Streptococcus/metabolismo , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/imunologia , Antígenos de Bactérias/química , Proteínas de Bactérias/química , Células Clonais , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Interações Hidrofóbicas e Hidrofílicas , Imunidade , Imunização , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Maleabilidade , Proteínas Recombinantes de Fusão/isolamento & purificação , Alinhamento de Sequência
19.
Appl Microbiol Biotechnol ; 99(2): 667-80, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25381907

RESUMO

Pseudomonas aeruginosa is an opportunistic pathogen that localizes to and colonizes mucosal tissue. Thus, vaccines that elicit a strong mucosal response against P. aeruginosa should be superior to other vaccination strategies. In this study, to stimulate rapid and enhanced mucosal immune responses, mannose-modified chitosan microspheres loaded with the recombinant outer membrane protein OprF190-342-OprI21-83 (FI) (FI-MCS-MPs) of P. aeruginosa were developed as a potent subunit vaccine for mucosal delivery. FI-MCS-MPs were successfully obtained via the tripolyphosphate ionic crosslinking method. Confocal and immunohistochemical analyses indicated that FI-MCS-MPs exhibited the ability to bind the macrophage mannose receptor (MMR, CD206) in vitro and in vivo. After intranasal immunization of mice with FI-MCS-MPs, FI-specific humoral immune responses were detected, measured as local IgM antibody titers in lung tissue slurry; IgA antibody titers in nasal washes, bronchoalveolar lavage (BAL), and intestinal lavage; and systemic IgA and IgG antibody titers in serum. FI-MCS-MPs induced early and high mucosal and systemic humoral antibody responses comparable to those in the group vaccinated with unmodified mannose. High levels of IFN-γ and IL-4 in addition to T lymphocyte subsets induced a mixed Th1/Th2 response in mice immunized with FI-MCS-MPs, resulting in the establishment of cellular immunity. Additionally, when immunized mice were challenged with P. aeruginosa via the nasal cavity, FI-MCS-MPs demonstrated 75 % protective efficacy. Together, these data indicate that mannose-modified chitosan microspheres are a promising subunit delivery system for vaccines against P. aeruginosa infection.


Assuntos
Quitosana/farmacologia , Imunidade nas Mucosas , Manose/farmacologia , Infecções por Pseudomonas/imunologia , Vacinas contra Pseudomonas/imunologia , Pseudomonas aeruginosa , Administração Intranasal , Animais , Anticorpos Antibacterianos/sangue , Formação de Anticorpos , Proteínas de Bactérias/imunologia , Sequência de Bases , Linhagem Celular , Quitosana/química , Feminino , Imunoglobulina A/sangue , Imunoglobulina A/química , Imunoglobulina G/sangue , Interferon gama/imunologia , Interleucina-4/imunologia , Lipoproteínas/imunologia , Macrófagos/química , Macrófagos/imunologia , Manose/química , Camundongos , Camundongos Endogâmicos BALB C , Microesferas , Dados de Sequência Molecular , Infecções por Pseudomonas/prevenção & controle , Vacinas contra Pseudomonas/química , Proteínas Recombinantes/imunologia , Linfócitos T/imunologia
20.
Res Vet Sci ; 98: 39-41, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25468796

RESUMO

Streptococcus dysgalactiae (S. dysgalactiae) GapC protein is a protective antigen that induces partial immunity against S. dysgalactiae infection in animals. To identify the conserved B-cell epitope of S. dysgalactiae GapC, a mouse monoclonal antibody 1E11 (mAb1E11) against GapC was generated and used to screen a phage-displayed 12-mer random peptide library (Ph.D.-12). Eleven positive clones recognized by mAb1E11 were identified, most of which matched the consensus motif TGFFAKK. Sequence of the motif exactly matched amino acids 97-103 of the S. dysgalactiae GapC. In addition, the epitope (97)TGFFAKK(103) showed high homology among different streptococcus species. Site-directed mutagenic analysis further confirmed that residues G98, F99, F100 and K103 formed the core of (97)TGFFAKK(103), and this core motif was the minimal determinant of the B-cell epitope recognized by the mAb1E11. Collectively, the identification of conserved B-cell epitope within S. dysgalactiae GapC highlights the possibility of developing the epitope-based vaccine.


Assuntos
Anticorpos Monoclonais/imunologia , Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Epitopos de Linfócito B/imunologia , Camundongos , Streptococcus/imunologia , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/metabolismo , Epitopos de Linfócito B/análise , Dados de Sequência Molecular , Especificidade da Espécie , Infecções Estreptocócicas/imunologia , Infecções Estreptocócicas/prevenção & controle , Infecções Estreptocócicas/veterinária , Vacinas Estreptocócicas/imunologia , Streptococcus/classificação , Fagos de Streptococcus/imunologia
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