scholarly journals Immunoproteomic Analysis Reveals Novel Candidate Antigens for the Diagnosis of Paracoccidioidomycosis Due to Paracoccidioides lutzii

2020 ◽  
Vol 6 (4) ◽  
pp. 357
Author(s):  
Anderson Messias Rodrigues ◽  
Paula Helena Kubitschek-Barreira ◽  
Breno Gonçalves Pinheiro ◽  
André Teixeira-Ferreira ◽  
Rosane Christine Hahn ◽  
...  
Keyword(s):  
Immune Responses ◽  
Point Of Care ◽  
Paracoccidioides Brasiliensis ◽  
Systemic Infection ◽  
Mass Spectrometry Analysis ◽  
Diagnostic Tools ◽  
Effective Vaccine ◽  
Spectrometry Analysis ◽  
Humoral Immune ◽  
Standard Serum

Paracoccidioidomycosis (PCM) is a life-threatening systemic infection caused by the fungal pathogen Paracoccidioides brasiliensis and related species. Whole-genome sequencing and stage-specific proteomic analysis of Paracoccidioides offer the opportunity to profile humoral immune responses against P. lutzii and P. brasiliensis s. str. infection using innovative screening approaches. Here, an immunoproteomic approach was used to identify PCM-associated antigens that elicit immune responses by combining 2-D electrophoresis of P. lutzii and P. brasiliensis proteomes, immunological detection using a gold-standard serum, and mass spectrometry analysis. A total of 16 and 25 highly immunoreactive proteins were identified in P. lutzii and P. brasiliensis, respectively, and 29 were shown to be the novel antigens for Paracoccidioides species, including seven uncharacterized proteins. Among the panel of proteins identified, most are involved in metabolic pathways, carbon metabolism, and biosynthesis of secondary metabolites in both immunoproteomes. Remarkably, six isoforms of the surface-associated enolase in the range of 54 kDa were identified as the major antigens in human PCM due to P. lutzii. These novel immunoproteomes of Paracoccidioides will be employed to develop a sensitive and affordable point-of-care diagnostic assay and an effective vaccine to identify infected hosts and prevent infection and development of human PCM. These findings provide a unique opportunity for the refinement of diagnostic tools of this important neglected systemic mycosis, which is usually associated with poverty.

scholarly journals Mucosal Immunization with Newcastle Disease Virus Vector Coexpressing HIV-1 Env and Gag Proteins Elicits Potent Serum, Mucosal, and Cellular Immune Responses That Protect against Vaccinia Virus Env and Gag Challenges

mBio ◽  
2015 ◽  
Vol 6 (4) ◽  
Author(s):  
Sunil K. Khattar ◽  
Vinoth Manoharan ◽  
Bikash Bhattarai ◽  
Celia C. LaBranche ◽  
David C. Montefiori ◽  
...  
Keyword(s):  
Immune Responses ◽  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Effective Vaccine ◽  
Gag Proteins ◽  
Humoral Immune ◽  
Vaccinia Viruses ◽  
Mucosal Immune Responses ◽  
Hiv 1

ABSTRACT Newcastle disease virus (NDV) avirulent strain LaSota was used to coexpress gp160 Env and p55 Gag from a single vector to enhance both Env-specific and Gag-specific immune responses. The optimal transcription position for both Env and Gag genes in the NDV genome was determined by generating recombinant NDV (rNDV)-Env-Gag (gp160 located between the P and M genes and Gag between the HN and L genes), rNDV-Gag-Env (Gag located between the P and M genes and gp160 between the HN and L genes), rNDV-Env/Gag (gp160 followed by Gag located between the P and M genes), and rNDV-Gag/Env (Gag followed by gp160 located between the P and M genes). All the recombinant viruses replicated at levels similar to those seen with parental NDV in embryonated chicken eggs and in chicken fibroblast cells. Both gp160 and Gag proteins were expressed at high levels in cell culture, with gp160 found to be incorporated into the envelope of NDV. The Gag and Env proteins expressed by all the recombinants except rNDV-Env-Gag self-assembled into human immunodeficiency virus type 1 (HIV-1) virus-like particles (VLPs). Immunization of guinea pigs by the intranasal route with these rNDVs produced long-lasting Env- and Gag-specific humoral immune responses. The Env-specific humoral and mucosal immune responses and Gag-specific humoral immune responses were higher in rNDV-Gag/Env and rNDV-Env/Gag than in the other recombinants. rNDV-Gag/Env and rNDV-Env/Gag were also more efficient in inducing cellular as well as protective immune responses to challenge with vaccinia viruses expressing HIV-1 Env and Gag in mice. These results suggest that vaccination with a single rNDV coexpressing Env and Gag represents a promising strategy to enhance immunogenicity and protective efficacy against HIV. IMPORTANCE A safe and effective vaccine that can induce both systemic and mucosal immune responses is needed to control HIV-1. In this study, we showed that coexpression of Env and Gag proteins of HIV-1 performed using a single Newcastle disease virus (NDV) vector led to the formation of HIV-1 virus-like particles (VLPs). Immunization of guinea pigs with recombinant NDVs (rNDVs) elicited potent long-lasting systemic and mucosal immune responses to HIV. Additionally, the rNDVs were efficient in inducing cellular immune responses to HIV and protective immunity to challenge with vaccinia viruses expressing HIV Env and Gag in mice. These results suggest that the use of a single NDV expressing Env and Gag proteins simultaneously is a novel strategy to develop a safe and effective vaccine against HIV.

scholarly journals Overexpression of Interleukin-33 in Recombinant Rabies Virus Enhances Innate and Humoral Immune Responses through Activation of Dendritic Cell-Germinal Center Reactions

Vaccines ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 34
Author(s):  
Zhizhong Mi ◽  
Ling Zhao ◽  
Ming Sun ◽  
Ting Gao ◽  
Yong Wang ◽  
...  
Keyword(s):  
Immune Responses ◽  
Rabies Virus ◽  
Germinal Center ◽  
Neutralizing Antibodies ◽  
Effective Vaccine ◽  
Interleukin 33 ◽  
Humoral Immune ◽  
Humoral Immune Responses ◽  
Long Acting ◽  
High Level

Rabies is a zoonotic infectious disease caused by rabies virus (RABV), and its mortality rate is as high as 100%. Globally, an average of 60,000 people die from rabies each year. The most effective method to prevent and limit rabies is vaccination, but it is currently expensive and inefficient, consisting of a 3-dose series of injections and requiring to be immunized annually. Therefore, it is urgent to develop a single dose of long-acting rabies vaccine. In this study, recombinant rabies virus (rRABV) overexpressing interleukin-33 (IL-33) was constructed and designated as rLBNSE-IL33, and its effect was evaluated in a mouse model. The results showed that rLBNSE-IL33 could enhance the quick production of RABV-induced immune antibodies as early as three days post immunization (dpi) through the activation of dendritic cells (DCs), a component of the innate immune system. Furthermore, rLBNSE-IL33 induced high-level virus-neutralizing antibodies (VNA) production that persisted for 8 weeks by regulating the T cell-dependent germinal center (GC) reaction, thus resulting in better protection against rabies. Our data suggest the IL-33 is a novel adjuvant that could be used to enhance innate and humoral immune responses by activating the DC-GC reaction, and thus, rLBNSE-IL33 could be developed as a safe and effective vaccine for animals.

SpyCatcher-SpyTagged ApxIA Toxoid and the Immune-Modulating Yeast Ghost Shells

2020 ◽  
Vol 16 (11) ◽  
pp. 1644-1657
Author(s):  
Seonhyung Lee ◽  
Beom-Ku Han ◽  
Yang-Hoon Kim ◽  
Ji-Young Ahn
Keyword(s):  
Immune Responses ◽  
Innate Immune ◽  
Mass Spectrometry Analysis ◽  
Cytotoxic Activities ◽  
Raw 264.7 Macrophages ◽  
Spectrometry Analysis ◽  
Cellular Immune Responses ◽  
Histocompatibility Complex ◽  
Tnf Α ◽  
Cellular Immune

Actinobacillus pleuropneumoniaesecretes the hemolytic cytotoxins ApxI, ApxII, ApxIII, and ApxIV, which cause pleurop- neumonia in swine. Of these, ApxI is the most toxic. ApxIA, a repeats-in-toxin toxin-like protein, has strong hemolytic and cytotoxic activities. This study aimed to develop an immune modulator ApxIA toxoid, with a Spytag/Spycatcher pair (SC::ST pair), in yeast ghost shells (YGSs). These YGSs were utilized as ApxIA toxoid delivery platforms for -glucan components that can be recognized by the innate immune system. The SC::ST pair was used to conjugate the ApxIA toxoid to YGSs. The YGS-SC::ST-ToxApxIA was successfully phagocytosed by RAW 264.7 macrophages cells, without any toxicity. Further investigation revealed that YGS-SC::ST-ToxApxIA led to defective immune responses, in addition to increased levels of cytokines IL-1β, TNF-α, and IL-10. A membrane proteomic analysis, to determine preferential major histocompatibility complex binding of ApxIA-derived peptides, was performed and four ApxIA peptides were successfully identified by liquid chromatography with tandem mass spectrometry analysis. The identified peptides may serve as poten- tial vaccine candidates in immunobiology studies of A. pleuropneumoniae. Our results indicate that YGS-SC::ST-ToxApxIA can prevent A. pleuropneumoniae pleuropneumonia (APP) by inducing both humoral and cellular immune responses.

scholarly journals Novel exosome biomarker candidates for Alzheimer´s disease unraveled through Mass Spectrometry analysis

2021 ◽  
Author(s):  
Tânia Soares Martins ◽  
Rui Marçalo ◽  
Cristóvão B. da Cruz e Silva ◽  
Dário Trindade ◽  
José Catita ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Multivariate Analysis ◽  
Cost Effective ◽  
Functional Enrichment ◽  
Mass Spectrometry Analysis ◽  
Precipitation Method ◽  
Molecular Diagnostic ◽  
Diagnostic Tools ◽  
Spectrometry Analysis ◽  
Wide Range

Abstract Background Exosomes are small extracellular vesicles (EVs) present in human biofluids that can carry disease-specific molecules. Blood-derived exosomes emerged as interesting peripheral sources of biomarkers for a wide range of diseases, and their potential is also being addressed in Alzheimer’s disease (AD) context. There is no effective cure or blood-based molecular diagnostic tools for a first clinical AD screening, which motivates research in this area to identify putative valuable blood-derived disease biomarkers. The ultimate goal is to produce a cost-effective and widely available alternative to the current molecular AD diagnosis that monitors a biomarker triplet in the CSF. Methods In the study here presented, EVs with exosome-like characteristics were isolated from the serum of Controls and AD cases through two distinct methods, precipitation- and column-based methods, followed by mass spectrometry analysis. The resulting proteomes from Controls and AD cases were characterized by Gene Ontology (GO) functional enrichment and multivariate analysis. Putative candidate targets identified were validated in distinct cohorts using antibody-based approaches. Results Both methodologies isolated particles with the expected morphology and size range. Although GO terms were similar for Controls and AD cases exosomes in both methodologies, the multivariate analysis revealed a clear segregation between Controls and AD cases obtained by the precipitation method. Nine significantly different abundant proteins were identified between Controls and AD cases and exosome levels of AACT and C4BPα, two Aβ-binding proteins, were validated in individuals from independent cohorts. Conclusions Serum-derived exosome proteomes were unraveled for the first time in the context of AD through two distinct isolation methodologies, holding disease discriminatory potential. The work carried out gives an important contribution to the identification of novel exosomal biomarker candidates potentially useful as blood-based tools for AD diagnosis.

Mass spectrometry analysis of the excretory-secretory (E-S) products of the model cestode Hymenolepis diminut a reveals their immunogenic properties and the presence of new E-S proteins in cestodes

2016 ◽  
Vol 61 (2) ◽  
Author(s):  
Justyna Bień ◽  
Rusłan Sałamatin ◽  
Anna Sulima ◽  
Kirsi Savijoki ◽  
David Bruce Conn ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Immune Responses ◽  
Drug Targets ◽  
Mass Spectrometry Analysis ◽  
Spectrometry Analysis ◽  
Biochemical Processes ◽  
Host Parasite Interactions ◽  
Host Parasite ◽  
Immunogenic Properties ◽  
S Proteins

Abstractis an important model species in studies of therapeutics, biochemical processes, immune responses and other aspects of cestodiasis. The parasite produces numerous excretory-secretory (E-S) proteins and a glycocalyx covering its body. Our study focused on the mass spectrometry analysis of the E-S material with an objective to determine if E-S contains any new proteins, in particular those that can be identified as: antigens, vaccine candidates and drug targets. These proteins might engage directly in host-parasite interactions. Adult parasites collected from experimentally infected rats were cultured

scholarly journals Evaluation of a Vaccine Formulation against Streptococcus pneumoniae Based on Choline-Binding Proteins

2014 ◽  
Vol 22 (2) ◽  
pp. 213-220 ◽  
Author(s):  
Eliane N. Miyaji ◽  
Cintia F. M. Vadesilho ◽  
Maria Leonor S. Oliveira ◽  
André Zelanis ◽  
David E. Briles ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
Binding Proteins ◽  
Choline Chloride ◽  
Protein A ◽  
Surface Protein ◽  
Mass Spectrometry Analysis ◽  
Effective Vaccine ◽  
Lethal Challenge ◽  
Content Type ◽  
Spectrometry Analysis

ABSTRACTStreptococcus pneumoniaehas proteins that are attached to its surface by binding to phosphorylcholine of teichoic and lipoteichoic acids. These proteins are known as choline-binding proteins (CBPs). CBPs are an interesting alternative for the development of a cost-effective vaccine, and PspA (pneumococcal surface protein A) is believed to be the most important protective component among the different CBPs. We sought to use CBPs eluted from pneumococci as an experimental vaccine. Since PspA shows variability between isolates, we constructed strains producing different PspAs. We used the nonencapsulated Rx1 strain, which produces PspA from clade 2 (PspA2), to generate apspA-knockout strain (Rx1 ΔpspA) and strains expressing PspA from clade 1 (Rx1pspA1) and clade 4 (Rx1pspA4). We grew Rx1, Rx1 ΔpspA, Rx1pspA1, and Rx1pspA4in Todd-Hewitt medium containing 0.5% yeast extract and washed cells in 2% choline chloride (CC). SDS-PAGE analysis of the proteins recovered by a CC wash showed few bands, and the CBPs PspA and PspC (pneumococcal surface protein C) were identified by mass spectrometry analysis. Subcutaneous immunization of mice with these full-length native proteins without adjuvant led to significantly higher rates of survival than immunization with diluent after an intranasal lethal challenge with two pneumococcal strains and also after a colonization challenge with one strain. Importantly, immunization with recombinant PspA4 (rPspA4) without adjuvant did not elicit significant protection.

scholarly journals Identification of Disease-Associated Cryptococcal Proteins Reactive With Serum IgG From Cryptococcal Meningitis Patients

2021 ◽  
Vol 12 ◽  
Author(s):  
A. Elisabeth Gressler ◽  
Daniela Volke ◽  
Carolina Firacative ◽  
Christiane L. Schnabel ◽  
Uwe Müller ◽  
...  
Keyword(s):  
Cryptococcal Meningitis ◽  
Mass Spectrometry Analysis ◽  
Healthy Individuals ◽  
Protein Antigens ◽  
Fungal Burden ◽  
Spectrometry Analysis ◽  
Humoral Immune ◽  
Human Proteins ◽  
Human Sera ◽  
Opportunistic Fungal Pathogen

Cryptococcus neoformans, an opportunistic fungal pathogen ubiquitously present in the environment, causes cryptococcal meningitis (CM) mainly in immunocompromised patients, such as AIDS patients. We aimed to identify disease-associated cryptococcal protein antigens targeted by the human humoral immune response. Therefore, we used sera from Colombian CM patients, with or without HIV infection, and from healthy individuals living in the same region. Serological analysis revealed increased titers of anti-cryptococcal IgG in HIV-negative CM patients, but not HIV-positive CM patients, compared to healthy controls. In contrast, titers of anti-cryptococcal IgM were not affected by CM. Furthermore, we detected pre-existing IgG and IgM antibodies even in sera from healthy individuals. The observed induction of anti-cryptococcal IgG but not IgM during CM was supported by analysis of sera from C. neoformans-infected mice. Stronger increase in IgG was found in wild type mice with high lung fungal burden compared to IL-4Rα-deficient mice showing low lung fungal burden. To identify the proteins targeted by human anti-cryptococcal IgG antibodies, we applied a quantitative 2D immunoproteome approach identifying cryptococcal protein spots preferentially recognized by sera from CM patients or healthy individuals followed by mass spectrometry analysis. Twenty-three cryptococcal proteins were recombinantly expressed and confirmed to be immunoreactive with human sera. Fourteen of them were newly described as immunoreactive proteins. Twelve proteins were classified as disease-associated antigens, based on significantly stronger immunoreactivity with sera from CM patients compared to healthy individuals. The proteins identified in our screen significantly expand the pool of cryptococcal proteins with potential for (i) development of novel anti-cryptococcal agents based on implications in cryptococcal virulence or survival, or (ii) development of an anti-cryptococcal vaccine, as several candidates lack homology to human proteins and are localized extracellularly. Furthermore, this study defines pre-existing anti-cryptococcal immunoreactivity in healthy individuals at a molecular level, identifying target antigens recognized by sera from healthy control persons.

scholarly journals The IMMUNE-ASSOCIATED NUCLEOTIDE-BINDING 9 protein is a regulator of basal immunity in Arabidopsis thaliana

2018 ◽  
Author(s):  
Yuanzheng Wang ◽  
Yansha Li ◽  
Tabata Rosas-Diaz ◽  
Carlos Caceres-Moreno ◽  
Rosa Lozano-Duran ◽  
...  
Keyword(s):  
Immune Responses ◽  
Pseudomonas Syringae ◽  
Expression Patterns ◽  
Nucleotide Binding ◽  
Negative Regulator ◽  
Mass Spectrometry Analysis ◽  
Crop Species ◽  
Spectrometry Analysis ◽  
Basal Immunity

AbstractA robust regulation of plant immune responses requires multitude of positive and negative regulators that act in concert. The immune-associated nucleotide-binding (IAN) gene family members are associated with immunity in different organisms, although no characterization of their function has been carried out to date in plants. In this work, we analyzed the expression patterns of IAN genes and found that IAN9 is repressed upon pathogen infection or treatment with immune elicitors. IAN9 encodes a plasma membrane-localized protein that genetically behaves as a negative regulator of immunity. A novel ian9 mutant generated by CRISPR/Cas9 shows increased resistance to Pseudomonas syringae, while transgenic plants overexpressing IAN9 show a slight increase in susceptibility. In vivo immunoprecipitation of IAN9-GFP followed by mass spectrometry analysis revealed that IAN9 associates with a previously uncharacterized C3HC4-type RING finger domain-containing protein that we named IAP1, for IAN9-associated protein 1, which also acts as a negative regulator of basal immunity. Interestingly, neither ian9 or iap1 mutant plants show any obvious developmental phenotype, suggesting that they display enhanced inducible immunity rather than constitutive immune responses. Since both IAN9 and IAP1 have orthologs in important crop species, they could be suitable targets to generate plants more resistant to diseases caused by bacterial pathogens without yield penalty.

scholarly journals Comparison of the Efficacy of Selected Bacterins against Edwardsiella Tarda in Immunized Japanese eel (Anguilla japonica)

2016 ◽  
Vol 29 (2) ◽  
pp. 62-69
Author(s):  
Md Mer Mosharraf Hossain ◽  
Kenji Kawai ◽  
Jim Duston ◽  
Syunichirou Oshima
Keyword(s):  
Immune Responses ◽  
Anguilla Japonica ◽  
Japanese Eel ◽  
Edwardsiella Tarda ◽  
Effective Vaccine ◽  
High Concentration ◽  
Humoral Immune ◽  
Humoral Immune Responses ◽  
Antibody Titers ◽  
Inactivated Vaccines

An effective vaccine against Edwardsiella tarda has not been reported in substitution for high concentration of formalin for the prevention of edwardsiellosis disease. In this study, the efficacy of inactivated E. tarda was evaluated and compared by intraperitoneal (IP) injection-immunization or challenge against Japanese eel (Anguilla japonica). Formalin, formalin with heat, citric acid, pressure and electric current were used for inactivation of the bacteria, and the relative percent survival (RPS) values of pressure (600 psi for 5 min) killed cells was determined. PKCinactivated vaccine showed 89-93% protection that was higher than others. PKC-inactivated vaccine at a concentration of 106 cells/fish was sufficient to induce high protection (RPS>89). Protection of the different-inactivated vaccines was evaluated at different time post immunization, and the peak of protection was observed at 9 days post-challenge. Fish immunized with PKC showed significantly (P<0.05) higher serum and mucus antibody titers elicit both systemic and mucosal adaptive immune responses, and induce specific humoral immune responses in eel. Coincident with higher protection, sera of fish immunized with the PKC vaccine had higher agglutination titers than FKC, FHKC, CAKC and ECKC. All these data strongly suggested that PKC vaccine is an effective strategy to protect eel against edwardsiellosis.Bangladesh J Microbiol, Volume 29, Number 2, Dec 2012, pp 62-69

scholarly journals Comparison of the efficacy of selected bacterins against Edwardsiella tarda in immunized Japanese eel (Anguilla japonica)

2013 ◽  
Vol 10 (2) ◽  
pp. 355-366 ◽  
Author(s):  
MMM Hossain ◽  
Kenji Kawai ◽  
Jim Duston ◽  
Syunichirou Oshima
Keyword(s):  
Immune Responses ◽  
Anguilla Japonica ◽  
Japanese Eel ◽  
Edwardsiella Tarda ◽  
Effective Vaccine ◽  
Inactivated Vaccine ◽  
High Concentration ◽  
Humoral Immune ◽  
Humoral Immune Responses ◽  
Inactivated Vaccines

An effective vaccine against Edwardsiella tarda has not been reported in substitution for high concentration of formalin for the prevention of edwardsiellosis disease. In this study, the efficacy of inactivated E. tarda was evaluated and compared by intraperitoneal (IP) injection-immunization or challenge against Japanese eel (Anguilla japonica). Formalin, formalin with heat, citric acid, pressure and electric current were used for inactivation of the bacteria, and the relative percent survival (RPS) values of pressure (600 psi for 5 min) killed cells was determined. PKC-inactivated vaccine showed-89-93 protection that was higher than others. PKC-inactivated vaccine at a concentration of 106 cells/fish was sufficient to induce high protection (RPS>89). Protection of the different-inactivated vaccines was evaluated at different time post immunization, and the peak of protection was observed at 9 days post-challenge. Fish immunized with PKC showed significantly (P<0.05) higher serum and mucus antibody titers elicit both systemic and mucosal adaptive immune responses, and induce specific humoral immune responses in eel. Coincident with higher protection, sera of fish immunized with the PKC vaccine had higher agglutination titers than FKC, FHKC, CAKC and ECKC. All these data strongly suggested that PKC vaccine is an effective strategy to protect eel against edwardsiellosis.DOI: http://dx.doi.org/10.3329/jbau.v10i2.14929 J. Bangladesh Agril. Univ. 10(2): 355-366, 2012

Sign in / Sign up

Export Citation Format

Share Document