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1.
Mem. Inst. Oswaldo Cruz ; 115: e190396, 2020. graf
Article in English | LILACS | ID: biblio-1101277

ABSTRACT

BACKGROUND Nanoparticles (NPs) are viable candidates as carriers of exogenous materials into cells via transfection and can be used in the DNA vaccination strategy against leptospirosis. OBJECTIVES We evaluated the efficiency of halloysite clay nanotubes (HNTs) and amine-functionalised multi-walled carbon nanotubes (NH2-MWCNTs) in facilitating recombinant LemA antigen (rLemA) expression and protecting Golden Syrian hamsters (Mesocricetus auratus) against Leptospira interrogans lethal infection. METHODS An indirect immunofluorescent technique was used to investigate the potency of HNTs and NH2-MWCNTs in enhancing the transfection and expression efficiency of the DNA vaccine in Chinese hamster ovary (CHO) cells. Hamsters were immunised with two doses of vaccines HNT-pTARGET/lemA, NH2-MWCNTs-pTARGET/lemA, pTARGET/lemA, and empty pTARGET (control), and the efficacy was determined in terms of humoral immune response and protection against a lethal challenge. FINDINGS rLemA DNA vaccines carried by NPs were able to transfect CHO cells effectively, inducing IgG immune response in hamsters (p < 0.05), and did not exhibit cytotoxic effects. Furthermore, 83.3% of the hamsters immunised with NH2-MWCNTs-pTARGET/lemA were protected against the lethal challenge (p < 0.01), and 66.7% of hamsters immunised with HNT-pTARGET/lemA survived (p < 0.05). MAIN CONCLUSIONS NH2-MWCNTs and HNTs can act as antigen carriers for mammalian cells and are suitable for DNA nanovaccine delivery.


Subject(s)
Animals , Female , Bacterial Proteins/administration & dosage , Transcription Factors/administration & dosage , Bacterial Vaccines/administration & dosage , Vaccines, DNA/administration & dosage , Leptospirosis/prevention & control , Antigens, Bacterial/administration & dosage , Bacterial Proteins/immunology , Transcription Factors/immunology , Bacterial Vaccines/immunology , Cricetinae , Fluorescent Antibody Technique, Indirect , Vaccines, DNA/immunology , Disease Models, Animal , Nanoparticles , Leptospira interrogans/immunology , Leptospirosis/immunology , Antibodies, Bacterial/immunology , Antigens, Bacterial/immunology
2.
Braz. j. infect. dis ; 23(4): 246-253, July-Aug. 2019. tab, graf
Article in English | LILACS | ID: biblio-1039236

ABSTRACT

Abstract Accurate and rapid diagnostic tools are important aspects of managing tuberculosis (TB) cases appropriately. However, the sensitivity and specificity of diagnostic kits based on immune response such as the tuberculin skin test (TST) and interferon gamma release assay (IGRA) are still debated. Thus, the exploration and assessment of specific biomarker-targeted antibodies are needed for the development of an accurate and rapid diagnostic tool. The present study was conducted in patients with a respiratory problem suspected to be TB at Dr. Soetomo Hospital, Surabaya, Indonesia. Among 102 patients tested by GeneXpert and AFB, 59 serum samples were from cases retrospectively determined to have active TB. A total of 102 serum of healthy controls (HC) was also collected. The PPD antigen and the recombinant CFP-10 and ESAT-6 proteins were prepared. Antibody responses against these proteins were evaluated by ELISA. All samples were also screened for the possibility of Mycobacterium avium-intracellulare complex (MAC) infection using Capilla MaC kit. The results showed that TB patients had a significantly higher concentration of IgG antibody in response to PPD than the HC. In addition, the receiver operating characteristic (ROC) curve analysis showed that PPD was acceptable for diagnostic purposes with an AUC value of 0.835 (95% CI 0.770-0.900, p < 0.0001). However, ESAT-6 and CFP-10 had low AUCs, and 32 samples from both groups showed a low concentration of IgA antibody against all antigens. The MAC detection results also showed that the concentration of IgA in the HC group was the highest. The current results indicate that PPD is a better antigen for antibody-based detection of TB than ESAT-6 and CFP-10. Based on the MAC detection assay, 53 people in the HC group were probably infected with rapidly growing nontuberculous mycobacteria (NTM), although antibody response to PPD was low.


Subject(s)
Humans , Male , Female , Child , Adolescent , Adult , Middle Aged , Aged , Young Adult , Bacterial Proteins/immunology , Tuberculin/immunology , Tuberculosis, Pulmonary/diagnosis , Tuberculosis, Pulmonary/microbiology , Antibody Formation/immunology , Mycobacterium tuberculosis/immunology , Antigens, Bacterial/immunology , Reference Values , Tuberculosis, Pulmonary/blood , Enzyme-Linked Immunosorbent Assay , Tuberculin Test , Case-Control Studies , Retrospective Studies , Sensitivity and Specificity , Statistics, Nonparametric , Indonesia
3.
Braz. j. infect. dis ; 22(6): 462-471, Nov.-Dec. 2018. tab, graf
Article in English | LILACS | ID: biblio-984018

ABSTRACT

ABSTRACT The Region of D eletion 2 (RD2) of Mycobacterium tuberculosis encodes reserved antigens that contribute to bacterial virulence. Among these antigens, Rv1983, Rv1986, Rv1987, and Rv1989c have been shown to be immunodominant in infected cattle; however, their diagnostic utility has not been evaluated in humans.In this study, we screened 87 overlapping synthetic peptides encoded by five RD2 proteins for diagnosing tuberculosis epitopes in 50 active tuberculosis (TB) cases, 31 non-tuberculosis patients and 36 healthy individuals. A pool of promising epitopes was then assessed for their diagnostic value in 233 suspected TB patients using a whole blood IFN-γ release assay.Only 10 peptides were recognized by more than 10% of active tuberculosis patients. The IFN-γ release responses to Rv1986-P9, P15, P16, Rv1988-P4, P11, and Rv1987-P11 were significantly higher in the active TB group than in the control groups (p < 0.05). The whole blood IFN-γ release assay based on these epitopes yielded a sensitivity of 51% and a specificity of 85% in diagnosing active tuberculosis, and the corresponding results using the T-SPOT.TB assay were 76% and 75%, respectively.In conclusion, these results suggest that the six epitopes from the RD2 of M. tuberculosis have potential diagnostic value in TB.


Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Young Adult , Bacterial Proteins/immunology , Tuberculosis/diagnosis , Epitopes, T-Lymphocyte/immunology , Mycobacterium tuberculosis/immunology , Antigens, Bacterial/immunology , Bacterial Proteins/blood , Tuberculosis/immunology , Tuberculosis, Pulmonary/diagnosis , Tuberculosis, Pulmonary/immunology , Case-Control Studies , Retrospective Studies , Sensitivity and Specificity , Epitopes, T-Lymphocyte/blood , Antigens, Bacterial/blood
4.
Braz. j. microbiol ; 49(2): 320-328, Apr.-June 2018. tab, graf
Article in English | LILACS | ID: biblio-889239

ABSTRACT

Abstract Detection of Salmonella is very important to minimize the food safety risk. In this study, the recombinant PagC protein and PagC antibody were prepared and coupled with immunomagnetic beads (IMBs) to capture Salmonella cells from pork and milk samples. And then the SYBR Green qualitative PCR was developed to detect the pathogenic Salmonella. The results showed that the PagC polyclonal antiserum is of good specificity and the capture rate of 0.1 mg IMBs for Salmonella tended to be stable at the range of 70-74% corresponding to the concentrations between 101 and 104 CFU/mL. The method developed demonstrated high specificity for the positive Salmonella samples when compared to non-specific DNA samples, such as Escherichia coli, Staphylococcus aureus, Yersinia enterocolitica, and Yersinia pseudotuberculosis. The limit of detection of this assay was 18 CFU/mL. Detection and quantitative enumeration of Salmonella in samples of pork or milk shows good recoveries of 54.34% and 52.07%. In conclusion, the polyclonal antibody of recombinant PagC protein is effective to capture Salmonella from detected samples. The developed pagC antibody IMBs-qPCR method showed efficiency, sensitivity and specificity for 30 Salmonella detection, enabling detection within 10 h, which is a promising rapid method to detect Salmonella in emergency.


Subject(s)
Animals , Salmonella/isolation & purification , Food Contamination , Immunomagnetic Separation/methods , Real-Time Polymerase Chain Reaction/methods , Food Microbiology/methods , Salmonella/genetics , Bacterial Proteins/immunology , Sensitivity and Specificity , Milk/microbiology , Meat/microbiology , Antibodies, Bacterial/immunology , Antibodies, Bacterial/metabolism
5.
Braz. j. microbiol ; 48(4): 774-781, Oct.-Dec. 2017. tab, graf
Article in English | LILACS | ID: biblio-889161

ABSTRACT

ABSTRACT Food-borne diseases, caused by the pathogenic bacteria, are highly prevalent in the world. Salmonella is one of the most important bacterial genera responsible for this. Salmonella Enteritidis (SE) is one of the non-typhoid Salmonellae that can be transmitted to human from poultry products, water, and contaminated food. In recent years, new and rapid detection methods such as enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) have been developed. In this study, recombinant FliC (rFliC) was produced to be used as an antigen. The immunization was conducted in mice with the purified recombinant FliC (rFliC). The mice were subcutaneously immunized with rFliC and elicited significant rFliC specific serum IgG antibodies. An indirect ELISA system was established for the detection of Salmonella Enteritidis. Our results confirmed that the recombinant flagellin can be one of the excellent indicators for the detection of Salmonella Enteritidis.


Subject(s)
Humans , Animals , Mice , Enzyme-Linked Immunosorbent Assay/methods , Flagellin/analysis , Salmonella enteritidis/isolation & purification , Antibodies, Bacterial/analysis , Antibodies, Bacterial/immunology , Antigens, Bacterial/analysis , Antigens, Bacterial/immunology , Bacterial Proteins/analysis , Bacterial Proteins/genetics , Bacterial Proteins/immunology , Flagellin/genetics , Flagellin/immunology , Mice, Inbred BALB C , Salmonella enteritidis/genetics , Salmonella enteritidis/immunology
6.
Mem. Inst. Oswaldo Cruz ; 112(12): 844-849, Dec. 2017. tab, graf
Article in English | LILACS | ID: biblio-894860

ABSTRACT

BACKGROUND Leprosy is a chronic infectious disease caused by the obligate intracellular bacillus Mycobacterium leprae. Because leprosy diagnosis is complex and requires professional expertise, new tools and methodologies are needed to detect cases in early stages and prevent transmission. The M. leprae genome contains mce1A, which encodes a putative mammalian cell entry protein (Mce1A). We hypothesised that the presence of Mce1A on the cell surface could be detected by the host's immune system. OBJECTIVE The aim of this study was to evaluate antibody responses against the Mce1A protein in leprosy patients, household contacts of patients, and the general population to present an addition tool for leprosy diagnosis. METHODS A cross-sectional study involving 89 volunteers [55 leprosy cases, 12 household contacts (HHC) and 22 endemic controls (EC)] was conducted at Couto Maia Hospital, in Salvador, Bahia (BA), Brazil. RESULTS The median anti-Mce1A IgA was significantly higher in multibacillary (MB) and paucibacillary (PB) cases than in EC (p < 0.0001). A similar trend was observed in IgM levels, which were significantly higher in both MB (p < 0.0001) and PB (p = 0.0006) groups compared to in EC individuals. The greatest differences were observed for IgG class-specific antibodies against Mce1A. The median levels of MB and PB were significantly higher compared to both controls HHC and EC (MB or PB vs EC, MB vs HHC p < 0.0001; PB vs HHC, p = 0.0013). Among leprosy cases, IgG enzyme-linked immunosorbent assay sensitivity and specificity were 92.7% and 97.1%, respectively. IgG positivity was confirmed in 92.1% and 94.1% of MB and PB patients, respectively. CONCLUSION This novel diagnostic approach presents an easy, non-invasive, and inexpensive method for leprosy screening, which may be applicable in endemic areas.


Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Aged , Bacterial Proteins/immunology , Immunoglobulin G/blood , Leprosy/diagnosis , Antibodies, Bacterial/blood , Mycobacterium leprae/immunology , Enzyme-Linked Immunosorbent Assay , Case-Control Studies , Family Characteristics , Sensitivity and Specificity
7.
Electron. j. biotechnol ; 28: 52-57, July. 2017. ilus, graf, tab
Article in English | LILACS | ID: biblio-1015847

ABSTRACT

Background: Gardnerella vaginalis is a bacterial vaginosis (BV)-associated vaginal bacterium that produces the toxin vaginolysin (VLY). VLY is a pore-forming toxin that is suggested to be the main virulence factor of G. vaginalis. The high recurrence rate of BV and the emergence of antibiotic-resistant bacterial species demonstrate the need for the development of recombinant antibodies as novel therapeutic agents for disease treatment. Single-chain variable fragments (scFvs) generated against VLY exhibited reduced efficacy to neutralize VLY activity compared to the respective full-length antibodies. To improve the properties of scFvs, monospecific dimeric scFvs were generated by the genetic fusion of two anti-VLY scFv molecules connected by an alpha-helix-forming peptide linker. Results: N-terminal hexahistidine-tagged dimeric scFvs were constructed and produced in Escherichia coli and purified using metal chelate affinity chromatography. Inhibition of VLY-mediated human erythrocyte lysis by dimeric and monomeric scFvs was detected by in vitro hemolytic assay. The circulating half-life of purified scFvs in the blood plasma of mice was determined by ELISA. Dimeric anti-VLY scFvs showed higher neutralizing potency and extended circulating half-life than parental monomeric scFv. Conclusions: The protein obtained by the genetic fusion of two anti-VLY scFvs into a dimeric molecule exhibited improved properties in comparison with monomeric scFv. This new recombinant antibody might implement new possibilities for the prophylaxis and treatment of the diseases caused by the bacteria G. vaginalis.


Subject(s)
Animals , Mice , Bacterial Proteins/immunology , Bacterial Toxins/immunology , Antibodies, Neutralizing/metabolism , Single-Chain Antibodies/metabolism , Bacterial Proteins/toxicity , Bacterial Toxins/toxicity , Enzyme-Linked Immunosorbent Assay , Gardnerella vaginalis , Vaginosis, Bacterial , Dimerization , Virulence Factors , Gene Fusion , Antibodies, Neutralizing/immunology , Single-Chain Antibodies/immunology , Half-Life
9.
Braz. oral res. (Online) ; 31: e39, 2017. tab, graf
Article in English | LILACS | ID: biblio-839507

ABSTRACT

Abstract The present study compared IgA specificity against oral streptococci in colostrum and saliva samples. Sixty-two mother-and-child pairs were included; samples of colostrum (C) and saliva (MS) were collected from the mothers and saliva samples were collected from babies (BS). The specificity of IgA against Streptococcus mutans and S. mitis were analyzed by western blot. Only 30% of babies’ samples presented IgA reactivity to S. mutans, while 74 and 80% of MS and C, respectively, presented this response. IgA reactivity to S. mutans virulence antigens (Ag I/II, Gtf and GbpB) in positive samples showed differences between samples for Gtf and especially for GbpB (p < 0.05), but responses to Ag I/II were similar (p > 0.05). The positive response of Gtf-reactive IgA was different between C (90%) and MS (58%) samples (p < 0.05), but did not differ from BS (p > 0.05). GbpB was the least detected, with 48 and 26% of C and MS, and only 5% of BS samples presenting reactivity (p > 0.05). Eight percent of MS and C samples presented identical bands to SM in the same time-point. In conclusion, the differences of IgA response found between C and MS can be due to the different ways of stimulation, proliferation and transportation of IgA in those secretions. The colostrum has high levels of IgA against S. mutans virulence antigens, which could affect the installation and accumulation process of S. mutans, mainly by supplying anti-GbpB IgA to the neonate.


Subject(s)
Humans , Female , Infant, Newborn , Saliva/immunology , Streptococcus mutans/immunology , Immunoglobulin A, Secretory/analysis , Immunoglobulin A, Secretory/immunology , Colostrum/immunology , Streptococcus mitis/immunology , Saliva/microbiology , Bacterial Proteins/analysis , Bacterial Proteins/immunology , Virulence , Enzyme-Linked Immunosorbent Assay , Glycoproteins/analysis , Glycoproteins/immunology , Blotting, Western , Analysis of Variance , Colostrum/microbiology , Glucosyltransferases/analysis , Glucosyltransferases/immunology , Mothers , Antibody Formation/immunology , Antigens, Bacterial/analysis , Antigens, Bacterial/immunology
10.
Braz. j. microbiol ; 47(2): 394-402, Apr.-June 2016. tab, graf
Article in English | LILACS | ID: lil-780824

ABSTRACT

Abstract Clostridium difficile has emerged as an increasingly important nosocomial pathogen and the prime causative agent of antibiotic-associated diarrhoea and pseudomembranous colitis in humans. In addition to toxins A and B, immunological studies using antisera from patients infected with C. difficile have shown that a number of other bacterial factors contribute to the pathogenesis, including surface proteins, which are responsible for adhesion, motility and other interactions with the human host. In this study, various clostridial targets, including FliC, FliD and cell wall protein 66, were expressed and purified. Phage antibody display yielded a large panel of specific recombinant antibodies, which were expressed, purified and characterised. Reactions of the recombinant antibodies with their targets were detected by enzyme-linked immunosorbent assay; and Western blotting suggested that linear rather than conformational epitopes were recognised. Binding of the recombinant antibodies to surface-layer proteins and their components showed strain specificity, with good recognition of proteins from C. difficile 630. However, no reaction was observed for strain R20291—a representative of the 027 ribotype. Binding of the recombinant antibodies to C. difficile M120 extracts indicated that a component of a surface-layer protein of this strain might possess immunoglobulin-binding activities. The recombinant antibodies against FliC and FliD proteins were able to inhibit bacterial motility.


Subject(s)
Humans , Bacterial Proteins/analysis , Clostridioides difficile/genetics , Clostridium Infections/microbiology , Antibodies, Bacterial/analysis , Bacterial Proteins/genetics , Bacterial Proteins/immunology , Recombinant Proteins/analysis , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Gene Expression , Blotting, Western , Clostridioides difficile/isolation & purification , Clostridioides difficile/immunology , Clostridium Infections/diagnosis , Ribotyping , Antibodies, Bacterial/genetics , Antibodies, Bacterial/immunology
11.
Mem. Inst. Oswaldo Cruz ; 110(8): 989-995, Dec. 2015. tab, graf
Article in English | LILACS | ID: lil-769835

ABSTRACT

Leptospirosis is a zoonotic disease caused by pathogenic spirochetes of theLeptospira genus. Vaccination with bacterins has severe limitations. Here, we evaluated the N-terminal region of the leptospiral immunoglobulin-like B protein (LigBrep) as a vaccine candidate against leptospirosis using immunisation strategies based on DNA prime-protein boost, DNA vaccine, and subunit vaccine. Upon challenge with a virulent strain ofLeptospira interrogans, the prime-boost and DNA vaccine approaches induced significant protection in hamsters, as well as a specific IgG antibody response and sterilising immunity. Although vaccination with recombinant fragment of LigBrep also produced a strong antibody response, it was not immunoprotective. These results highlight the potential of LigBrep as a candidate antigen for an effective vaccine against leptospirosis and emphasise the use of the DNA prime-protein boost as an important strategy for vaccine development.


Subject(s)
Animals , Cricetinae , Female , Antigens, Bacterial/immunology , Bacterial Proteins/immunology , Bacterial Vaccines/immunology , Leptospira/immunology , Leptospirosis/prevention & control , Vaccination/methods , Adjuvants, Immunologic , Biopsy , Chlorocebus aethiops , Conserved Sequence , Enzyme-Linked Immunosorbent Assay , Immunity, Humoral/immunology , Immunoglobulin A/genetics , Immunoglobulin A/immunology , Immunoglobulin G/immunology , Immunoglobulins/genetics , Immunoglobulins/immunology , Kidney/pathology , Leptospirosis/immunology , Lung/pathology , Mesocricetus , Survival Analysis , Vero Cells , Vaccines, DNA/immunology , Vaccines, Synthetic/immunology , Vaccines, Synthetic/microbiology
12.
Braz. j. infect. dis ; 19(6): 636-642, Nov.-Dec. 2015.
Article in English | LILACS | ID: lil-769619

ABSTRACT

ABSTRACT Mycobacterium tuberculosis is the etiologic agent of tuberculosis, one of the world's greatest cause of morbidity and mortality due to infectious disease. Many evolutionary mechanisms have contributed to its high level of adaptation as a host pathogen. Prior to become dormant, a group of about 50 genes related to metabolic changes are transcribed by the DosR regulon, one of the most complex and important systems of host-pathogen interaction. This genetic mechanism allows the mycobacteria to persist during long time periods, establishing the so-called latent infection. Even in the presence of a competent immune response, the host cannot eliminate the pathogen, only managing to keep it surrounded by an unfavorable microenvironment for its growth. However, conditions such as immunosuppression may reestablish optimal conditions for bacterial growth, culminating in the onset of active disease. The interactions between the pathogen and its host are still not completely elucidated. Nonetheless, many studies are being carried out in order to clarify this complex relationship, thus creating new possibilities for patient approach and laboratory screening.


Subject(s)
Humans , Antigens, Bacterial/immunology , Bacterial Proteins/immunology , Host-Pathogen Interactions/immunology , Latent Tuberculosis/microbiology , Mycobacterium tuberculosis/physiology , Protein Kinases/immunology , Antigens, Bacterial/genetics , Bacterial Proteins/genetics , Immune Evasion , Immunologic Tests , Latent Tuberculosis/diagnosis , Mycobacterium tuberculosis/immunology , Protein Kinases/genetics
13.
Rev. méd. Chile ; 143(3): 304-309, mar. 2015. ilus, tab
Article in Spanish | LILACS | ID: lil-745627

ABSTRACT

Background: Facioscapulohumeral muscular dystrophy is the third most common muscular dystrophy with an estimated prevalence of 1 per 20.000 and a normal life expectancy in the majority of patients. However, approximately 15% of patients become wheelchair bound in the course of their life. It is a hereditary autosomal dominant disease with high (95%) penetrance by the age of 20, but with variable degree of phenotypic expression even in the same family group. Symptoms frequently start in the second decade of life, with facial and scapular weakness. Aim: To report the clinical features of seven patients with the disease, seen at a public hospital. Material and Methods: Analysis of seven patients with genetic study seen in a public Hospital in Santiago. Results: The age of patients fluctuated from 18 to 61 years and four were females. The mean age at onset of symptoms was 29 years and four had a family history of the disease. The usual presenting complaint was arm or shoulder asymmetric weakness. Four patients had bone pain. Facial involvement was present in four. A genetic study was done in five patients, the other two patients were relatives, confirming the contraction or lower number of repetitions in D4Z4 region. After 12 years of follow up only 2 patients older than 60 years cannot work and one female patients is in a semi dependent state at the age of 30. Conclusions: The clinical workup in the diagnosis and the timely indication of genetic studies are highlighted, to avoid unnecessary and invasive procedures. The variability in the phenotypic expression in a similar genetic defect is discussed and the genetic or epigenetic mechanisms of this muscular dystrophy are described.


Subject(s)
Animals , Female , Humans , Male , Mice , Bacterial Proteins/immunology , Gene Expression Regulation, Bacterial/immunology , Lipoproteins/immunology , Pneumonia, Pneumococcal/immunology , Streptococcus pneumoniae/immunology , /immunology , Bacterial Proteins/genetics , Disease Models, Animal , Gene Expression Regulation, Bacterial/genetics , Immunologic Deficiency Syndromes/genetics , Immunologic Deficiency Syndromes/immunology , Immunologic Deficiency Syndromes/pathology , Interleukin-1 Receptor-Associated Kinases/genetics , Interleukin-1 Receptor-Associated Kinases/immunology , Lipoproteins/genetics , Mice, Knockout , Macrophages/immunology , Macrophages/pathology , NF-kappa B/genetics , NF-kappa B/immunology , Pneumonia, Pneumococcal/genetics , Pneumonia, Pneumococcal/pathology , Streptococcus pneumoniae/genetics , /genetics , /immunology , /genetics , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/immunology
14.
Ciênc. saúde coletiva ; 20(2): 441-448, 02/2015. tab
Article in Portuguese | LILACS | ID: lil-742219

ABSTRACT

Objetivou-se analisar as internações por condições sensíveis à atenção primária (ICSAP) específicas em mulheres e os fatores que determinam ou influenciam a ocorrência dessas internações (fatores socioeconômicos, sociodemográficos e controle de saúde) por meio de um inquérito de morbidade hospitalar realizado com amostra de 429 mulheres internadas em hospitais conveniados ao Sistema Único de Saúde. O percentual de ICSAP foi 49,42% (n = 212), com destaque para as internações específicas do sexo feminino 19,35% (n = 83). Associaram ao risco de internar por CSAP: idade superior a 60 anos, baixa escolaridade, internação prévia, realização de controle regular de saúde, falta de vínculo com a Estratégia Saúde da Família (ESF) e ser gestante. As causas evidentes foram as condições relacionadas à gravidez, ao parto e ao puerpério e às inflamações nos órgãos pélvicos femininos. Os resultados sugerem falhas no atendimento ambulatorial que deveria ser oportuno e resolutivo no contexto da saúde da mulher.


The scope of this paper was to analyze female-specific sensitive hospitalization occurring in primary care conditions and factors that determine or affect the occurrence of such hospitalizations (social, economic and demographic factors; health control). Analysis was performed by surveys on hospital morbidity with a sample of 429 females attended in Unified Health System (SUS) contracted hospitals. The sensitive hospitalizations percentage in primary care reached 49.42% (n = 212), highlighting female-specific hospitalization at 19.35% (n = 83). Hospitalization risks comprised elderly people over sixty, low schooling, previous hospitalizations, normal health control, lack of association with the Family Health Strategy and pregnancy. Evident causes were related to conditions of pregnancy, childbirth, post-partum and inflammations of the female pelvic organs. Results suggested flaws in outpatient attendance that should be adequate and provide solutions in women’s health.


Subject(s)
Humans , Infant , Bacterial Proteins/immunology , Carrier Proteins/immunology , Diphtheria-Tetanus-Pertussis Vaccine/adverse effects , Diphtheria-Tetanus-Pertussis Vaccine/immunology , Haemophilus Vaccines/adverse effects , Haemophilus Vaccines/immunology , Immunoglobulin D/immunology , Lipoproteins/immunology , Pneumococcal Vaccines/adverse effects , Pneumococcal Vaccines/immunology , Poliovirus Vaccine, Inactivated/adverse effects , Poliovirus Vaccine, Inactivated/immunology , Antibodies, Bacterial/immunology , Antibodies, Viral/immunology , Diphtheria-Tetanus-Pertussis Vaccine/administration & dosage , Haemophilus Vaccines/administration & dosage , Immunization Schedule , Netherlands , Pneumococcal Vaccines/administration & dosage , Poliovirus Vaccine, Inactivated/administration & dosage , Vaccination , Vaccines, Conjugate , Vaccines, Combined/administration & dosage , Vaccines, Combined/adverse effects , Vaccines, Combined/immunology
15.
Mem. Inst. Oswaldo Cruz ; 109(1): 29-37, 02/2014. tab, graf
Article in English | LILACS | ID: lil-703641

ABSTRACT

Rheumatoid arthritis (RA) is an autoimmune disease characterised by the destruction of articular cartilage and bone damage. The chronic treatment of RA patients causes a higher susceptibility to infectious diseases such as tuberculosis (TB); one-third of the world’s population is latently infected (LTBI) with Mycobacterium tuberculosis (Mtb). The tuberculin skin test is used to identify individuals LTBI, but many studies have shown that this test is not suitable for RA patients. The goal of this work was to test the specific cellular immune responses to the Mtb malate synthase (GlcB) and heat shock protein X (HspX) antigens of RA patients and to correlate those responses with LTBI status. The T-helper (Th)1, Th17 and Treg-specific immune responses to the GlcB and HspX Mtb antigens were analysed in RA patients candidates for tumour necrosis factor-α blocker treatment. Our results demonstrated that LTBI RA patients had Th1-specific immune responses to GlcB and HspX. Patients were followed up over two years and 14.3% developed active TB. After the development of active TB, RA patients had increased numbers of Th17 and Treg cells, similar to TB patients. These results demonstrate that a GlcB and HspX antigen assay can be used as a diagnostic test to identify LTBI RA patients.


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Antigens, Bacterial/immunology , Arthritis, Rheumatoid/immunology , Bacterial Proteins/immunology , Latent Tuberculosis/diagnosis , Malate Synthase/immunology , Mycobacterium tuberculosis/immunology , T-Lymphocytes, Regulatory/immunology , Analysis of Variance , Arthritis, Rheumatoid/complications , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Immunity, Cellular/immunology , /blood , Longitudinal Studies , Latent Tuberculosis/complications , Latent Tuberculosis/immunology , Leukocytes, Mononuclear/immunology , Th1 Cells/immunology , /immunology , Transforming Growth Factor beta/analysis , Tumor Necrosis Factor-alpha/immunology
16.
Article in English | WPRIM | ID: wpr-191843

ABSTRACT

Clostridium (C.) difficile is a common cause of nosocomial diarrhea in horses. Vancomycin and metronidazole have been used as standard treatments but are only moderately effective, which highlights the need for a novel alternative therapy. In the current study, we prepared antiserum of equine origin against both C. difficile toxins A and B as well as whole-cell bacteria. The toxin-neutralizing activities of the antibodies were evaluated in vitro and the prophylactic effects of in vivo passive immunotherapy were demonstrated using a conventional mouse model. The data demonstrated that immunized horses generated antibodies against both toxins A and B that possessed toxin-neutralizing activity. Additionally, mice treated with the antiserum lost less weight without any sign of illness and regained weight back to a normal range more rapidly compared to the control group when challenged orally with 10(7) C. difficile spores 1 day after serum injection. These results indicate that intravenous delivery of hyperimmune serum can protect animals from C. difficile challenge in a dose-dependent manner. Hence, immunotherapy may be a promising prophylactic strategy for preventing C. difficile infection in horses.


Subject(s)
Animals , Antibodies, Bacterial/blood , Bacterial Proteins/immunology , Bacterial Toxins/immunology , Clostridium Infections/microbiology , Clostridioides difficile/immunology , Enterotoxins/immunology , Female , Horse Diseases/microbiology , Horses , Immune Sera/immunology , Immunization, Passive/veterinary , Mice , Mice, Inbred C57BL , Spores, Bacterial/immunology
17.
West Indian med. j ; 62(1): 12-20, Jan. 2013. ilus, tab
Article in English | LILACS | ID: biblio-1045581

ABSTRACT

The rationale of this study was to use several immunological assays to investigate the reactivity of immunoglobulin binding protein (IBP) to immunoglobulins from various avian and mammalian species. The IBP studied were Staphylococcal protein A (SpA), Streptococcal protein G (SpG), Peptostreptococcal protein L (SpL) and recombinant protein LA (SpLA). The various immunological techniques used were double immunodiffusion (Ouchterlony technique) that tested positive high protein reactivities, direct and competitive enzyme-linked immunosorbent assays (ELISAs) that tested moderate and low positive protein binding capacities, respectively. In addition to sandwich ELISAs, immunoblot analyses and Ig-purification by SpA-affinity chromatography, which were sensitive tests and helpful in the screening and confirmatory tests were also used. The Ouchterlony technique showed that compared to the other proteins, SpLA had the highest range of reactivity with animal sera and purified immunoglobulins while SpL was least reactive. With the direct ELISA, SpL reacted with the raccoon sera, rabbit IgG and with IgY from bantam hens and pigeons. While with the direct ELISA, SpA reacted with sera from skunk, coyote, raccoon, mule, donkey and human. The sandwich ELISA revealed high reactivity of both SpG and SpLA with mammalian sera titres ranging from 1:32 (raccoon serum) to 1:1024 (mule and donkey sera).These results suggest that IBP can be used for the detection of immunoglobulin using various immunological assays and this is important for the diagnosis of infectious diseases in animal and bird populations studied and in the purification of immunoglobulins.


El fundamento de este estudio radica en el uso de varios ensayos inmunológicos para investigar la reactividad de la proteína de unión de la inmunoglobulina (IBP) frente a las inmunoglobulinas de varias especies aviarias y mamíferas. Las proteínas IBP estudiadas fueron la proteína estafilocócica A (SpA), la proteína estreptocócica G (SpG), la proteína peptoestreptocócica L (SpL), y la proteína recombinante LA (SpLA). Las varias técnicas inmunológicas usadas fueron: la inmunodifusión doble (técnica de Ouchterlony) para examinar las reactividades positivas de la proteína alta; el ensayo por inmunoabsorción ligado a enzimas(ELISA), de tipo directo y competitivo, para examinar la capacidad de realizar uniones positivas de proteína moderada y baja, respectivamente, además del ensayo ELISA 'Sándwich', los análisis inmunoblot, yla purificación de IgG, mediante cromatografía de afinidad, los cuales fueron pruebas sensibles y útiles en el tamizaje y las pruebas de confirmación. La técnica de Ouchterlony mostró que - en comparación con otras proteínas - la SpLA tenía el grado más alto de reactividad con los sueros animales y las inmunoglobulinas purificadas, mientras que la SpL fue la menos reactiva. Con el ELISA directo, la SpL reaccionó con los sueros de mapache, la IgG de conejo, así como con la IgY de palomas y gallinas de Bantam, en tanto con el ELISA directo, la SpA reaccionó con sueros de mofeta, coyote, mapache, mula, asno y seres humanos. ELISA "sándwich" reveló una alta reactividad tanto de SpG como de SpLA, con títulos séricos mamíferos que iban desde 1:32 (suero de mapache) hasta 1:1024 (sueros de mula y de asno). Estos resultados sugieren que la proteína de unión IBP puede usarse en la detección de la inmunoglobulina usando varios ensayos inmunológicos, lo cual es importante para el diagnóstico de enfermedades infecciosas en las poblaciones animales y aviarias bajo estudio, así como para la purificación de inmunoglobulinas.


Subject(s)
Humans , Animals , Bacterial Proteins/immunology , Birds/immunology , Immunoglobulins/biosynthesis , Chromatography, Affinity , Immunoenzyme Techniques/methods , Mammals/immunology , Recombinant Proteins/immunology , Carrier Proteins/immunology , Communicable Diseases/diagnosis
18.
Journal of Infection and Public Health. 2013; 6 (3): 166-172
in English | IMEMR | ID: emr-142717

ABSTRACT

Dialysis patients are more likely than the general population to develop active tuberculosis [TB]. In these patients, the availability of a highly sensitive and specific test to diagnose latent TB will ensure earlier treatment and decreased progression to active disease. In the current study, the Quanti-FERON-TB Gold In-Tube [QFT-G] test was compared with the tuberculin skin test [TST] for the diagnosis of latent tuberculosis infection [LTBI] among 200 hemodialysis patients and 15 confirmed TB disease cases in a tertiary care center in Saudi Arabia. Among the LTBI cases, 26 [13%] were TST positive, and 65 [32.5%] were positive by the QTF-G test, with an overall agreement between the 2 tests of 75.5% [k = 0.34] being observed. Among the confirmed tuberculosis disease cases, none were positive by TST, and 10 [66.7%] were positive by the QTF-G test, resulting in an overall agreement of 33.3% [k = 0]. A comparison between the TST and the QTF-G test was performed based on the sensitivity, specificity, and area under the curve [AUC] obtained for the tests. The QTF-G test was more sensitive and less specific than the TST in predicting the confirmed TB disease cases. When we tested the correspondence of the AUC values between the 2 diagnostic modalities, the obtained p-value was 0.0003. In conclusion, the AUCs of the examined diagnostic modalities are significantly different in predicting LTBI and tuberculosis


Subject(s)
Humans , Male , Female , Tuberculin Test , Renal Dialysis/adverse effects , Mycobacterium tuberculosis/immunology , Bacterial Proteins/immunology , Recombinant Proteins/immunology , Renal Insufficiency/complications , Sensitivity and Specificity
19.
Braz. j. med. biol. res ; 45(12): 1183-1194, Dec. 2012. ilus, mapas, tab
Article in English | LILACS | ID: lil-659642

ABSTRACT

In the last several years, the use of dendritic cells has been studied as a therapeutic strategy against tumors. Dendritic cells can be pulsed with peptides or full-length protein, or they can be transfected with DNA or RNA. However, comparative studies suggest that transfecting dendritic cells with messenger RNA (mRNA) is superior to other antigen-loading techniques in generating immunocompetent dendritic cells. In the present study, we evaluated a new therapeutic strategy to fight tuberculosis using dendritic cells and macrophages transfected with Hsp65 mRNA. First, we demonstrated that antigen-presenting cells transfected with Hsp65 mRNA exhibit a higher level of expression of co-stimulatory molecules, suggesting that Hsp65 mRNA has immunostimulatory properties. We also demonstrated that spleen cells obtained from animals immunized with mock and Hsp65 mRNA-transfected dendritic cells were able to generate a mixed Th1/Th2 response with production not only of IFN-γ but also of IL-5 and IL-10. In contrast, cells recovered from mice immunized with Hsp65 mRNA-transfected macrophages were able to produce only IL-5. When mice were infected with Mycobacterium tuberculosis and treated with antigen-presenting cells transfected with Hsp65 mRNA (therapeutic immunization), we did not detect any decrease in the lung bacterial load or any preservation of the lung parenchyma, indicating the inability of transfected cells to confer curative effects against tuberculosis. In spite of the lack of therapeutic efficacy, this study reports for the first time the use of antigen-presenting cells transfected with mRNA in experimental tuberculosis.


Subject(s)
Animals , Male , Mice , Antigen-Presenting Cells/immunology , Bacterial Proteins/administration & dosage , /administration & dosage , Mycobacterium tuberculosis/immunology , RNA, Messenger/immunology , Tuberculosis Vaccines/administration & dosage , Tuberculosis/immunology , Bacterial Proteins/adverse effects , Bacterial Proteins/immunology , /adverse effects , /immunology , Mice, Inbred BALB C , RNA, Messenger/adverse effects , Spleen/immunology , Transfection , Tuberculosis Vaccines/adverse effects , Tuberculosis Vaccines/immunology , Tuberculosis/prevention & control
20.
Mem. Inst. Oswaldo Cruz ; 107(supl.1): 90-94, Dec. 2012. ilus, mapas, tab
Article in English | LILACS | ID: lil-659746

ABSTRACT

Leprosy is a slowly evolving disease that occurs mainly in adults. In this study, the Mamaría Village, state of Portuguesa was selected because it had one of the highest prevalence rates (13.25%) of leprosy cases in 1997. Between 1998-2004, 20.2% of the 89 cases registered in this village were less than 15 years old and 61.8% were males. Pau-cibacillary (PB) lesions were the predominant clinical forms identified, although also multibacillary (MB) forms were found. Additionally, 76% of the patients were bacteriologically negative. At the time of diagnosis, 75% of the patients presented with grade 0 disabilities, 23% with grade 1 and 2% with grade 2. Serum samples were collected from 18 PB and 15 MB patients, in addition to 14 family contacts, at the beginning and end of treatment. All the groups were re-evaluated during a three-year period (2008-2011). The proteins used for evaluation were ML0405, ML2331 and LID-1. These mycobacterial proteins were highly specific for Mycobacterium leprae and the IgG responses decreased in both MB and PB patients during multidrug treatment. Our results suggest that these antigens could be used as markers for successful treatment of non-reactional lepromatous patients.


Subject(s)
Adolescent , Adult , Female , Humans , Male , Young Adult , Bacterial Proteins/blood , Immunoglobulin G/blood , Leprosy, Multibacillary/diagnosis , Leprosy, Paucibacillary/diagnosis , Mycobacterium leprae/immunology , Antibodies, Bacterial/blood , Bacterial Proteins/immunology , Leprosy, Multibacillary/epidemiology , Leprosy, Paucibacillary/epidemiology , Recombinant Proteins/blood , Recombinant Proteins/immunology , Venezuela/epidemiology
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