RESUMO
Giardiasis is a parasitic disease caused by Giardia lamblia (G. lamblia) that affects people worldwide. Still, few studies report on the immunoregulatory effects of the biomolecules of colostrum during interactions with G. lamblia. This study aimed to assess the concentrations of melatonin and cortisol hormones, the percentage of Treg cells, and the levels of cytokines IL-10 and TGF-ß in colostrum from mothers who tested positive for the parasite. This cross-sectional study analyzed colostrum samples from 25 puerperal. The samples were tested using an ELISA to determine if they were seropositive for G. lamblia and the type of antibody present (IgM and IgG). Based on the results, the samples were divided into three groups: a control group (N = 10) with no reaction to either IgM or IgG, a group seropositive for IgG (IgG+/IgM-; N = 8), and a group seropositive for IgM (IgM+/IgG-; N = 7). The concentrations of melatonin and cortisol were measured using the ELISA method. Additionally, cytokines IL-10 and TGF-ß and immunophenotyping were analyzed using flow cytometry. In the group that tested positive for IgM anti-G. lamblia, the concentration of melatonin was lower. However, in the colostrum from mothers who tested positive for IgG anti-G. lamblia, the level of this hormone had increased. The cortisol levels were similar between the groups, regardless of seropositivity. There was a higher percentage of Treg cells in the colostrum from mothers who tested positive for IgM anti-G. lamblia. TGF-ß levels also increased in the colostrum of mothers who tested positive for IgM anti-G. lamblia. In the seronegative group for G. lamblia, there was a positive correlation between melatonin concentration and the percentage of Treg cells. These data suggest that the increase in regulatory cells and cytokines and the reduction in melatonin in colostrum from mothers with recent giardia infection may contribute to the evolution and manifestation of the disease.
Assuntos
Colostro , Giardia lamblia , Giardíase , Melatonina , Linfócitos T Reguladores , Fator de Crescimento Transformador beta , Melatonina/metabolismo , Melatonina/imunologia , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo , Humanos , Feminino , Giardíase/imunologia , Giardíase/parasitologia , Giardia lamblia/imunologia , Adulto , Colostro/imunologia , Colostro/química , Estudos Transversais , Fator de Crescimento Transformador beta/metabolismo , Fator de Crescimento Transformador beta/imunologia , Interleucina-10/metabolismo , Interleucina-10/imunologia , Imunoglobulina M/imunologia , Imunoglobulina G/imunologia , Imunoglobulina G/sangue , Hidrocortisona , Gravidez , Adulto JovemRESUMO
Discriminate the severity level of COVID-19 disease is still a challenge. Here we investigate the capability of micro-infrared absorption spectroscopy (micro-FTIR) to probe COVID-19 severity level and predict hyperinflammation, correlating the assigned vibrational data to relevant biomolecules related to the immune system. Saliva of 184 patients was analysed by ELISA assay (Hepcidin) and micro-FTIR. Vibrational bands related to IgM and IgA can discriminate healthy from Severe individuals (sensitivity ≥ 0.749, specificity ≥ 0.945) and are less effective in discriminating Mild or Moderate individuals from the Severe group (sensitivity ≥ 0.628, specificity ≥ 0.867). Analysis of the second derivative of spectra probed increased levels of IL-6 in the saliva a key additional information for the degree of severity prediction. Because the model discriminates all the groups regarding the Severe group, it predicts an intense state of inflammation based on FTIR analysis. It is a powerful tool for predicting hyperinflammation conditions related to SARS-CoV-2 infection and may be an ally in implementing drugs or therapeutic approaches to manage COVID-19 in the Severe stage in healthcare facilities.
Assuntos
COVID-19 , Inflamação , SARS-CoV-2 , Saliva , Índice de Gravidade de Doença , Humanos , COVID-19/diagnóstico , Saliva/química , Saliva/virologia , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Feminino , Masculino , SARS-CoV-2/isolamento & purificação , SARS-CoV-2/imunologia , Adulto , Pessoa de Meia-Idade , Interleucina-6/análise , Idoso , Imunoglobulina A/análise , Imunoglobulina M/análise , Imunoglobulina M/imunologiaRESUMO
INTRODUCCIÓN: La comprensión del comportamiento de la respuesta humoral en COVID-19 continúa siendo un desafío para la producción de vacunas que proporcionen inmunidad más duradera. OBJETIVO: Describir la respuesta humoral natural inducida por SARS- CoV-2 en personal de salud con base en el perfil epidemiológico y clínico. MATERIALES Y MÉTODOS: Estudio transversal en personal de salud de hospitales públicos de referencia del Departamento de Alto Paraná, Paraguay. Se incluyeron 962 participantes, mediante muestreo no probabilístico de tipo consecutivo, aplicación de cuestionario y toma de muestras sanguíneas. Se buscaron anticuerpos por ensayo inmunocromatográfico para detección de IgM e IgG contra SARS- CoV-2 y por el método ELISA de captura de IgG específicos contra la proteína spike (SARS-CoV-2) y se evaluaron factores asociados a la seropositividad. RESULTADOS: La seroprevalencia global fue 36,5% (IC 95%: 33,4 - 39,5); 59,3% (n: 571) de los encuestados refirió haber tenido síntomas compatibles al COVID-19 entre el inicio de la pandemia y la fecha de toma de muestra, de estos 44% (n: 251) resultó seropositivo; 10,4% (n: 100) manifestó no haber tenido síntomas en el periodo estudiado, pero tuvo un resultado positivo. Los factores asociados a la seropositividad fueron: presencia de síntomas (p 90 días). CONCLUSIONES: Las características clínicas fueron mayormente asociadas con la seropositividad y la seropreva- lencia en los sintomáticos varió de acuerdo con el tiempo transcurrido desde el inicio de los síntomas y la serología.
BACKGROUND: Understanding the behavior of humoral response in COVID-19 continues to be a challenge to produce vaccines that provide long-lasting immunity. AIM: To describe the natural humoral response induced by SARS-CoV-2 among healthcare workers based on epidemiological and clinical profiles. METHODS: Cross-sectional study in healthcare workers from public hospitals in the Department of Alto Paraná, Paraguay, 962 participants were recruited through consecutive sampling, using a questionnaire and blood sampling. Antibodies were determined by immunochromatography assay for detection of IgM and IgG and by SARS-CoV-2 IgG anti-spike capture ELISA method and factors associated with seropositivity were evaluated. RESULTS: The overall seropositivity was 36.5% (95% CI: 33.4 - 39.5); 59.3% (n: 571) of respondents reported symptoms compatible with COVID-19 since the start of the pandemic and the date of blood draw, 44% (n: 251) of them tested positive; 10.4% (n: 100) who reported no history of symptoms tested positive. The factors associated with seropositivity were the presence of symptoms (p 90 days). CONCLUSIONS: Clinical characteristics were mostly associated with seropositivity and sero prevalence in symptomatic participants varied according to the time elapsed from the onset of symptoms to serology.
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Pessoal de Saúde , SARS-CoV-2/imunologia , COVID-19/imunologia , COVID-19/epidemiologia , Paraguai , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Ensaio de Imunoadsorção Enzimática , Estudos Soroepidemiológicos , Estudos Transversais , Análise Multivariada , Inquéritos e Questionários , Cromatografia de Afinidade , Vacinação , Imunidade HumoralRESUMO
The diagnosis of dengue disease, caused by the dengue virus (DENV) (a flavivirus), often requires serologic testing during acute and early convalescent phases of the disease. Some symptoms of DENV infection, such as nonspecific fever, are similar to those caused by infection with SARS-CoV-2, the virus that causes COVID-19. In studies with few COVID-19 cases, positive DENV immunoglobulin M (IgM) results were reported with various serologic tests, indicating possible cross-reactivity in these tests for DENV and SARS-CoV-2 infections (1,2). DENV antibodies can cross-react with other flaviviruses, including Zika virus. To assess the potential cross-reactivity of SARS-CoV-2, DENV, and Zika virus IgM antibodies, serum specimens from 97 patients from Puerto Rico and 12 U.S.-based patients with confirmed COVID-19 were tested using the DENV Detect IgM Capture enzyme-linked immunosorbent assay (ELISA) (InBios International).* In addition, 122 serum specimens from patients with confirmed dengue and 121 from patients with confirmed Zika virus disease (all from Puerto Rico) were tested using the SARS-CoV-2 pan-Ig Spike Protein ELISA (CDC). Results obtained for DENV, Zika virus IgM, and SARS-CoV-2 antibodies indicated 98% test specificity and minimal levels of cross-reactivity between the two flaviviruses and SARS-CoV-2. These findings indicate that diagnoses of dengue or Zika virus diseases with the serological assays described in this report are not affected by COVID-19, nor do dengue or Zika virus diseases interfere with the diagnosis of COVID-19.
Assuntos
Anticorpos Antivirais/sangue , Vírus da Dengue/imunologia , Imunoglobulina M/imunologia , SARS-CoV-2/imunologia , Testes Sorológicos , Zika virus/imunologia , COVID-19/diagnóstico , Reações Cruzadas/imunologia , Dengue/diagnóstico , Ensaio de Imunoadsorção Enzimática , Humanos , Porto Rico , Sensibilidade e Especificidade , Estados Unidos , Infecção por Zika virus/diagnósticoRESUMO
Serological cross-reactivity has proved to be a challenge to diagnose Zika virus (ZIKV) infections in dengue virus (DENV) endemic countries. Confirmatory testing of ZIKV IgM positive results by plaque reduction neutralization tests (PRNTs) provides clarification in only a minority of cases because most individuals infected with ZIKV were previously exposed to DENV. The goal of this study was to evaluate the performance of a ZIKV/DENV DUO IgM antibody capture ELISA (MAC-ELISA) for discriminating between DENV and ZIKV infections in endemic regions. Our performance evaluation included acute and convalescent specimens from patients with real-time reverse transcription polymerase chain reaction (RT-PCR)-confirmed DENV or ZIKV from the Sentinel Enhanced Dengue Surveillance System in Ponce, Puerto Rico. The ZIKV/DENV DUO MAC-ELISA specificity was 100% for DENV (N = 127) and 98.4% for ZIKV (N = 275) when specimens were tested during the optimal testing window (days post-onset of illness [DPO] 6-120). The ZIKV/DENV DUO MAC-ELISA sensitivity of RT-PCR confirmed specimens reached 100% for DENV by DPO 6 and for ZIKV by DPO 9. Our new ZIKV/DENV DUO MAC-ELISA was also able to distinguish ZIKV and DENV regardless of previous DENV exposure. We conclude this novel serologic diagnostic assay can accurately discriminate ZIKV and DENV infections. This can potentially be useful considering that the more labor-intensive and expensive PRNT assay may not be an option for confirmatory diagnosis in areas that lack PRNT capacity, but experience circulation of both DENV and ZIKV.
Assuntos
Anticorpos Antivirais/imunologia , Dengue/diagnóstico , Ensaio de Imunoadsorção Enzimática/métodos , Imunoglobulina M/imunologia , Infecção por Zika virus/diagnóstico , Reações Cruzadas , Dengue/imunologia , Dengue/transmissão , Vírus da Dengue/imunologia , Doenças Endêmicas , Feminino , Humanos , Masculino , Testes Sorológicos/métodos , Proteínas não Estruturais Virais , Zika virus/imunologia , Infecção por Zika virus/imunologia , Infecção por Zika virus/transmissãoRESUMO
Cryptococcosis is an opportunistic disease caused by the fungus Cryptococcus neoformans and Cryptococcus gattii. It starts as a pulmonary infection that can spread to other organs, such as the brain, leading to the most serious occurrence of the disease, meningoencephalitis. The humoral response has already been described in limiting the progression of cryptococcosis where the B-1 cell seems to be responsible for producing natural IgM antibodies, crucial for combating fungal infections. The role of the B-1 cell in C. neoformans infection has been initially described, however the role of the humoral response of B-1 cells has not yet been evaluated during C. gattii infections. In the present study we tried to unravel this issue using XID mice, a murine model deficient in the Btk protein which compromises the development of B-1 lymphocytes. We use the XID mice compared to BALB/c mice that are sufficient for the B-1 population during C. gattii infection. Our model of chronic lung infection revealed that XID mice, unlike the sufficient group of B-1, had early mortality with significant weight loss, in addition to reduced levels of IgM and IgG specific to GXM isolated from the capsule of C. neoformans. In addition to this, we observed an increased fungal load in the blood and in the brain. We described an increase in the capsular size of C. gattii and the predominant presence of cytokines with a Th2 profile was also observed in these animals. Thus, the present study strongly points to a higher susceptibility of the XID mouse to C. gattii, which suggests that the presence of B-1 cells and anti-GXM antibodies is fundamental during the control of infection by C. gattii.
Assuntos
Criptococose/etiologia , Cryptococcus gattii , Suscetibilidade a Doenças/imunologia , Hospedeiro Imunocomprometido , Doenças por Imunodeficiência Combinada Ligada ao Cromossomo X/complicações , Animais , Biomarcadores , Contagem de Colônia Microbiana , Criptococose/metabolismo , Cryptococcus gattii/imunologia , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imunoglobulina M/sangue , Imunoglobulina M/imunologia , Camundongos , Doenças por Imunodeficiência Combinada Ligada ao Cromossomo X/genética , Doenças por Imunodeficiência Combinada Ligada ao Cromossomo X/imunologiaRESUMO
BACKGROUND: In 2015-2016, a large Zika virus (ZIKV) outbreak occurred in the Americas. Although the exact ZIKV antibody kinetics after infection are unknown, recent evidence indicates the rapid waning of ZIKV antibodies in humans. Therefore, we aimed to determine the levels of ZIKV antibodies more than three years after a ZIKV infection. METHODS: We performed ZIKV virus neutralization tests (VNT) and a commercial ZIKV non-structural protein 1 (NS1) IgG ELISA in a cohort of 49 participants from Suriname who had a polymerase-chain-reaction-confirmed ZIKV infection more than three years ago. Furthermore, we determined the presence of antibodies against multiple dengue virus (DENV) antigens. RESULTS: The ZIKV seroprevalence in this cohort, assessed with ZIKV VNT and ZIKV NS1 IgG ELISA, was 59.2% and 63.3%, respectively. There was, however, no correlation between these two tests. Furthermore, we did not find evidence of a potential negative influence of DENV immunity on ZIKV antibody titers. CONCLUSIONS: ZIKV seroprevalence, assessed with two commonly used serological tests, was lower than expected in this cohort of participants who had a confirmed previous ZIKV infection. This can have implications for future ZIKV seroprevalence studies and possibly for the duration of immunological protection after a ZIKV infection.
Assuntos
Anticorpos Neutralizantes/análise , Infecção por Zika virus/imunologia , Zika virus/imunologia , Adulto , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/análise , Anticorpos Antivirais/imunologia , Estudos de Coortes , Reações Cruzadas/imunologia , Dengue/virologia , Vírus da Dengue/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Humanos , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Masculino , Pessoa de Meia-Idade , Testes de Neutralização/métodos , Estudos Soroepidemiológicos , Testes Sorológicos/métodos , Suriname , Zika virus/patogenicidade , Infecção por Zika virus/metabolismo , Infecção por Zika virus/virologiaRESUMO
Dengue is a major public health problem in tropical and sub-tropical regions worldwide. Since the Zika epidemic and the increased co-circulation of other arboviruses, the serology-based diagnosis of dengue has become more problematic due to the high antigenic resemblance, especially among the flavivirus family. Therefore, a more comprehensive understanding of the diversity, specificity and temporal evolution of the antibody response following dengue infection is needed. In order to close this knowledge gap, we used a high-density peptide microarray of 9,072 linear peptides covering the entire proteome diversity of dengue, Zika, yellow fever and chikungunya viruses. The IgM and IgG antibody responses were measured against the designed microarray in symptomatic dengue infected individuals from an arbovirus endemic area in Peru and in overseas travelers returning to Belgium, as representatives of multiple-exposed and primary infections, respectively. Serum samples were collected longitudinally across four time points over the period of six months in Peru and over two time points in travelers. We show that epitopes eliciting the strongest flavivirus cross-reactive antibodies, in both primary and secondary infections were concentrated in the capsid, E, NS1, NS3 and NS5 proteins. The IgG antibody responses against NS1 and NS3 followed a rise-and-fall pattern, with peak titers between two to four weeks after onset of illness. The response to the E and NS5 proteins increased rapidly in the acute phase and was maintained at stable levels until at least 6 months after illness. A more scattered IgM antibody reactivity across the viral proteome was observed in the acute phase of the disease and that persisted through the 6-month window. The magnitude, breadth (i.e. number of unique epitopes targeted) and depth (i.e. number of epitope variants recognized) of the IgG response was higher in secondary infections compared to primary infections. For IgM antibodies, the magnitude of the response was higher in primary infected individuals whereas the breadth and depth of the response was lower in this group compared with the endemic subjects. Finally, through this arboviral proteome-wide epitope mapping, we were able to identify IgM and IgG dengue-specific epitopes which can be useful serological markers for dengue diagnosis and serostatus determination.
Assuntos
Anticorpos Antivirais/imunologia , Vírus da Dengue/imunologia , Dengue/imunologia , Epitopos/imunologia , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Adolescente , Adulto , Anticorpos Antivirais/sangue , Bélgica , Reações Cruzadas/imunologia , Dengue/sangue , Vírus da Dengue/genética , Ensaio de Imunoadsorção Enzimática , Epitopos/química , Feminino , Flavivirus/imunologia , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , Pessoa de Meia-Idade , Peru , Testes Sorológicos , Viagem , Adulto JovemRESUMO
Mayaro virus (MAYV), which causes mayaro fever, is endemic to limited regions of South America that may expand due to the possible involvement of Aedes spp. mosquitoes in its transmission. Its effective control will require the accurate identification of infected individuals, which has been restricted to nucleic acid-based tests due to similarities with other emerging members of the Alphavirus genus of the Togaviridae family; both in structure and clinical symptoms. Serological tests have a more significant potential to expand testing at a reasonable cost, and their performance primarily reflects that of the antigen utilized to capture pathogen-specific antibodies. Here, we describe the assembly of a synthetic gene encoding multiple copies of antigenic determinants mapped from the nsP1, nsP2, E1, and E2 proteins of MAYV that readily expressed as a stable chimeric protein in bacteria. Its serological performance as the target in ELISAs revealed a high accuracy for detecting anti-MAYV IgM antibodies. No cross-reactivity was observed with serum from seropositive individuals for dengue, chikungunya, yellow fever, Zika, and other infectious diseases as well as healthy individuals. Our data suggest that this bioengineered antigen could be used to develop high-performance serological tests for MAYV infections.
Assuntos
Infecções por Alphavirus/diagnóstico , Alphavirus/imunologia , Epitopos/imunologia , Infecções por Togaviridae/diagnóstico , Aedes/virologia , Alphavirus/patogenicidade , Infecções por Alphavirus/imunologia , Infecções por Alphavirus/transmissão , Infecções por Alphavirus/virologia , Animais , Ensaio de Imunoadsorção Enzimática , Epitopos/genética , Epitopos/ultraestrutura , Feminino , Genes Sintéticos/genética , Genes Sintéticos/imunologia , Humanos , Imunoglobulina M/imunologia , Masculino , Testes Sorológicos , América do Sul/epidemiologia , Togaviridae/isolamento & purificação , Togaviridae/patogenicidade , Infecções por Togaviridae/imunologia , Infecções por Togaviridae/transmissão , Infecções por Togaviridae/virologiaRESUMO
Hypogammaglobulinemia is the most frequently observed immune defect in chronic lymphocytic leukemia (CLL). Although CLL patients usually have low serum levels of all isotypes (IgG, IgM and IgA), standard immunoglobulin (Ig) preparations for replacement therapy administrated to these patients contain more than 95% of IgG. Pentaglobin is an Ig preparation of intravenous application (IVIg) enriched with IgM and IgA (IVIgGMA), with the potential benefit to restore the Ig levels of all isotypes. Because IVIg preparations at high doses have well-documented anti-inflammatory and immunomodulatory effects, we aimed to evaluate the capacity of Pentaglobin and a standard IVIg preparation to affect leukemic and T cells from CLL patients. In contrast to standard IVIg, we found that IVIgGMA did not modify T cell activation and had a lower inhibitory effect on T cell proliferation. Regarding the activation of leukemic B cells through BCR, it was similarly reduced by both IVIgGMA and IVIgG. None of these IVIg preparations modified spontaneous apoptosis of T or leukemic B cells. However, the addition of IVIgGMA on in vitro cultures decreased the apoptosis of T cells induced by the BCL-2 inhibitor, venetoclax. Importantly, IVIgGMA did not impair venetoclax-induced apoptosis of leukemic B cells. Overall, our results add new data on the effects of different preparations of IVIg in CLL, and show that the IgM/IgA enriched preparation not only affects relevant mechanisms involved in CLL pathogenesis but also has a particular profile of immunomodulatory effects on T cells that deserves further investigation.