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Introduction: The attenuation of BCG has led to the loss of not only immunogenic proteins but also lipid antigens. Methods: Thus, we compared the macrophage and T-cell responses to nonpolar lipid extracts harvested from BCG and Mycobacterium tuberculosis (Mtb) to better understand the role of BCG lipids in the already known diminished responses of the vaccine strain. Results: Relative to Mtb, nonpolar lipid extract from BCG presented a reduced capacity to trigger the expression of the genes encoding TNF, IL-1b, IL-6 and IL-10 in RAW 264.7 macrophages. Immunophenotyping of PBMCs isolated from healthy individuals revealed that lipids from both BCG and Mtb were able to induce an increased frequency of CD4+ and CD8+ T cells, but only the lipid extract from Mtb enhanced the frequency of CD4-CD8-double-negative, γσ+, CD4+HLA-DR+, and γσ+HLA-DR+ T cells relative to the nonstimulated control. Interestingly, only the Mtb lipid extract was able to increase the frequency of CD4+ memory (CD45RO+) T cells, whereas the BCG lipid extract induced a diminished frequency of CD4+ central memory (CD45RO+CCR7-) T cells after 48 h of culture compared to Mtb. Discussion: These findings show that the nonpolar lipids of the BCG bacilli presented diminished ability to trigger both proinflammatory and memory responses and suggest a potential use of Mtb lipids as adjuvants to increase the BCG vaccine efficacy.
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Mycobacterium bovis , Tuberculose , Humanos , Vacina BCG , Linfócitos T CD8-Positivos , Células T de Memória , Linfócitos T CD4-Positivos , Macrófagos , Antígenos HLA-DR , LipídeosRESUMO
The objective of this study was to evaluate the seasonal effect of months of the year upon the physiological and productive responses of crossbred dairy cows raised in an Amazonian climate. Twenty lactating cows were evaluated, fed on Brachiaria decumbens pasture, with free access to water and mineral supplementation. Data from climate variables air temperature (AT), relative humidity (RH), rainfall (RA) and temperature and humidity index (THI) were recorded dur ing the months of January to April 2019. The physiological data collected were: respiratory rate (RR, mov/min), heart rate (HR, beats/min), rectal temperature (RT, ºC), udder surface temperature (UST, ºC), body surface temperature (BST, ºC), dorsum surface temperature (DST, ºC), front surface temperature (FST, ºC) and rear shin temperature (RST). Milk production (MP) was also measured. There was a significant difference (P<0.05) of RST and RR with values ranging from 34.8 to 35.5°C and 32.0 to 36.2 mov/min, respectively. There were no significant difference (P>0.05) for BST, and the values for MP and THI were 3.8; 3.8; 4.6; 4.1 kg and 77.7; 79.7; 80.6; 80.1, respectively. It was concluded that there was a seasonal effect of the months of the year evaluated on the respiratory rate of animals, however, it did not change the MP and the THI. The Amazon environment is conducive to causing thermal stress in lactating cows raised on pasture, requiring the use of shading to facilitate the ability of these animals to dissipate heat.(AU)
O objetivo desse estudo foi avaliar os efeitos sazonais dos meses do ano sobre as respostas fisiológicas e produtivas de vacas leiteiras mestiças criadas em clima amazônico. Foram avaliadas 20 vacas lactantes, alimentadas com pasto de Brachiaria decumbens, com acesso livre a água e suplementação mineral. Foram registrados dados das variáveis climáticas temperatura do ar (TA), umidade relativa do ar (UR), precipitação pluviométrica (PP) e índice de temperatura e Umidade (ITU) durante os meses de janeiro a abril de 2019. Os dados fisiológicos coletados foram: frequência respiratória (FR, mov/min), frequência cardíaca (FC, bat./min), temperatura retal (TR, ºC), temperatura superficial do úbere (TSU, ºC), temperatura superfície corporal (TSC, ºC), temperatura superficial do dorso (TSD, ºC), temperatura superficial da fronte (TSF, ºC) e temperatura superficial da canela (TSCA, °C). Também foi mensurada a produção de leite (PL, kg). Houve diferença significativa (P<0,05) da TSCA e FR com valores variando de 34,8 a 35,5°C e 32,0 a 36,2 mov/min, respectivamente. Não houve diferença significativa (P>0,05) para TSC e os valores observados para PL e ITU foram 3,8; 3,8; 4,6; 4,1 kg e 77,7; 79,7; 80,6; 80,1, respectivamente. Houve efeito sazonal dos meses avaliados sobre a taxa respiratória dos animais, no entanto, isso não alterou a PL nem o ITU. O ambiente amazônico é propício a causar estresse térmico em vacas lactantes mantidas a pasto, sendo necessário o uso de sombreamento para facilitar a capacidade de dissipação de calor corporal desses animais.(AU)
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Animais , Feminino , Bovinos/fisiologia , Modalidades Fisiológicas , Estações do Ano , Pastagens , Resposta ao Choque Térmico/fisiologia , BrachiariaRESUMO
BACKGROUND: Leishmaniases are neglected tropical diseases that inflict great burden to poor areas of the globe. Intense research has aimed to identify parasite genetic signatures predictive of infection outcomes. Consistency of diagnostic tools based on these markers would greatly benefit from accurate understanding of Leishmania spp. population genetics. We explored two chromosomal loci to characterize a population of L. braziliensis causing human disease in Northeast Brazil. METHODOLOGY/PRINCIPAL FINDINGS: Two temporally distinct samples of L. braziliensis were obtained from patients attending the leishmaniasis clinic at the village of Corte de Pedra: (2008-2011) primary sample, N = 120; (1999-2001) validation sample, N = 35. Parasites were genotyped by Sanger's sequencing of two 600 base pairs loci starting at nucleotide positions 3,074 and 425,451 of chromosomes 24 and 28, respectively. Genotypes based on haplotypes of biallelic positions in each locus were tested for several population genetic parameters as well as for geographic clustering within the region. Ample geographic overlap of genotypes at the two loci was observed as indicated by non-significant Cusick and Edward's comparisons. No linkage disequilibrium was detected among combinations of haplotypes for both parasite samples. Homozygous and heterozygous genotypes displayed Hardy-Weinberg equilibrium (HWE) at both loci in the two samples when straight observed and expected counts were compared by Chi-square (p>0.5). However, Bayesian statistics using one million Monte-Carlo randomizations disclosed a less robust HWE for chromosome 24 genotypes, particularly in the primary sample (p = 0.04). Fixation indices (Fst) were consistently lower than 0.05 among individuals of the two samples at both tested loci, and no intra-populational structuralization could be detected using STRUCTURE software. CONCLUSIONS/SIGNIFICANCE: These findings suggest that L. braziliensis can maintain stable populations in foci of human leishmaniasis and are capable of robust genetic recombination possibly due to events of sexual reproduction during the parasite's lifecycle.
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Leishmania braziliensis , Leishmaniose Cutânea , Leishmaniose , Teorema de Bayes , Brasil/epidemiologia , Genótipo , Humanos , Leishmania braziliensis/genética , Leishmaniose/parasitologia , Leishmaniose Cutânea/epidemiologia , Leishmaniose Cutânea/parasitologiaRESUMO
BACKGROUND: Leptospirosis is an important public health problem affecting vulnerable urban slum populations in developing country settings. However, the complex interaction of meteorological factors driving the temporal trends of leptospirosis remain incompletely understood. METHODS AND FINDINGS: From March 1996-March 2010, we investigated the association between the weekly incidence of leptospirosis and meteorological anomalies in the city of Salvador, Brazil by using a dynamic generalized linear model that accounted for time lags, overall trend, and seasonal variation. Our model showed an increase of leptospirosis cases associated with higher than expected rainfall, lower than expected temperature and higher than expected humidity. There was a lag of one-to-two weeks between weekly values for significant meteorological variables and leptospirosis incidence. Independent of the season, a weekly cumulative rainfall anomaly of 20 mm increased the risk of leptospirosis by 12% compared to a week following the expected seasonal pattern. Finally, over the 14-year study period, the annual incidence of leptospirosis decreased significantly by a factor of 2.7 (8.3 versus 3.0 per 100,000 people), independently of variations in climate. CONCLUSIONS: Strategies to control leptospirosis should focus on avoiding contact with contaminated sources of Leptospira as well as on increasing awareness in the population and health professionals within the short time window after low-level or extreme high-level rainfall events. Increased leptospirosis incidence was restricted to one-to-two weeks after those events suggesting that infectious Leptospira survival may be limited to short time intervals.
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Leptospira , Leptospirose , Clima , Humanos , Umidade , Incidência , Leptospirose/epidemiologia , Conceitos MeteorológicosRESUMO
Despite highly variable efficacy, BCG (Bacillus Calmette-Guérin) is the only vaccine available to prevent the tuberculosis (TB). Genomic heterogeneity between attenuated BCG strains and virulent Mycobacterium tuberculosis might help to explain this vaccine's impaired capacity to induce long-term protection. Here, we investigate the lipid-related genes absent in attenuated BCG strains in order to correlate changes in both lipid metabolism and cell-wall lipid content to vaccine impairment. Whole genome sequences of M. tuberculosis H37Rv and the six most used BCG strains worldwide were aligned and the absent regions functionally categorized. Genomes of the BCG strains showed a total of 14 non-homologous lipid-related genes, including those belonging to mce3 operon, as well as the gene echaA1, which encodes an enoyl-CoA hydratase, and the genes encoding phospholipases PlcA, PlcB and PlcC. Taken together, the depletion of these M. tuberculosis H37Rv genomic regions were associated with marked alterations in lipid-related genes of BCG strains. Such alterations may indicate a dormant-like state and can be determining factors to the vaccine's inability to induce long-term protection. These lipids can be further evaluated as an adjuvant to boost the current BCG-based vaccine.
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Current diagnostic tests for tuberculosis (TB) are not able to predict reactivation disease progression from latent TB infection (LTBI). The main barrier to predicting reactivation disease is the lack of our understanding of host biomarkers associated with progression from latent infection to active disease. Here, we applied an immune-based gene expression profile by NanoString platform to identify whole blood markers that can distinguish active TB from other lung diseases (OPD), and that could be further evaluated as a reactivation TB predictor. Among 23 candidate genes that differentiated patients with active TB from those with OPD, nine genes (CD274, CEACAM1, CR1, FCGR1A/B, IFITM1, IRAK3, LILRA6, MAPK14, PDCD1LG2) demonstrated sensitivity and specificity of 100%. Seven genes (C1QB, C2, CCR2, CCRL2, LILRB4, MAPK14, MSR1) distinguished TB from LTBI with sensitivity and specificity between 82 and 100%. This study identified single gene candidates that distinguished TB from OPD and LTBI with high sensitivity and specificity (both > 82%), which may be further evaluated as diagnostic for disease and as predictive markers for reactivation TB.
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Regulação da Expressão Gênica , Tuberculose Latente , Mycobacterium tuberculosis/metabolismo , RNA Mensageiro/sangue , Tuberculose Pulmonar , Adolescente , Adulto , Feminino , Humanos , Tuberculose Latente/sangue , Tuberculose Latente/diagnóstico , Masculino , Pessoa de Meia-Idade , Tuberculose Pulmonar/sangue , Tuberculose Pulmonar/diagnósticoRESUMO
The cell wall of wild-type (WT) Mycobacterium tuberculosis (Mtb), an etiologic agent of tuberculosis (TB) and a Mtb strain disrupted in a 13-gene operon mce1 (Δmce1) varies by more than 400 lipid species. Here, we examined Mtb lipid-induced response in murine macrophage, as well as in human T-cell subpopulations in order to gain an insight into how changes in cell wall lipid composition may modulate host immune response. Relative to WT Mtb cell wall lipids, the non-polar lipid extracts from Δmce1 enhanced the mRNA expression of lipid-sense nuclear receptors TR4 and PPAR-γ and dampened the macrophage expression of genes encoding TNF-α, IL-6, and IL-1ß. Relative to untreated control, WT lipid-pre-stimulated macrophages from healthy individuals induced a higher level of CD4-CD8- double negative T-cells (DN T-cells) producing TNF-α. Conversely, compared to WT, stimulation with Δmce1 lipids induced higher mean fluorescence intensity (MFI) in IL-10-producing DN T cells. Mononuclear cells from TB patients stimulated with WT Mtb lipids induced an increased production of TNF-α by CD8+ lymphocytes. Taken together, these observations suggest that changes in mce1 operon expression during a course of infection may serve as a strategy by Mtb to evade the host pro-inflammatory responses.
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Proteínas de Bactérias/genética , Parede Celular/imunologia , Ativação Linfocitária/imunologia , Ativação de Macrófagos/imunologia , Mycobacterium tuberculosis/imunologia , Tuberculose/imunologia , Adolescente , Adulto , Animais , Parede Celular/química , Feminino , Humanos , Lipídeos/imunologia , Macrófagos/imunologia , Masculino , Camundongos , Pessoa de Meia-Idade , Óperon , Células RAW 264.7 , Linfócitos T/imunologia , Adulto JovemRESUMO
Key measures to halt the spread of tuberculosis (TB) include early diagnosis, effective treatment, and monitoring disease management. We sought to evaluate the use of serum immunoglobulin levels against antigens present in cell envelope of Mycobacterium tuberculosis to monitor TB treatment response in children and adolescents with pulmonary (PTB) or extrapulmonary TB (EPTB). Blood samples were collected prior to and one, two, and six months following treatment initiation. Serum immunoglobulin levels against cardiolipin, sulfatide, mycolic acid and Mce1A protein were measured by ELISA. Serum from 53 TB patients and 12 healthy participants were analyzed. After six months of successful treatment, there was a significant decrease (p < 0.0001) in IgM levels against cardiolipin, sulfatide, mycolic acid and Mce1A protein and IgG levels against Mce1A protein when compared to baseline immunoglobulin levels. There was no significant variation in antibody levels during follow-up between participants with PTB and EPTB, confirmed and unconfirmed TB diagnosis, and HIV infection status. Antibody levels in control participants without TB did not decrease during follow-up. These results suggest that immunoglobulin responses to mycobacterial cell wall products may be a useful tool to monitor treatment response in children and adolescents with PTB or EPTB.
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Antituberculosos/uso terapêutico , Monitoramento de Medicamentos , Ensaio de Imunoadsorção Enzimática , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Mycobacterium tuberculosis/efeitos dos fármacos , Tuberculose Pulmonar/tratamento farmacológico , Adolescente , Fatores Etários , Proteínas de Bactérias/imunologia , Biomarcadores/sangue , Cardiolipinas/imunologia , Estudos de Casos e Controles , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Mycobacterium tuberculosis/imunologia , Ácidos Micólicos/imunologia , Valor Preditivo dos Testes , Estudos Prospectivos , Sulfoglicoesfingolipídeos/imunologia , Fatores de Tempo , Resultado do Tratamento , Tuberculose Pulmonar/diagnóstico , Tuberculose Pulmonar/imunologia , Tuberculose Pulmonar/microbiologiaRESUMO
Cutaneous leishmaniasis (CL) is caused by the bite of the infected sand fly, which inoculates parasites of Leishmania spp and triggers an immune response. An exacerbated cutaneous inflammatory response is crucial for controlling parasite burden but can also promote tissue damage. This study aimed to characterize the populations of natural killer (NK), CD57+, CD4+, and CD8+ T cells, CD20+ B cells, as well as CD68+ macrophages, in biopsies of ulcerated CL lesions, and quantify the production of perforin+, grazyme B+, interleukin 1 beta (IL-1ß+) and Tumor Necrosis Factor (TNF-α+ cells). We then correlated these parameters with necrosis, inflammation and the number of amastigotes. CD4+ T cells were positively correlated to the extent of inflammation, B cells and IL-1ß+ were associated with the extent of necrosis, CD68+ macrophages and perforin were correlated with the number of amastigotes, and CD57+ NK cells was correlated to CD68+ macrophages and amastigotes. In sum, the finding suggests that the production of cytotoxic granules and cytokines by inflammatory cells contributes to tissue damage in CL lesions.