RESUMO
SETTING: Mexico City, Mexico. OBJECTIVE: To identify proteins synthetised by Mycobacterium tuberculosis in hypoxic culture, which resemble more closely a granuloma environment than aerobic culture, and to determine if they are recognised by antibodies from patients with active pulmonary tuberculosis (PTB). DESIGN: Soluble extracts from M. tuberculosis H37Rv cultured under aerobic or hypoxic conditions were analysed using two-dimensional polyacrylamide gel electrophoresis, and proteins over-expressed under hypoxia were identified by mass spectrometry. The presence of immunoglobulin (Ig) G, IgA and IgM antibodies against these proteins was determined in the serum of 42 patients with active PTB and 42 healthy controls. RESULTS: We selected three M. tuberculosis H37Rv proteins (alpha-crystallin protein [Acr, Rv2031c], universal stress protein Rv2623 and isocitrate lyase [ICL, RV0467]) that were over-expressed under hypoxia. Titres of anti-Acr and anti-ICL IgA antibodies were higher in patients than in healthy controls, with an area under the receiver operating characteristic curve of 0.71 for anti-ICL IgA antibodies. CONCLUSION: ICL could be used in combination with other M. tuberculosis antigens to improve the sensitivity and specificity of current serological TB diagnostic methods.
Assuntos
Anticorpos Antibacterianos/sangue , Imunoglobulina A/sangue , Isocitrato Liase/imunologia , Tuberculose Pulmonar/diagnóstico , alfa-Cristalinas/imunologia , Adulto , Idoso , Antígenos de Bactérias/sangue , Estudos de Casos e Controles , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , México , Pessoa de Meia-Idade , Mycobacterium tuberculosis/isolamento & purificação , Sensibilidade e Especificidade , Tuberculose Pulmonar/sangue , Adulto JovemRESUMO
Candida albicans causes opportunistic systemic infections with high mortality (30%-50%). Despite significant nephrotoxicity, amphotericin (AmB) is still used for the treatment of this serious fungal infection. Therefore, alternative treatments are urgently needed. Dialyzable leukocyte extracts have been used successfully to treat patients with mucocutaneous candidiasis, but their effectiveness in systemic candidiasis has not been evaluated. In this study, low-dose AmB (0.1 mg/kg) plus 10 pg of murine dialyzable spleen extracts (mDSE) were tested in a systemic candidiasis mouse model. Survival, tissue fungal burden, kidney damage, kidney cytokines, and serum levels of IL-6 and hepcidin were evaluated. Our results showed that the combined treatment of low-dose AmB plus mDSE improved survival and reduced kidney fungal burden and histopathology; these effects correlated with increased kidney concentration of IFN- γ and TGF- ß 1, decreased levels of TNF- α , IL-6, and IL-10, as well as high levels of systemic IL-6 and hepcidin. Low-dose AmB and mDSE synergized to clear the infectious agent and reduced tissue damage, confirming the efficacy of a low dose of AmB, which might decrease the risk of drug toxicity. Further studies are necessary to explore these findings and its implications in future therapeutic approaches.
Assuntos
Anfotericina B/administração & dosagem , Antifúngicos/administração & dosagem , Candidíase/tratamento farmacológico , Linfocinas/administração & dosagem , Baço/metabolismo , Animais , Candidíase/mortalidade , Candidíase/patologia , Citocinas/biossíntese , Modelos Animais de Doenças , Feminino , Hepcidinas/biossíntese , Interleucina-6/biossíntese , Rim/metabolismo , Rim/microbiologia , CamundongosRESUMO
BALB/c mice with pulmonary tuberculosis (TB) develop a T helper cell type 1 that temporarily controls bacterial growth. Bacterial proliferation increases, accompanied by decreasing expression of interferon (IFN)-γ, tumour necrosis factor (TNF)-α and inducible nitric oxide synthase (iNOS). Activation of dendritic cells (DCs) is delayed. Intratracheal administration of only one dose of recombinant adenoviruses encoding granulocyte-macrophage colony-stimulating factor (AdGM-CSF) 1 day before Mycobacterium tuberculosis (Mtb) infection produced a significant decrease of pulmonary bacterial loads, higher activated DCs and increased expression of TNF-α, IFN-γ and iNOS. When AdGM-CSF was given in female mice B6D2F1 (C57BL/6J X DBA/2J) infected with a low Mtb dose to induce chronic infection similar to latent infection and corticosterone was used to induce reactivation, a very low bacilli burden in lungs was detected, and the same effect was observed in healthy mice co-housed with mice infected with mild and highly virulent bacteria in a model of transmissibility. Thus, GM-CSF is a significant cytokine in the immune protection against Mtb and gene therapy with AdGM-CSF increased protective immunity when administered in a single dose 1 day before Mtb infection in a model of progressive disease, and when used to prevent reactivation of latent infection or transmission.
Assuntos
Adenoviridae/genética , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Fator Estimulador de Colônias de Granulócitos e Macrófagos/imunologia , Tuberculose Pulmonar/terapia , Animais , Modelos Animais de Doenças , Feminino , Terapia Genética , Imunoterapia , Pulmão/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Tuberculose Pulmonar/genética , Tuberculose Pulmonar/imunologia , Tuberculose Pulmonar/transmissãoRESUMO
It has been shown recently that different genotypes of Mycobacterium tuberculosis induce distinct immune responses in the host, as reflected by variations in cytokine and iNOS expression. Because these molecules are probably regulated by multiple factors in vivo this complex phenomenon was partially analysed by assessing cytokine and iNOS expression by real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) in an in vitro model of bone marrow-derived macrophages infected with three different M. tuberculosis genotypes: Canetti, H37 Rv and Beijing. Although the three genotypes induced production of iNOS and the different cytokines tested at 24 h post-infection, macrophages infected with the Beijing isolate expressed the highest levels of mRNA for iNOS, interleukin (IL)-1beta, tumour necrosis factor (TNF)-alpha, IL-12 cytokines and lower levels of IL-10 compared with cells infected with other genotypes. This expression pattern has been associated with infection control, but during infection in vivo with the Beijing genotype it is lost upon progression to chronic phase. The failure to control infection is likely to be influenced by cytokines produced by other cell types and bacterial molecules expressed during the course of disease. Results presented in this work show that each genotype has the ability to induce different levels of cytokine expression that could be related to its pathogenesis during infection.
Assuntos
Citocinas/imunologia , Macrófagos/imunologia , Tuberculose/imunologia , Animais , Células da Medula Óssea/imunologia , Células Cultivadas , Genótipo , Interleucina-1/imunologia , Interleucina-10/imunologia , Interleucinas/imunologia , Camundongos , Mycobacterium tuberculosis/genética , Óxido Nítrico Sintase/imunologia , Óxido Nítrico Sintase Tipo II , Fagocitose/imunologia , RNA Mensageiro/imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Fator de Crescimento Transformador beta/imunologia , Tuberculose/genéticaRESUMO
BACKGROUND: Atopic dermatitis is a skin inflammatory disease which has been associated to high levels of IgE, eosinophiles and change of T lymphocytes. The transfer factor is an immunomodulator active substance and decreases the number of inflammatory cells and the severity of the symptoms of atopic dermatitis. OBJECTIVE: To determine the efficacy of the transfer factor as treatment of moderate and severe atopic dermatitis. MATERIAL AND METHODS: Articles related to treatment with transfer factor in the atopic dermatitis were looked up in Medline and EMBASE, and the ones referring to controlled studies in patients with moderate and severe atopic dermatitis in accord to SCORAD. RESULTS: We found seven articles with 121 patients and 88 controls demonstrating significant decrease in the symptoms of the SCORAD index, decreased IgE, and eosinophils in patients treated with transfer factor. CONCLUSIONS: The transfer factor is a choice treatment for moderate and severe atopic dermatitis.
Assuntos
Dermatite Atópica/tratamento farmacológico , Fator de Transferência/uso terapêutico , Dermatite Atópica/imunologia , Humanos , Índice de Gravidade de DoençaRESUMO
Iron is known to play an important role in different bacterial infections and, in particular, in their development. One example is infection with Mycobacterium tuberculosis where iron contributes to growth and survival of the bacteria within the host cell. The majority of studies performed on tuberculosis have focused on the direct effect of iron on bacterial growth; however, little is known about how iron modifies the mycobacterial-host interaction. In order to address this, we have investigated the effect of iron on intracellular growth of M. tuberculosis in J774 macrophages and the molecular mechanisms that are affected during this interaction. We observed that iron modifies intracellular growth of the mycobacteria and that their growth kinetics was modified from that observed for the extracellular situation in the presence of iron. Similarly, when iron was present during the infection, there was a reduced release of tumour necrosis factor-alpha and it was related to a higher number of bacilli inside the host cell and low expression of interleukin-1 (IL-1) and IL-6 mRNA. Hence, this work demonstrates that iron, besides promoting mycobacterial growth, also regulates the relationship between macrophage and bacteria.
Assuntos
Citocinas/biossíntese , Ferro/farmacologia , Macrófagos/imunologia , Macrófagos/microbiologia , Mycobacterium tuberculosis/patogenicidade , Citocinas/genética , Mediadores da Inflamação/metabolismo , Interleucina-1/biossíntese , Interleucina-1/genética , Interleucina-6/biossíntese , Interleucina-6/genética , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Mycobacterium tuberculosis/efeitos dos fármacos , Mycobacterium tuberculosis/crescimento & desenvolvimento , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase Tipo II , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/genéticaRESUMO
Problems of logistics, compliance and drug resistance point to an urgent need for immunotherapeutic strategies capable of shortening the current six month antibiotic regimens used to treat tuberculosis. One potential immunotherapeutic agent is transfer factors. Transfer factors (TF) are low molecular weight dialysable products from immune cells which transmit the ability to express delayed-type hypersensitivity (DTH) and cell mediated immunity from sensitized donors to nonimmune recipients. In this study we determined the efficiency of TF as immunotherapy to treat experimental tuberculosis. When BALB/c mice are infected via the trachea with Mycobacterium tuberculosis H37Rv there is an initial phase of partial resistance dominated by Th-1 type cytokines plus tumour necrosis factor-alpha (TNFalpha) and the inducible isoform of nitric oxide synthase (iNOS), followed by a phase of progressive disease characterized by increasing expression of IL-4, diminished expression of TNFalpha and iNOS, and low DTH. Animals in this late progressive phase of the disease (day 60) were treated with different doses of TF (one injection per week) obtained from spleen cells when the peak of immune protection in this animal model is reached (day 21), or with different doses of TF from peripheral leucocytes of PPD + healthy subjects. We show here that the treatment with murine or human TF restored the expression of Th-1 cytokines, TNFalpha and iNOS provoking inhibition of bacterial proliferation and significant increase of DTH and survival. This beneficial effect was dose dependent. Interestingly, murine TF in combination with conventional chemotherapy had a synergistic effect producing significant faster elimination of lung bacteria loads than chemotherapy alone.
Assuntos
Adjuvantes Imunológicos/administração & dosagem , Hipersensibilidade Tardia/imunologia , Imunoterapia Ativa/métodos , Fator de Transferência/administração & dosagem , Tuberculose Pulmonar/terapia , Animais , Antituberculosos/uso terapêutico , Ensaio de Imunoadsorção Enzimática/métodos , Interleucina-4/genética , Interleucina-4/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Modelos Animais , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico Sintase Tipo II , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Pele/imunologia , Fator de Transferência/imunologia , Tuberculose Pulmonar/tratamento farmacológico , Tuberculose Pulmonar/imunologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Mycoplasma infection affects the host's immune system in different ways. In this work, a kinetic approach was used to try to determine the mechanisms by which Mycoplasma cause these effects. Experiments were performed using Balb/c mice infected with Mycoplasma pulmonis and several immunological parameters were determined. It was found that at days 10 and 15 post-infection, there were significant changes in the percentages of CD4+ and CD8 + cells, in both peripheral blood and the thymus. Significant sequential increases in concentrations of both IFN-gamma and IL-4 were detected in sera, such that at day 15, there was a peak in IFN-gamma, concentration and at day 38, IL-4 concentration also peaked. By day 46, both IFN-gamma and IL-4 fell to control levels despite continued infection. Delayed hypersensitivity (DTH) was reduced in infected animals compared to non-infected controls. A small recovery in DTH was observed at day 30, which was reduced again by day 40. Altogether, the results show features of a transitional shift from Th1 to Th2 in animals that are ultimately immunologically incompetent (in both cellular and humoral immunity). It appears to be this state of incompetence that allows the microorganism to survive and thus provides an explanation for the chronic state of the disease, which is a characteristic of Mycoplasma infection.
Assuntos
Adjuvantes Imunológicos/fisiologia , Mycoplasma/imunologia , Animais , Relação CD4-CD8 , Bovinos , Ensaio de Unidades Formadoras de Colônias , Citocinas/sangue , Hipersensibilidade Tardia/imunologia , Interferon gama/sangue , Interferon gama/imunologia , Interleucina-4/sangue , Interleucina-4/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Mitógenos/farmacologia , Tamanho do Órgão/fisiologia , Fito-Hemaglutininas , Pneumonia por Mycoplasma/imunologia , Pneumonia por Mycoplasma/patologia , Soroalbumina Bovina/imunologia , Baço/citologia , Baço/imunologia , Timidina/metabolismo , Timo/citologia , Timo/imunologiaAssuntos
Anticorpos Antibacterianos/imunologia , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/imunologia , Especificidade de Anticorpos , Focalização Isoelétrica , Fragmentos Fab das Imunoglobulinas , Fragmentos Fab das Imunoglobulinas/imunologia , Hanseníase/imunologia , Imunoglobulina G , Mycobacterium leprae/imunologia , Mycobacterium tuberculosis/imunologia , Tuberculose Pulmonar/imunologiaRESUMO
BACKGROUND: The atopic dermatitis is a chronic skin disease that appears in patients with a personal or family history of allergic asthma and rhinitis. It is associated to the specific activation of a gene group. In most instances, the response to the conventional treatment is adequate. The are cases, though, know as refractory, where that is not the case. The study of two therapeutic alternatives, Transfer Factor (TF) and Cyclosporin A (CyA), was elaborated for this type of patients. MATERIAL AND METHODS: Patients with severe refractory AD were studied, being admitted to the Allergic Service to the ISSSTE Lic. Adolfo López Mateos, ISSSTE, between September 1997 and june 1998. They were randomly divided in two groups. The first one was subjected to CyA, on a 4 mg/kg/day dosage, with monthly surveillance of kidney and hepatic functions and blood pressure twice a week. Group two was subjected to TF, as follows: one unit every third day for the first week, two units per week for the next three weeks and one monthly unit to complete six months. Initial and final clinical and immunologic testing was performed on both groups (eosinophils, total IgE, CD4 and CD8). RESULTS: Six patients included group A, and 12 patients in group B. Both groups showed a significant statistic reduction in the total eosinophils count, without an statistic difference between them. None showed changes in the total IgE. CyA reduced the CD4 levels, while the TF increased the levels of CD8 cells, both with a p < 0.05. Both groups showed clinical improvement satistically significant, but no differences with a p > 0.05 appeared between them. Tolerance to the treatments was adequate, and there was not need to suspend the treatment in any case. Only three patients showed hypertricosis and other one presented headaches, with CyA. CONCLUSION: Both treatments showed therapeutic benefits in the treatment of patients with severe refractory AD, with similar immunologic improvement. Both drugs present different action mechanisms, so their joint application could offer clinical benefit to the patient (synergetic action), cost reduction, and long term treatments with reduced adverse effects.