RESUMO
Influenza infections are responsible for significant morbidity and mortality each year, with the highest infection rates found in the elderly population. The main strategy to reduce the impact of influenza infections in the elderly population is vaccination. However, the efficacy of influenza vaccines that are licensed for use in the elderly is relatively low (17-53%). The complex age-related changes that occur in both innate and adaptive immunity are thought to hamper the immune response to influenza immunization and to reduce protection against infection in the elderly. For the development of improved vaccines that overcome the limitations of an aged immune system, it is crucial to understand the mechanisms that lead to immune dysfunction. Here, we review the recent progress in unravelling the mechanisms behind the age-related immune dysfunction in elderly, as well as the recent developments in improving influenza vaccines and identification of new correlates of protection.
Assuntos
Envelhecimento/imunologia , Vacinas contra Influenza/imunologia , Influenza Humana/prevenção & controle , Humanos , Sistema Imunitário/fisiologia , Vacinas contra Influenza/administração & dosagemRESUMO
Lung inflammation is often associated with sustained neutrophil migration into the lung tissue, causing undesired side effects, i.e. substantial damage of lung tissue and fibrin deposition, which hamper complete recovery. The need for additional anti-inflammatory treatment strategies focused attention on the function of cellular adhesion molecules (CAMs) on the leukocyte membrane, which guide migration of leukocytes across the endothelium to the site of inflammation. Recent data indicate that neutrophil migration in the lung is mediated by unique pathways, involving different CAM as compared to other organs. These pulmonary characteristics of neutrophil migration enable specific targeting of CAM for anti-inflammatory treatment of pneumonia. In addition, the recent interest in intracellular signaling revealed that most CAM not only function in adhesion, but also play an important role in initiation of intracellular signal transduction and vice versa may be modulated by intracellular signaling molecules (ISMs). In this review, the mechanisms of neutrophil migration in the lung and the role of ISM with respect to CAM function are described in the context of potential anti-inflammatory intervention strategies.
Assuntos
Pulmão/imunologia , Neutrófilos/fisiologia , Animais , Bovinos , Moléculas de Adesão Celular/fisiologia , Movimento CelularRESUMO
Neutrophil emigration from the pulmonary vasculature, is mediated by cellular adhesion molecules (CAM) expressed on the outer membranes of endothelial cells and neutrophils. Although beta(2)-integrin-dependent migration is a major mechanism of neutrophil migration, which was demonstrated by extensive invasion of neutrophils in pulmonary tissue of calves suffering from a genetic deficit in expression of beta(2)-integrins, termed bovine leukocyte adhesion deficiency (LAD), the role of alternative CAM is still unclear. We investigated whether an alternate CAM for beta(2)-integrin function, i.e. the alpha(4)-integrin, was expressed on peripheral blood neutrophils of calves. As we detected basal but significant expression, the effect of naturally acquired pulmonary infection on the expression of either integrin was determined, as an indication for its function in the migration process. In our experiments, basal expression of alpha(4)-integrins on peripheral blood neutrophils from clinically healthy calves was detected. On neutrophils of calves, experiencing field outbreaks of enzootic bronchopneumonia, higher expression of the alpha(4)-integrin was detected, which returned to normal after successful treatment of the disease. In addition, its level of expression was linearly related to plasma acute phase protein (haptoglobin) concentrations, which is a sensitive parameter for severity of respiratory inflammation. Increased expression of the alpha(4)-integrin on peripheral blood neutrophils during pulmonary inflammation indicates a role for this CAM in neutrophil migration in the lung.
Assuntos
Regulação da Expressão Gênica , Integrina alfa4/metabolismo , Neutrófilos/metabolismo , Pneumonia/patologia , Pneumonia/veterinária , Animais , Infecções Bacterianas/imunologia , Infecções Bacterianas/veterinária , Bovinos , Inflamação/metabolismo , Pneumonia/sangue , Sistema Respiratório/metabolismo , Sistema Respiratório/patologia , Viroses/imunologia , Viroses/veterináriaRESUMO
Vaccination against influenza induces homologous as well as cross-specific hemagglutination inhibiting (HI) responses. Induction of cross-specific HI responses may be essential when the influenza strain does not match the vaccine strain, or even to confer a basic immune response against a pandemic influenza virus. We carried out a clinical study to evaluate the immunological responses after seasonal vaccination in healthy adults 18-60 years of age, receiving the yearly voluntary vaccination during the influenza season 2006/2007. Vaccinees of different age groups were followed for laboratory confirmed influenza (LCI) and homologous HI responses as well as cross-specific HI responses against the seasonal H1N1 strain of 2008 and pandemic H1N1 virus of 2009 (H1N1pdm09) were determined. Homologous HI titers that are generally associated with protection (i.e. seroprotective HI titers ≥40) were found in more than 70% of vaccinees. In contrast, low HI titers before and after vaccination were significantly associated with seasonal LCI. Cross-specific HI titers ≥40 against drifted seasonal H1N1 were found in 69% of vaccinees. Cross-specific HI titers ≥40 against H1N1pdm09 were also significantly induced, especially in the youngest age group. More specifically, cross-specific HI titers ≥40 against H1N1pdm09 were inversely correlated with age. We did not find a correlation between the subtype of influenza which was circulating at the age of birth of the vaccinees and cross-specific HI response against H1N1pdm09. These data indicate that the HI titers before and after vaccination determine the vaccination efficacy. In addition, in healthy adults between 18 and 60 years of age, young adults appear to be best able to mount a cross-protective HI response against H1N1pdm09 or drifted seasonal influenza after seasonal vaccination.
Assuntos
Testes de Inibição da Hemaglutinação , Vacinas contra Influenza/administração & dosagem , Influenza Humana/prevenção & controle , Adolescente , Adulto , Anticorpos Antivirais/sangue , Estudos de Coortes , Proteção Cruzada , Feminino , Seguimentos , Humanos , Imunidade Humoral , Vírus da Influenza A Subtipo H1N1/imunologia , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Vírus da Influenza A Subtipo H1N1/patogenicidade , Vacinas contra Influenza/imunologia , Influenza Humana/imunologia , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Pandemias , Vacinação , Adulto JovemRESUMO
Seasonal influenza causes more morbidity and mortality in older adults than in young adults, apparently because of a decline in immune function with increasing age, known as immunosenescence. In this study, we compared the capacity of dendritic cells (DCs) from healthy older adults (≥65 years) with DCs from healthy young adults (20-40 years) to initiate a T cell response against influenza. DCs from older adults were impaired in the induction of influenza-specific CD8+ T cells as compared to DCs from young adults, which was demonstrated by a decreased proliferation, an impaired production of IFN-γ and a reduced expression of the degranulation marker CD107a by CD8+ T cells. Importantly, DCs from older adults produced significantly less TNF-α, showed a decreased expression of HLA class I and had a lower maturation state after influenza virus infection. Supplementing TNF-α increased the expression of HLA class I and of maturation markers and enhanced the induction of the influenza-specific CD8+ T cell response. Together, these findings indicate that the impaired influenza-specific CD8+ T cell response in older adults is associated with a reduced production of TNF-α and with a lower DC maturation. We suggest that the production of TNF-α is a determining factor in the DC-mediated CD8+ T cell response against influenza.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Células Dendríticas/imunologia , Influenza Humana/imunologia , Fator de Necrose Tumoral alfa/metabolismo , Adulto , Fatores Etários , Idoso , Linfócitos T CD8-Positivos/virologia , Células Dendríticas/metabolismo , Feminino , Genes MHC Classe I , Humanos , Imunidade Celular , Vírus da Influenza A Subtipo H3N2 , Interferon gama/imunologia , Masculino , Proteínas Recombinantes/administração & dosagem , Fator de Necrose Tumoral alfa/administração & dosagem , Adulto JovemRESUMO
Influenza vaccine efficacy does not always correlate with humoral immune responses. Recent reports indicate that the cellular immune response also contributes to protection, however robust assays are lacking. We standardized and validated assays for detection of human influenza-specific cellular responses in four international laboratories. The production of granzyme B as marker of T cell-mediated cytotoxicity and release of Th1 and Th2 cytokines were evaluated. The granzyme B and cytokine assays were specific, accurate, precise, and robust. Replicate stimulations with PBMC from the same donors showed an intra-laboratory robustness (coefficient of variation) for quantitation of granzyme B of 33% and for cytokines - including IFN-gamma, TNF-alpha, IL-2, IL-10, IL-4, IL-13, GM-CSF and including the log IFN-gamma/IL-10 ratio - of 52%. The inter-laboratory robustness for detection of granzyme B was 29% and for detection of all cytokines was 49%. The assays can now be used for determining cell-mediated immunity and explored as correlates of protection. Moreover, the precision and robustness of these cellular assays allow the reliable detection of cellular responses even in small study populations.
Assuntos
Vacinas contra Influenza/imunologia , Influenza Humana/prevenção & controle , Citocinas/metabolismo , Granzimas/biossíntese , Humanos , Imunoensaio/normas , Sensibilidade e Especificidade , Linfócitos T Citotóxicos/imunologiaRESUMO
Virus infection induces an adaptive immune response by T cells that is specific for defined viral epitopes. The epitope-specific analysis of T cells has become an important tool for investigating the anti viral response following infection or vaccination. In this review, the inherent differences in the procedures to identify the epitopes are discussed. Specifically, the screening of lymphocytes for epitope specific responses and the usage of mass spectrometry for sequencing of viral epitopes are evaluated.
Assuntos
Antígenos Virais/imunologia , Epitopos/imunologia , Linfócitos T Citotóxicos/imunologia , Animais , Humanos , Vacinas Virais/imunologiaRESUMO
Identification of peptides presented in major histocompatibility complex (MHC) class I molecules after viral infection is of strategic importance for vaccine development. Until recently, mass spectrometric identification of virus-induced peptides was based on comparative analysis of peptide pools isolated from uninfected and virus-infected cells. Here we report on a powerful strategy aiming at the rapid, unambiguous identification of naturally processed MHC class I-associated peptides, which are induced by viral infection. The methodology, stable isotope tagging of epitopes (SITE), is based on metabolic labeling of endogenously synthesized proteins during infection. This is accomplished by culturing virus-infected cells with stable isotope-labeled amino acids that are expected to be anchor residues (i.e. residues of the peptide that have amino acid side chains that bind into pockets lining the peptide-binding groove of the MHC class I molecule) for the human leukocyte antigen allele of interest. Subsequently these cells are mixed with an equal number of non-infected cells, which are cultured in normal medium. Finally peptides are acid-eluted from immunoprecipitated MHC molecules and subjected to two-dimensional nanoscale LC-MS analysis. Virus-induced peptides are identified through computer-assisted detection of characteristic, binomially distributed ratios of labeled and unlabeled molecules. Using this approach we identified novel measles virus and respiratory syncytial virus epitopes as well as infection-induced self-peptides in several cell types, showing that SITE is a unique and versatile method for unequivocal identification of disease-related MHC class I epitopes.