RESUMO
A review of the British Society for Histocompatibility and Immunogenetics (BSHI) Guideline 'HLA matching and donor selection for haematopoietic progenitor cell transplantation' published in 2016 was undertaken by a BSHI appointed writing committee. Literature searches were performed and the data extracted were presented as recommendations according to the GRADE nomenclature.
Assuntos
Seleção do Doador/normas , Transplante de Células-Tronco Hematopoéticas/normas , Histocompatibilidade , Doadores de Tecidos , Sistema ABO de Grupos Sanguíneos/análise , Adulto , Algoritmos , Aloenxertos , Substituição de Aminoácidos , Incompatibilidade de Grupos Sanguíneos , Consultores , Transplante de Células-Tronco de Sangue do Cordão Umbilical/normas , Feminino , Técnicas de Genotipagem , Antígenos HLA/análise , Antígenos HLA/genética , Antígenos HLA/imunologia , Haplótipos/genética , Teste de Histocompatibilidade , Humanos , Isoanticorpos/imunologia , Masculino , Troca Materno-Fetal , Gravidez , Receptores Imunológicos , Sociedades Médicas/normas , Obtenção de Tecidos e ÓrgãosRESUMO
Lipoxins (LXs) are endogenously produced anti-inflammatory agents that modulate leukocyte trafficking and stimulate nonphlogistic macrophage phagocytosis of apoptotic neutrophils, thereby promoting the resolution of inflammation. Previous data suggest a role for altered protein phosphorylation and cytoskeletal rearrangement in LX-stimulated phagocytosis but the exact mechanisms remain unclear. In this study we examine the effects of LXA4 on the protein phosphorylation pattern of THP-1 cells differentiated into a macrophage-like phenotype. THP-1 cells stimulated with LXA4 (1 nM) exhibit dephosphorylation of a 220-kDa protein. Using mass spectrometry, this protein was identified as MYH9, a nonmuscle myosin H chain II isoform A, which is involved in cytoskeleton rearrangement. THP-1 cells treated with LXA4 adopt a polarized morphology with activated Cdc42 localized toward the leading edge and MYH9 localized at the cell posterior. Polarized distribution of Cdc42 is associated with Akt/PKB-mediated Cdc42 activation. Interestingly, the annexin-derived peptide Ac2-26, a recently described agonist for the LXA4 receptor, also stimulates macrophage phagocytosis, MYH9 dephosphorylation, and MYH9 redistribution. In addition, we demonstrate that LXA4 stimulates the phosphorylation of key polarity organization molecules: Akt, protein kinase Czeta, and glycogen synthase kinase-3beta. Inhibition of LXA4-induced Akt and protein kinase Czeta activity with specific inhibitors prevented LXA4-stimulated phagocytosis of both apoptotic polymorphonuclear neutrophils and lymphocytes, highlighting a potential use for LXA4 in the treatment of autoimmune diseases. Furthermore, phosphorylation and subsequent inactivation of glycogen synthase kinase-3beta resulted in an increase in phagocytosis similar to that of LXA4. These data highlight an integrated mechanism whereby LXA4 regulates phagocytosis through facilitative actin cytoskeleton rearrangement and cell polarization.
Assuntos
Apoptose/imunologia , Leucócitos/citologia , Leucócitos/imunologia , Lipoxinas/fisiologia , Macrófagos/metabolismo , Miosina não Muscular Tipo IIA/metabolismo , Fagocitose/imunologia , Proteína cdc42 de Ligação ao GTP/metabolismo , Sequência de Aminoácidos , Células Cultivadas , Quinase 3 da Glicogênio Sintase/metabolismo , Glicogênio Sintase Quinase 3 beta , Humanos , Dados de Sequência Molecular , Miosina não Muscular Tipo IIA/química , Miosina não Muscular Tipo IIA/genética , Fosforilação , Proteína Quinase C/metabolismo , Proteínas Proto-Oncogênicas c-akt/fisiologiaRESUMO
BACKGROUND: Phagocytic removal of apoptotic cells is an important regulatory event in development, tissue homoeostasis, and inflammation. There are several methodologic problems with most in vitro studies of the molecular mechanisms of apoptotic cell phagocytosis. First, cell loss occurs during rigorous washing of adherent macrophages required to ensure removal of noningested particles. Second, discrimination of adherent or internalised apoptotic cells is difficult. Third, microscopic quantification is time consuming and has the potential for significant interobserver error. Fourth, subsequent analysis of phagocyte populations is difficult. METHODS: We used a flow cytometric method that allows quantification of phagocytosis of fluorescently labelled apoptotic cells with the use of multiparameter flow cytometric analysis. RESULTS: Phagocytosis of apoptotic cells was validated by use of inhibitors (cytochalasins) or low temperature and counterstaining with cell surface markers for the phagocytic targets to exclude binding to the phagocytic surface. Populations of phagocytic macrophages were sorted, and the presence of internalized apoptotic material was validated by microscopy. CONCLUSIONS: The technique we used in this study allows observer-independent analysis of phagocytosis of apoptotic cells by macrophages. Importantly, phagocytic or nonphagocytic populations could be subjected to further characterization with the use of flow cytometry with additional fluorochrome reagents and can be re-cultured to study underlying regulatory mechanisms.
Assuntos
Apoptose/fisiologia , Citometria de Fluxo/métodos , Macrófagos/fisiologia , Fagocitose/fisiologia , Antígenos de Superfície/imunologia , Citocalasinas/farmacologia , Citometria de Fluxo/instrumentação , Fluoresceínas , Corantes Fluorescentes , Humanos , Macrófagos/citologia , Neutrófilos/citologia , Neutrófilos/fisiologia , Reprodutibilidade dos TestesRESUMO
Understanding the cellular and molecular mechanisms that determine whether inflammation resolves or progresses to scarring and tissue destruction should lead to the development of effective therapeutic strategies for inflammatory diseases. Apoptosis of neutrophil granulocytes is an important determinant of the resolution of inflammation, providing a mechanism for down-regulation of function and triggering clearance by macrophages without inducing a pro-inflammatory response. However, if the rate of cell death by apoptosis is such that the macrophage clearance capacity is exceeded, apoptotic cells may progress to secondary necrosis, resulting in the release of harmful cellular contents and in damage to the surrounding tissue. There are many possible ways in which the rate and capacity of the macrophage-mediated clearance of apoptotic cells may be enhanced or suppressed. Ligation of human macrophage surface CD44 by bivalent monoclonal antibodies rapidly and profoundly augments the capacity of macrophages to phagocytose apoptotic neutrophils in vitro. The molecular mechanism behind this effect and its potential significance in vivo is a current focus of research.