RESUMO
BACKGROUND AND OBJECTIVES: Buffy coat-derived granulocytes have been described as an alternative to the apheresis product from donors pretreated with dexamethasone and granulocyte colony-stimulating factor (G-CSF). The latter is - dependent on the local and national settings - obtained following a demanding and time-consuming procedure, which is undesirable in critically ill septic patients. In contrast, buffy coat-derived products have a large volume and are often heavily contaminated with red cells and platelets. We developed a new pooled buffy coat-derived product with high purity and small volume, and performed a comprehensive functional characterization of these granulocytes. MATERIALS AND METHODS: We pooled ten buffy coats following the production of platelet concentrates. Saline 0·9% was added to decrease the viscosity and the product was split into plasma, red cells and a 'super' buffy coat. Functional data of the granulocytes were compared to those obtained with granulocytes from healthy controls and G-CSF/dexamethasone-pretreated donors. RESULTS: Buffy coat-derived granulocytes showed adhesion, chemotaxis, reactive oxygen species production, degranulation, NETosis and in vitro killing of Staphylococcus aureus, Escherichia coli and Aspergillus species comparable to control and G-CSF/dexamethasone-derived granulocytes. Candida killing was superior compared to G-CSF/dexamethasone-derived granulocytes. Immunophenotyping was normal; especially no signs of activation in the buffy coat-derived granulocytes were seen. Viability was reduced. Buffy coats are readily available in the regular blood production process and would take away the concerns around the apheresis product. CONCLUSION: The product described appears a promising alternative for transfusion purposes.
Assuntos
Buffy Coat/citologia , Dexametasona/farmacologia , Fator Estimulador de Colônias de Granulócitos/farmacologia , Granulócitos/efeitos dos fármacos , Adulto , Antígenos de Superfície/metabolismo , Remoção de Componentes Sanguíneos , Doadores de Sangue , Plaquetas/citologia , Adesão Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Quimiotaxia/efeitos dos fármacos , Granulócitos/citologia , Granulócitos/imunologia , Granulócitos/metabolismo , Humanos , Imunofenotipagem , Contagem de Leucócitos , Masculino , NADPH Oxidases/metabolismo , Espécies Reativas de Oxigênio/metabolismoRESUMO
The Killer Immunoglobulin-like Receptor (KIR) proteins constitute a family of highly homologous surface receptors involved in the regulation of the innate cytotoxicity of natural killer (NK) cells. Within the human genome, 17 KIR genes are present, many of which show large variation across the population owing to the high number of allelic variants and copy number variation (CNV). KIR genotyping and CNV determination were used to map the KIR locus in a large cohort of >400 Caucasian individuals. Gene order and structure was determined by sequence-specific polymerase chain reaction of the intergenic regions. In this way, we could show that KIR3DL1 and KIR2DS4 gene variants are linked and that--contrary to current views--the gene KIR2DS5 is only present in the telomeric half of the KIR locus. Our study revealed novel insights in the highly organized distribution of KIR genes. Novel recombination hotspots were identified that contribute to the diversity of KIR gene distribution in the Caucasian population. Next-generation sequencing of the KIR intergenic regions allowed for a detailed single-nucleotide polymorphism analysis, which demonstrated several gene-specific as well as haplotype-specific nucleotides for a more accurate genotyping of this notoriously complex gene cluster.
Assuntos
DNA Intergênico , Receptores KIR3DL1/genética , Receptores KIR/genética , Variações do Número de Cópias de DNA , Ordem dos Genes , Genoma Humano , Haplótipos , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Regiões Promotoras Genéticas , Recombinação GenéticaRESUMO
The human FCGR2/3 locus, containing five highly homologous genes encoding the major IgG receptors, shows extensive copy number variation (CNV) associated with susceptibility to autoimmune diseases. Having genotyped >4000 individuals, we show that all CNV at this locus can be explained by nonallelic homologous recombination (NAHR) of the two paralogous repeats that constitute the majority of the locus, and describe four distinct CNV regions (CNRs) with a highly variable prevalence in the population. Apart from CNV, NAHR events also created several hitherto unidentified chimeric FCGR2 genes. These include an FCGR2A/2C chimeric gene that causes a decreased expression of FcγRIIa on phagocytes, resulting in a decreased production of reactive oxygen species in response to immune complexes, compared with wild-type FCGR2A. Conversely, FCGR2C/2A chimeric genes were identified to lead to an increased expression of FCGR2C. Finally, a rare FCGR2B null-variant allele was found, in which a polymorphic stop codon of FCGR2C is introduced into one FCGR2B gene, resulting in a 50% reduction in protein expression. Our study on CNRs and the chimeric genes is essential for the correct interpretation of association studies on FCGR genes as a determinant for disease susceptibility, and may explain some as yet unidentified extreme phenotypes of immune-mediated disease.
Assuntos
Receptores de IgG/genética , Alelos , Variações do Número de Cópias de DNA/genética , Estudos de Associação Genética , Predisposição Genética para Doença , Recombinação Homóloga , Humanos , Proteínas Mutantes Quiméricas/genética , Proteínas Mutantes Quiméricas/metabolismo , Polimorfismo de Nucleotídeo ÚnicoAssuntos
Bactérias/imunologia , Citocinas/imunologia , Fungos/imunologia , Doença Granulomatosa Crônica/imunologia , Inflamação/imunologia , Linhagem Celular , Doença Granulomatosa Crônica/genética , Humanos , Inflamação/genética , Monócitos/imunologia , Mutação/genética , NADPH Oxidases/genética , Neutrófilos/imunologiaRESUMO
In this study we present evidence that the mouse and rat sialoadhesin (originally named sheep erythrocyte receptor) on macrophages can function as a lymphocyte adhesion molecule. Lymphocytes were shown to bind to the splenic marginal zone, and lymph node subcapsular sinus and medulla in a frozen section assay. Selective depletion experiments showed that binding was mediated by macrophages. Adhesion was blocked by preincubation of the sections with monoclonal antibodies against mouse or rat sialoadhesin. Binding was temperature dependent, divalent cation independent, and involved sialic acid residues on the lymphocyte, as it could be inhibited by prior neuraminidase treatment or addition of the ganglioside GD1a. Binding to sialoadhesin was confirmed using the purified receptor and was observed among T cells, T blasts, B cells, and B blasts. Isolated macrophages or dendritic cells showed little binding. Sialoadhesin provides the first example of a macrophage-restricted lymphocyte adhesion molecule.
Assuntos
Moléculas de Adesão Celular/análise , Macrófagos/química , Glicoproteínas de Membrana , Receptores Imunológicos/análise , Animais , Western Blotting , Moléculas de Adesão Celular/metabolismo , Células Cultivadas , Técnicas Imunoenzimáticas , Linfócitos/citologia , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Especificidade de Órgãos , Ratos , Ratos Endogâmicos , Receptores Imunológicos/metabolismo , Lectina 1 Semelhante a Ig de Ligação ao Ácido SiálicoAssuntos
Dosagem de Genes , Receptores KIR3DL1/genética , Receptores KIR3DL2/genética , Espondilartrite/genética , Espondilite Anquilosante/genética , Adulto , Estudos de Coortes , Feminino , Humanos , Masculino , Países Baixos/epidemiologia , Espondilartrite/epidemiologia , Espondilite Anquilosante/epidemiologia , Adulto JovemRESUMO
Brain function requires precisely orchestrated connectivity between neurons. Establishment of these connections is believed to require signals secreted from outgrowing axons, followed by synapse formation between selected neurons. Deletion of a single protein, Munc18-1, in mice leads to a complete loss of neurotransmitter secretion from synaptic vesicles throughout development. However, this does not prevent normal brain assembly, including formation of layered structures, fiber pathways, and morphologically defined synapses. After assembly is completed, neurons undergo apoptosis, leading to widespread neurodegeneration. Thus, synaptic connectivity does not depend on neurotransmitter secretion, but its maintenance does. Neurotransmitter secretion probably functions to validate already established synaptic connections.
Assuntos
Encéfalo/embriologia , Encéfalo/fisiologia , Proteínas do Tecido Nervoso/fisiologia , Neurotransmissores/metabolismo , Sinapses/fisiologia , Proteínas de Transporte Vesicular , Animais , Apoptose , Encéfalo/citologia , Diferenciação Celular , Divisão Celular , Deleção de Genes , Cones de Crescimento/fisiologia , Camundongos , Camundongos Knockout , Proteínas Munc18 , Degeneração Neural , Proteínas do Tecido Nervoso/genética , Vias Neurais , Junção Neuromuscular/embriologia , Junção Neuromuscular/fisiologia , Neurônios/citologia , Neurônios/fisiologia , Técnicas de Patch-Clamp , Sinapses/ultraestrutura , Transmissão Sináptica , Vesículas Sinápticas/metabolismo , Vesículas Sinápticas/ultraestruturaRESUMO
Whereas, neutrophils have long been considered to mainly function as efficient innate immunity killers of micro-organisms at infected sites, they are now recognized to also be involved in modulation of adaptive immune responses. Immature and mature neutrophils were reported to have the capacity to suppress T cell-mediated immune responses as so-called granulocyte-myeloid-derived suppressor cells (g-MDSCs), and thereby affect the clinical outcome of cancer patients and impact the chronicity of microbial infections or rejection reactions in organ transplantation settings. These MDSCs were at first considered to be immature myeloid cells that left the bone marrow due to disease-specific signals. Current studies show that also mature neutrophils can exert suppressive activity. In this study we investigated in a robust T cell suppression assay whether immature CD11b+ myeloid cells were capable of MDSC activity comparable to mature fully differentiated neutrophils. We compared circulating neutrophils with myeloid cell fractions from the bone marrow at different differentiation stages. Our results indicate that functional MDSC activity is only becoming detectable at the final stage of differentiation, depending on the procedure of cell isolation. The MDSC activity obtained during neutrophil maturation correlated with the induction of the well-known highly mobile and toxic effector functions of the circulating neutrophil. Although immature neutrophils have been suggested to be increased in the circulation of cancer patients, we show here that immature neutrophils are not efficient in suppressing T cells. This suggests that the presence of immature neutrophils in the bloodstream of cancer patients represent a mere association or may function as a source of mature neutrophils in the tumor environment but not a direct cause of enhanced MDSC activity in cancer.
Assuntos
Proliferação de Células , Tolerância Imunológica , Ativação Linfocitária , Neutrófilos/imunologia , Linfócitos T/imunologia , Humanos , Células Supressoras Mieloides/citologia , Células Supressoras Mieloides/imunologia , Neutrófilos/citologia , Linfócitos T/citologiaRESUMO
Lutheran/basal cell adhesion molecule (Lu/BCAM) is a transmembrane adhesion molecule expressed by erythrocytes and endothelial cells that can interact with the extracellular matrix protein laminin-α5. In sickle cell disease, Lu/BCAM is thought to contribute to adhesion of sickle erythrocytes to the vascular wall, especially during vaso-occlusive crises. On healthy erythrocytes however, its function is unclear. Here we report that Lu/BCAM is activated during erythrocyte aging. We show that Lu/BCAM-mediated binding to laminin-α5 is restricted by interacting, in cis, with glycophorin-C-derived sialic acid residues. Following loss of sialic acid during erythrocyte aging, Lu/BCAM is released from glycophorin-C and allowed to interact with sialic acid residues on laminin-α5. Decreased glycophorin-C sialylation, as observed in individuals lacking exon 3 of glycophorin-C, the so-called Gerbich phenotype, was found to correlate with increased Lu/BCAM-dependent binding to laminin-α5. In addition, we identified the sialic acid-binding site within the third immunoglobulin-like domain within Lu/BCAM that accounts for the interaction with glycophorin-C and laminin-α5. Last, we present evidence that neuraminidase-expressing pathogens, such as Streptococcus pneumoniae, can similarly induce Lu/BCAM-mediated binding to laminin-α5, by cleaving terminal sialic acid residues from the erythrocyte membrane. These results shed new light on the mechanisms contributing to increased adhesiveness of erythrocytes at the end of their lifespan, possibly facilitating their clearance. Furthermore, this work may contribute to understanding the pathology induced by neuraminidase-positive bacteria, because they are especially harmful to patients suffering from sickle cell disease and are associated with the occurrence of vaso-occlusive crises.
Assuntos
Moléculas de Adesão Celular/metabolismo , Adesão Celular , Envelhecimento Eritrocítico , Glicoforinas/metabolismo , Sistema do Grupo Sanguíneo Lutheran/metabolismo , Ácido N-Acetilneuramínico/metabolismo , Anemia Falciforme/sangue , Sítios de Ligação , Humanos , Laminina/metabolismo , NeuraminidaseRESUMO
One of the actions of glucocorticoids (GC) in multiple sclerosis (MS) is an inhibitory effect on demyelination. This can be caused by a reduction in the number of infiltrating macrophages and/or by an effect on the phagocytosis of myelin. Here we investigate the effect of GC on the phagocytosis of myelin. Contrary to what was expected, we found that incubation of human monocytes with dexamethasone (DEX) for 48 h augmented (approximately threefold) the phagocytosis of myelin. This enhancement of phagocytosis by human monocytes was not restricted to myelin. Phagocytosis of various particles mediated by different macrophage receptors was increased by DEX. We found that not only the phagocytosis of Staphylococcus aureus bacteria was augmented, but also the killing of these bacteria was at least twice as effective after culture with DEX. Tumor necrosis factor alpha production of human monocyte-derived macrophages induced by lipopolysaccharide and S. aureus was suppressed by DEX. Together our results show that DEX promotes the phagocytosis of particles by human monocytes and thereby may contribute to tissue repair after immune-mediated tissue damage or infection. These data shed a new light on the clinical application of GC.
Assuntos
Dexametasona/farmacologia , Macrófagos/efeitos dos fármacos , Monócitos/efeitos dos fármacos , Fagocitose/efeitos dos fármacos , Staphylococcus aureus/imunologia , Células Cultivadas , Humanos , Macrófagos/citologia , Macrófagos/imunologia , Macrófagos/microbiologia , Monócitos/citologia , Monócitos/imunologia , Monócitos/microbiologia , Fagocitose/imunologia , Fator de Necrose Tumoral alfa/biossínteseRESUMO
CD163 is a member of the group B scavenger receptor cysteine-rich (SRCR) superfamily. This study describes aspects of the tissue distribution, the regulation of expression, and signal transduction after cross-linking of this receptor at the cell surface of macrophages. CD163 showed an exclusive expression on resident macrophages (e.g., red pulp macrophages, alveolar macrophages). The expression was inducible on monocyte-derived macrophages by glucocorticoids but not by interleukin-4 (IL-4), granulocyte-macrophage colony-stimulating factor (GM-CSF), and interferon-gamma. The combination of IL-4 or GM-CSF with glucocorticoids resulted in a further increase. Subcellular analysis of alveolar macrophages by immunoelectron microscopy showed a plasma membrane localization of the antigen. Cross-linking of CD163 with monoclonal antibody induced a protein tyrosine kinase-dependent signal that resulted in (1) slow-type calcium mobilization, (2) inositol triphosphate production, and (3) secretion of IL-6 and GM-CSF. The data suggest a function for the SRCR-superfamily receptor CD163 in the regulation of inflammatory processes by macrophages.
Assuntos
Antígenos CD , Antígenos de Diferenciação Mielomonocítica/metabolismo , Ativação de Macrófagos , Macrófagos/metabolismo , Receptores de Superfície Celular , Transdução de Sinais , Animais , Antígenos de Diferenciação Mielomonocítica/genética , Reagentes de Ligações Cruzadas , Citocinas/biossíntese , Humanos , Immunoblotting , Líquido Intracelular , Macrófagos/imunologia , Macrófagos Alveolares/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Receptores Imunológicos/genética , Receptores Imunológicos/metabolismo , Receptores Depuradores , Frações Subcelulares/metabolismo , Distribuição Tecidual , Transfecção , Células Tumorais Cultivadas , Células U937RESUMO
OBJECTIVE: Ankylosing spondylitis (AS) is an autoimmune disease that mainly affects the sacroiliac joints and the spine of the lower back. The disease is strongly associated with HLA-B27. Additional genes, single-nucleotide polymorphisms, and molecular components have been identified to be associated with AS, but the exact mechanism that drives disease development remains poorly understood. The killer cell immunoglobulin-like receptors (KIRs) are regulators of cytotoxicity of natural killer cells and T cell subsets and may be relevant in binding to HLA-B27 and the development of AS. We undertook this study to identify possible associations of KIR genotype with susceptibility to AS and disease characteristics including the presence of the HLA-B27 allele, disease severity, and uveitis. METHODS: We performed complete genotyping of the KIR locus in 303 Caucasian AS patients, 119 randomly selected healthy Caucasian controls, and 50 HLA-B27-positive healthy Caucasian controls by multiplex ligation-dependent probe amplification assay for detection of gene presence and copy number. RESULTS: We did not observe a significant association of any specific KIR gene or haplotype with susceptibility to AS or any other clinical manifestation. Disease severity, as measured by fulfilling the criteria for treatment with tumor necrosis factor blocking therapy, was linked to a lower number of genes for the functional variant of KIR3DL1 (P = 0.007). CONCLUSION: Our exploratory study indicates that KIR genes are not a major risk factor for susceptibility to AS. However, the data do suggest a role for KIRs in progression of the disease, whereby KIR3DL1 has a protective effect against the more severe manifestations of AS.
Assuntos
Alelos , Predisposição Genética para Doença , Receptores KIR3DL1/genética , Espondilite Anquilosante/genética , Adulto , Progressão da Doença , Feminino , Frequência do Gene , Genótipo , Haplótipos , Humanos , Masculino , Polimorfismo de Nucleotídeo ÚnicoRESUMO
Synthesis of parasite specific IgE plays a critical role in the defence against helminth infections. We report here that IgE from serum from Schistosoma mansoni infected mice and Haemonchus contortus infected sheep recognizes complex-type N-glycans from Arabidopsis thaliana, which contain R-GlcNAcbeta1-->4(Fucalpha1-->3)GlcNAcbeta1-Asn (core alpha1-->3-Fuc) and Xylbeta1-->2Manbeta1-->4GlcNAcbeta1-R (core beta1-->2-Xyl) modifications, and honeybee phospholipase A2, which carries N-glycans that contain the core alpha1-->3-Fuc epitope. Evidence is presented that core alpha1-->3-fucosylated N-glycans bind a substantial part of the parasite specific IgE in serum of H. contortus infected sheep. These results suggest that the core alpha1-->3-Fuc antigen may contribute to induction of a Th2 response leading to the production of IgE. In addition we show here that N-glycans carrying core alpha1-->3-Fuc and beta1-->2-Xyl antigens are synthesized by many parasitic helminths and also by the free living nematode Caenorhabditis elegans. Since N-glycans containing the core alpha1-->3-Fuc have also been implicated in honeybee and plant induced allergies, this conserved glycan might represent an important common IgE epitope.
Assuntos
Epitopos/imunologia , Fucose/imunologia , Hemoncose/imunologia , Haemonchus/química , Imunoglobulina E/imunologia , Polissacarídeos/imunologia , Animais , Arabidopsis/química , Ligação Competitiva , Western Blotting , Caenorhabditis elegans/química , Relação Dose-Resposta a Droga , Ensaio de Imunoadsorção Enzimática , Fucose/química , Haemonchus/imunologia , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Camundongos , Polissacarídeos/química , Schistosoma mansoni/química , Ovinos/imunologia , Ovinos/parasitologia , Especificidade por SubstratoRESUMO
Liposomes can be used as vehicles for intracellular delivery of drugs into phagocytic cells. Clodronate and propamidine, delivered into macrophages in this way, will kill these cells as a result of intracellular accumulation and irreversible metabolic damage. The so-called liposome-mediated macrophage 'suicide' approach, which is based on this principle, is now frequently applied in studies aimed at unravelling macrophage function. In the present study, the mechanism of phagocytic cell death induced by liposome encapsulated drugs was investigated 'in vitro'. Peritoneal macrophages and macrophages of the RAW 264 cell line were cultured in the presence of the liposome encapsulated drugs clodronate, propamidine and several forms of ethylenediaminetetraacetic acid (EDTA). The results obtained suggest that apoptotic death is induced in phagocytic cells both by liposomally delivered clodronate and by liposomally delivered propamidine. Although intracellular EDTA did induce apoptosis in a minority of the experiments, the results support earlier findings that EDTA does not deplete macrophages as effectively as clodronate and propamidine.
Assuntos
Apoptose/efeitos dos fármacos , Benzamidinas/administração & dosagem , Ácido Clodrônico/administração & dosagem , Sistemas de Liberação de Medicamentos , Macrófagos Peritoneais/efeitos dos fármacos , Benzamidinas/farmacologia , Linhagem Celular Transformada , Ácido Clodrônico/farmacologia , Ácido Edético/farmacologia , Líquido Intracelular/efeitos dos fármacos , Lipossomos/imunologiaRESUMO
The perivascular (PVM) and meningeal (MM) macrophages form a distinct population of resident CNS cells, selectively expressing the mature macrophage marker ED2 in the rat. In order to elucidate the role of the PVM and MM in rats during normal functioning of the brain and pathology, we have developed a strategy employing a single intraventricular injection of clodronate liposomes. This resulted in a complete depletion of the PVM and MM. Clodronate liposomes did not deplete the microglial cells. In other parts of the body, a temporal and mild depletion effect was observed, which was restored within 1 week. Detailed analysis of the elimination and repopulation kinetics of the PVM and MM revealed a slow repopulation of the CNS, starting at 14 days post depletion. This selective depletion method of the PVM and MM will enable us to get direct insight in their functions during normal and pathologic conditions of the CNS.
Assuntos
Barreira Hematoencefálica/imunologia , Macrófagos/citologia , Meninges/citologia , Meninges/imunologia , Analgésicos não Narcóticos/farmacologia , Animais , Contagem de Células , Sistema Nervoso Central/irrigação sanguínea , Sistema Nervoso Central/citologia , Sistema Nervoso Central/imunologia , Ácido Clodrônico/farmacologia , Cinética , Lipossomos/farmacologia , Masculino , Meninges/irrigação sanguínea , Ratos , Ratos WistarRESUMO
Reactive oxygen species (ROS) are thought to be involved in the pathogenesis of multiple sclerosis (MS) and experimental allergic encephalomyelitis (EAE). In this study we showed that the phagocytosis of myelin by macrophages triggers the production of ROS. We also demonstrated that ROS play a crucial role in the myelin phagocytosis. Blocking the ROS production with NADPH oxidase inhibitors (100 microM DPI or 10 mM Apocynin) essentially prevented the phagocytosis of myelin. Furthermore, scavenging of ROS with catalase (H2O2) or mannitol (OH-) decreased the phagocytosis of myelin by macrophages, whereas superoxide dismutase (O2-) did not show this effect. In addition, Lipoic acid (LA), a non-specific scavenger of ROS, also decreased the phagocytosis of myelin by macrophages. In our results, we demonstrate for the first time that ROS appear to play a regulatory role in the phagocytosis of myelin.
Assuntos
Macrófagos/fisiologia , Bainha de Mielina/fisiologia , Fagocitose/fisiologia , Espécies Reativas de Oxigênio/fisiologia , Animais , Sequestradores de Radicais Livres/farmacologia , Indicadores e Reagentes/metabolismo , Macrófagos/metabolismo , Masculino , NADPH Oxidases/antagonistas & inibidores , Nitroazul de Tetrazólio/metabolismo , Fagocitose/efeitos dos fármacos , Ratos , Ratos Endogâmicos , Espécies Reativas de Oxigênio/metabolismo , Rodaminas/metabolismo , Ácido Tióctico/farmacologiaRESUMO
The role of carbohydrate structures in the interaction of lymphocytes and endothelial cells is well established. Here the influence of sialic acid in the entrance and localization of lymphocytes in the lymphoid white pulp area of the spleen was studied by injecting sialidase in vivo. A role for sialic acid molecules on stromal elements of the spleen was determined. Although the identity of the cells that bear sialidase sensitive receptors could not be established, a role for marginal zone macrophages could be ruled out by macrophage depletion studies.
Assuntos
Receptores de Retorno de Linfócitos/química , Ácidos Siálicos/fisiologia , Baço/citologia , Animais , Adesão Celular/fisiologia , Movimento Celular/fisiologia , Endotélio Linfático/citologia , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Ácido N-Acetilneuramínico , Neuraminidase/sangueRESUMO
In previous studies it has been demonstrated that sialoadhesin is a macrophage-restricted adhesion receptor for lymphocytes and myeloid cells. It is under normal circumstances expressed by subpopulations of macrophages in lymphoid and haemopoietic tissues. In this study different immunoelectronmicroscopical techniques are used to investigate the ultrastructural localisation of sialoadhesin within the lymph node and spleen of rodents. The results show that sialoadhesin is selectively expressed by a subset of macrophages in peripheral lymphoid tissues. Sialoadhesin was localised predominantly on the plasma membrane and in particular in areas of intimate contact with lymphocytes, thereby visualizing putative local interaction between these cells. Interestingly, sialoadhesin was also detected in intracellular vesicles that were apparently taken up by macrophages. These findings are consistent with the putative role of sialoadhesin in local cell-cell interactions in lymphoid tissues. Surprisingly, sialoadhesin was also found at contact points of macrophages with other macrophages, sinus-lining cells and reticulum cells, suggesting that sialoadhesin also mediates interactions with these cell types.
Assuntos
Moléculas de Adesão Celular/análise , Macrófagos/química , Glicoproteínas de Membrana/análise , Receptores Imunológicos/análise , Animais , Membrana Celular/química , Tecido Linfoide/citologia , Macrófagos/ultraestrutura , Masculino , Camundongos , Ratos , Ratos Wistar , Lectina 1 Semelhante a Ig de Ligação ao Ácido SiálicoRESUMO
The capacity of immune-complex (IC) trapping was examined using purified horse radish peroxidase (HRP)-anti-HRP (PAP) on frozen sections of mouse spleen in vitro. We investigated the trapping mechanisms by applying the IC with or without fresh mouse serum added on the spleen sections of naive as well as immunized mice. When the PAP was applied alone, it mainly located on the macrophages in red pulp. In the splenic white pulp of immunized mice, PAP was trapped on follicular dendritic cells (FDC) in a small area of the germinal center whereas it scarcely bound to the splenic white pulp of non-immunized mice. An antibody against mouse Fc receptor (2.4G2) blocked the trapping but antibodies against mouse complement receptor (8C12, Mac-1 and 7G6) did not. When the PAP was applied mixed with fresh mouse serum, it bound on FDC in the primary follicles in the spleen of non-immunized mice. The density and area of IC trapping increased in the spleen of immunized mice. IC trapping in the presence of fresh mouse serum was blocked by the antibodies 8C12 and 7G6 but not by 2.4G2 or Mac-1.