RESUMO
Apoptosis is a noninflammatory, programmed form of cell death. One mechanism underlying the non-phlogistic nature of the apoptosis program is the swift phagocytosis of the dying cells. How apoptotic cells attract mononuclear phagocytes and not granulocytes, the professional phagocytes that accumulate at sites of inflammation, has not been determined. Here, we show that apoptotic human cell lines of diverse lineages synthesize and secrete lactoferrin, a pleiotropic glycoprotein with known antiinflammatory properties. We further demonstrated that lactoferrin selectively inhibited migration of granulocytes but not mononuclear phagocytes, both in vitro and in vivo. Finally, we were able to attribute this antiinflammatory function of lactoferrin to its effects on granulocyte signaling pathways that regulate cell adhesion and motility. Together, our results identify lactoferrin as an antiinflammatory component of the apoptosis milieu and define what we believe to be a novel antiinflammatory property of lactoferrin: the ability to function as a negative regulator of granulocyte migration.
Assuntos
Apoptose/fisiologia , Movimento Celular/fisiologia , Granulócitos/fisiologia , Lactoferrina/metabolismo , Animais , Antígeno CD11b/metabolismo , Linhagem Celular , Forma Celular , Meios de Cultivo Condicionados/química , Granulócitos/citologia , Humanos , Selectina L/metabolismo , Neutrófilos/citologia , Neutrófilos/metabolismo , Transdução de Sinais/fisiologiaRESUMO
BACKGROUND: Several genes exhibit copy number variation (CNV), including FCGR3B which encodes the IgG receptor FcγRIIIb. Engagement of Fcγ receptors by IgG complexes may contribute to the pathogenesis of idiopathic pulmonary fibrosis (IPF). OBJECTIVES: To investigate whether FCGR3B CNV is associated with susceptibility to IPF. METHODS: In a case-control study we compared FCGR3B copy number in 142 patients with IPF and in 221 controls by real-time quantitative PCR using CD36 as gene copy control. RESULTS: Significantly increased FCGR3B:CD36 ratio was evident in the IPF cohort compared to controls (p = 0.009). Association of FCGR3B copy number with IPF susceptibility was further confirmed by a likelihood ratio statistical approach (p = 0.003). FCGR3B copy number assignment based on FCGR3B:CD36 ratios revealed significant skewing in the distribution of FCGR3B copy number between IPF patients and controls. In the IPF cohort, there was increased frequency of >2 FCGR3B copies compared to controls (0.30 vs. 0.19; χ(2) = 9.27; d.f. 2; p = 0.0097). The presence of >2 FCGR3B copies was associated with higher risk of IPF (p = 0.01, OR: 1.914, 95% CI: 1.17-3.12). CONCLUSIONS: These findings support an association of FCGR3B copy number with susceptibility to IPF and propose a novel role for Fcγ receptors in IPF disease pathogenesis.
Assuntos
Variações do Número de Cópias de DNA , Fibrose Pulmonar Idiopática/genética , Receptores de IgG/genética , Idoso , Idoso de 80 Anos ou mais , Progressão da Doença , Feminino , Proteínas Ligadas por GPI/genética , Proteínas Ligadas por GPI/metabolismo , Predisposição Genética para Doença , Humanos , Fibrose Pulmonar Idiopática/metabolismo , Masculino , Pessoa de Meia-Idade , Neutrófilos/metabolismo , Receptores de IgG/metabolismoRESUMO
Eosinophilic granulocytes are innate effector cells that are important in immune responses against helminth parasitic infections and contribute towards the pathology associated with allergic inflammatory conditions, including allergic rhinitis and asthma. Their recruitment to inflammatory sites occurs in response to chemotactic and activation signals, such as eotaxin and interleukin-5, and is a tightly controlled process. However, the mechanisms that counterbalance these positive chemoattractive processes, thereby preventing excessive eosinophil infiltration, have received little attention. Here, we show that, lactoferrin (LTF), a pleiotropic 80-kDa glycoprotein with iron-binding properties, acts as a powerful inhibitor of eosinophil migration. Irrespective of its source (milk or neutrophil derived), LTF inhibits eotaxin-stimulated eosinophil migration with no effects on eosinophil viability. Transferrin, a closely related cationic glycoprotein, failed to produce an analogous effect. Furthermore, the iron-saturation status of LTF did not influence the observed inhibitory effect on migration, proving that LTF exerts its effect on eosinophil chemotaxis independent of its iron-chelating activity. These results highlight LTF as one of the few molecules reported to negatively regulate eosinophil migration. Thus, through its ability to inhibit eosinophil migration, LTF has potential as an effective therapeutic in the control of eosinophil infiltration in atopic inflammatory conditions.
Assuntos
Quimiotaxia de Leucócito/efeitos dos fármacos , Eosinófilos/citologia , Eosinófilos/efeitos dos fármacos , Lactoferrina/farmacologia , Quimiocina CCL11/farmacologia , Humanos , Ferro/metabolismo , Lactoferrina/metabolismo , Ligação Proteica/efeitos dos fármacosRESUMO
An excess of neutrophils in the alveoli and lung interstitium has been described in idiopathic pulmonary fibrosis (IPF). Engagement of neutrophil Fcγ receptors with IgG complexes may contribute to the pathogenesis of IPF. The neutrophil FcγRIIIb receptor occurs in two codominantly expressed allelic variants, NA1 and NA2, which exhibit different binding affinities for IgG1 and IgG3 subclasses. The aim of this study was to investigate whether FcγRIIIb genotype is associated with IPF susceptibility or disease progression. In a case-control study we compared the distribution of FcγRIIIb NA1/2 polymorphisms in 142 patients with IPF and in 218 controls using allele-specific PCR amplification. Significant skewing in the distribution of FcγRIIIb genotypes was observed between patients with IPF and control subjects. In the IPF cohort, there was higher frequency of the NA1/NA1 genotype (0.19 vs. 0.07), and lower NA2/NA2 frequency (0.31 vs. 0.50; χ(2) = 17.71, df = 2, P < 0.001). The overall frequency of the NA1 allele was increased in IPF patients compared to controls (0.44 vs. 0.29; P < 0.0001, odds ratio [OR] = 1.93, 95% confidence interval [CI] = 1.42-2.64). Heterozygotes and homozygotes of the NA1 allele were at higher risk of developing IPF (OR = 2.19, 95% CI = 1.40-3.41, P = 0.0005), whereas the NA2 allele was protective against IPF (OR = 0.34, 95% CI = 0.17-0.65, P = 0.0014). There was no association of FcγRIIIb genotype with disease progression as assessed by serial lung function measurements. FcγRIIIb NA1/2 polymorphisms are associated with IPF disease susceptibility. These results support a role for immunological mechanisms contributing to IPF pathogenesis.
Assuntos
Fibrose Pulmonar Idiopática/genética , Polimorfismo Genético , Receptores de IgG/genética , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Distribuição de Qui-Quadrado , Progressão da Doença , Feminino , Volume Expiratório Forçado , Proteínas Ligadas por GPI/genética , Frequência do Gene , Predisposição Genética para Doença , Heterozigoto , Homozigoto , Humanos , Fibrose Pulmonar Idiopática/imunologia , Fibrose Pulmonar Idiopática/fisiopatologia , Pulmão/fisiopatologia , Masculino , Pessoa de Meia-Idade , Razão de Chances , Fenótipo , Reação em Cadeia da Polimerase , Capacidade de Difusão Pulmonar , Medição de Risco , Fatores de Risco , Escócia , Capacidade Pulmonar Total , Capacidade VitalRESUMO
An expanding body of evidence demonstrates that cells undergoing apoptosis send out a selection of molecular navigational signals including proteins, lipids and nucleotides that serve to recruit phagocytes to the dying targets which are subsequently engulfed and removed. This homeostatic process is essentially non-phlogistic, contrasting markedly with the acute inflammatory responses elicited in phagocytes by damaging or infectious agents. The "professional" scavengers of apoptotic cells are mononuclear phagocytes-the macrophages-and sites of high-rate apoptosis are clearly characterized by macrophages associated with the apoptotic cells. By contrast, members of the other class of professional phagocytes-the granulocytes-are not recruited to sites of apoptosis as a direct consequence of the cell-death program. Indeed, recent work indicates that apoptotic cells release a mixture of migratory cues to leukocytes in order to selectively attract mononuclear phagocytes but not granulocytes through functional balancing of positive and negative signals. Here we discuss these molecular mechanisms which not only serve as migratory cues but also may activate responding phagocytes to engulf apoptotic cells effectively. Finally, we speculate upon new therapeutic opportunities these mechanisms offer for a range of pathological conditions, including inflammatory disorders and cancer.