RESUMO
BACKGROUND AND OBJECTIVE: Prophylactic platelet transfusions are administered to prevent bleeding in haemato-oncological patients. However, bleeding still occurs, despite these transfusions. This practice is costly and not without risk. Better predictors of bleeding are needed, and flow cytometric evaluation of platelet function might aid the clinician in identifying patients at risk of bleeding. This evaluation can be performed within the hour and is not hampered by low platelet count. Our objective was to assess a possible correlation between bleeding and platelet function in thrombocytopenic haemato-oncological patients. MATERIALS AND METHODS: Inclusion was possible for admitted haemato-oncology patients aged 18 years and above. Furthermore, an expected need for platelet transfusions was necessary. Bleeding was graded according to the WHO bleeding scale. Platelet reactivity to stimulation by either adenosine diphosphate (ADP), cross-linked collagen-related peptide (CRP-xL), PAR1- or PAR4-activating peptide (AP) was measured using flow cytometry. RESULTS: A total of 114 evaluations were available from 21 consecutive patients. Platelet reactivity in response to stimulation by all four studied agonists was inversely correlated with significant bleeding. Odds ratios (OR) for bleeding were 0·28 for every unit increase in median fluorescence intensity (MFI) [95% confidence interval (CI) 0·11-0·73] for ADP; 0·59 [0·40-0·87] for CRP-xL; 0·59 [0·37-0·94] for PAR1-AP; and 0·43 [0·23-0·79] for PAR4-AP. The platelet count was not correlated with bleeding (OR 0·99 [0·96-1·02]). CONCLUSION: Agonist-induced platelet reactivity was significantly correlated to bleeding. Platelet function testing could provide a basis for a personalized transfusion regimen, in which platelet transfusions are limited to those at risk of bleeding.
Assuntos
Plaquetas/efeitos dos fármacos , Coagulantes/administração & dosagem , Hemorragia/tratamento farmacológico , Leucemia Mieloide Aguda/complicações , Adulto , Idoso , Antineoplásicos/uso terapêutico , Feminino , Citometria de Fluxo , Hemorragia/etiologia , Hemorragia/prevenção & controle , Humanos , Leucemia Mieloide Aguda/sangue , Leucemia Mieloide Aguda/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Ativação Plaquetária , Contagem de Plaquetas , Testes de Função Plaquetária , Transfusão de Plaquetas/efeitos adversosRESUMO
Human immunodeficiency virus type-1 (HIV-1)-associated dementia (HAD) is a neurodegenerative disease characterized by HIV infection and replication in brain tissue. HIV-1-infected monocytes overexpress inflammatory molecules that facilitate their entry into the brain. Prostanoids are lipid mediators of inflammation that result from cyclooxygenase-2 (COX-2) activity. Because COX-2 is normally induced during inflammatory processes, the aim of this study was to investigate whether COX-2 expression is up-regulated during monocyte-brain endothelium interactions. In vitro cocultures of HIV-infected macrophages and brain endothelium showed an up-regulation of COX-2 expression by both cell types. This up-regulation occurs via an interleukin-1beta (IL1beta)-dependent mechanism in macrophages and via an IL-1beta-independent mechanism in endothelial cells. Thus, interactions between HIV-infected monocytes and brain endothelium result in COX-2 expression and, as such, might contribute to the neuropathogenesis of HIV infection.
Assuntos
Complexo AIDS Demência/enzimologia , Encéfalo/irrigação sanguínea , Comunicação Celular/fisiologia , Endotélio Vascular/enzimologia , HIV-1 , Isoenzimas/biossíntese , Macrófagos/enzimologia , Prostaglandina-Endoperóxido Sintases/biossíntese , Complexo AIDS Demência/sangue , Complexo AIDS Demência/patologia , Encéfalo/virologia , Técnicas de Cocultura , Ciclo-Oxigenase 2 , Endotélio Vascular/citologia , Humanos , Interleucina-1/biossíntese , Isoenzimas/genética , Macrófagos/citologia , Macrófagos/virologia , Proteínas de Membrana , Monócitos/citologia , Monócitos/enzimologia , Monócitos/virologia , Prostaglandina-Endoperóxido Sintases/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Regulação para CimaRESUMO
HIV-1 is a hematogenously spread virus that most likely gains entry into the brain within blood-derived macrophages. Indeed, productive viral replication selectively occurs within perivascular and parenchymal blood-derived macrophages and microglia and HIV-infected macrophages have increased potential to bind and transmigrate through the blood-brain barrier. Once inside the brain, HIV-infected macrophages secrete a variety of pro-inflammatory mediators that display neuromodulatory and neurotoxic activities in several in vitro models for HIV-1 encephalitis. The final outcome regarding neuronal function and cell loss is regulated through intercellular interactions between these virus-infected cells and astrocytes. In this regard, both HIV-induced intracellular events in macrophages and interactions between HIV-infected macrophages and brain cells are reviewed as factors that might lead to neuronal injury in in vitro model systems for HIV-1 encephalitis.
Assuntos
Complexo AIDS Demência/patologia , Complexo AIDS Demência/fisiopatologia , Encéfalo/fisiopatologia , Encéfalo/virologia , HIV-1/fisiologia , Macrófagos/virologia , Neurônios/fisiologia , Neurônios/virologia , Replicação Viral , Animais , Astrócitos/virologia , Humanos , Microglia/virologia , Modelos BiológicosRESUMO
Although neurotrophic factors are currently considered as treatment for neurodegenerative diseases, little is still known about their presence in the central nervous system under pathological conditions. We investigated the expression of the neurotrophic molecules NGF, bFGF, BDNF and IGF-1 in brain tissue of patients suffering from AIDS dementia complex. In contrast to IGF-1 and BDNF, NGF and bFGF mRNA levels were significantly elevated. Strong NGF immunoreactivity was found in perivascular areas and was colocalized with infiltrating macrophages, whereas intense bFGF staining was found in cells with characteristic astrocytic morphology. These data suggest that the induction of NGF and bFGF alone appears to be insufficient as a compensatory mechanism to prevent ADC.
Assuntos
Complexo AIDS Demência/genética , Complexo AIDS Demência/imunologia , Fator 2 de Crescimento de Fibroblastos/genética , Fatores de Crescimento Neural/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos , Elementos Antissenso (Genética) , Astrócitos/química , Astrócitos/imunologia , Astrócitos/virologia , Química Encefálica/imunologia , Fator Neurotrófico Derivado do Encéfalo/análise , Fator Neurotrófico Derivado do Encéfalo/genética , Fator Neurotrófico Derivado do Encéfalo/imunologia , Córtex Cerebral/imunologia , Córtex Cerebral/patologia , Córtex Cerebral/virologia , Feminino , Fator 2 de Crescimento de Fibroblastos/análise , Fator 2 de Crescimento de Fibroblastos/imunologia , Expressão Gênica/imunologia , Soronegatividade para HIV , Humanos , Fator de Crescimento Insulin-Like I/análise , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/imunologia , Macrófagos/química , Macrófagos/imunologia , Macrófagos/virologia , Masculino , Pessoa de Meia-Idade , Fatores de Crescimento Neural/análise , Fatores de Crescimento Neural/imunologia , RNA Mensageiro/análise , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
The chemokine receptor CCR5 and to a lesser extent CCR2b and CCR3 have been shown to serve as coreceptors for HIV-1 entry into macrophages. Individuals that are homozygous for a defective CCR5 allele (DeltaCCR5) are highly, but not fully, resistant to infection with HIV-1. Here, we want to emphasize the importance of DeltaCCR5 in in vitro as well as in vivo studies. We provide data that suggest that CCR5 polymorphism may affect the onset of AIDS dementia complex in vivo and data that show that HIV-1 replication is influenced by the DeltaCCR5 allele in vitro. Knowing the CCR5 genotype of an individual will help to better interpret research results and may even provide new information about mechanisms of disease.
Assuntos
Complexo AIDS Demência/etiologia , HIV-1/fisiologia , Polimorfismo Genético , Receptores CCR5/genética , Complexo AIDS Demência/genética , Complexo AIDS Demência/virologia , Produtos do Gene tat/metabolismo , Genótipo , Heterozigoto , Humanos , Macrófagos/virologia , Reação em Cadeia da Polimerase , RNA Viral/análise , Replicação Viral , Produtos do Gene tat do Vírus da Imunodeficiência HumanaRESUMO
One of the strongest predictors for HIV-1-associated dementia is the presence of monocytic infiltration in perivascular areas of the brain. Therefore, macrophages have been suggested to play a major role in the development of this disease. This review focuses on possible mechanisms through which the macrophage may enhance disease progression by mediating neuronal damage.
Assuntos
Complexo AIDS Demência/etiologia , HIV-1 , Macrófagos , Doenças Neurodegenerativas/etiologia , Astrócitos , Barreira Hematoencefálica , Humanos , Neurotoxinas/toxicidade , Proteínas dos Retroviridae/toxicidadeRESUMO
In human immunodeficiency virus (HIV)-1-associated dementia (HAD), consequences of interactions between infiltrating monocytes and brain endothelial cells are not yet fully understood. This study investigated whether the blood-brain barrier is affected in brain tissue of patients suffering from HAD and whether it was possible to find a correlation with the presence or absence of monocytic cells, which have been suggested to play a major role in HAD. Immunohistochemical analysis for zonula occludens 1, a tight junction protein, and CD68, a macrophage marker, revealed that loss of tight junction immunoreactivity was highly correlated with monocyte infiltration and with HAD. This suggests that the presence of perivascular macrophages cells is associated with breakdown of the blood-brain barrier thereby facilitating infiltration of more monocytic cells hence enhancing disease progression.
Assuntos
Complexo AIDS Demência/patologia , HIV-1 , Proteínas de Membrana/análise , Monócitos/patologia , Fosfoproteínas/análise , Junções Íntimas/patologia , Complexo AIDS Demência/complicações , Complexo AIDS Demência/metabolismo , Síndrome da Imunodeficiência Adquirida/complicações , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Barreira Hematoencefálica , Encéfalo/metabolismo , Encéfalo/patologia , Humanos , Imuno-Histoquímica , Proteínas de Membrana/deficiência , Fosfoproteínas/deficiência , Estudos Retrospectivos , Junções Íntimas/metabolismo , Proteína da Zônula de Oclusão-1RESUMO
MS is a demyelinating disease characterized by infiltration of monocytes and lymphocytes into the brain parenchyma, destruction of oligodendrocytes and loss of myelin. Since chemokines play a major role in the migration of monocytes and T cells, we here investigated the expression of the CC chemokines MIP-1alpha, MIP-1beta, and RANTES in brain tissue from MS patients using reverse transcriptase-polymerase chain reaction techniques. Both MIP-1beta as well as RANTES were found to be significantly elevated in brain tissue of MS patients. In addition, MIP-1alpha was also increased, although not significantly. Immunohistochemistry revealed that, whereas RANTES was mainly localized in reactive astrocytes, MIP-1alpha and MIP-1beta immunoreactivity was predominantly found in perivascular and parenchymal macrophages, containing myelin degradation products. Thus, chemokines appear to be associated with MS and an increased chemokine expression may further enhance disease progression by attracting more leucocytes into the brain parenchyma and by activation of effector functions of astrocytes and microglial cells.
Assuntos
Quimiocina CCL5/genética , Quimiocina CCL5/metabolismo , Proteínas Inflamatórias de Macrófagos/genética , Proteínas Inflamatórias de Macrófagos/metabolismo , Esclerose Múltipla/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Sequência de Bases , Encéfalo/imunologia , Encéfalo/metabolismo , Encéfalo/patologia , Estudos de Casos e Controles , Quimiocina CCL3 , Quimiocina CCL4 , Primers do DNA/genética , Feminino , Expressão Gênica , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/genética , Esclerose Múltipla/patologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
The structure and function of neurons are changed not only during development of the central nervous system but also in certain neurological disorders, such as Alzheimer's disease and human immunodeficiency virus type 1 (HIV-1) -associated dementia. Immunological activation and altered production of neurotoxins and neurotrophins by brain macrophages are thought to play an important role in neuronal structure and function. This review describes the clinical and pathological features of both Alzheimer's disease and HIV-1-associated dementia and tries to interpret the role of the macrophage and astrocytes therein. The consequences of activation of macrophages by amyloid-beta in Alzheimer's disease and HIV infection of macrophages in HIV-1-associated dementia and the similarities between these diseases will be discussed. Although the neuropathology of Alzheimer's disease and HIV-1-associated dementia differs, Alzheimer's disease is a cortical dementia and HIV-1-associated dementia is a subcortical dementia, the process of macrophage activation and the resulting pathways leading to neurotoxicity seem very similar. In both Alzheimer's disease and HIV-1-associated dementia, interaction of macrophages and astrocytes appear to play an important role.
Assuntos
Complexo AIDS Demência/etiologia , Complexo AIDS Demência/imunologia , Doença de Alzheimer/etiologia , Doença de Alzheimer/imunologia , Ativação de Macrófagos/imunologia , Complexo AIDS Demência/patologia , Doença de Alzheimer/patologia , HumanosRESUMO
The presence of perivascular monocytic infiltration is a major hallmark of HIV-1-associated dementia. Since CC chemokines are chemoattractant cytokines that are able to attract T cells and monocytes/macrophages to sites of inflammation, and since infiltrating monocytes/macrophages remain in close contact with the brain endothelium, we investigated whether interactions between HIV-1-infected macrophages and brain endothelium result in an altered chemokine production. We found an increased mRNA expression of monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein (MIP)-1 alpha and MIP-1 beta, and RANTES by macrophages after HIV-1 infection. Interactions between HIV-infected macrophages and brain microvascular endothelial cells resulted in an additional upregulation of chemokine mRNA expression, during cell-cell contact as well as in a trans-well system. Since IL-1 beta can function as a modulator of chemokine expression we investigated if interleukin-1 beta could be involved in the regulation of chemokine induction. Coculturing of HIV-infected macrophages and endothelial cells resulted in immune-activation as indicated by increased mRNA expression of IL-1 beta. Subsequently, addition of a neutralizing antibody against IL-1 beta resulted in altered chemokine expression by macrophages, but not by endothelial cells. Thus, IL-1 beta appears to play a major role in the regulation of chemokines during cellular interactions in HIV-associated dementia, but other factors may also be involved.
Assuntos
Circulação Cerebrovascular/imunologia , Quimiocinas/genética , Endotélio Vascular/imunologia , Endotélio Vascular/virologia , Infecções por HIV/imunologia , Macrófagos/imunologia , Macrófagos/virologia , Anticorpos/farmacologia , Elementos Antissenso (Genética) , Células Cultivadas , Quimiocina CCL2/genética , Quimiocina CCL2/imunologia , Quimiocina CCL4 , Quimiocina CCL5/genética , Quimiocina CCL5/imunologia , Quimiocinas/imunologia , Endotélio Vascular/citologia , Expressão Gênica/imunologia , Humanos , Interleucina-1/genética , Interleucina-1/imunologia , Proteínas Inflamatórias de Macrófagos/genética , Proteínas Inflamatórias de Macrófagos/imunologia , Macrófagos/citologia , Microcirculação/imunologia , Microcirculação/virologia , Monócitos/citologia , Monócitos/imunologia , Monócitos/virologia , Testes de Neutralização , RNA Mensageiro/análise , SolubilidadeRESUMO
Feline immunodeficiency virus (FIV) isolates differ in the ability to replicate in Crandell feline kidney (CRFK) cells. The difference in tropism between two variants of the Dutch isolate FIV-UT113 was studied by using molecular clones which contained the envelope genes of the variants in a background of the FIV-14 Petaluma sequence. Virus produced from clone pPET-113Th replicated in thymocytes, whereas virus from pPET-113Cr propagated in both thymocytes and CRFK cells, thereby reflecting the phenotypes of the parental variants. Exchange of envelope gene fragments showed that a 464-bp surface protein (SU)-encoding fragment encompassing the third variable region (V3) determines CRFK cell tropism. Sequence analysis of the exchanged fragments demonstrated two amino acid changes that led to an increase of the overall charge of the V3 domain: a G-->R transition at position 397 and a E-->K change at position 407. Mutational analysis of these residues revealed that the E-->K shift was responsible for the change in tropism, while the G-->R mutation improved the replication kinetics in CRFK cells. Mapping of a tropism determinant for FIV to a region which is also a major neutralization domain is reminiscent of human immunodeficiency virus type 1, in which a similar colocation was found.
Assuntos
Vírus da Imunodeficiência Felina/fisiologia , Mutação , Proteínas do Envelope Viral/fisiologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Gatos , Linhagem Celular , Clonagem Molecular , Primers do DNA , Desoxirribonuclease HindIII/metabolismo , Vírus da Imunodeficiência Felina/genética , Dados de Sequência Molecular , Testes de Neutralização , Proteínas do Envelope Viral/genética , Replicação Viral/genética , Replicação Viral/fisiologiaRESUMO
Oxidative stress is suggested to be involved in several neurodegenerative diseases. One mechanism of oxidative damage is mediated by peroxynitrite, a neurotoxic reaction product of superoxide anion and nitric oxide. Expression of two cytokines and two key enzymes that are indicative of the presence of reactive oxygen intermediates and peroxynitrite was investigated in brain tissue of AIDS patients with and without AIDS dementia complex and HIV-seronegative controls. RNA expression of IL-1beta, IL-10, inducible nitric oxide synthase, and superoxide dismutase (SOD) was found to be significantly higher in demented compared with nondemented patients. Immunohistochemical analysis showed that SOD was expressed in CD68-positive microglial cells while inducible nitric oxide synthase was detected in glial fibrillary acidic protein (GFAP)-positive astrocytes and in equal amounts in microglial cells. Approximately 70% of the HIV p24-Ag-positive macrophages did express SOD, suggesting a direct HIV-induced intracellular event. HIV-1 infection of macrophages resulted in both increased superoxide anion production and elevated SOD mRNA levels, compared with uninfected macrophages. Finally, we show that nitrotyrosine, the footprint of peroxynitrite, was found more intense and frequent in brain sections of demented patients compared with nondemented patients. These results indicate that, as a result of simultaneous production of superoxide anion and nitric oxide, peroxynitrite may contribute to the neuropathogenesis of HIV-1 infection.