RESUMO
BACKGROUND: Flowcytometric identification of basophils is a prerequisite for measuring activation of basophils with IgE-dependent or IgE-independent stimuli. Aim of this study was to compare different marker combinations in a simultaneous multicolor flowcytometric measurement. METHODS: Ten patients with a grass pollen allergy and three controls were included in the study. Basophilic cells were gated by using anti-CCR3, anti-IgE, anti-CRTH2, anti-CD203c, and anti-CD3. Cells were activated by a monoclonal anti-FcεRI antibody, N-formyl-methionyl-leucyl-phenylalanine (fMLP), and the allergen extract Phleum pratense. The activation marker anti-CD63 was used. RESULTS: The highest relative number of basophils was found with anti-CCR3+ cells, anti-IgE+ and anti-IgE+ /anti-CD203c+ cells, the lowest with CRTH2+/CD203c+/CD3- cells. A very good and good concordance of CCR3+ cells was seen with CCR3+/CD3- cells and CRTH2+/CD203c+/CD3- cells in all experiments. The contamination of the CCR3+ population with CD3+ cells and the contamination of the IgE+-population with CCR3- cells and CD203- cells were the lowest compared to all other marker combinations. CONCLUSIONS: As the highest relative number of basophils was identified by anti-CCR3 followed by the anti-IgE and anti-IgE/antiCD203c positive population in most cases, these markers can generally be recommended for identification of basophils. If a basophil population with very high purity is needed, anti-IgE should be chosen.
Assuntos
Anticorpos Anti-Idiotípicos/química , Basófilos/imunologia , Imunoglobulina E/sangue , Imunofenotipagem/métodos , Hipersensibilidade Respiratória/diagnóstico , Adulto , Alérgenos/química , Alérgenos/imunologia , Anticorpos Monoclonais/farmacologia , Teste de Degranulação de Basófilos , Basófilos/efeitos dos fármacos , Basófilos/patologia , Complexo CD3/genética , Complexo CD3/imunologia , Estudos de Casos e Controles , Células Cultivadas , Feminino , Citometria de Fluxo/métodos , Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , N-Formilmetionina Leucil-Fenilalanina/farmacologia , Diester Fosfórico Hidrolases/genética , Diester Fosfórico Hidrolases/imunologia , Extratos Vegetais/química , Extratos Vegetais/imunologia , Extratos Vegetais/farmacologia , Pólen/química , Pólen/imunologia , Pirofosfatases/genética , Pirofosfatases/imunologia , Receptores CCR3/genética , Receptores CCR3/imunologia , Receptores Imunológicos/genética , Receptores Imunológicos/imunologia , Receptores de Prostaglandina/genética , Receptores de Prostaglandina/imunologia , Hipersensibilidade Respiratória/imunologia , Hipersensibilidade Respiratória/patologia , Tetraspanina 30/genética , Tetraspanina 30/imunologiaRESUMO
BACKGROUND AND OBJECTIVE: This study was aimed to analyse the curative effects of Pinellia ternata, Citrus reticulata, and their combination on airway hyperresponsiveness (AHR) to inhaled methacholine, pulmonary eosinophilic infiltration, Th2 cytokine production, and IgE and histamine production in a murine model of asthma. METHODS: For this purpose, BALB/c mice were systemically sensitized to ovalbumin (OVA) followed intratracheally, intraperitoneally, and by aerosol allergen challenges for 12 weeks. We examined the development of pulmonary eosinophilic accumulation, control of Th2 cytokine, immunoglobulin E (IgE), and histamine productions in a murine model of asthma. RESULTS: Our data suggest that the therapeutic mechanism by which Pinellia ternata, Citrus reticulata, and their combinational prescription effectively treats asthma is based on reductions of eosinophil infiltration, eotaxin receptor (CCR3), histamine, OVA-specific IgE productions in serum, and Th2 cytokines (IL-5, IL-13) by marked reductions of IL-5 and IL-13 mRNA expression in lung tissue. CONCLUSIONS: These findings provide evidence that Pinellia ternata, Citrus reticulata, and their combination play a regulatory role in allergic inflammation and offer therapeutic approaches as novel CCR3 antagonists for treatment asthma. However, it is not clear whether pharmacological activities of prescription composed of two herbs are potentiated due to synergistic effect or additive effect.