RESUMO
BACKGROUND: The immune processes involved in the development of alloantibodies against the human platelet antigens in alloimmune disorders remain unclear. Antibody recognition of the platelet antigens on their respective platelet glycoproteins has been shown to be dependent on glycoprotein conformation. Furthermore, the post-translational modification of glycoproteins adds complexity to the alloantigenic determinants. METHODS: Nine anti-HPA-3a sera along with several control sera were tested for reactivity to an 11-mer peptide straddling the HPA-3a/b polymorphism. Sera found to specifically recognize the 3a peptide were further assessed by platelet pre-exposure and immunoblotting. RESULTS: Three of the nine antisera were found to specifically recognize an 11-mer synthetic 3a peptide by ELISA. Further analysis of all anti-HPA-3a sera by Western blot showed that only those reactive to the 3a peptide were able to bind both reduced and non-reduced GPIIb. CONCLUSION: The results presented in this study provide the first known evidence for the identification of an antibody population capable of recognizing a linear and non-glycosylated form of the HPA-3a epitope.
Assuntos
Antígenos de Plaquetas Humanas/imunologia , Doenças Autoimunes/imunologia , Isoanticorpos/imunologia , Peptídeos/imunologia , Polimorfismo Genético/imunologia , Formação de Anticorpos/genética , Antígenos de Plaquetas Humanas/química , Antígenos de Plaquetas Humanas/genética , Doenças Autoimunes/sangue , Epitopos/química , Epitopos/genética , Epitopos/imunologia , Feminino , Humanos , Isoanticorpos/sangue , Isoanticorpos/química , Masculino , Peptídeos/química , Peptídeos/genéticaRESUMO
The synthetic chalcone derivative 1-(2,4-dichlorophenyl)-3-(3-(6,7-dimethoxy-2-chloroquinolinyl))-2-propen-1-one (ClDQ) was evaluated for its anti-inflammatory, analgesic and immunomodulatory efficacy in-vitro and in-vivo. ClDQ concentration-dependently inhibited the production of nitric oxide (NO) (IC50 4.3 microM) and prostaglandin E(2) (PGE(2)) (IC50 1.8 microM) in RAW 264.7 macrophages stimulated with lipopolysaccharide. Human mononuclear cell proliferation was significantly inhibited by 10 microM ClDQ. Oral administration of ClDQ (10-30 mg kg(-1)) in the 24-h zymosan-stimulated mouse air-pouch model produced a dose-dependent reduction of cell migration as well as NO and PGE(2) levels in exudates. ClDQ (20 mg kg(-1), p.o.) inhibited ear swelling and leucocyte infiltration in the delayed-type hypersensitivity response to 2,4-dinitrofluorobenzene in mice. In the rat adjuvant-arthritis model, this compound reduced joint inflammation as well as PGE(2) and cytokine levels. In addition, ClDQ displayed analgesic effects in the phenylbenzoquinone-induced abdominal constriction model in mice and in the late phase of the nociceptive response to formalin. Our findings indicated the potential interest of ClDQ in the modulation of some immune and inflammatory conditions.
Assuntos
Inflamação/prevenção & controle , Piridazinas/síntese química , Traumatismos Abdominais/induzido quimicamente , Traumatismos Abdominais/prevenção & controle , Administração Oral , Animais , Artrite Experimental/induzido quimicamente , Artrite Experimental/imunologia , Plaquetas/efeitos dos fármacos , Plaquetas/enzimologia , Linhagem Celular , Ciclo-Oxigenase 1/metabolismo , Ciclo-Oxigenase 2/metabolismo , Dinitrofluorbenzeno , Dinoprostona/metabolismo , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Hipersensibilidade a Drogas/imunologia , Hipersensibilidade a Drogas/prevenção & controle , Feminino , Formaldeído , Fosfolipases A2 do Grupo II , Fosfolipases A2 do Grupo IV , Humanos , Inflamação/induzido quimicamente , Lipopolissacarídeos/farmacologia , Ativação Linfocitária/efeitos dos fármacos , Ativação Linfocitária/imunologia , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Microssomos/efeitos dos fármacos , Microssomos/enzimologia , Nitritos/metabolismo , Dor/induzido quimicamente , Dor/prevenção & controle , Medição da Dor/métodos , Fosfolipases A/antagonistas & inibidores , Fosfolipases A/metabolismo , Piridazinas/farmacologia , Ratos , Ratos Endogâmicos Lew , Tromboxano B2/metabolismo , ZimosanRESUMO
OBJECTIVE AND DESIGN: The synthetic chalcone derivative 1-(2,3,4-trimethoxyphenyl)-3-(3-(2-chloroquinolinyl))-2-propen-1-one (TQ) was evaluated for its immunomodulatory and anti-inflammatory efficacy in vitro and in vivo. MATERIAL AND SUBJECTS: Human neutrophils and lymphocytes from healthy volunteers and RAW 264.7 murine macrophages. Swiss mice and Lewis rats were randomly divided into groups of six animals. TREATMENT: TQ was orally administered in all in vivo assays (10-30 mg/kg). METHODS: Elastase, superoxide and LTB(4) release were assayed in human neutrophils, NO/PGE(2) production and NF-kappaB activation in RAW 264.7, and (3)H thymidine incorporation in human lymphocytes. Zymosan-stimulated air pouches, DNFB-DTH, PBQ-induced writhings and formalin-induced pain were assayed in mice. Adjuvant-induced arthritis was tested in rats. Dunnett's t-test was employed for statistical analysis. RESULTS: Human T-cell proliferation, neutrophil functions and NO/PGE(2) production in murine macrophages were inhibited by TQ (IC(50) in the microM range), which showed anti-inflammatory, immunomodulatory and analgesic effects. CONCLUSIONS: Our findings indicate the potential interest of TQ in the modulation of some immune and inflammatory responses probably by NF-kappaB inhibition.