Modificação induzida por Î²2-glicoproteína I na resposta oxidativa de polimorfonucleares humanos durante a fagocitose

Pereira, Elisângela Monteiro

doi:10.11606/D.9.2018.tde-11012018-141639

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Master's Dissertation

DOI

https://doi.org/10.11606/D.9.2018.tde-11012018-141639

Document

Master's Dissertation

Author

Pereira, Elisângela Monteiro (Catálogo USP)

Full name

Elisângela Monteiro Pereira

E-mail

Institute/School/College

Faculdade de Ciências Farmacêuticas

Knowledge Area

Clinical Analysis

Date of Defense

2005-08-19

Published

São Paulo, 2005

Supervisor

Gomes, Ligia Ferreira (Catálogo USP)

Committee

Gomes, Ligia Ferreira (President)
Colepicolo, Pio
Negro, Sonia Jancar

Title in Portuguese

Modificação induzida por β₂-glicoproteína I na resposta oxidativa de polimorfonucleares humanos durante a fagocitose

Keywords in Portuguese

Beta2-Glicoproteína I
Biologia celular
Bioquímica clínica
Citometria de fluxo
Espécies reativas de nitrogênio
Espécies reativas de oxigênio
Glicoproteínas (Fisiopatologia)
Polimorfonucleares
Quimiluminescência
Radicais livres (Fisiopatologia)

Abstract in Portuguese

β₂-glicoproteína I (β₂GPI) é encontrada (200µg/mL) no plasma, 60% livre e 40% em lipoproteínas. Esta proteína de fase aguda, com afinidade por superfícies negativas pode ser clivada pela plasmina. Fragmentos são purificados como dímeros ou multímeros de β₂GPI. Formas monomérica e dimérica de β₂GPI foram purificadas de soro humano e identificadas por SDS-PAGE, imunoblot e ELISA Somente a forma monomérica foi detectada no teste ELISA Os efeitos de ambas as formas sobre o burst respiratório de polimorfonucleares humanos (PMN), estimulados in vitro com zymosan opsonizado, foram estudados por quimioluminescência amplificada por luminol ou lucigenina e citometria de fluxo, pela oxidação do OCFH. A forma monomérica inibiu a quimioluminescência amplificada por luminol (-43.6 x -7.33 AEU/s), mas não por lucigenina, e aumentou a oxidação de DCFH e a produção de Óxido Nítrico (^•NO). É provável que o ^•No, via peroxinitrito, medeie os efeitos de β₂GPI sobre o burst respiratório de PMN.

Title in English

Modification induced by β₂-glycoprotein I in the oxidative response of human polymorphonuclear cells during phagocytosis

Keywords in English

Beta2-Glycoprotein I
Cell biology
Chemiluminescence
Clinical biochemistry
Flow cytometry
Free radicals (Physiopathology)
Glycoproteins (Physiopathology)
Polymorphonuclear cells
Reactive nitrogen species
Reactive oxygen species

Abstract in English

Circulating blood contains approximately 200µg/mL of β ₂-glycoprotein I (β₂GPI), either free (60%) or lipoprotein bound (40%). This acute phase protein, with affinity for negative surfaces, can be cleaved by plasmin. Fragments purify as dimeric or multimeric (β₂GPI. Both (β₂GPI forms were purified from human sera and identified by SDS-PAGE, immunoblotting and ELISA. ELISA reactivity was dependent on the monomeric status of (β₂GPI. The effects of dimeric and monomeric (β₂GPI upon respiratory burst of human polimorphonuclear neutrophils (PMN) stimulated in vitro with opsonized zymosan were studied. Respiratory burst was evaluated by luminol- or lucigenin-amplified chemiluminescence, or by DCFH oxidation (flow-cytometry assay). The monomeric, but not the dimeric form, inhibited the luminol chemiluminescence of zymosan-stimulated PMNs (-43.6 x -7.33 AEU/s). Lucigenin chemiluminescence was insensitive to (β₂-GPI. Monomeric (β₂GPI increases both DCFH oxidation and nitric oxide production. Nitric oxide, probably through peroxynitrite reactions, mediates (β₂GPI effects upon PMNs respiratory burst.

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Publishing Date

2018-01-11

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