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In Vitro Phagocytosis and Production of Hydrogen Peroxide (H2O2) by Human Peripheral Blood Mononuclear Cells (PBMC) Obtained from Jaundice Patients

Abstracts
2002 Digestive Disease Week

# 104837 Abstract ID: 104837 In Vitro Phagocytosis and Production of Hydrogen Peroxide (H2O2) by Human Peripheral Blood Mononuclear Cells (PBMC) Obtained from Jaundice Patients
Marisa Treglia-Dal-lago, José Jukemura, Marcel C Machado, José A Barbuto, Sao Paulo, Brazil

Jaundice patients present several alterations in immune function. Here we describe phagocytosis and H2O2 production by PBMC from these patients. Peripheral blood was obtained from patients before and after surgery, with relief of cholestasis. Bilirubin was measured at the time of blood collections. PBMC were separated by density gradient centrifugation and cultured for one hour in peroxidase and phenol red containing medium. In the presence or not of phorbol myristate acetate (PMA 30 ng/mL), or for one week, in serum supplemented medium. Phagocytosis was evaluated by the ingestion of E.coli by PBMC (10:1 ratio) co-incubated for 30 min at 37C. H2O2 release was evaluated by phenol oxidation and transformed in nmoles/mL according to standard curve. PBMC from the eight patients tested presented low phagocytosis (as compared to normal controls). One week after surgery PBMC from all patients showed an increase in this activity. Similarly, cell obtained from 47 patients before surgery did not release H2O2, while after relief of cholestasis, 9/47 did. When cultured in fetal calf serum-containing medium, PBMC from 9/9 patients produced H2O2 after PMA stimulation. Culture in the presence of patients`sera inhibited this recovey. Together, these data suggest that jaundiced patients have a reversible decrease in monocyte function. Support: CNPq.




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