American Society of Hematology

Fig. 5.

$Fig. 5. Apo A-I inhibits the production of TNF-α and IL-1β in THP-1 cells activated by membranes of stimulated HUT-78 cells. / (A) THP-1 cells were activated by membranes of stimulated HUT-78 cells in the presence of increasing concentrations of Apo A-I purchased from Sigma. After 48 hours, TNF-α and IL-1β were measured in culture supernatants. Results represent mean ± SD, n = 3. (B,C) THP-1 cells were activated by membranes of stimulated HUT-78 cells in the presence of increasing concentrations of proteins electroeluted from preparative SDS-PAGE of delipidated HDL: panel B, Mr = 28 000, and panel C, Mr = 18 000. (D) THP-1 cells were activated by membranes of stimulated HUT-78 cells in the presence of increasing concentrations of apo A-I (Mr = 28 000) isolated by gel filtration on Superdex S75. After 48 hours, TNF-α (closed squares) and IL-1β (closed circles) were measured in culture supernatants. Results represent mean ± SD, n = 3, 100% being the amount of cytokine produced in the absence of inhibitor. (E) Isolated fractions that were tested for inhibitory activity were analyzed for their contents in apo A-I by Western blot: (a) 28 000 electroeluted band tested in panel B, (b) 18 000 electroeluted band tested in panel C, and (c) 28 000 protein recovered from Superdex S75 gel filtration and tested in panel D.$

Apo A-I inhibits the production of TNF-α and IL-1β in THP-1 cells activated by membranes of stimulated HUT-78 cells.

(A) THP-1 cells were activated by membranes of stimulated HUT-78 cells in the presence of increasing concentrations of Apo A-I purchased from Sigma. After 48 hours, TNF-α and IL-1β were measured in culture supernatants. Results represent mean ± SD, n = 3. (B,C) THP-1 cells were activated by membranes of stimulated HUT-78 cells in the presence of increasing concentrations of proteins electroeluted from preparative SDS-PAGE of delipidated HDL: panel B, M_r = 28 000, and panel C, M_r = 18 000. (D) THP-1 cells were activated by membranes of stimulated HUT-78 cells in the presence of increasing concentrations of apo A-I (M_r = 28 000) isolated by gel filtration on Superdex S75. After 48 hours, TNF-α (closed squares) and IL-1β (closed circles) were measured in culture supernatants. Results represent mean ± SD, n = 3, 100% being the amount of cytokine produced in the absence of inhibitor. (E) Isolated fractions that were tested for inhibitory activity were analyzed for their contents in apo A-I by Western blot: (a) 28 000 electroeluted band tested in panel B, (b) 18 000 electroeluted band tested in panel C, and (c) 28 000 protein recovered from Superdex S75 gel filtration and tested in panel D.

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