American Society of Hematology

Fig. 2.

$Fig. 2. Pro–IL-1β is present in P1 and P2 fractions; processing on interaction with alloreactive T cells. / P1 and P2 fractions (lanes 3 and 4) were obtained by sequential ultracentrifugation of PNS from mature DCs cultured 5 hours alone (A) or with alloreactive CD8+ T cells (at a DC/T cell ratio of 1:10, panel B), treated with proteinase K and solubilized in sample buffer. Lane 1: cytosol (TCA-concentrated, 1/20 of total, 100 μg); lane 2: PNS (1/20 of total). Samples were blotted and hybridized with anti–IL-1β mAb. Note the appearance of the ICE-specific 29-kd IL-1β band, of approximately the same intensity as the 35-kd, in P1 from DCs cultured with alloreactive T cells (B, lane 3). The low-molecular-weight band (17 kd) in lanes 3B and 4B is erratic and probably due to nonspecific endoproteases activated during the preparation of the samples, as already reported.10 One representative experiment of 5 is shown.$

Pro–IL-1β is present in P1 and P2 fractions; processing on interaction with alloreactive T cells.

P1 and P2 fractions (lanes 3 and 4) were obtained by sequential ultracentrifugation of PNS from mature DCs cultured 5 hours alone (A) or with alloreactive CD8⁺ T cells (at a DC/T cell ratio of 1:10, panel B), treated with proteinase K and solubilized in sample buffer. Lane 1: cytosol (TCA-concentrated, 1/20 of total, 100 μg); lane 2: PNS (1/20 of total). Samples were blotted and hybridized with anti–IL-1β mAb. Note the appearance of the ICE-specific 29-kd IL-1β band, of approximately the same intensity as the 35-kd, in P1 from DCs cultured with alloreactive T cells (B, lane 3). The low-molecular-weight band (17 kd) in lanes 3B and 4B is erratic and probably due to nonspecific endoproteases activated during the preparation of the samples, as already reported.10 One representative experiment of 5 is shown.

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