Fig. 4.
Fig. 4. Comparison of ubiquitin serum levels with LPS-stimulated whole blood TNF-α production and neutralization of the inhibitory activity for TNF-α production in patients' serum with antiubiquitin antibody. / (A) Ubiquitin serum concentrations in volunteers (n = 12) and trauma patients (n = 10) during 14 days after trauma. Data represent means ± SEM. (B) LPS-induced whole blood TNF-α secretion. Volunteers and trauma patients are the same as described in panel A. Whole blood cultures were incubated for 4 hours in the presence of 100 ng/mL LPS. Data represent means ± SEM. (C-D) Antiubiquitin antibodies neutralize the inhibitory activity of ubiquitin on LPS-induced whole blood (C) and PBMNCs (D) TNF-α production. TNF-α production (%) indicates percent of the TNF-α secretion in cultures without exogenous ubiquitin and without antibodies. Data represent means ± SEM from 3 different whole blood and PBMNC cultures obtained from healthy volunteers. Cultures without (■) or with 500 ng/mL exogenous ubiquitin (▪) in the presence of LPS (100 ng/mL for 4 hours). Control indicates cultures without addition of antibody. AS: antiubiquitin AS diluted 1:103, 1:102, and 1:10 in the cell cultures. UbP4D1 and UbN-19: diluted 1:103 in the cell cultures. (E-F) Effect of antiubiquitin antibody on the inhibitory activity of trauma (E) and sepsis (F) patients' serum on LPS-induced TNF-α production of whole blood and PBMNCs. Whole blood (■) and PBMNCs (▪) were cultured with 100 ng/mL LPS for 4 hours. TNF production (%) indicates percent of the TNF-α secretion in cultures containing additional healthy volunteers' serum (30%, vol/vol, in the cell culture mixture) without antibodies. Data represent means ± SEM from 4 different cultures obtained from healthy volunteers. C indicates control, healthy volunteers' serum; TS, trauma patients' serum (n = 4, 30%, vol/vol, in the cell culture mixture); SS, sepsis patients' serum (n = 4, 30%, vol/vol, in the cell culture mixture). Antiubiquitin AS diluted 1:102 and 1:10 in the cell cultures. UbP4D1 and UbN19 were diluted 1:103 in the cell cultures.

Comparison of ubiquitin serum levels with LPS-stimulated whole blood TNF-α production and neutralization of the inhibitory activity for TNF-α production in patients' serum with antiubiquitin antibody.

(A) Ubiquitin serum concentrations in volunteers (n = 12) and trauma patients (n = 10) during 14 days after trauma. Data represent means ± SEM. (B) LPS-induced whole blood TNF-α secretion. Volunteers and trauma patients are the same as described in panel A. Whole blood cultures were incubated for 4 hours in the presence of 100 ng/mL LPS. Data represent means ± SEM. (C-D) Antiubiquitin antibodies neutralize the inhibitory activity of ubiquitin on LPS-induced whole blood (C) and PBMNCs (D) TNF-α production. TNF-α production (%) indicates percent of the TNF-α secretion in cultures without exogenous ubiquitin and without antibodies. Data represent means ± SEM from 3 different whole blood and PBMNC cultures obtained from healthy volunteers. Cultures without (■) or with 500 ng/mL exogenous ubiquitin (▪) in the presence of LPS (100 ng/mL for 4 hours). Control indicates cultures without addition of antibody. AS: antiubiquitin AS diluted 1:103, 1:102, and 1:10 in the cell cultures. UbP4D1 and UbN-19: diluted 1:103 in the cell cultures. (E-F) Effect of antiubiquitin antibody on the inhibitory activity of trauma (E) and sepsis (F) patients' serum on LPS-induced TNF-α production of whole blood and PBMNCs. Whole blood (■) and PBMNCs (▪) were cultured with 100 ng/mL LPS for 4 hours. TNF production (%) indicates percent of the TNF-α secretion in cultures containing additional healthy volunteers' serum (30%, vol/vol, in the cell culture mixture) without antibodies. Data represent means ± SEM from 4 different cultures obtained from healthy volunteers. C indicates control, healthy volunteers' serum; TS, trauma patients' serum (n = 4, 30%, vol/vol, in the cell culture mixture); SS, sepsis patients' serum (n = 4, 30%, vol/vol, in the cell culture mixture). Antiubiquitin AS diluted 1:102 and 1:10 in the cell cultures. UbP4D1 and UbN19 were diluted 1:103 in the cell cultures.

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