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. 2009 Jan 14;58(8):1307–1317. doi: 10.1007/s00262-008-0644-9

Fig. 3.

Fig. 3

Capturing of human IL-10 by immunoadhesins. a IL-10 non-producing human melanoma cell line ZA and COS-1 cells were transfected with plasmid vectors and culture supernatants from transfected cells were collected. The culture supernatants were incubated with 1 ng/ml of rhIL-10 for 30 min and unbound rhIL-10 was measured by an IL-10 ELISA kit. The data were shown as recovery rates (%) of rhIL-10. The recovery rates were calculated using the rhIL-10 levels detected in the supernatants obtained from untransfected cells as 100%. Control: supernatant from untransfected cells, #0 backbone pVAX1, #1 IL10R1 extracellular domain only, #2 IL10R1/IgG1Fc with no hinge, monomer, #3 IL10R1/IgG1Fc with mutated hinge, monomer, #4 IL10R1/IgG1Fc with wild type hinge, dimer. **P < 0.01, ***P < 0.001, compared to control. b Different concentrations of rhIL-10 (0.1, 1, 10 ng/ml) were incubated with COS-1 culture supernatants that contained different types of IL-10 immunoadhesin. After 30 min of incubation, unbound rhIL-10 was measured by an IL-10 ELISA kit. Control: supernatant from untransfected cells, #0 backbone pVAX1, #1 IL10R1 extracellular domain only, #2 IL10R1/IgG1Fc with no hinge, monomer, #3 IL10R1/IgG1Fc with mutated hinge, monomer, #4 IL10R1/IgG1Fc with wild type hinge, dimer. ***P < 0.001, compared to control. c IL-10 producing human melanoma cell lines JB and H88 were transfected with IL-10 immunoadhesin plasmid vectors and then incubated for 3 days. Endogenously produced human IL-10 in the culture supernatants were measured by an ELISA kit. The data were shown as IL-10 detection rates (%). The IL-10 detection rates were calculated using the IL-10 levels in the supernatants obtained from untransfected cells (control: JB 90 pg/ml, H88 620 pg/ml) as 100%. Control supernatant from untransfected cells, #0 backbone pVAX1, #1 IL10R1 extracellular domain only, #2 IL10R1/IgG1Fc with no hinge, monomer, #3 IL10R1/IgG1Fc with mutated hinge, monomer, #4 IL10R1/IgG1Fc with wild type hinge, dimer. **P < 0.01, ***P < 0.001, compared to control