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. Author manuscript; available in PMC: 2013 Mar 1.
Published in final edited form as: Am J Transplant. 2011 Dec 7;12(3):640–648. doi: 10.1111/j.1600-6143.2011.03869.x

Figure 1. Indirect pathway T-cell analysis.

Figure 1

(A) The tvDTH responses to donor antigen (Teff responses) are significantly different between subject groups (p < 0.0001). PBMC from patients (n = 45) were co-injected into the mouse footpad with cell-free donor antigen. Swelling was measured before and 24 h after each injection. Net swelling was determined by subtracting the swelling response obtained from the injection of the same PBMC in PBS. Patient groups: TWIN, identical twin donor (n = 2); TOL, clinically tolerant, no immunosuppression for >1 year (n = 11); MONO, prednisone monotherapy (n = 7); SI, standard immunosuppression (n = 18) and CR, chronic rejection (n = 7). (B) Antibody blocking of indirect pathway T-cell responses. PBMC from patients with CR (n = 5) were injected into the footpad with cell-free donor antigen and blocking antibodies to IFNγ or IL-17 (right side), whereas PBMC from patients in the TOL group (n = 5) were injected with cell-free donor antigen and antibodies to block TGFβ or IL-10 (left side). The net swelling response to donor antigen was significantly elevated in the TOL patients when TGFβ was neutralized (p < 0.01) but not when IL10 was neutralized whereas blocking of either IFNγ or IL17 significantly decreased the swelling response to donor antigen in the CR patients tested (p < 0.01). Net swelling response for each subject with donor antigen and a given test antibody were compared to that with PBMC + donor antigen and isotype control Ab (−) using a paired t-test. (C) Indirect pathway Treg responses are significantly different between subject groups (p = 0.005). PBMC from patients in the same groups as described in (A) were injected with recall antigen (TT/DT or EBV) alone and with recall plus cell-free donor antigen into the mouse footpad. Net swelling was determined as in (A). The percentage inhibition of recall alone response in the presence of donor antigen was calculated as described in the methods. All panels: Individual results from each subject (individual symbols) and group median (horizontal line) with quartiles are shown. Each patient was tested one or two times. Significant differences between groups using pair-wise comparisons (Dunn’s correction for multiple comparisons shown in solid lines); *p < 0.05, **p < 0.01 and ***p < 0.001; (Mann–Whitney U test unadjusted for multiple comparisons shown with dotted lines) #p = 0.04, ##p = 0.03 and ###p = 0.008.