Figure 2
Figure 2. Antigen-specific “nonresponsiveness” persists after subsequent RBC transfusion in the presence of inflammation. (A) Six weeks after hGPA or wild-type FVB RBC transfusion, recipients were challenged with hGPA RBCs in the presence of poly(I:C), with anti-hGPA levels assessed by flow cross-matching after the final transfusion. Compilation of 3 experiments, 29 recipients total. Adjusted mean fluorescence intensity (MFI) was 119 after poly(I:C) hGPA transfusion in the group initially not transfused, 103 in the group initially transfused with hGPA RBCs, and 1 in the group initially transfused with FVB RBCs. P < .0001 between the group initially not transfused and the hGPA group; P = .6 between the hGPA and FVB transfused groups. No anti-hGPA was produced in any group 2 weeks after the initial transfusion. (B) FVB recipients were either not transfused or transfused with hGPA RBCs, then transfused again 6 weeks later with HOD or HOD × hGPA F1 RBCs in the presence of poly(I:C), with anti-HOD assessed by flow cross-matching after the final transfusion. Compilation of 4 experiments, 40 recipients total. Mean adjusted MFI was 56 after poly(I:C) HOD or HOD × hGPA F1 in the group initially not transfused compared with 44 in the group initially transfused with hGPA RBCs (P = .65). No anti-hGPA or anti-HOD was detected in either group after the initial hGPA transfusion, and no anti-hGPA was detected after the final transfusion. (C) B10.BR recipients were transfused with mHEL or control C57BL/6 RBCs and then boosted 6 weeks later with mHEL RBCs in the presence of poly(I:C), with anti-HEL assessed by ELISA 2 weeks after the first and last transfusions. Compilation of 2 experiments with 18 recipients. Mean optical density (OD) was 0.45 before and 0.33 after poly(I:C) mHEL transfusion in the group initially transfused with mHEL RBCs; OD was 0.1 before and 1.1 after poly(I:C) mHEL transfusion in the group initially transfused with C57BL/6 RBCs (P = .004 between groups after the last transfusion). (D) B10.BR recipients were transfused with mHEL or control C57BL/6 RBCs and then injected subcutaneously 6 weeks later with HEL protein emulsified in complete Freund's adjuvant (CFA), with anti-HEL assessed 2 weeks after this immunization by flow cross-matching. Compilation of 2 experiments, 21 recipients total. Adjusted MFI was 3 after HEL/CFA treatment in the group initially transfused with mHEL RBCs compared with 38 in the group initially transfused with C57BL/6 RBCs (P = .0006).

Antigen-specific “nonresponsiveness” persists after subsequent RBC transfusion in the presence of inflammation. (A) Six weeks after hGPA or wild-type FVB RBC transfusion, recipients were challenged with hGPA RBCs in the presence of poly(I:C), with anti-hGPA levels assessed by flow cross-matching after the final transfusion. Compilation of 3 experiments, 29 recipients total. Adjusted mean fluorescence intensity (MFI) was 119 after poly(I:C) hGPA transfusion in the group initially not transfused, 103 in the group initially transfused with hGPA RBCs, and 1 in the group initially transfused with FVB RBCs. P < .0001 between the group initially not transfused and the hGPA group; P = .6 between the hGPA and FVB transfused groups. No anti-hGPA was produced in any group 2 weeks after the initial transfusion. (B) FVB recipients were either not transfused or transfused with hGPA RBCs, then transfused again 6 weeks later with HOD or HOD × hGPA F1 RBCs in the presence of poly(I:C), with anti-HOD assessed by flow cross-matching after the final transfusion. Compilation of 4 experiments, 40 recipients total. Mean adjusted MFI was 56 after poly(I:C) HOD or HOD × hGPA F1 in the group initially not transfused compared with 44 in the group initially transfused with hGPA RBCs (P = .65). No anti-hGPA or anti-HOD was detected in either group after the initial hGPA transfusion, and no anti-hGPA was detected after the final transfusion. (C) B10.BR recipients were transfused with mHEL or control C57BL/6 RBCs and then boosted 6 weeks later with mHEL RBCs in the presence of poly(I:C), with anti-HEL assessed by ELISA 2 weeks after the first and last transfusions. Compilation of 2 experiments with 18 recipients. Mean optical density (OD) was 0.45 before and 0.33 after poly(I:C) mHEL transfusion in the group initially transfused with mHEL RBCs; OD was 0.1 before and 1.1 after poly(I:C) mHEL transfusion in the group initially transfused with C57BL/6 RBCs (P = .004 between groups after the last transfusion). (D) B10.BR recipients were transfused with mHEL or control C57BL/6 RBCs and then injected subcutaneously 6 weeks later with HEL protein emulsified in complete Freund's adjuvant (CFA), with anti-HEL assessed 2 weeks after this immunization by flow cross-matching. Compilation of 2 experiments, 21 recipients total. Adjusted MFI was 3 after HEL/CFA treatment in the group initially transfused with mHEL RBCs compared with 38 in the group initially transfused with C57BL/6 RBCs (P = .0006).

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