Figure 5.
Anti-HOD mAb IgG2c leads to enhanced T-cell proliferation and increases RBC consumption by DCs. To evaluate RBC consumption, B6 mice were passively immunized followed by a DiO+ HOD RBC transfusion. Spleens were harvested 18 to 24 hours posttransfusion and leukocytes were stained to delineate cell subsets. The percent erythrophagocytosis of total Thy1.2−TER119− leukocytes was determined (A) and the DiO MFI of individual APC subsets was calculated (B). To assess whether passive immunization modulated T-cell responses, B6 mice were adoptively transferred with 1 × 105 purified CD4+ OTII T cells labeled with CellTrace-far-red (FR). The next day, recipient mice were passively immunized with PBS, nonspecific anti-K mAb IgG2c, or anti-HOD mAb IgG2c followed by a HOD RBC transfusion. CellTrace-FR dilution (C) and absolute number (D) was determined for CD4+Va2+Vb5+Thy1.1+ OTII T cells 3 days posttransfusion. Each experiment was performed 3 times with 3 to 5 mice per group. Representative data are shown. Data were analyzed with a 1-way ANOVA with Dunnett’s multiple comparisons test to the control PBS group. *P ≤ .05, **P ≤ .01, ****P ≤ .0001.

Anti-HOD mAb IgG2c leads to enhanced T-cell proliferation and increases RBC consumption by DCs. To evaluate RBC consumption, B6 mice were passively immunized followed by a DiO+ HOD RBC transfusion. Spleens were harvested 18 to 24 hours posttransfusion and leukocytes were stained to delineate cell subsets. The percent erythrophagocytosis of total Thy1.2TER119 leukocytes was determined (A) and the DiO MFI of individual APC subsets was calculated (B). To assess whether passive immunization modulated T-cell responses, B6 mice were adoptively transferred with 1 × 105 purified CD4+ OTII T cells labeled with CellTrace-far-red (FR). The next day, recipient mice were passively immunized with PBS, nonspecific anti-K mAb IgG2c, or anti-HOD mAb IgG2c followed by a HOD RBC transfusion. CellTrace-FR dilution (C) and absolute number (D) was determined for CD4+Va2+Vb5+Thy1.1+ OTII T cells 3 days posttransfusion. Each experiment was performed 3 times with 3 to 5 mice per group. Representative data are shown. Data were analyzed with a 1-way ANOVA with Dunnett’s multiple comparisons test to the control PBS group. *P ≤ .05, **P ≤ .01, ****P ≤ .0001.

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