Nuclear Antigens and Auto/Alloantibody Responses: Friend or Foe in Transplant Immunology
Figure 3
Alloantibody response during the rejection phase after OLT. The alloantibody response was measured by flow cytometry on a single cell suspension of DA rat splenocytes. Briefly, 50 μL aliquots containing splenocytes was incubated with 50 μL of diluted naive or post-OLT sera (1 : 16, 1 : 64, 1 : 256) for 45 min at 4°C. The washed cells were reacted with 50 μL of a mixture of FITC-conjugated goat antibody specific for the Fc portion of rat IgG ( dilution) (Jackson ImmunoResearch Laboratories, West Grove, PA, USA) in PBS containing 1% BSA and 0.02% NaN3. After staining, the cells were washed, fixed, and analyzed using an LSRII flow cytometer (BD Biosciences, San Jose, CA, USA). Histograms (representative of three individuals) show the percentage of DA splenocytes recognized by alloantibody (IgG) in the post-OLT sera at day 7 after OLT.
