T-regulatory cell inflammation is potently increased by lymphatic fluid stasis and lymphedema. A.

Flow cytometry analysis for T-regulatory (T-reg) cells in single cell suspensions prepared from upper extremity soft tissues 1.5 cm distal to the axillary wound in animals treated with axillary lymph node dissection (ALND) or axillary incision without lymphadenectomy (sham; n = 5–7/group) 3 or 6 weeks after surgery. B. Flow cytometry analysis for T-regulatory (T-reg) cells in single cell suspensions prepared from tail tissue 2 cm distal to the wound of lymphedema or control mice (n = 5–7/group) 6 weeks after surgery. C. Flow cytometry of splenic single cell suspensions for CD4+/CD25- cells after treatment with control or CD25 neutralizing antibodies. Note no significant decrease in the overall number of CD4+ cells. Representative dot plot is shown to the right. D. Flow cytometry of splenic single cell suspensions for CD4+/CD25+/Foxp3+ (T-regs) cells after treatment with control or CD25 neutralizing antibodies. Note no significant decrease in the overall number of CD4+ cells. Representative dot plot is shown to the right.