Regulatory Immunotherapy in Bone Marrow Transplantation

Figure 3

Donor CD4⁺CD25⁺ T cells, which expand in vivo after a nonmyeloablative HSCT conditioning regimen, secrete IL-10 but no IL-4 or IL-5. (a) Representative analysis of in vivo expanded donor H-2K^b+CD4⁺CD25⁺ T cells sorted from recipient mouse spleen at day 6 after allogeneic bone marrow transplantation. (b) Cytokine profile of sorted CD4⁺CD25⁺ T cells. H-2K^b+CD4⁺CD25⁺ and H-2K^b+CD4⁺CD25⁻ splenocytes were sorted from spleens of wild-type C57BL/6 mice (untreated, UNT) or from wild-type BALB/c hosts conditioned with TLI/ATS, transplanted with bone marrow and spleen cells from a wild-type C57BL/6 donor, and harvested on day 6 after transplant. A total of 4 × 10⁴ cells/well were incubated in triplicate wells for 72 hrs with or without 10 ng/mL myristate acetate (PMA) + 2 𝜇M ionomycin. Supernatants were analyzed by 22-plex Milliplex Map mouse cytokine/chemokine kit (Millipore, Mass, USA). Data represent means ± SEM of triplicate wells in 3 experiments, 𝑛=4-6 mice per experiment (TLI/ATS group) and 𝑛= 5–7 mice per experiment (Untreated group). TLI/ATS: total lymphoid irradiation/anti- thymocyte setum.

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