Supplementary MaterialsSupplemental data jci-130-129965-s079

Supplementary MaterialsSupplemental data jci-130-129965-s079. during acute GVHD. Host T cells were present in all skin and colon acute GVHD specimens studied, yet were largely absent in blood. We observed acute skin GVHD in the presence of 100% host T cells. Analysis demonstrated that a subset of sponsor T cells in peripheral cells were proliferating (Ki67+) and generating the proinflammatory cytokines IFN- and IL-17 in situ. Comparatively, the majority of antigen-presenting cells (APCs) in cells in acute GVHD were donor derived, and donor-derived APCs were observed directly adjacent to sponsor T cells. A humanized mouse ABT-639 model shown that sponsor skin-resident T cells could be triggered by donor monocytes to generate a GVHD-like dermatitis. Therefore, sponsor tissue-resident T cells may play a previously unappreciated pathogenic part in acute GVHD. = 3. To determine whether T cells in pores and skin after HSCT were sponsor or donor derived, we performed high-throughput sequencing of the gene to identify clonal populations of memory space T cells (18). Unique T cell clones were recognized by their CDR3 sequences. In all 3 patients, the majority of T cell clones in pores and skin after HSCT were identical to sponsor pores and skin T cell clones before HSCT (Number 1C). The 20 most abundant T cell clones in sponsor pores and skin after HSCT and the similar frequency of those clones in sponsor pores and skin before HSCT or donor Rabbit Polyclonal to SH3GLB2 infusion product, respectively, are demonstrated in Number 1D. Correlation between rate of recurrence of T cell clones in sponsor pores and skin before HSCT and pores and skin after HSCT was high (patient 1-0.6464, patient 2-0.8740, patient 3-0.5867) (Supplemental Number 1A). In contrast, higher rate of recurrence of clones in donor cells did not correlate with increased frequency in pores and skin after HSCT (individual 1-0.0041, patient 2-0.0142, patient 3-0.0012) (Supplemental Number 1, A and B). Of the top 100 most frequent clones in sponsor pores and skin after HSCT in each patient, only 0, 1, and 16, respectively, were donor derived (Supplemental Number 1A). Thus, T cell clonality data paralleled the results ABT-639 from FISH-IF and STR analysis. Host T cells are present in pores and skin during acute GVHD. Given that pores and skin T cells survived HSCT through 30 6 days, a peak time point for onset of acute GVHD (19), and that the main cells affected by GVHD are the same cells containing large populations of tissue-resident T cells, we hypothesized that sponsor T cells would be present in pores and skin and gut during acute GVHD. Supplemental Table 2 details retrospective patient medical data. Chemoimmunotherapeutics received by each patient before transplant are detailed in Supplemental Table 3 and Supplemental Table 4. Pores and skin biopsies from 26 male individuals with acute GVHD who received female donor transplants were labeled via FISH-IF to determine the quantity and percentage of sponsor ABT-639 and donor T cells (Number 2, A and B). Host T cells were observed in pores and skin during acute GVHD of all patients studied, regardless of the conditioning routine (myeloablative, median 39%, range 4%C100%) (nonmyeloablative, median 58%, range 3%C78%) (= 0.24, Mann-Whitney test, 2-tailed) (Figure 2B). Host T cells were observed throughout the pores and skin, including within the epidermis and at the dermal-epidermal junction, the primary sites of damage in acute pores and skin GVHD (Supplemental Number 2). Open in a separate window ABT-639 Number 2 Host T cells are present in pores and skin during acute GVHD.(A) Example of FISH-IF from FFPE pores and skin during acute GVHD. X chromosome, reddish; Y chromosome, green; CD3, yellow; DAPI nuclear stain, blue. Solid level pub: 50 m; dotted level pub: 10 m. Good dotted line shows dermal-epidermal junction. Red arrow points to donor T cell; green arrow points to sponsor T cell. (B) Percentage of sponsor T cell chimerism in pores and skin during acute GVHD, determined by FISH-IF. Solid reddish squares, all myeloablative-conditioned individuals; open reddish squares, breakdown of.