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. Author manuscript; available in PMC: 2019 Aug 1.
Published in final edited form as: Biol Blood Marrow Transplant. 2018 Mar 27;24(8):1581–1589. doi: 10.1016/j.bbmt.2018.03.019

Figure 2. Donor Activated NK Cells Detected by Flow Cytometry Chimerism and Elevated CD69 Expression on NK cells in the Blood of 4 of 8 Patients Evaluated.

Figure 2

All results in figures A-D are from plots gated on CD56+CD3- lymphocytes (NK cells).

For 6 patients it was possible to evaluate chimerism in blood and bone marrow samples using donor-specific anti-HLA mAbs. In (A) and (B), rectangle gates include cells with the donor HLA type; numbers shown are the frequency of donor cells in the total NK cell population. (A) Circulating donor cells were detected by HLA antibodies in day 7 blood of 5 of the 6 patients with an identifying HLA antibody. (B) The 6th patient, patient 08-004, had insufficient cells to perform flow cytometry chimerism analysis on day 7; flow chimerism analysis of subsequent samples detected donor cells by anti-HLA antibody as late as day 56 in the blood and bone marrow.

In 8 patients, total NK cells in the blood (both donor and recipient) were evaluated for elevated expression of CD69. In (C) and (D), numbers shown are the percent of NK cells. (C) Representative plots from subject 08-006 (Pt 9) showing expression of CD69 vs NKG2A on total NK cells in the blood on days 7, 14, 28 and 56 post-infusion. (D) Fraction of total NK cells expressing CD69 in the blood of the 4 patients with elevated CD69 expression.