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. 2018 Jan 24;9:24. doi: 10.3389/fimmu.2018.00024

Figure 6.

Figure 6

CD69 on natural killer (NK) cells is induced by a heat-labile soluble factor derived from infected monocytes. Human peripheral blood mononuclear cells (PBMCs) [(A) (top panel), (B–E)] or purified NKp46+CD3− NK cells [(A) (bottom panel)] were stimulated with supernatants (SN) of various cell-Leishmania-cocultures for 20 h. CD69 expression of NKp46+CD3− NK cells was determined by flow cytometry. Unless otherwise indicated (C), SNs were added (A,B,D,E) at a final concentration of 60% (vol/vol) and used untreated (A–D), sterile filtered (E), or heat-inactivated (E). [(A) top panel] Mean ± SEM of 59/59/50/49 (SN PBMC), 4/3/4/3 (SN NK), 4/4/4/2 (SN NK + Mo), 4/4/4/4 (SN Mo), or 8/8/6 donors (SN Leishmania only). [(A) bottom panel] Mean ± SEM of 7/7/6/5 (SN PBMC), 3/3/2/3 (SN NK), or 6/6/6/6 (SN Mo) donors for the four stimulations. (B) Mean ± SEM of 24/23/20/15 donors for the four stimulations. (C) Mean ± SEM of 11/4/3/4 (Leishmania spp.), 9/9/7/8 (20% SN), 10/11/7/7 (40% SN), or 9/9/7/9 (80% SN) donors for the four stimulations. (D) Mean ± SEM of seven (SN viable), seven (SN pfa-fixed), and four [SN freeze–thaw lysate (ft-lysate)] donors. (E) Mean ± SEM of 10/10/8/10 (SN untreated), 9/9/7/9 (SN filtered), and 9/9/6/8 (SN heat-inactivated) donors for the four stimulations. (F) PBMCs or PBMCs depleted of monocytes were cocultured with Leishmania spp. promastigotes (multiplicity of infection 10) for 20 h. The concentration of interleukin (IL)-12p40 in culture SNs was determined by ELISA (values below detection limit are marked by triangles). Mean ± SEM of 6/6/5/5 (PBMC) and 6/5/5/6 (PBMC w/o monocytes) donors for the four stimulations. *,#p < 0.05, **p < 0.01, and ***p < 0.001 two-tailed Mann–Whitney U test.