Figure 1.

NK cell repertoires in peripheral blood of controls and patients. (A) Analysis of KIR2D expression in peripheral blood NK and T cells. Gates to identify total lymphocyte (a), as well as CD3⁺ (b), CD4⁺ and CD8⁺ (c), CD16⁺CD56⁺ (d), KIR2DS1⁺, L1⁺ and L1⁺S1⁺ (e), and KIR2DL2/S2⁺ and L3⁺ (f) subsets were set in appropriate dotplots.³⁵ These gates were logically combined to assigned KIR2D expression to CD3⁺CD4⁺, CD3⁺CD8⁺ and CD3⁻CD16⁺CD56⁺ (NK) lymphocytes in a representative patient with the KIR2DL1⁺S1⁺L2/S2⁺L3⁺ genotype. (B) Special analyses were implemented for patients with KIR2DL3 alleles that were non-reactive with anti-KIR2DL3 antibody clone 180701 (presumably KIR2DL3*005 or *015 alleles).³⁶ As described, KIR2DL3*005 was cross-reactive with anti-KIR2DL1/S1 antibody clone EB6 but not with KIR2DL1 antibody clone 143211. In these patients, the possibility of three genetic backgrounds required cytometric analysis to be adapted: (1) for KIR2DL2⁻ individuals (a, b and c; n = 13) all cells stained with CD158b/j (clone GL183) were assigned as L3⁺ (yellow and orange), disregarding alleged-alleles *005 and/or *015 were homozygous (a, n = 3 ), or heterozygous with others KIR2DL3 alleles (orange) which are reactive with the CD158b antibody (b, n = 5 and c, n = 5); (2) for KIR2DL2⁺ individuals (d and e; n = 6) cells co-reacting with anti-CD158b/j and CD158a/h antibodies were assigned as L3⁺ (yellow), and cells that were non-reactive with CD158a/h antibody as L2⁺ (dark blue); (3) for KIR2DS1⁺/KIR2DL3*005 individuals (c and e; n = 6 ), S1⁺ cells were considered only as those that reacted with CD158a/h but not with CD158a nor CD158bj antibodies (violet and red, in c and e, respectively). (C) NK cell repertoires in peripheral blood of controls and patients. Left, no differences in the number (cells/μL) of total NK cells (CD3⁻CD16⁺CD56⁺) or NK cells expressing KIR2D receptors (KIR2D⁺) or not (KIR2D⁻) between controls and MGUS or myeloma (SMM+MM) patients were detected by using CD158a/h⁺CD158bj antibodies. Right, distribution of peripheral blood NK cell subsets (percent of total NK cells) expressing KIR2DL1, and/or KIR2DL2/S2 and/or KIR2DL3, in total patients (MGUS+SMM+MM) with different KIR2D genotypes. KIR2DL1⁻L2⁺L3⁻ showed only KIR2DL⁻ or KIR2DL2⁺ clones.