Version 29 (16 August 2019) Journal of

Supplemental Material

Supplemental Table 1. Boolean subset comparison across groups (Wilcoxon Rank Test Result)

1. HIV+ CCR7, CD27, and CD45RA expression Category P-value CD8 CD16+ CCR7 CD27 CD45RA vs. CD16- + + + 0.000 + + - 0.000 + - + 0.000 + - - 0.013 - + + 0.001 - + - 0.000 - - + 0.000 - - - 0.038

2. HIV+ CD57, NKG2A, and NKG2D expression Category P-value CD8 CD16+ CD57 NKG2A NKG2D vs. CD16- + + + 0.000 + + - 0.001 + - + 0.000 + - - 0.000 - + + 0.281 - + - 0.130 - - + 0.000 - - - 0.000

3.HIV+ Perforin and CD161 expression Category P-value CD8 CD16+ vs. CD16- NKG2A NKG2D + + 0.000 + - 0.000 - + 0.000 - - 0.000

4. HIV+ Transcription Factors (Eomesodermin, Helios, and T-bet) expression Category P-value CD8 CD16+ FDR of CD8 CD16+ P-value CD8 CD16+ FDR of CD8 CD16+ Eomes Helios T-bet vs. CD16- vs. CD16- vs. NK cell vs. NK cell + + + 0.000 0.001 0.019 0.0338 + + - 0.028 0.045 0.000 0.0015 + - + 0.007 0.014 0.001 0.0023 + - - 0.112 0.138 0.000 0.0015 - + + 0.001 0.002 0.000 0.0015 - + - 0.880 0.880 0.496 0.5669 - - + 0.059 0.079 0.049 0.0713 - - - 0.001 0.002 0.597 0.6368

5. HIV+ Killer Immunoglobulin (KIR) Expression Category P-value CD8+CD16+ FDR of CD8+CD16+ P-value CD8+CD16+ FDR of CD8+CD16+ P-value CD8+CD16+ FDR of CD8+CD16+ 2DL1/DS1 2DL2/3/DS2 KIR3DL1 vs. CD8+CD16- vs. CD8+CD16- vs. CD8+CD45- vs. CD8+CD45- vs. NK cell vs. NK cell + + + 0.208 0.357 0.462 0.504 0.001 0.005 + + - 0.793 0.827 0.248 0.372 0.016 0.038 + - + 0.916 0.916 0.401 0.481 0.001 0.005 + - - 0.462 0.504 0.401 0.481 0.027 0.059 - + + 0.248 0.372 0.141 0.260 0.012 0.032 - + - 0.093 0.186 0.001 0.005 0.294 0.392 - - + 0.003 0.012 0.012 0.032 0.294 0.392 - - - 0.005 0.017 0.001 0.005 0.001 0.005

6. CD8+CD16+ Killer Immunoglobulin receptor (KIR) Expression Category P-value HIV- vs. 2DL1/DS1 2DL2/3/DS2 KIR3DL1 HIV+ + + + 0.058 + + - 0.142 + - + 0.270 + - - 0.426 - + + 0.298 - + - 0.178 - - + 0.951 - - - 0.006

1 Version 29 (16 August 2019) Journal of Immunology

Supplemental Table 2. expression RT-PCR reagents Gene symbol Gene Name Gene ID Assay ID CD226 CD226 molecule 10666 Hs00170832_m1 EOMES eomesodermin 8320 Hs00172872_m1 NCR1 natural cytotoxicity triggering receptor 1 9437 Hs00183118_m1 IKZF2 IKAROS family zinc finger 2 (Helios) 22807 Hs00212361_m1 KLRF1 killer cell lectin-like receptor subfamily F, member 1 51348 Hs00212979_m1 LAIR1 leukocyte-associated immunoglobulin-like receptor 1 3903 Hs00253790_m1 ITGAM integrin, alpha M (complement component 3 receptor 3 subunit) 3684 Hs00355885_m1 KIR2DL4;LOC100287534 killer cell immunoglobulin-like receptor, two domains, long cytoplasmic tail, 4 3805 Hs00427106_m1 killer cell immunoglobulin-like receptor 2DL4-like 100287534 KIR3DL2 killer cell immunoglobulin-like receptor, three domains, long cytoplasmic tail, 2 3812 Hs00601497_gH KIR3DX1 killer cell immunoglobulin-like receptor, three domains, X1 90011 Hs00736082_g1 KIR3DL1;KIR3DS1 killer cell immunoglobulin-like receptor, three domains, long cytoplasmic tail, 1 3811 Hs00744448_s1 killer cell immunoglobulin-like receptor, three domains, short cytoplasmic tail, 1 3813 IL7R interleukin 7 receptor 3575 Hs00902334_m1 KLRG1 killer cell lectin-like receptor subfamily G, member 1 10219 Hs00929964_m1 IL2RG interleukin 2 receptor, gamma 3561 Hs00953624_m1 IKZF1 IKAROS family zinc finger 1 (Ikaros) 10320 Hs00958474_m1 IL18R1 interleukin 18 receptor 1 8809 Hs00977691_m1 IL18RAP interleukin 18 receptor accessory 8807 Hs00977695_m1 GZMA granzyme A (granzyme 1, cytotoxic T--associated serine esterase 3) 3001 Hs00989184_m1 IFNG interferon, gamma 3458 Hs00989291_m1 TLR6 toll-like receptor 6 10333 Hs01039989_s1 LAIR2 leukocyte-associated immunoglobulin-like receptor 2 3904 Hs01102514_m1 LILRB1 leukocyte immunoglobulin-like receptor, subfamily B (with TM and ITIM domains), member 1 10859 Hs01848117_s1 KIR2DL3;KIR2DL2;KIR2DS2 killer cell immunoglobulin-like receptor, two domains, long cytoplasmic tail, 3 3804 Hs03407415_gH killer cell immunoglobulin-like receptor, two domains, long cytoplasmic tail, 2 3803 killer cell immunoglobulin-like receptor, two domains, short cytoplasmic tail, 2 100132285 KIR2DS2;KIR2DL5A;KIR2DL2;KIR2DL3;KIR2DS4 killer cell immunoglobulin-like receptor, two domains, short cytoplasmic tail, 2 100132285 Hs04190776_gH killer cell immunoglobulin-like receptor, two domains, long cytoplasmic tail, 2 3803 killer cell immunoglobulin-like receptor, two domains, long cytoplasmic tail, 3 3804 killer cell immunoglobulin-like receptor, two domains, long cytoplasmic tail, 5A 57292 killer cell immunoglobulin-like receptor, two domains, short cytoplasmic tail, 4 3809 KIR2DS2;KIR2DS1;KIR2DS3;KIR2DL1;KIR2DS4 killer cell immunoglobulin-like receptor, two domains, short cytoplasmic tail, 2 100132285 Hs04190778_mH killer cell immunoglobulin-like receptor, two domains, long cytoplasmic tail, 1 3802 killer cell immunoglobulin-like receptor, two domains, short cytoplasmic tail, 1 3806 killer cell immunoglobulin-like receptor, two domains, short cytoplasmic tail, 3 3808 killer cell immunoglobulin-like receptor, two domains, short cytoplasmic tail, 4 3809 KIR2DS3;KIR2DS5 killer cell immunoglobulin-like receptor, two domains, short cytoplasmic tail, 3 3808 Hs04190781_mH killer cell immunoglobulin-like receptor, two domains, short cytoplasmic tail, 5 3810 KIR2DS3;KIR2DL2;KIR2DS2;KIR2DS5;KIR2DL3 killer cell immunoglobulin-like receptor, two domains, short cytoplasmic tail, 3 3808 Hs04190782_gH killer cell immunoglobulin-like receptor, two domains, long cytoplasmic tail, 2 3803 killer cell immunoglobulin-like receptor, two domains, long cytoplasmic tail, 3 3804 killer cell immunoglobulin-like receptor, two domains, short cytoplasmic tail, 2 100132285 killer cell immunoglobulin-like receptor, two domains, short cytoplasmic tail, 5 3810 KLRC2;KLRC3 killer cell lectin-like receptor subfamily C, member 2 3822 Hs04192492_gH killer cell lectin-like receptor subfamily C, member 3 3823 TNFSF10 tumor necrosis factor (ligand) superfamily, member 10 8743 Hs00921974_m1 GZMK granzyme K (granzyme 3; tryptase II) 3003 Hs00157878_m1 IL12RB1 interleukin 12 receptor, beta 1 3594 Hs00538167_m1 IL12RB2 interleukin 12 receptor, beta 2 3595 Hs01548202_m1 TLR8 toll-like receptor 8 51311 Hs00152972_m1 LAMP1 lysosomal-associated 1 3916 Hs00174766_m1 NCR2 natural cytotoxicity triggering receptor 2 9436 Hs00183113_m1 ICOS inducible T-cell co-stimulator 29851 Hs00359999_m1 PDCD1 programmed cell death 1 5133 Hs01550088_m1 TLR1 toll-like receptor 1 7096 Hs00413978_m1 CD33 CD33 molecule 945 Hs01076281_m1 CXCR1 chemokine (C-X-C motif) receptor 1 3577 Hs00174146_m1 TLR3 toll-like receptor 3 7098 Hs01551078_m1 IL10 interleukin 10 3586 Hs00961622_m1 TBX21 T-box 21 30009 Hs00203436_m1 CXCR3 chemokine (C-X-C motif) receptor 3 2833 Hs00171041_m1 KLRK1;KLRC4-KLRK1 killer cell lectin-like receptor subfamily K, member 1 22914 Hs00183683_m1 KLRC4-KLRK1 readthrough 100528032 IRF7 interferon regulatory factor 7 3665 Hs01014809_g1 KLRC1 killer cell lectin-like receptor subfamily C, member 1 3821 Hs00970273_g1 CCL3 chemokine (C-C motif) ligand 3 6348 Hs00234142_m1 LILRB2;LILRB1 leukocyte immunoglobulin-like receptor, subfamily B (with TM and ITIM domains), member 2 10288 Hs00601427_g1 leukocyte immunoglobulin-like receptor, subfamily B (with TM and ITIM domains), member 1 10859 CCR7 chemokine (C-C motif) receptor 7 1236 Hs01013469_m1 MKI67 identified by monoclonal antibody Ki-67 4288 Hs01032443_m1 B3GAT1 beta-1,3-glucuronyltransferase 1 ( P) 27087 Hs00218629_m1 BIRC3 baculoviral IAP repeat containing 3 330 Hs00154109_m1 CCL2 chemokine (C-C motif) ligand 2 6347 Hs00234140_m1 CCL4 chemokine (C-C motif) ligand 4 6351 Hs99999148_m1 CCL5 chemokine (C-C motif) ligand 5 6352 Hs00174575_m1 BCL2 B-cell CLL/lymphoma 2 596 Hs99999018_m1 CD244 CD244 molecule, receptor 2B4 51744 Hs00900271_m1 CD38 CD38 molecule 952 Hs01120071_m1 CD69 CD69 molecule 969 Hs00934033_m1 CLEC2B C-type lectin domain family 2, member B 9976 Hs00192860_m1 CTSB cathepsin B 1508 Hs00947433_m1 CXCR4 chemokine (C-X-C motif) receptor 4 7852 Hs00237052_m1 CXCR6 chemokine (C-X-C motif) receptor 6 10663 Hs00174843_m1 DPP4 dipeptidyl-peptidase 4 1803 Hs00175210_m1 ENTPD1 ectonucleoside triphosphate diphosphohydrolase 1 953 Hs00969559_m1 FASLG (TNF superfamily, member 6) 356 Hs00181225_m1 CD160 CD160 molecule 11126 Hs00199894_m1 GAPDH glyceraldehyde-3-phosphate dehydrogenase 2597 Hs99999905_m1 GNLY granulysin 10578 Hs00246266_m1 GZMB granzyme B (granzyme 2, cytotoxic T-lymphocyte-associated serine esterase 1) 3002 Hs01554355_m1 HLADRA major histocompatibility complex, class II, DR alpha 3122 Hs00219575_m1 IL16 interleukin 16 3603 Hs00189606_m1 IL21R interleukin 21 receptor 50615 Hs00222310_m1 IL2Ra interleukin 2 receptor, alpha 3559 Hs00907777_m1 IL6ST interleukin 6 signal transducer (gp130, oncostatin M receptor) 3572 Hs00174360_m1 KLRD1 killer cell lectin-like receptor subfamily D, member 1 3824 Hs00233844_m1 LGALS9 lectin, galactoside-binding, soluble, 9 3965 Hs00371321_m1 KLRF1 killer cell lectin-like receptor subfamily F, member 1 51348 Hs00212979_m1 NCR3 natural cytotoxicity triggering receptor 3 259197 Hs00394809_m1 POLR2A polymerase (RNA) II (DNA directed) polypeptide A, 220kDa 5430 Hs00172187_m1 PRDM1 PR domain containing 1, with ZNF domain 639 Hs00153357_m1 PRF1 perforin 1 (pore forming protein) 5551 Hs00169473_m1 PSMG2 proteasome (prosome, macropain) assembly chaperone 2 56984 Hs00220315_m1 SLAMF5 CD84 molecule 8832 Hs01547121_m1 SLAMF7 SLAM family member 7 57823 Hs00221793_m1 SMAD4 SMAD family member 4 4089 Hs00232068_m1 SPATA2 spermatogenesis associated 2 9825 Hs00195835_m1 TGFB1 transforming growth factor, beta 1 7040 Hs00998133_m1 NKG7 natural killer cell group 7 sequence 4818 Hs01120688_g1 TGFBR1 transforming growth factor, beta receptor 1 7046 Hs00610318_m1 TIA1 TIA1 cytotoxic granule-associated RNA binding protein 7072 Hs00254561_m1 TNF tumor necrosis factor 7124 Hs00174128_m1 TNFAIP3 tumor necrosis factor, alpha-induced protein 3 7128 Hs00234713_m1 TNFRSF9 tumor necrosis factor receptor superfamily, member 9 3604 Hs00155512_m1 TNFSF13B tumor necrosis factor (ligand) superfamily, member 13b 10673 Hs00198106_m1

2 Version 29 (16 August 2019) Journal of Immunology

A. Initial CD8 CD16+ survey Donor 1 Donor 2

60.5 24.2 250K 52.5 14.4 250K 5 250K 105 105 10 105 42.4

200K 200K 200K 75.9 4 4 4 A 4

A 10 A 10 10 10

150K 150K 150K 37.2

5.18 C- C-

C- 3 1b FITC 03 103 10 103 100K 100K 100K

2 SS 59.4 2 2 SS 2 SS 10 10 10 10 50K 50K 92.8 50K 44.5 0 0 0 0 CRa

CD38 APC 10.5 4.78 CD38 APC 32.1 0.974 0 0 0 T CD8 PE-Cy7 2 3 4 5 2 3 4 5 0 50K 100K 150K 200K 250K 0 102 103 104 105 0 102 103 104 105 0 10 10 10 10 0 10 10 10 10 0 102 103 104 105 0 102 103 104 105 FSC-A Aqua CD3 PPCy5.5 CD16 PB PD-1 PE CD16 PB PD-1 PE B. Homing Panel

250K 250K 250K 43.9 105 5 250K 10 250K 250K 250K

200K 200K 200K 200K 97.6 4 200K A 10 200K 10 4 5.5 200K A A

150K 150H K 150K 150K A 150K 150K 150K 99.7 C- 103 10 3 C- 100K 100K 100K C- 61.1 100K

C- 100K 100K 2 100K 2

SC- 10

SS 10 50K 50K 50K 8 PE-Cy7 50K SS SS 50K F 0 50K 50K 0 0 0 0 SS 42.8 0 36.6 6.12 0 50K 100K 150K 200K 250K 0 50K 100K 150K 200K 250K 0 102 103 104 105 0 102 103 104 105 43.9 0 CD 0 102 103 104 105 0 0 2 3 4 5 2 3 4 5 2 3 4 5 0 10 10 10 10 0 102 103 104 105 0 10 10 10 10 0 10 10 10 10 FSC-A FSC-A Aqua CD3 AH7 CD16 PE-Cy5 CD8 PE-Cy7 250K 250K 250K 250K

A 200K 200K 200K 200K

150K

C- 150K 150K 150K

100K 100K 100K 100K SS

50K 50K 50K 50K 74.8 54.1 1.07 57.8 0 0 0 0 2 3 4 5 2 3 4 5 2 3 4 5 0 10 10 10 10 0 10 10 10 10 0 102 103 104 105 0 10 10 10 10 CCR5 BV421 CXCR3 FITC PD-1 AF647 TRAIL C. NK cell receptor, perforin, and IL-7 receptor Panel

250K 250K 250K 34.3 250K 250K 250K 105 105 250K 3.39

200K 200K 200K 200K 200K 200K 200K 104 104 A A A A A A H 150K 150K 150K 150K 150K 150K 150K 97.4

3 6 PB 3 C- C- C- C- C- 69.4 C- 10 10 100K 100K 100K 100K 100K 100K 100K SC- 2 2 50K 50K 50K 10 10 SS SS SS SS SS SS F 50K 50K 50K 50K 0 0 84 CD1 20 0 0 0 0 13.8 0.0863 3.06 0 0 0 2 3 4 5 2 3 4 5 2 3 4 5 2 3 4 5 2 3 4 5 0 50K 100K 150K 200K 250K 0 50K 100K 150K 200K 250K 0 10 10 10 10 CD8 PE-Cy7 0 10 10 10 10 0 10 10 10 10 0 10 10 10 10 2 3 4 5 0 102 103 104 105 0 10 10 10 10 0 10 10 10 10 FSC-A FSC-A Aqua CD3 AH7 CD8 PE-Cy7 IL-7R AF647 Perforin FITC NKp46 PE CD161 PPCy5.5

D. NK cell receptor

250K 250K 250K 5 5 10 44.4 10 250K 47.3 3.11 105 200K 200K 200K 104 10 4 5.5 200K 104 A A A H 96.4 150K 150K 150K 88.8 59.7 3 3 150K 10 10 6 PB 3 C- C- C- 10 100K 100K 100K 7 FITC

SC- 2 100K 102 10

8 PE-Cy7 2 SS SS SS F 10 50K 50K 50K 0

CD1 0 0 50K CD5 CD 0 0 0 84.1 46.9 2.68 0 0 50K 100K 150K 200K 250K 0 50K 100K 150K 200K 250K 0 10 3 10 4 105 0 102 103 104 105 0 10 2 10 3 10 4 10 5 2 3 4 5 0 102 103 104 105 0 10 10 10 10 FSC-A FSC-A Aqua CD3-AH7 CD8 PE-Cy7 NKG2A APC NKG2D PerCP

E. Differentiation/Maturation Panel

250K 250K 250K 250K 5 5 10 45.9 10 5 24.7 7.87 2.14 10 200K 200K 200K 200K 4 4 10 10 104 A A A 150K H 150K 150K 150K 3 103 103 10

C- 62.7 94.1 C- C- C- 93.4 100K 100K 100K 100K 16 PB

102

8 PE-Cy7 2 2 SS SS SS 50K FS 50K 50K 10 10 45RA APC 0 50K

0 CD 0 54.5 0 0 0 CD 62.2 5.25 0 0 50K 100K 150K 200K 250K 0 50K 100K 150K 200K 250K 0 102 103 104 105 0 102 103 104 105 0 102 103 104 105 CD 2 3 4 5 0 102 103 104 105 0 10 10 10 10 FSC-A FSC-A Aqua CD3 AH7 CD8 PE-Cy7 CCR7 FITC CD27 PerCP

F. Transcription Factor Panel

250K 250K 5 48.4 5 4.6 250K 10 10 250K 250K

200K 200K 4 200K 4 10 200K 200K 10 A A A A 150K 150K 3 150K 150K 150K 98.3 10

C- 3 C- C- C- 68.4 10 100K 100K APC-C7 100K 100K 100K 10 2 FSC-H SS 2 SS SS SS 50K 50K 10 0 50K 50K 50K 0

69.7 80.3 70.5 0 0 CD8 PP-Cy5.5 CD16 0 0 0 0 50K 100K 150K 200K 250K 0 50K 100K 150K 200K 250K 0 102 10 3 104 10 5 0 102 10 3 104 10 5 2 3 4 5 2 3 4 5 0 102 103 104 105 0 10 10 10 10 0 10 10 10 10 FSC-A FSC-A CD3 AmCyan CD8 PP-Cy5.5 Eomes PE T-bet FITC Helios eF450

G. Killer Immunoglobulin like Receptor Panel

250K 250K 250K 250K 105 5 10.9 10 250K 250K 22.7 250K 250K

200K 200K 200K 200K 87.5 4 97.2 10 4 200K 10 200K 200K 200K

3 A A

150K 150K 150K 150K A

92.4 10 A

A 150K H A H 150K 150K 150K 46.8 21.3 103 64.1 C- 100K 100K 100K 100K 2 C- C-

10 C- 100K C- C- 100K 100K 100K 0 2 35.9 10 57.7

SC- 41.9 50K SC- 50K 50K 50K 50K 50K 50K SS

SS 50K SS

0 SS SS SS F F 0 0 0 0 0 0 0 2 3 4 5 2 3 4 5 2 3 4 5 2 3 4 5 0 2 3 4 5 2 3 4 5 2 3 4 5 0 50K 100K 150K 200K 250K 0 50K 100K 150K 200K 250K 0 10 10 10 10 0 10 10 10 10 0 10 10 10 10 CD57 APC 0 10 10 10 10 0 10 10 10 10 0 10 10 10 10 0 102 103 104 105 0 10 10 10 10 FSC-A FSC-A Aqua CD14/19 CD8 APC-H7 CD3 CD45 BV785 CD16 PB KIR3DL1 DL2/DS2/DL3 2DL1/S1 PE-Cy5 PE-CF594 A700 PE PP-Cy5.5 7 250K 250K 250K 250K 5 250K 250K 10 5 250K 16.2 10 87.5 200K 200K 200K 200K 4 200K 200K 97.2 10 200K 104 A A A A H A H 27.3 150K 150K 150K 150K 150K 3 150K 26.8 92.4 10 150K 21.3 3

C- 10 C- C- C- C- 100K 100K 100K 100K 100K 2 100K 10 100K SC- SC- 0 64.3 2 3.41 10 50K 50K SS SS SS SS SS F

50K F 50K 50K 50K 50K 0 0 0 0 0 0 0 0 2 3 4 5 2 3 4 5 2 3 4 5 2 3 4 5 2 3 4 5 0 102 103 104 105 0 10 10 10 10 0 50K 100K 150K 200K 250K 0 50K 100K 150K 200K 250K 0 10 10 10 10 0 10 10 10 10 0 10 10 10 10 0 10 10 10 10 0 102 103 104 105 CD8 APC-H7 FSC-A FSC-A Aqua CD14/19 CD3 CD56 PE-CY CD16 PB KIR3DL1 DL2/DS2/DL3 2DL1/S1 PE-Cy5 PE-CF594 A700 PE PP-Cy5.5

Supplemental Figure 1. Flow cytometry panels. Successive gating strategy used to characterize subpopulations of FcγRIIIa-expressing cell subsets using 7 flow cytometry panels. Gating strategies began by first identifying small through forward and side scatter profiles, single cells, and dead cell exclusion using an amine reactive dye. An initial survey identified an expanded FcγRIIIa+CD3+CD8+ cell subset within HIV+ donors, with differential PD-1 expression, as represented here (A). Five additional flow cytometry panels were designed to further characterize this subset. Homing markers were characterized in CD16 positive versus negative CD8 T cells, with CCR5, CXCR3, PD-1 and TRAIL gated individually within each CD16 expression subset (B). Additionally, CD16+ CD8 T cells were screened for surface and intracellular NK cell, innate-like markers through individual gating of IL-7R, Perforin, NKp46, and CD161 (C), single gating on NKG2D, and all combinations of NKG2A and CD57, individually or together (D). CD16+ CD8 T cells were also characterized for their differentiation and maturation markers through single gating on CD27, or all combinations of CD45RA and CCR7 (E). Transcription factors T-bet, Eomes, and Helios were characterized alone or in combination within CD16+ CD8 T cells, CD16- CD8 T cells, as well as CD56dim FcγRIIIa+ NK cells (F). Finally, surface KIR3DL1, KIRDL2/DS2/DL3, and KIR2DL1/S1 expression was measured in FcRγIIIa+ CD8 T cells and NK cells. CD8 T cells were gated on as CD3+CD8+ T cells, enriched for late-stage effector phenotype by gating on CD57+CD45+ cells, and further gated as CD16+. NK cells were identified by their CD3 negativity, followed by gating on CD56dimCD16+ cells. All KIR gates were drawn as individual gates (G).

3 Version 29 (16 August 2019) Journal of Immunology

Color Key

0123 4

IRF7 SMAD4 CTSB CD226 SLAMF7 FASLG TNFSF10 CD38 EOMES LGALS9 BIRC3 IL16 SLAMF5 IL21R IL6ST TNFRSF9 KIR3DL1.KIR3DS1 KLRC1 KIR2DS2.KIR2DL5A B3GAT1..CD57. KLRF1 IFNG TGFB1 CCL4 PSMG2 CLEC2B HLADRA GZMB PRF1 KLRD1 POLR2A PRDM1 KLRG1 CXCR4 IKZF1 IL2RG TGFBR1 CD244 CCL3 ITGAM IKZF2 KLRC2L.KLRC3 KIR2DS2.KIR2DS1. CD69 GZMA KLRK1.KLRC4.KLRK1 NKG7 CCL5 IL12RB1 TLR1 TNFSF13B SPATA2 CD160 TLR3 TIA1 TNF IL18R1 TLR6 BCL2 LAIR1 LAIR2 NCR1 LILRB1 IL18RAP DPP4 CXCR3 PDCD1 ICOS GZMK CCR7 IL7R

NK cells

CD45RA-CD57- CD8 T cells CD45RA+CD57+ CD45RA+CD57+ CD16+ CD8 T cellsCD16- CD8 T cells Supplemental Figure 2. Hierarchecal clustering of in sorted cell subsets. Two-way hierarchical clustering of gene expression in sorted subsets of CD8 T cells; CD45RA-CD57-,

CD45RA+CD57+FcγRIIIA-, CD45RA+CD57+FcγRIIIA+, as well as CD56dim FcγRIIIA+ NK cells. Data is normalized 2^DCt values.

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