Supplementary Table S 1 Hscores of ICAM1 VCAM1

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Supplementary Table S 1. H-scores of ICAM-1, VCAM-1 and IP-10 expression in tumors of

Supplementary Table S 1. H-scores of ICAM-1, VCAM-1 and IP-10 expression in tumors of melanoma patients before and after ipilimumab or Ipi-Bev treatment. Treatment Patient P 1 P 4 P 9 Ipi-Bev P 20 P 27 P 28 P 31 P 2 Ipilimumab P 3 P 4 P 5 P 6 Tumor Pre Post Pre Post Pre Post Post ICAM-1 Cancer EC 140 0 120 90 270 210 180 0 270 20 210 240 180 140 0 300 210 160 60 140 150 180 60 300 180 260 0 70 20 160 100 180 80 80 40 0 0 110 0 50 0 VCAM-1 Cancer EC 0 0 0 80 90 0 0 20 0 0 0 0 90 0 0 0 0 IP-10 Cancer 180 30 120 100 200 80 100 50 0 0 100 300 0 200 90 200 200 N/A 240 EC 0 0 0 0 0 0 H-scores were calculated by multiplying the percentage of positive cells with their intensity (1, 2 or 3). Cases with concomitant upregulation of ICAM-1 and VCAM-1 in EC and IP-10 in tumor cells were highlighted.

Supplementary Table S 2. CXCR 3 expression on circulating CD 4 + and CD

Supplementary Table S 2. CXCR 3 expression on circulating CD 4 + and CD 8+ T cells before and after Ipi-Bev treatment. Patient Serum P 12 (CR) P 8 (PR) P 13 (PR) P 27 (PR) P 37 (SD) PBI-2 (SD) Pre Post Post/Pre Pre Post/Pre CD 4+CXCR 3+/CD 4+ (%) 2. 7 63. 1 23. 2 43. 0 39. 2 0. 9 33. 3 43. 7 1. 3 39. 7 42. 0 1. 1 53. 9 52. 5 1. 0 36. 0 43. 3 1. 2 CD 8+CXCR 3+/CD 8+ (%) 10. 1 91. 6 9. 1 74. 2 65. 2 0. 9 71. 2 61. 1 0. 9 72. 1 71. 1 1. 0 67. 2 65. 9 1. 0 97. 0 85. 6 0. 9 Patient Serum P 10 (PD) P 14 (PD) PMGH 2 (PD) PBI-7 (PD) PBI-13 (PD) Pre Post Post/Pre Pre Post/Pre CD 4+CXCR 3+/CD 4+ (%) 61. 7 53. 4 0. 9 57. 9 65. 5 1. 1 41. 8 55. 2 1. 3 28. 6 35. 4 1. 2 14. 3 13. 0 0. 9 CD 8+CXCR 3+/CD 8+ (%) 60. 2 58. 8 1. 0 80. 3 75. 9 0. 9 70. 1 75. 4 1. 1 84. 3 69. 0 0. 8 30. 0 24. 8 0. 8

Supplementary Table S 3. Pre-treatment serum levels of cytokines/chemokines were not associated with response

Supplementary Table S 3. Pre-treatment serum levels of cytokines/chemokines were not associated with response (CR/PR vs. SD/PD/Uneval) to Ipi-Bev. BOR Wilcoxon Cytokines/ SD/PD/Uneval CR/PR rank-sum p Chemokines N Min Median Max -value EGF 35 0. 3 134. 7 509. 3 8 23. 7 170. 0 282. 7 0. 89 EOTAXIN 35 3. 2 79. 3 196. 5 8 49. 0 74. 0 177. 8 0. 87 FGF-2 35 1. 2 48. 2 131. 6 8 1. 2 52. 5 80. 9 0. 68 GM-CSF 35 1. 2 7. 6 26. 1 8 2. 2 8. 7 28. 8 0. 36 GRO 35 3. 2 551. 9 1648. 6 8 231. 7 517. 6 693. 3 0. 55 G-CSG 35 3. 1 21. 1 114. 4 8 21. 1 44. 0 97. 6 0. 17 IFNg 35 0. 5 3. 2 18. 0 8 0. 5 3. 2 15. 0 0. 78 IL-8 35 1. 2 18. 2 296. 5 8 3. 1 12. 8 110. 7 0. 89 IL-1 a 35 0. 4 6. 3 307. 8 8 1. 9 15. 1 48. 2 0. 63 IFNa 2 35 0. 8 7. 3 94. 9 8 3. 2 10. 9 19. 7 0. 74 IP-10 35 11. 6 351. 8 1563. 1 8 244. 3 381. 7 2273. 2 0. 61 MCP-1 35 3. 2 411. 5 755. 9 8 173. 7 340. 2 695. 8 0. 57 MDC 35 3. 2 845. 4 2171. 9 8 483. 1 783. 8 1228. 9 0. 53 MIP-1 b 35 3. 2 25. 5 102. 2 8 2. 4 29. 6 57. 6 0. 96 TNFa 35 0. 1 4. 0 24. 1 8 0. 8 3. 0 15. 3 0. 60 VEGF 35 3. 2 71. 5 331. 7 8 3. 2 77. 1 142. 5 0. 79

Supplementary Table S 4. Pre-treatment serum levels of cytokines/chemokines were not associated with disease

Supplementary Table S 4. Pre-treatment serum levels of cytokines/chemokines were not associated with disease control (CR/PR/SD vs. PD/Uneval) in Ipi-Bev-treated patients. DCR Wilcoxon Cytokines/ PD/Uneval CR/PR/SD rank-sum p Chemokines N Min Median Max -value EGF 13 0. 3 149. 7 506. 5 30 23. 7 138. 7 509. 3 0. 84 EOTAXIN 13 45. 8 80. 2 196. 5 30 3. 2 75. 7 177. 8 0. 56 FGF-2 13 16. 8 60. 1 127. 8 30 1. 2 41. 5 131. 6 0. 28 GM-CSF 13 4. 2 7. 6 26. 1 30 1. 2 7. 4 28. 8 0. 27 GRO 13 55. 1 541. 2 1648. 6 30 3. 2 553. 7 1288. 3 0. 78 G-CSG 13 8. 0 37. 0 114. 4 30 3. 1 22. 1 111. 1 0. 29 IFNg 13 0. 5 3. 2 18. 0 30 0. 5 3. 2 15. 0 0. 41 IL-8 13 1. 7 17. 4 243. 1 30 1. 2 18. 5 296. 5 0. 64 IL-1 a 13 1. 2 13. 7 307. 8 30 0. 4 4. 9 49. 2 0. 10 IFNa 2 13 4. 3 15. 5 94. 9 30 0. 8 6. 3 26. 3 0. 01 IP-10 13 120. 9 363. 1 1552. 3 30 11. 6 351. 7 2273. 2 0. 96 MCP-1 13 168. 5 449. 0 755. 9 30 3. 2 359. 2 705. 7 0. 07 MDC 13 460. 5 770. 7 1149. 3 30 3. 2 883. 3 2171. 9 0. 22 MIP-1 b 13 10. 7 35. 4 86. 4 30 2. 4 24. 9 102. 2 0. 16 TNFa 13 1. 4 4. 1 14. 1 30 0. 1 3. 0 24. 1 0. 35 VEGF 13 33. 8 94. 6 211. 1 30 3. 2 65. 0 331. 7 0. 25

Supplementary Table S 5. Fold-changes of serum cytokines/chemokines were not associated with response (CR/PR

Supplementary Table S 5. Fold-changes of serum cytokines/chemokines were not associated with response (CR/PR vs. SD/PD/Uneval) to Ipi-Bev. BOR Wilcoxon Cytokines/ SD/PD/Uneval CR/PR rank-sum p Chemokines N Min Median Max -value EGF 35 0. 06 18. 83 8 0. 38 1. 03 2. 37 0. 79 EOTAXIN 35 0. 71 1. 10 37. 85 8 0. 59 1. 06 1. 42 0. 59 FGF-2 35 0. 49 1. 00 50. 11 8 0. 28 0. 91 2. 62 0. 27 GM-CSF 35 0. 35 1. 00 4. 15 8 0. 32 0. 88 1. 95 0. 57 GRO 35 0. 56 1. 29 194. 8 8 0. 73 1. 23 2. 68 0. 55 G-CSG 35 0. 15 1. 00 8. 13 8 0. 38 1. 04 3. 63 0. 50 IFNg 35 0. 09 1. 00 36. 98 8 0. 15 0. 92 41. 41 0. 84 IL-8 35 0. 04 1. 72 19. 50 8 0. 14 1. 17 15. 51 0. 61 IL-1 a 35 0. 43 1. 16 16. 14 8 0. 92 2. 82 14. 51 0. 18 IFNa 2 35 0. 26 1. 24 10. 11 8 0. 27 1. 98 10. 45 0. 69 IP-10 35 0. 42 1. 29 30. 68 8 1. 08 1. 74 6. 00 0. 19 MCP-1 35 0. 15 0. 95 142. 3 8 0. 63 1. 00 2. 56 0. 96 MDC 35 0. 33 0. 89 142. 2 8 0. 62 0. 94 1. 59 0. 63 MIP-1 b 35 0. 21 1. 11 14. 70 8 0. 68 1. 18 4. 04 0. 61 TNFa 35 0. 36 1. 70 14. 29 8 0. 87 1. 47 6. 07 0. 86 VEGF 35 0. 07 0. 62 12. 17 8 0. 11 0. 82 1. 91 0. 55

Supplementary Table S 6. Fold-changes of serum cytokines/chemokines were not associated with disease control

Supplementary Table S 6. Fold-changes of serum cytokines/chemokines were not associated with disease control (CR/PR/SD vs. PD/Uneval) in Ipi-Bev-treated patients. DCR Wilcoxon Cytokines/ PD/Uneval CR/PR/SD rank-sum p Chemokines N Min Median Max -value EGF 13 0. 23 1. 00 18. 83 30 0. 06 1. 08 3. 50 0. 74 EOTAXIN 13 0. 76 1. 07 1. 34 30 0. 59 1. 10 37. 85 0. 50 FGF-2 13 0. 57 1. 00 3. 11 30 0. 28 1. 00 50. 11 0. 55 GM-CSF 13 0. 55 1. 06 3. 05 30 0. 32 0. 95 4. 15 0. 81 GRO 13 0. 78 1. 29 6. 54 30 0. 56 1. 28 194. 8 0. 99 G-CSG 13 0. 60 1. 00 6. 41 30 0. 15 1. 00 8. 13 0. 52 IFNg 13 0. 32 1. 00 4. 71 30 0. 09 1. 00 41. 41 0. 96 IL-8 13 0. 72 19. 50 30 0. 04 1. 51 18. 40 0. 46 IL-1 a 13 0. 43 1. 27 7. 30 30 0. 60 1. 21 16. 14 0. 34 IFNa 2 13 0. 72 1. 17 4. 93 30 0. 26 1. 35 10. 45 0. 21 IP-10 13 0. 42 1. 20 6. 33 30 0. 65 1. 59 30. 68 0. 38 MCP-1 13 0. 62 0. 92 1. 63 30 0. 15 0. 99 142. 3 0. 93 MDC 13 0. 45 0. 85 1. 29 30 0. 33 0. 94 142. 2 0. 14 MIP-1 b 13 0. 72 1. 15 2. 79 30 0. 21 1. 15 14. 70 0. 89 TNFa 13 0. 49 1. 70 6. 62 30 0. 36 1. 68 14. 29 0. 57 VEGF 13 0. 27 0. 60 3. 44 30 0. 07 0. 64 12. 17 0. 65

Supplementary Table S 7. Response (CR/PR vs. SD/PD/Uneval) to Ipi-Bev by cytokine expression category.

Supplementary Table S 7. Response (CR/PR vs. SD/PD/Uneval) to Ipi-Bev by cytokine expression category. Comparison Fishers’ exact p-value Response by FLT-31 0. 99 Response by fractalkine 0. 99 Response by IL-2 0. 99 Response by IL-3 0. 11 Response by IL-4 0. 42 Response by IL-5 0. 40 Response by IL-6 0. 44 Response by IL-7 0. 44 Response by IL-9 0. 69 Response by IL-10 0. 99 Response by IL-13 0. 69 Response by IL-15 0. 44 Response by IL-17 0. 99 Response by IL-12 p 40 0. 46 Response by IL-12 p 70 0. 99 Response by IL-1 b 0. 70 Response by IL-1 ra 0. 22 Response by MCP-3 0. 69 Response by MIP-1 a 0. 99 Response by TGFa 0. 99 Response by TNFb 0. 70 Response by SIL-2 ra 0. 99 Response by s. CD 40 L 0. 31

Supplementary Table S 8. Disease control (CR/PR/SD vs. PD/Uneval) in Ipi-Bev-treated patients by cytokine

Supplementary Table S 8. Disease control (CR/PR/SD vs. PD/Uneval) in Ipi-Bev-treated patients by cytokine expression category. Comparison Disease-control by FLT-31 Disease-control by fractalkine Disease-control by IL-2 Disease-control by IL-3 Disease-control by IL-4 Disease-control by IL-5 Disease-control by IL-6 Disease-control by IL-7 Disease-control by IL-9 Disease-control by IL-10 Disease-control by IL-13 Disease-control by IL-15 Disease-control by IL-17 Disease-control by IL-12 p 40 Disease-control by IL-12 p 70 Disease-control by IL-1 b Disease-control by IL-1 ra Disease-control by MCP-3 Disease-control by MIP-1 a Disease-control by TGFa Disease-control by TNFb Disease-control by SIL-2 ra Disease-control by s. CD 40 L Fishers’ exact p-value 0. 14 0. 49 0. 16 0. 99 0. 32 0. 73 0. 99 0. 74 0. 33 0. 74 0. 99 0. 31 0. 33 0. 32 0. 33 0. 99 0. 02 0. 99 0. 29 0. 33 0. 57 0. 99

Supplementary Table S 9. Pre-treatment cytokine levels according to immune related adverse events (ir.

Supplementary Table S 9. Pre-treatment cytokine levels according to immune related adverse events (ir. AEs). Wilcoxon ranksum p-values N Mean Std Min Median Max IFNa 2 Severe ir. AE 0. 01 No 31 14. 2 16. 6 0. 8 11. 0 94. 9 Yes 12 6. 2 4. 6 3. 2 4. 8 18. 0 IL-1 a Severe ir. AE 0. 46 No 31 23. 2 55. 0 0. 4 7. 5 307. 8 Yes 12 13. 9 15. 8 1. 2 4. 4 49. 2 TNFa Severe ir. AE 0. 21 No 31 5. 6 5. 4 0. 8 4. 1 24. 1 Yes 12 3. 8 4. 0 0. 1 3. 2 15. 3 GRO Severe ir. AE 0. 17 No 31 617. 7 360. 9 3. 2 580. 9 1648. 6 Yes 12 460. 3 208. 0 212. 7 413. 6 963. 3 IP-10 IL-8 Severe ir. AE No Yes 0. 22 31 12 515. 9 512. 3 406. 1 597. 5 11. 6 120. 9 373. 2 303. 2 1563. 1 2273. 2 0. 30 31 12 41. 2 19. 6 69. 5 29. 0 1. 2 2. 5 19. 9 8. 7 296. 5 105. 8

 Supplementary Table S 10. Relationship between cytokine fold-changes according to ir. AEs. N

Supplementary Table S 10. Relationship between cytokine fold-changes according to ir. AEs. N IFNa 2 IL-1 a TNFa GRO IP-10 IL-8 Severe ir. AE No Yes Mean 31 12 2. 1 1. 9 Std 2. 4 1. 1 Min 0. 3 0. 7 Median 1. 2 1. 8 Max Wilcoxon rank-sum p-values 0. 51 10. 4 4. 0 0. 56 31 12 3. 1 3. 0 4. 3 3. 8 0. 4 0. 5 1. 2 1. 4 16. 1 13. 5 0. 43 31 12 3. 1 2. 2 3. 3 1. 9 0. 4 0. 8 1. 3 14. 3 7. 1 0. 36 31 12 7. 8 1. 3 34. 7 0. 6 0. 7 1. 3 1. 2 194. 8 2. 8 0. 70 31 12 3. 1 2. 3 5. 6 1. 9 0. 4 0. 5 1. 6 30. 7 6. 0 0. 34 31 12 3. 1 4. 7 4. 5 6. 3 0. 0 0. 3 1. 8 18. 4 19. 5

Supplementary Table S 11. Clinical responses and survival of patients whose plasma samples were

Supplementary Table S 11. Clinical responses and survival of patients whose plasma samples were used in antibody response analyses. Patient P 6 P 8 Clinical Response Partial response Survival (months) 52 51 Alive Yes

Supplementary Table S 12. Effects of ipilimumab alone and Ipi-Bev on circulating cytokine/chemokine levels.

Supplementary Table S 12. Effects of ipilimumab alone and Ipi-Bev on circulating cytokine/chemokine levels. Therapy Cytokines / Chemokines IFNa 2 FC <1. 5 FC ≥ 1. 5 IL-1 a FC <1. 5 FC ≥ 1. 5 TNFa FC <1. 5 FC ≥ 1. 5 GRO FC <1. 5 FC ≥ 1. 5 IP-10 FC <1. 5 FC ≥ 1. 5 IL-8 FC <1. 5 FC ≥ 1. 5 Ipi-Bev Fisher’s Exact p-value Ipi N % 25 16 58. 1 37. 2 14 7 60. 9 30. 4 20 18 46. 5 41. 9 4 1 17. 4 4. 3 18 25 41. 9 58. 1 13 10 56. 5 43. 5 26 17 60. 5 39. 5 16 7 69. 6 30. 4 21 22 48. 8 51. 2 15 8 65. 2 34. 8 21 22 48. 8 51. 2 9 13 39. 1 56. 5 0. 78 0. 36 0. 31 0. 59 0. 3 0. 61