Immunity Feedback and Clinical Outcome in Colon Cancer Patients Undergoing Chemoimmunotherapy with Gemcitabine + FOLFOX followed by Subcutaneous Granulocyte Macrophage Colony-Stimulating Factor and Aldesleukin (GOLFIG-1 Trial)

The curves compare lymphocyte (A) and eosinophil counts (B) in patients receiving the GOLFIG regimen () in a group of 20 patients with metastatic colorectal carcinoma receiving the standard FOLFOX-4 chemotherapy () or in a group of 10 patients receiving the GOLF chemotherapy without cytokine administration (). Higher lymphocyte and eosinophil counts in patients receiving chemoimmunotherapy (P < 0.05) were shown. C and D, a progressive increase in the number of T lymphocytes expressing a CD3⁺CCR7⁺ (), CD3⁺CD45Ro⁺ (), CD8⁺CD27⁺ (), or CD3⁺CD62L⁺ () phenotype in the PBMCs of 20 patients receiving GOLFIG chemoimmunotherapy.

Curves showing the percentage of lymphocytes expressing a CD4⁺CD25⁺FoxP3⁺ phenotype in the PBMCs of 20 patients during treatment with the GOLFIG regimen (○). PBMCs derived from 10 patients treated with FOLFOX regimen (▪) and four healthy donors (⧫) were used as controls. There was a significant and stable decrement in the number of T_reg cells in the patients after treatment (P < 0.05), with the levels becoming similar to those observed in the PBMCs of healthy donors.

Immunohistochemical analysis using anti-FoxP3 (A, B, E, F, I, J, M, N, Q, and R) and anti-CCR7 (C, D, G, H, K, L, O, P, S, and T) polyclonal antibodies in tumor specimens of colon cancer patients before (first and third column) and after (second and fourth column) 12 cycles of chemotherapy. A to D, tumor samples of a representative control patient who underwent FOLFOX chemotherapy; E to T, tumor samples of patients who underwent GOLFIG regimen. The pictures show a significant reduction of FoxP3⁺ lymphocytes (brown) and a significant increase of CCR7⁺ lymphocytes (green) in the tumor specimens derived from patients who underwent GOLFIG chemoimmunotherapy compared with those derived from patients who underwent FOLFOX chemotherapy (P < 0.05). Score 0 of FoxP3⁺ lymphocytes was shown in F, J, M, and N; score 1 of CCR7⁺ lymphocytes was shown in C, G, O, and S; score 2 was shown in A, D, E, I, and K; score 3 was shown in L and R; score 4 was shown in B, H, P, and T; and score 5 was shown Q. Original magnifications, ×100 (A-D, F-H, J-L, and O-S) and ×200 (E, I, M, N, P, and T).