Cross regulation by IL-10 and IL-2/IL-12 of the helper T cells and the cytolytic activity of lymphocytes from malignant effusions of lung cancer patients.
OBJECTIVE
Our previous report demonstrated that there was impairment of local cellular immunity with elevated interleukin-10 (IL-10) and undetectable IL-12 in neoplastic pleural effusion. These findings suggest that the local immune reactions favor the T-helper type 2 (Th2) pathway instead of Th1 pathway. The present study was designed to examine whether local cellular immunity could be manipulated by IL-2 and/or IL-12 treatment, and to determine their effect on the helper T-cell pathways and the cytolytic activity of the effusion-associated lymphocytes (EALs).


METHODS
Using malignant pleural effusions obtained from four patients suffering from adenocarcinoma of lung, we separated the tumor cells from the EALs with Ficol-Hypaque centrifugation, followed by Percoll density centrifugation. To test whether the cytolytic function of lymphocytes could be enhanced by culturing with IL-2 and/or IL-12, lymphocytes were incubated with recombinant IL-2 with/without IL-12 for 6 days. Following this, the tumoricidal activity was assessed in an overnight 5'chromium-release assay. Autologous tumor cells for measuring specific antitumor activity, Daudi cells susceptible to lymphokine-activated killer cells, and NK-susceptible K562 cells were used as target cells.


RESULTS
After treatment in vitro with IL-2, IL-12, or IL-2 plus IL-12, the Th pathway shifted from Th2 to Th1 type (increased gamma-interferon production). To further study the effect of cytokine treatment on the cytolytic activity of EALs, it was found that after 6-day culturing, the EALs failed to kill any of the three tumor targets, whereas the 6-day cultured peripheral blood lymphocytes (PBLs) gave low level of cytotoxicity against all three tumor targets. Stimulation with IL-2 alone partially restored the immunocompetence of EALs to kill the tumor targets. Stimulation with IL-12 alone showed no significant effect on their cytolytic activity. However, IL-12 synergized with IL-2 to increase the cytolytic activity of EALs and PBLs against autologous tumor targets. This synergistic effect was not found for Daudi cells and K562 cells.


CONCLUSIONS
These results suggest that EALs activated with IL-12 in the presence of a low concentration of IL-2, which converted the EALs from Th2 pathway to Th1 pathway, could be an alternative source of antitumor effectors for adoptive immunotherapy of cancer.
Chen
Y M
YM
Chest Department, Veterans General Hospital-Taipei, Taiwan, ROC.
Yang
W K
WK
Ting
C C
CC
Tsai
W Y
WY
Yang
D M
DM
Whang-Peng
J
J
Perng
R P
RP
D000230:Adenocarcinoma / Q000276:immunology / Q000473:pathology*; D002469:Cell Separation; D002478:Cells, Cultured; D002498:Centrifugation; D003102:Colloids; D003287:Contrast Media; D003602:Cytotoxicity, Immunologic / Q000276:immunology*; D003973:Diatrizoate; D005362:Ficoll; D006801:Humans; D007108:Immune Tolerance; D007121:Immunocompetence; D016753:Interleukin-10 / Q000276:immunology*; D018664:Interleukin-12 / Q000276:immunology* / Q000627:therapeutic use; D007376:Interleukin-2 / Q000276:immunology* / Q000627:therapeutic use; D015979:Killer Cells, Lymphokine-Activated / Q000276:immunology; D007694:Killer Cells, Natural / Q000276:immunology; D008175:Lung Neoplasms / Q000276:immunology / Q000473:pathology*; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology; D016066:Pleural Effusion, Malignant / Q000276:immunology / Q000473:pathology*; D011205:Povidone; D011994:Recombinant Proteins; D012822:Silicon Dioxide; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology*; D018417:Th1 Cells / Q000276:immunology; D018418:Th2 Cells / Q000276:immunology
D003102:Colloids; D003287:Contrast Media; D007376:Interleukin-2; D011994:Recombinant Proteins; D003973:Diatrizoate; D016753:Interleukin-10; D018664:Interleukin-12; D005362:Ficoll; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone