Induction of anti-tumour lymphocytes in cancer patients after brief exposure to supernatants from cultures of anti-CD3-stimulated allogeneic lymphocytes

Abstract

Summary The present study investigated the ability of supernatants collected from cultures of healthy donor-derived peripheral blood mononuclear cells (HD-PBMCs) stimulated with anti-CD3 monoclonal antibody (MAb) (allogeneic CD3 supernatants; ACD3S) to induce, upon brief exposure, tumour-reactive cytotoxic lymphocytes in cancer patients' PBMCs. ACD3S enhanced natural killer (NK) and lymphokineactivated killer (LAK) cell-mediated cytotoxicity. ACD3S contained increased levels of interleukins (IL) 1, 2, 6, 7 and 12, as well as of granulocyte-macrophage colony-stimulating factor (GM-CSF), gamma-interferon (IFN-y) and tumour necrosis factor-a (TNF-a). MAbs against these cytokines significantly reduced the ACD3S-induced cytotoxicity. ACD3S-induced cytotoxicity was not inhibited by anti-CD4, CD8 and MHC class MAbs, but was markedly reduced in the presence of MAb against CD18. In contrast to HD-PBMC, ACD3S derived from cancer patients' lymphocytes exhibited lower levels of the above-mentioned cytokines and exerted reduced biological activity. In conclusion, ACD3S are able to activate, upon short-term incubation, tumour-reactive lymphocytes from cancer patients' PBMCs that lyse a variety of tumour targets, including autologous tumours. ACD3S contain high levels of certain cytokines that positively influence the induction of autologous tumour-reactive lymphocytes. Such supernatants can be collected easily from healthy donors and stored until use in clinical trials for adoptive cellular therapy of cancer. They may also be indicated in the construction of cytokine cocktails that have the ability to induce antitumour cytotoxicity.

Combined treatment with Bevacizumab and standard chemotherapy restores abnormal immune parameters in advanced colorectal cancer patients

“…Previous reports by our group [26,33], have demonstrated reduced levels of these monokines in patients with various types of cancer and deficiencies in functional activities of antigen presenting cells in cancer patients have also been reported by others. In particular Ninomiya et al [34] have demonstrated that monocytes from patients with hepatocellular carcinoma expressed low levels of HLA-DR, induced decreased amounts of IL-12 and were less potent in stimulating MLR responses.…”

“…PBMCs stimulation with anti-CD3 mAb was performed as recently reported [24,26]. In brief, PBMCs (2×10 6 cells/ml) were activated in 25-cm 2 flasks (Costar), precoated with anti-CD3 mAb in 5 ml of culture medium.…”

Compromised anti-tumor responses in tumor necrosis factor-α knockout mice

“…Unlike CTL, NK cells kill their targets spontaneously and without restriction by MHC. On the other hand, LAK cells generated in cultures with IL-2, which can be of T or NK cell origin [12,13], lyse tumor cells including those that are NK resistant, in a non-MHC-restricted fashion [13]. The mechanisms of cytotoxicity utilized by all three types of effectors mentioned are classified into secretory and nonsecretory pathways.…”

Chemotherapy ± Cetuximab Modulates Peripheral Immune Responses in Metastatic Colorectal Cancer

“…Furthermore, deficiencies in functional activities of antigen-presenting cells in cancer patients have also been previously reported [14,31]. Monocytes from patients with hepatocellular carcinoma express low levels of HLA-DR, decreased amounts of IL-12 and were less potent in stimulating auto-MLR responses [32].…”

“…This was performed as recently reported [12,14]. In brief, PBMCs (2 × 10 6 cells/ml) were activated in 25 cm 2 flasks (Costar), precoated with anti-CD3 mAb in 5 ml of culture medium.…”