Cytotherapy 2013; Volume 15, Issue 4, Supplement, Page S19.
SP Chin1, VK Wai-Ping2, ZL Qi3, KY Then4, SK Cheong5
1Elixir Biotech, Cyberjaya, Selangor, Malaysia, 2The University of Hong Kong, Hong Kong, 3University Tunku Abdul Rahman,Bandar Sungai Long, Selangor, Malaysia.
Background: Cytokine-induced killer (CIK) cells are a group of immune cells that include natural killer (NK) cells, T cells and natural killer T (NKT) cells which can be generated from peripheral blood mononuclear cells (PBMC). NKT cells can be identified by the presence of CD3+ and CD56+ surface markers and are able to target cancer cells directly or indirectly in the absence of antibodies or MHC specific proteins allowing a rapid and non-restricted immune reaction. Treatment with CIK also replenishes immune cells lost during chemotherapy. Studies have shown that 5×10 CIK cells with at least 30% fraction of NKT cells were associated with better outcomes. However most critically-ill cancer patients are unable to tolerate large volume apheresis to obtain sufficient PBMC for CIK expansion.
Objective: We explored a new protocol to expand the CIK cells by using only 60ml of peripheral venous blood from stage III-IV cancer patients undergoing chemotherapy and radiotherapy. The benefit of this approach is the usage of smaller amount of blood through simple venipuncture as compared to the more invasive apheresis method.
Methodology: 60ml of peripheral venous blood was withdrawn from 9 patients (M=5, F=4) with mean age of 62±11 years old and 1 healthy donor (male, 45 years) using simple venipuncture technique. The samples were isolated and induced using interferon-gamma (IFN-γ), anti-CD3 antibody, recombinant human interleukin-2 (IL-2) for up to 21 days.
Results: We successfully isolated and expanded CIK from all 10 blood samples. The number of CIK was significantly increased from Day 1 to Day 21 of cell expansion (0.12±0.08 x10 vs. 7.46±1.85 x10 cells, p<0.001) with average of 84±49 fold increase. The expansion number and rate was similar to the healthy donor. In addition, the proportion of NKT cells (CD16+ and CD56+) was 43.7±11.0%.
Conclusion: Using our protocol, we have successfully isolated and expanded CIK to significant and sufficient number of cells using only 60ml of peripheral venous blood. This method may allow more critically-ill cancer patients to consider adjunctive cellular immunotherapy.