Introduction
Cytokine-induced killer cells were first discovered in 1991 [1] and are a heterogeneous population of CD8+ T cells, which were generated from human peripheral blood lymphocytes (PBLs) and simply expanded ex vivo via incubation with an anti-CD3 antibody, interferon-γ (IFN-γ), and interleukin (IL)-2. They can kill tumor cells mediated by FasL and perforin [2].
CIK cells can lyse cancer cells in an MHC-unrestricted manner through activating NK cell receptors such as DNAX accessory molecule-1, NKp46, NKG2D, and NKp30 [3-5]. In addition to the direct killing effect of CIK on cancer cells, they can also regulate the immune function by secreting various cytokines. A lot of studies have indicated that after stimulation by tumor cells, the levels of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-α, IFN-γ, and IL-2 secreted by CIK cells are significantly upregulated [6], and these cytokines further enhance systemic antitumor activity and induce a Th1 immune response.
Figure 1: Possible mechanisms of CIK cell cytotoxicity. Being a mixture of cells, CIK includes T cells (CD3+CD56−), NK‐T cells (CD3+CD56+), and NK cell population (CD3−CD56+).
Mechanism of Action
There has been less information about the cytotoxicity of CIK cells and its exact mechanism has not been completely elucidated. Blocking monoclonal antibodies against leukocyte function‐associated antigen‐1 and intercellular adhesion molecule‐1 inhibits the cytotoxic effect of CIK cells. As a result, CIK cells have a cell‐to‐cell contact mediated by tumor cell lysis. [7].
The molecule that seems to play the most important role in tumor recognition by CIK cells is probably the natural killer group 2 member D (NKG2D) receptor. NKG2D is a member of the c-type lectin-activating receptor family that is evolutionarily conserved and is located within the NK gene complex on human chromosome 12p12-p13 [8]
Figure 2: Double T/NK specificity of CIK cells and mechanism of action. CIK cells are T-EMRA lymphocytes that maintain the original TCR specificity and acquire NK-like cytotoxicity. In both cases, DNAM1 and LFA1 are required for target binding while NK-like cytotoxicity is mediated by CD56, NKp30 and NKG2D molecules.
Cancer Types
In cooperation with the Tungs’ Taichung Metro Harbor Hospital, and various private hospitals in Malaysia, our group participated in application of various cell-therapies (DC, CIK, DC-CIK) in a variety of solid cancer treatment projects, that were approved by both the Ministry of Health and Welfare of Taiwan and the Ministry of Health of Malaysia.
The cell therapy applications include 22 types of stage 4 solid cancers, including the cancer types listed in the table below [ref: MOHW, Taiwan].
- Adrenal Cancer
- Biliary Tract Cancer
- Bladder Cancer
- Brain Malignant Tumor
- Breast Cancer
- Cervical Cancer
- Colorectal Cancer
- Endometrial Cancer
- Esophageal Cancer
- Gastric Cancer
- Germ Cell Cancer
- Head & Neck Cancer
- Kidney Cancer
- Liver Cancer
- Lungs Cancer
- Neuroendocrine Cancer
- Ovarian Cancer
- Pancreatic Cancer
- Prostate Cancer
- Soft Tissue Sarcoma
- Skin Cancer
- Thyroid Cancer
Treatment Flow
Application
For autologous use only. Patients will receive at least 2 cycles of CIK cells infusion with one-week intervals between each cycle. 5~7 x109 cells were suspended in 240mL normal saline and administered via intravenous infusion within 30-45 minutes.
The immune cell therapy program can be combined with other treatment methods, including anti-inflammatory drugs (NSAID) and antipyretic analgesics to relieve the patient’s discomfort. Or combined use of radiation therapy, targeted therapy, chemotherapy, immune checkpoint inhibitor therapy, and so on to improve the overall anti-cancer efficacy. All concomitant treatment methods must be evaluated by the operating specialist’s professional clinical experience, and the relevant adverse reactions of concomitant treatment should be observed at any time.
All patients were followed up after discharge, including blood routine examination and PET CT/CT screening every 3 months for the first 2 years, 6 months for the next 3 years, and yearly thereafter from the fifth year.
Possible Side Effects
According to the results of clinical trial literature, the most common adverse reactions of CIK cell therapy are mostly mild symptoms such as fever and pain. Although the frequency is incredibly low, the symptoms will resolve spontaneously without treatment. However, each patient may have special reactions (idiosyncrasy) due to individual differences and special constitutions. If nausea, vomiting, diarrhea, or shock occurs after the cell reinfusion, the cell reinfusion should be stopped and treated according to the specialist’s instructions.
| Adverse Effect | Descriptions |
|---|---|
| Grade 1 | The operating physician needs to place the patient in a comfortable space and observe until the adverse event resolves spontaneously or gives appropriate physical support (ice compress, drinking warm water, sleep, etc.). |
| Grade 2 | The operating physician immediately places the patient and observes whether the adverse event tends to ease or become more serious. If it tends to be relieved, physical support is given to accelerate the resolution of the adverse event; if it tends to be serious, evaluate the severity of the patient and give appropriate drug support (antipyretic, analgesic, steroid, etc.). Continue observation and drug support until the adverse event stabilizes or resolves. Suspend this course of treatment and enter the observation period and reassess whether to continue treatment after the cause is found out. |
| Grade 3 | The operating physician must immediately take emergency measures and administer medication support (antipyretic, analgesic, steroids, pressure boosters, electrolyte infusion… etc.) according to the patient’s symptoms. Arrange to be hospitalized and continue treatment and slow medication reduction until the symptoms are completely resolved. The patient withdraws from this course of treatment, terminates the cell reinfusion, and arranges for routine medical care for the patient. |
| Grade 4 | The operating physician must immediately be sent to the emergency department to take first aid measures and provide life support (intratracheal intubation, steroids, pressure boosters, electrolyte infusion, etc.) according to the patient’s symptoms. Arrange to the intensive care unit as soon as possible and continue treatment until the symptoms are completely stable or resolved. The patient withdraws from this course of treatment, terminates the cell reinfusion, and arranges for routine medical care for the patient. |
Laboratory Processing
This picture illustrates the production of CIK cells in the laboratory. The patient’s PBMC cells will be harvested by apheresis in hospital and sent directly to our laboratory in Malaysia via a professional courier service (World Courier). Once received, the PBMC sample will be prepared for isolation and performed cell counting. The cells are then incubated at 370C for 24 hours, under 5% CO2, 95% relative humidity using serum-free medium containing 1000 U/ml IFN-g. After 24 hours, anti-CD3 antibody (at a final concentration of 100 ng/ml), 1 ng/ml IL-1a and 1000 U/ml recombinant human IL-2 is added to the medium. After 14 days of culture, autologous CIK cells are suspended in 240 ml normal saline. The cells are then transferred into a clinical grade cell culture bag. The cell products were tested according to the current Chinese Pharmacopoeia, such as microbial contamination testing being carried out to ensure the reliability, PCR based method for the mycoplasma detection to rule out contamination and quantification of bacterial endotoxin by kinetic turbidimetric methods. After passing the QC, the cell product will be cryopreserved and shipped to the Hospital. After recovery in hospital, the cells are ready for infusion.
Good Tissue Practice Lab
The facility is designed with regulations that are stricter than any Good Tissue Laboratory (GTP) regulations to avoid risks of environmental contamination including multiple gowning procedures (primary and secondary changing rooms, biologically safety areas, and limited access to qualified persons. The facility complies with the FDA high standard secondary dressing facility.
SITE SPECIFICATIONS
It has a cleanliness level of 10,000, a “positive pressure, constant humidity, constant temperature” high-standard dust-free independent laboratory, and a complete control mechanism, which can provide various process facilities for cell therapy.
CLEANLINESS
With a cleanliness of 10,000 and a complete control mechanisms, it can provide various process facilities required for cell therapy.
STRICT REQUIREMENTS
Adhere to the use of high-quality manufacturer-sourced reagents, high-end equipment, and full-process serum-free culture to improve the effectiveness and safety of human cell tissue operations. Strict operation procedures, manufacturing procedures, storage conditions, effective time and records are established for the acceptance/feeding of human cell tissues, acceptance or return, distribution, and destruction or disposal.
Comprehensive Support
- Patient Educational Materials
- Doctor’s Educational Materials
- Free Pre-treatment Assessment
- Comprehensive Auxiliary support for treatments in Malaysia
- Post Treatment Monitoring
Additional Reading
Presentations
Scientific Publications
Vamos life-cell therapies
The Vamos group of companies is engaged in life-cell therapies since 2016. We provide cutting-edge biotechnology products to both hospitals and patients seeking the most effective cancer treatments. Our suite of T-cell based immunotherapies and services are highly accurate, personalized, and focused on providing the most effective treatments based on highest quality standards and at the lowest cost possible.
Vamos Services
Our group provides the following services:
- Production and supply of autologous CAR-T cells for CAR-T treatments provided by hospitals in Malaysia and internationally.
- Medical tourism and auxiliary services related to hosting of international patients taking CAR-T treatments in Malaysia.
Facts about Vamos
- Our group is the only provider of CAR-T therapy in Malaysia for blood cancers.
- Our CAR-T therapy is approved by the Malaysian Ministry of Health.
- All patients that participated in the clinical trial achieved complete remission are live a life free of cancer (with exception of 3 patients that passed away before receiving the actual CAR-T treatment).
- 6 of the top hospitals in Malaysia provide CAR-T treatments with our products.
- So far, we have treated more than 100 number of patients with stage 4 cancer, and the success rate of our CAR-T therapy is over 90%.
- Up to date, all patients treated with our CAR-T therapy have been treated successfully and without experienced any severe side effects.
- The quality of our CAR-T therapy is second to none, while the cost of our therapy is possibly the lowest cost in healthcare worldwide.
- In addition to CAR-T therapy (for both blood and solid cancers), we also provide DC therapy, CIK therapy, and DC-CIK therapy for solid cancers. Together with our clinical trial partner hospital, our group has conducted more than 2,000 DC, CIK & DC-CIK treatments.
References
- Schmidt-Wolf IG, Negrin RS, Kiem HP, Blume KG, Weissman IL. Use of a SCID mouse/human lymphoma model to evaluate cytokine-induced killer cells with potent antitumor cell activity. J Exp Med (1991) 174(1):139–49. doi:10.1084/jem.174.1.139
- Lee HK, Kim YG, Kim JS, Park EJ, Kim B, Park KH, et al. Cytokine-induced killer cells interact with tumor lysate-pulsed dendritic cells via CCR5 signaling. Cancer Lett (2016) 378(2):142–9. doi:10.1016/j.canlet.2016.05.020
- Verneris MR, Karimi M, Baker J, Jayaswal A, Negrin RS. Role of NKG2D signaling in the cytotoxicity of activated and expanded CD8+ T cells. Blood (2004) 103(8):3065–72. doi:10.1182/blood-2003-06-2125
- Schmidt TL, Negrin RS, Contag CH. A killer choice for cancer immunotherapy. Immunol Res (2014) 58(2–3):300–6. doi:10.1007/s12026-014-8507-2
- Pievani A, Belussi C, Klein C, Rambaldi A, Golay J, Introna M. Enhanced killing of human B-cell lymphoma targets by combined use of cytokine-induced killer cell (CIK) cultures and anti-CD20 antibodies. Blood (2011) 117(2):510–8. doi:10.1182/blood-2010-06-290858
- Linn YC, Wang SM, Hui KM. Comparative gene expression profiling of cytokine-induced killer cells in response to acute myloid leukemic and acute lymphoblastic leukemic stimulators using oligonucleotide arrays. Exp Hematol (2005) 33(6):671–81. doi:10.1016/j.exphem.2005.03.005
- Solmaz Shirjang, Nazila Alizadeh, Behzad Mansoori, Ata Mahmoodpoor, Hossein Samadi Kafil, Mohammad Hojjat‐Farsangi, Mehdi Yousefi. Promising immunotherapy: Highlighting cytokine‐induced killer cells. J Cell Biochem. 2018;1-21. doi: 10.1002/jcb.28250
- Martino Introna and Fabio Correnti. Innovative Clinical Perspectives for CIK Cells in Cancer Patients. Int. J. Mol. Sci. 2018, 19(2), 358. https://doi.org/10.3390/ijms19020358