Phenotype Of Cytokine-induced Killer Cells Research Articles

Inhibition Human Nasopharyngeal Carcinoma Cells TNF-α Gene Transfected Cytokine-Induced Killer Cells Based on Nanomaterial Polyphthalamide Monoamine Dendrimer Nanomaterial.

The present study aims to investigate the possibility of TNF-α gene transfection into CIK (cytokine-induced killer) cells using the nanomaterial PAMAM and the inhibitory effects of these cells on the growth of the human nasopharyngeal carcinoma cell line CNE-2. The pEGFP-N1-TNF-α recombinant plasmid was constructed and used to transfect the CIK cells using the nanomaterial PAMAM. Subsequently, the transfection efficiency was measured. The ELISA method was used to analyze the CIK cell culture supernatant. TNF-α concentration in fluid, and CIK cell phenotype was analyzed by the flow cytometry. The MTT assay was used to detect the inhibitory activity of CIK cells on the growth of nasopharyngeal carcinoma cell line CNE-2 after transfection. The CIK cells were transfected with the nanomaterial PAMAM using the successfully constructed recombinant plasmid pEGFP-N1-TNF-α. The growth characteristics and phenotypic characteristics of the transfected CIK cells were not changed, and an increase in the TNF-α secretion was observed, indicating that the CIK cells can significantly inhibit CNE-2 cell growth (P < 0005).

Adoptive cell therapy of hematological malignancies using cytokine-induced killer cells retargeted with monoclonal antibodies

Background & Aim Cytokine-Induced Killer (CIK) cells are a population of effector cells that represent a promising tool for adoptive cell therapy. From a technical and methodological point of view, they are easily expandable ex-vivo and safe, and demonstrated efficacy against hematological malignancies. CIK cells exert cytotoxicity against a broad range of tumor histotypes but not against normal tissues and hematopoietic precursors. We recently reported that they have a relevant expression of FcγRIIIa (CD16a), which can be exploited in combination with clinical-grade monoclonal antibodies (mAbs) to redirect their cytotoxicity in an antigen-specific manner, to improve their antitumor activity. Indeed, the engagement of CD16a on CIK cells leads to a potent antibody-dependent cell-mediated cytotoxicity (ADCC) against ovarian cancer both in vitro and in vivo. Based on this observation, we investigated whether CIK cells could be specifically retargeted against B-cell malignancies by combination with anti-CD20 mAbs, namely Rituximab® and Obinutuzumab®. Methods, Results & Conclusion CIK cells were obtained from peripheral blood mononucleated cells of healthy donors, and stimulated in vitro with IFN-γ, CD3 mAb (OKT3) and IL-2 for 14 days; fresh IL-2 was provided every 3-4 days. CIK cell phenotype was analyzed by multicolor flow cytometry; cytotoxic activity was assessed by calcein AM-release assay against EHEB (CLL), Raji (Burkitt lymphoma), RCK-8, TMB-8, Karpas 422 (DLBCL) tumor cell lines, and primary samples obtained from CLL and DLBCL patients after written informed consent. The combination with either the mAbs significantly increased CIK cell cytotoxicity not only against lymphoma cell lines, but also against the primary tumor samples. Depletion of NK cells from CIK cell bulk cultures did not affect target cell lysis, thus confirming that the antibody-mediated increase of cytotoxicity was mainly accountable to the CIK cell fraction. Taken together, these data indicate that the combination treatment with CIK cells and anti-CD20 mAbs could represent a novel approach for the treatment of hematological malignancies, alternative to the use of chimeric antigen receptor (CAR)-T cells.

CIK cell cytotoxicity is a predictive biomarker for CIK cell immunotherapy in postoperative patients with hepatocellular carcinoma.

Adjuvant cytokine-induced killer (CIK) cell immunotherapy has shown potential in improving the prognosis of hepatocellular carcinoma (HCC) patients after curative resection. However, whether an individual could obtain survival benefit from CIK cell treatment remains unknown. In the present study, we focused on the characteristics of CIK cells and aimed to identify the best predictive biomarker for adjuvant CIK cell treatment in patients with HCC after surgery. This study included 48 patients with HCC treated with postoperative adjuvant CIK cell immunotherapy. The phenotype activity and cytotoxic activity of CIK cells were determined by flow cytometry and xCELLigence™ Real-Time Cell Analysis (RTCA) system, respectively. Correlation analysis revealed that the cytotoxic activity of CIK cells was significantly negative correlated with the percentage of CD3+ CD4+ cell subsets, but significantly positive correlated with CD3-CD56+ and CD3+ CD56+ cell subsets. Survival analysis showed that there were no significant associations between patients' prognosis and the phenotype of CIK cells. By contrast, there was statistically significant improvement in recurrence-free survival (RFS) and overall survival (OS) for patients with high cytotoxic activity of CIK cells as compared with those with low cytotoxic activity of CIK cells. Univariate and multivariate analyses indicated that CIK cell cytotoxicity was an independent prognostic factor for RFS and OS. In conclusion, a high cytotoxic activity of CIK cells can serve as a valuable biomarker for adjuvant CIK cell immunotherapy of HCC patients after surgery.

Cytokine-induced killer cells and supernatant on the role of klebsiella pneumoniae

Objective To investigate the ability of cytokine-induced killer (CIK) cells and supernatant against klebsiella pneumoniae in vitro.Methods CIK cells were prepared in vitro.The phenotype of CIK cells was analyzed by flow cytometer in 1,7,14,21 day.Both microdilution method was used to detect the minimum inhibitory concentration (MIC) values of CIK cells against klebsiella pneumoniae (final inoculation concentration of bacteria was 1.0 × 105 cfu/ml,target ratio was 40∶ 1,20∶ 1,10∶ 1,5∶1),the supernatant,and amikacin (AK),piperacillin (PIPC) joint after,to determine their antimicrobial activity in vitro.Results In the culture section 14-21 day,CD3 + CD56 + cell phenotype proportion remained at a high level more than 45％.Absorbance (A) values for each test showed that different concentrations of CIK cells killing effect were no statistically significant (F =3.065,P ＞ 0.05) at 24 h.The supernatant group showed no significant bacterial growth.The MIC values of amikacin (AK),piperacillin (PIPC) were 2,512 mg/L respectively.CIK cells respectively,and AK,PIPC joint after,the latter failed to reduce the MIC values,but the supernatant reduced the MIC values of both to the minimize inoculum concentrations of 0.25 mg/L and 64.00 mg/L.Conclusion CIK cells showed no direct killing effect against klebsiella pneumoniae in vitro,and no synergistic reaction combined with antibiotics,but the supernatant has antibacterial effects to klebsiella pneumoniae. Key words: Cytokine-induced killer cells; Klebsiella pneumoniae; Minimum inhibitory concentration