BACKGROUND AND OBJECTIVEThe CIK cell is one of the most important means of the adoptive cellular immunotherapy, and it is a hotspot of which to simplify its culture procedure and to promote its inhibition rate. The aim of this study is to observe the biological function of the CIK cells cultivated by the recombinant human fibronectin (RN) and to establish an effective and simple way of cells expansion.

METHODSWe separated the mononuclear cells (PBMCs) in 50 mL peripheral blood from 10 healthy persons with density gradient centrifugation in the lymphocyte-separating medium, and the PBMCs were divided into two groups, of which were cultivated by RN-introduced and conventional method separately. Then we estimated the proliferation ability, and analyzed the immunologic type, IFN-gamma, IL-4, perforin and granzyme B of them with flow cytometry. Besides that, we tested the inhibition rate of CIKs cells to four kinds of human lung cancer cell lines in vitro by MTT assay.

RESULTSThe RN-induced group had a higher proliferation rate that was 2.0-3.5 times of the conventional group, and there was an obvious statistical difference between the two (P < 0.05). The proliferation rates of CD3+CD16+CD56+T cells in each group were 3 778 and 2 068 times of the initial number, respectively. There was also a higher percentage of CD3+CD8+ T cells in RN-induced group (P < 0.05), while the percentage of CD3+CD4+T cells had no significant statistical difference (P > 0.05). We found a similar inhibition rate of the CIK cells to all this human lung cancer cell lines (P > 0.05). The cells which secreted IFN-gamma increased, while the cells which secreted IL-4 did not. The cells which secreted granzyme B and perforin were positive.

CONCLUSIONIt is an effective and simple way to cultivate the CIK cells with RN, which should be adopted.

CD3 Complex ; metabolism ; CD4-Positive T-Lymphocytes ; immunology ; CD56 Antigen ; metabolism ; Cell Line, Tumor ; Cells, Cultured ; Cytokine-Induced Killer Cells ; drug effects ; metabolism ; Fibronectins ; genetics ; metabolism ; pharmacology ; Flow Cytometry ; Humans ; Immunotherapy, Adoptive ; methods ; Interleukin-4 ; metabolism ; Lung Neoplasms ; therapy ; Receptors, IgG ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; pharmacology ; T-Lymphocytes ; immunology

BACKGROUND AND OBJECTIVEThe actual evaluation of immunological function is significant for studing the tumor development and devising a treatment in time. The aim of this study is to evaluate the immunological function of advanced lung cancer patients systematically, and to discuss the clinical significance.

METHODSThe nucleated cell amounts of advanced lung cancer patients and the healthy individuals were counted. The immune cell subsets and the levels of IL-4, INF-gamma, perforin and granzyme in CD8+T cells by the flow cytometry were measured. The proliferation activity and the inhibition ratio of immune cells to several tumor cell lines were evaluated by MTT assay.

RESULTSThe absolute amounts and subsets of T, B, NK cells of advanced lung cancer patients were lower than the healthy individuals (P < 0.05); However, the proportion of regulatory T cells of advanced lung cancer patients (4.00 +/- 1.84)% was lower than the healthy individuals (1.27 +/- 0.78)% (P < 0.05). The positive rates of IFN-gamma perforin, granzyme in CD8+T cells decreased while them in IL-4 did not in the advanced lung cancer patients compared to the healthy control group (P < 0.05). The proliferation activity of immune cells, the positive rate of PPD masculine and the inhibition ratio to tumor cells in the advanced lung cancer patients was lower than the healthy subsets obviously (P < 0.05).

CONCLUSIONThere was a significant immune depression in the advanced lung cancer patients compared to the healthy individuals.

Adult ; Aged ; CD8-Positive T-Lymphocytes ; immunology ; metabolism ; Cell Line, Tumor ; Female ; Flow Cytometry ; Granzymes ; metabolism ; Humans ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Lung Neoplasms ; immunology ; pathology ; Male ; Middle Aged ; Perforin ; metabolism

Objective: To analyze the interactions between different structural types of volatile oil components (VOCs) and skin lipid molecules, and investigate the mechanism of volatile oil in Chinese materia medica (VOCMM) as penetration enhancers. Methods: In this study, 210 different structural types of VOCs were selected from the VOCMM penetration enhancer database, and the molecular docking experiments were conducted with three main lipid molecules of skin: ceramide 2 (CER2), cholesterol (CHL), and free fatty acid (FFA). Each VOC was docked individually with each lipid molecule. Cluster analysis was used to explore the relationship between the binding energy of VOCs and their molecular structures. Nine specific pathogen-free (SPF) Sprague Dawley (SD) rats were randomly divided into Control, Nootkatone, and 3-Butylidenephthalide groups for in vitro percutaneous experiments, with three rats in each group. The donor pool solutions were 3% gastrodin, 3% gastrodin + 3% nootkatone, and 3% gastrodin + 3% 3-butylidenephthalide, respectively. The penetration enhancing effects of VOCs with higher binding energy were evaluated by comparing the 12-hour cumulative percutaneous absorption of gastrodin (Q12, µg/cm²). Result: (i) Most of the VOCs were non-hydrogen bonded to the hydrophobic parts of CHL and FFA, and hydrogen bonded to the head group of CER2. Among them, sesquiterpene oxides showed the most pronounced binding affinity to CER2. The VOCs with 2 − 4 rings (including carbon rings, benzene rings, and heterocycles) demonstrated stronger binding affinity for three skin lipid molecules compared with the VOCs without intramolecular rings (P < 0.01). (ii) According to the cluster analysis, most of the VOCs that bond well to CER2 had 2 − 3 intramolecular rings. The non-oxygenated VOCs were bonded to CER2 in a hydrophobic manner. The oxygenated VOCs were mostly bonded to CER2 by hydrogen bonding. (iii) The results of Franz diffusion cell experiment showed that the Q12 of Control group was 260.60 ± 25.09 µg/cm2, and the transdermal absorption of gastrodin was significantly increased in Nootkatone group (Q12 = 5 503.00 ± 1 080.00 µg/cm², P < 0.01).  The transdermal absorption of gastrodin was also increased in 3-Butylidenephthalide  group (Q12 = 495.40 ± 56.98 µg/cm², P > 0.05). (iv) The type of oxygen-containing functional groups in VOCs was also an influencing factor of binding affinity to CER2. Conclusion The interactions between different types of VOCs with different structures in the VOCMM and three skin lipid molecules in the stratum corneum were investigated at the molecular level in this paper. This research provided theoretical guidance and data support for the screening of volatile oil-based penetration enhancers,  and a simple and rapid method for studying the penetration-enhancing mechanism of volatile oils.