Immunopotentiation effects of apigenin on NK cell proliferation and killing pancreatic cancer cells.

Apigenin is a kind of flavonoid with many beneficial biological effects. It not only has direct cytotoxicity to tumor cells, but also can boost the antitumor effect of immune cells by modulating immune system. The purpose of this study was to investigate the proliferation of NK cells treated with apigenin and its cytotoxicity to pancreatic cancer cells in vitro, and explore its potential molecular mechanism. In this study, the effect of apigenin on NK cell proliferation and killing pancreatic cancer cells were measured by CCK-8 assay. Perforin, granzyme B (Gran B), CD107a, and NKG2D expressions of NK cells induced with apigenin were detected by flow cytometry (FCM). The mRNA expression of Bcl-2, Bax and protein expression of Bcl-2, Bax, p-ERK, and p-JNK in NK cells were evaluated by qRT-PCR and western blotting analysis, respectively. The results showed that appropriate concentration of apigenin could significantly promote the proliferation of NK cells in vitro and enhance the killing activity of NK cells against pancreatic cancer cells. The expressions of surface antigen NKG2D and intracellular antigen perforin and Gran B of NK cells were upregulated after treating with apigenin. Bcl-2 mRNA expression was increased, while Bax mRNA expression was decreased. Similarly, the expression of Bcl-2, p-JNK, and p-ERK protein was upregulated, and the expression of Bax protein was downregulated. The molecular mechanism of the immunopotentiation effects of apigenin may be that it up-regulates Bcl-2 and down-regulates Bax expression at the gene and protein levels to facilitate NK cell proliferation, and up-regulates the expression of perforin, Gran B, and NKG2D through the activation of JNK and ERK pathways to enhance NK cell cytotoxicity.

Preparation and Performance of Ultra-Fine Polypropylene Antibacterial Fibers via Melt Electrospinning.

Polypropylene (PP) fibers are employed commonly as the raw material of technical textiles (nonwovens), and the research focuses on fine-denier fibers and their functionalities. In this work, antibacterial PP masterbatches with different dosage (1-5 wt.%) of nano-ZnO particles as the antibacterial agent were prepared via a twin-screw extruder. The as-prepared PP masterbatches were electrospun on a home-made electrospinning device to afford ultra-fine PP fibers. The morphologies of as-spun ultrathin PP fibers with 16 µm of average diameter were observed by SEM. The structure and element distribution were characterized by means of energy-dispersive spectroscopy (EDS) and Fourier-transfer infrared spectroscopy (FTIR), respectively. There was some zinc obviously distributed on the surface when a dosage of ZnO more than 1 wt.% was used, which contributed to the antibacterial activity. The crystallinity of PP fibers was not affected strongly by the dosage of ZnO based on the differential scanning calorimetry (DSC) heating curves, while thermal decomposition improved with the increase in ZnO content, and the mechanical strength decreased predictably with the increase in inorganic ZnO content.

In Situ Electrospinning Iodine-Based Fibrous Meshes for Antibacterial Wound Dressing.

For effective application of electrospinning and electrospun fibrous meshes in wound dressing, we have in situ electrospun poly(vinyl pyrrolidone)/iodine (PVP/I), PVP/poly(vinyl pyrrolidone)-iodine (PVPI) complex, and poly(vinyl butyral) (PVB)/PVPI solutions into fibrous membranes by a handheld electrospinning apparatus. The morphologies of the electrospun fibers were examined by SEM, and the hydrophobicity, gas permeability, and antibacterial properties of the as-spun meshes were also investigated. The flexibility and feasibility of in situ electrospinning PVP/I, PVP/PVPI, and PVB/PVPI membranes, as well as the excellent gas permeabilities and antibacterial properties of the as-spun meshes, promised their potential applications in wound healing.

Wnt pathway activator TWS119 enhances the proliferation and cytolytic activity of human γδT cells against colon cancer.

γδT cells are a distinct T-cell subset that display unique characteristics regarding T-cell receptor gene usage, tissue tropism and antigen recognition. Adoptive γδT cell transfer therapy has recently been gaining importance as an efficient approach in cancer immunotherapy. However, exploiting γδT cell response for tumour immunotherapy is a challenge due to cell numbers, activities and differentiation states that minimize the clinical therapeutic effects. Previous studies have indicated that the wnt/ß-catenin signalling pathway plays a crucial role in the differentiation, survival and enhancement of the immune response of T lymphocytes. In this study, we sought to evaluate whether the activation of the wnt/ß-catenin pathway through inhibition of glycogen synthase kinase-3ß (GSK-3ß) using 4,6-disubstituted pyrrolopyrimidine (TWS119) could be an efficient strategy to improve the proliferation, differentiation and cytolytic activity of γδT cells against colon cancer cells. Remarkably, we found that TWS119 significantly enhanced the proliferation and survival of γδT cells via activation of the mammalian target of rapamycin (mTOR) pathway, upregulation of the expression of the anti-apoptotic protein Bcl-2 and inhibition of cleaved caspase-3 in addition to the Wnt pathway. Our results also showed that enhancement of the cytolytic activity of γδT cells against human colon cancer cells by TWS119 was chiefly associated with upregulation of the expression of perforin and granzyme B in vitro and in vivo. Additionally, TWS119 can induce the expression of CD62L or CCR5 to generate a population of CD62L+γδT or CCR5+γδT cells in a dose-dependent manner. These findings suggested that TWS119 could be a useful complementary agent for improving γδT cell-based immunotherapy.

Enhancement of CD3AK cell proliferation and killing ability by α-Thujone.

Thujone is a monoterpene ketone natural substance found mainly in wormwood and sage. Previous studies have shown that Thujone has various pharmacological effects, such as anti-tumor, analgesic, and insecticide. The effect of α-Thujone to human immune cells is still unknown. Our study focuses on investigating the effects and mechanism of α-Thujone to CD3AK (anti- CD3 antibody induced activated killer) cells proliferation and cytotoxicity to colon cancer cell lines. With cell proliferation and FCM assay, it is found that α-Thujone could significantly enhance CD3AK cell proliferation and expression of CD107a in a dose-dependent manner. The cytotoxicity to colon cancer cells detected by CCK-8 assay is also improved. The expressions of TNF-α and FasL detected with ELISA assay were not significantly changed. Mechanically, the study shows that α-Thujone could enhance the expression of p-ERK1/2 and p-Akt. In addition, α-Thujone has no cytotoxicity to HCT116 and SW620 cells proliferation. In a word, α-Thujone enhances CD3AK cell proliferation and cytotoxicity via the improvement of expression of CD107a, p-Akt and p-ERK1/2.

Enhancement effect of dihydroartemisinin on human γδ T cell proliferation and killing pancreatic cancer cells.

γδ T cells play important roles in innate immunity against tumors and infections. Inhibitory effect of dihydroartemisinin on growth of cancer cells has been found in recent years. In this study, we investigated the effect of dihydroartemisinin on human γδ T cell proliferation by MTT assay and killing activity against pancreatic cancer cells SW1990, BxPC-3 and PANC-1 by LDH release assay in vitro. Intracellular molecule alterations were verified by flow cytometry. The results suggested that appropriate concentration of dihydroartemisinin favored the expansion of γδ T cells and enhanced γδ T cell mediated killing activity against pancreatic cancer cells. Up-regulation of intracellular perforin, granzyme B expression and IFN-γ production may be the important mechanism of dihydroartemisinin on increased antitumor activity of γδ T cells.

[Effects of different stimulatory factors on functions of CIK cells].

This study was aimed to investigate the effects of different stimulatory factors on proliferation and function of cytokine induced killer (CIK) cells. Peripheral blood mononuclear cells (PBMNC) were separated by Ficoll-Hypacue gradient. According to supplement of different stimulatory factors (CD28 mAb, IL-15 and IL-21), the experiment was divided into five groups:control group (CIK), CB28+IL-15+IL-21 group, IL-15+IL-21 group, CD28+IL-15 group and CD28+IL-21 group. Effects of different stimulatory factors on the proliferation of CIK cells were assayed by an automated hematology analyzer. Changes of granzyme B,perforin and CD107a were detected by flow cytometry. IL-10, IL-12, INF-γ and TNF-α were quantified by ELISA. Cytotoxicities on lung cancer cell line A549, breast adenocarcinoma cell line MFC-7 and human melanoma cell line HME1 were examined by lactate dehydrogenase release method. The results showed that there were significant differences among different groups. The highest proliferation index on days 10 was observed in group CD28mAb, IL-15 and IL-21(255.3 ± 6.3), which was higher than control group, IL-21+IL-15 group and CD28 mAb+IL-21 group (166.6 ± 13.5, 199.4 ± 15.0 and 228.8 ± 16.6) (P < 0.05). The expression of perforin in CD28 mAb+IL-15 group was higher than the other groups. The expression of perforin,GranB and CD107a of costimulatory groups was higher than control group. The cytotoxicities of CD28 mAb+IL-15 group on A549, MFC-7 and HME1 cells (82.2%, 59.3% and 70.6%) were much higher than that of control group (60.9%, 49.6% and 48.4%) (P < 0.05). The highest IFN-γsecretion was found in CD28 mAb, IL-15 and IL-21 groups. It is concluded that there are significant difference of proliferative capacity, cytokine secretion and cytotoxicity after being activated by different stimulatory factors. Adding corresponding stimulatory factors into the culture system displays a great value for target cells culture.

[Association of traumatic severity with change in lymphocyte subsets in the early stage after trauma].

OBJECTIVE: To investigate the association of traumatic severity with changes in lymphocyte subsets in the early stage after trauma. METHODS: Sixty-three male patients admitted within 4 hours after trauma were enrolled. According to injury severity score (ISS), the patients were divided into two groups: mild trauma group (ISS<16, n=35) and severe trauma group (ISS≥16, n=28). At admission, the patients peripheral blood were extracted to detect T lymphocytes subsets, blood routine test, blood biochemical and arterial blood gas analysis which were used to calculate the acute physiology and chronic health evaluation II (APACHEII) scores. The correlation of lymphocyte subsets and ISS score, and the correlation of lymphocyte subsets and APACHEII score were both analyzed statistically. Another 20 cases of healthy male adults were enrolled as the control group. RESULTS: Compared with the healthy control group, CD3(+) T cell contents in blood were decreased obviously in mild trauma group and severe trauma group (0.648±0.112, 0.647±0.110 vs. 0.708±0.082, both P<0.05); CD4(+) T cells contents in severe group were decreased significantly (0.317±0.086 vs. 0.389±0.064, P<0.05), and natural killer (NK) cells were significantly increased (0.217±0.107 vs. 0.158±0.068, P<0.05). B cells content in severe group was decreased significantly than that of mild group (0.114±0.060 vs. 0.155±0.075, P<0.05). There were no significant difference in CD8(+) and CD4/CD8 ratio among the healthy control group, mild trauma group and severe trauma group (CD8(+): 0.260±0.074, 0.260±0.091, 0.271±0.105; CD4/CD8 ratio: 1.69±0.75, 1.56±0.83, 1.34±0.65, all P>0.05). Except that there were negative correlation between CD3(+) T cells and the ISS scores (r=-0.42, P=0.03), the other lymphocyte subsets showed no correlation with the ISS scores and the APACHEII scores (mild trauma group with ISS scores: CD3(+) r=-0.10, CD4(+) r=-0.31, CD8(+) r=0.18, B cells r=0.20, NK cells r=-0.04; mild trauma group with APACHEII scores: CD3(+) r=0.04, CD4(+) r=-0.07, CD8(+) r=0.06, B cells r=-0.10, NK cells r=0.05, severe trauma group with ISS scores: CD4(+) r=-0.12, CD8(+) r=-0.17, B cells r=0.02, NK cells r=0.31,all P>0.05;severe trauma group with APACHEII scores:CD3(+) r=-0.24, CD4(+) r=0.11, CD8(+) r=-0.26, B cells r=0.15, NK cells r=0.08, all P>0.05). CONCLUSIONS: CD3(+) and CD4(+) T cells decreased and NK cells increased significantly in blood in the early stage after severe trauma. CD3(+) T cells are independent indexes which reflect body injury. Therefore, it is necessary to monitor the changes of immune cells dynamically after severe trauma.

The enhanced effect of lupeol on the destruction of gastric cancer cells by NK cells.

Lupeol, a triterpene, was reported to possess beneficial effects as a therapeutic and preventive agent for a range of disorders. Many studies have confirmed that lupeol possesses strong activities such as antioxidative, antiinflammatory, antiarthritic, antimutagenic, and antimalarial, both in vitro and in vivo, and at its effective therapeutic doses exhibit no toxicity to normal cells and tissues. Lupeol was observed to inhibit the proliferation of gastric tumour cells in a dose-dependent manner, as assessed by MTT assay, and induce the proliferation of NK cells, as assessed by flow cytometry and Western blotting. The killing effect of NK cells on gastric tumour cells was assessed by LDH. Our experiment demonstrated that lupeol at appropriate concentrations could promote the proliferation of NK cells, inhibit the proliferation of gastric cancer cell lines BGC823, N87 and HGC27, and increase the killing effect of NK cells on gastric cancer cells. We speculated that lupeol might increase the expression of PFP, IFN-γ, and CD107a via the activation of PI3K/Akt and Wnt/ß-catenin signalling pathways. Lupeol could serve as a potential agent against gastric cancer; however, further in-depth in vivo studies are still required.

The effect of phloretin on human γδ T cells killing colon cancer SW-1116 cells.

OBJECTIVE: To explore the effect and mechanism of Phloretin on human γδ T cells killing colon cancer SW-1116 cells. METHODS: γδ T cells were amplified in vitro from human peripheral blood mononuclear cells through isopentenyl pyrophosphate method (IPP). After cocultured different concentrations of Phloretin with γδ T cells or SW-1116 cells for 48h respectively, MTT assay was used to test the growth curve of these two cells; Flow cytometry to test the expression of Granzyme B (GraB), perforin (PFP), CD107a and IFN-γ of γδ T cells; Lactate dehydrogenase (LDH) release assay to test the cytotoxic activity of the γδ T cells on SW-1116 cells; and Western blot to test the Wnt3a expression of the γδ T cells. RESULTS: After cultured with IPP for ten days, the percentage of γδ T cells increased from 3.31±3.00% to 78.40±10.30%. Compared to the control group, when the concentration of Phloretin increased from 2.35µg/ml to 18.75µg/ml, it could significantly proliferate the γδ T cell growth (P<0.05) and inhibit the growth of SW-1116 cells in dose-response, and the expression of GraB, PFP, CD107a and Wnt3a significantly increased (P<0.05). Significant positive relationships were observed among CD107a and PFP, GraB, cytotoxicity (P<0.05). The percentage of IFN-γ producing γδ T cells treated with Phloretin was significantly higher than control group. CONCLUSION: Phloretin can enhance the killing effect of γδ T cells on SW-1116 cells; the mechanism may be that Phloretin could proliferate the γδ T cell growth, increase the expression of PFP and GraB, activate the Wnt signaling pathway, and produce higher level of IFN-γ. Indeed CD107a expression probably correlates quite well with antitumor activity.

[Inhibitory effect of toll-like receptor 7 agonist on K562 cells].

The aim of this study was to investigate the inhibitory effect of Toll-like receptor 7 (TLR7) agonist gardiquimod on K562 cells. Human γδT cells from peripheral blood cells were amplified by isopentenyl pyrophosphate. The proliferation capacity of γδT cells and K562 cells were measured with MTT assay after treatment with different concentrations of gardiquimod. Cytotoxicity of γδT cells on K562 cells was detected by CCK-8 kit, and the intracellular expression of TLR7, cell cycle and apoptosis of K562 cells before and after treatment with gardiquimod were measured by flow cytometry. The results demonstrated that gardiquimod could significantly stimulate the proliferation of γδT cells, and inhibit proliferation of K562 cells under the concentration of 11.0 µg/ml for 48 h. The expression of TLR7 increased after treatment with gardiquimod. No apoptosis was observed, but there were significant changes in cell cycle, moreover the K562 cells treated with gardiquimod were more killed by γδT cells. It is concluded that the gardiquimod can inhibit the proliferation of K562 cells and enhance their sensitivity to killing activity of human γδT cells.

[Co-stimulation of multiple activating factors on proliferation and phenotype of T lymphocytes in peripheral blood in vitro].

AIM: To observe the costimulation of multiple activating factors effects on the proliferation and phenotype of T lymphocytes in vitro. METHODS: Peripheral blood mononuclear cells (PBMCs) were separated by fractionation on Ficoll-Hypaque gradient. According to adding different cytokines (CD3 mAb, CD28 mAb, IFN-γ, IL-1α, IL-2 and IL-15), the experiments were divided into seven groups. Effects of different cytokines on the proliferation of PBMC were counted by automated hematology analyzer five categories. The phenotypes (CD3, CD4, CD8, CD28, CD16, CD56(+);CD16, CD3(+);CD8(+);, CD3(+);CD4(+);, CD3(+); CD56(+);, CD45RO) expressing on the surface of costimulatory cells were detected by flow cytometry, and the cytotoxicity of costimulatory cells on SGC-7901, SW-1990 and SW-116 cell lines was examined by lactate dehydrogenase release method. RESULTS: The proliferation has significant difference when adding different cytokines into PBMCs culture system, the highestest proliferation multiples group is the one contains cytokines CD3, CD28, IFN-γ, IL-2, IL-1α, IL-15 and IL-21, which proliferation multiple is 255.3±6.3 at the tenth day of cell culture, obviously higher than the other culture systems which only contains CD3, IFN-γ and IL-2 (166.6±5.5) (P<0.05). Part of cells'phenotype changed when adding different activating factors. Without IL-15, the proportion of CD16(+);CD56(+);(NK) cells and CD3(+);CD56(+); cells was higher than the other groups; CD45RO(+); memory cells is most evident when delayed adding IL-15 and IL-21 for three days. The cytotoxicity of PBMCs cultured for ten days with different activating factors had significant difference, the highest was the one which delayed adding IL-15 and IL-21 for three days (76.2%, 60.3% and 70.6%, respectively.), higher than the cell culture groups containing CD3, IFN-γ and IL-2 (54.9%, 44.6% and 50.4%, respectively) (P<0.05). The cultured cells had the strongest cytotoxicity on SGC-7901 gastric adenocarcinoma cells. CONCLUSION: The PBMCs' proliferation, phenotype and cytotoxicity had significant difference after being activated by different stimulating factors, adding matching stimulating factors into the culture system have great value on cell-directed culture.

Effects of pituitary adenylate cyclase activating polypeptide on CD4(+)/CD8(+) T cell levels after traumatic brain injury in a rat model.

BACKGROUND: The effect of pituitary adenylate cyclase activating polypeptide (PACAP) during traumatic brain injury (TBI) and whether it can modulate secondary injury has not been reported previously. The present study evaluated the potential protective effects of ventricular infusion of PACAP in a rat model of TBI. METHODS: Male Sprague Dawley rats were randomly divided into 3 treatment groups (n=6, each): sham-operated, vehicle (normal saline)+TBI, and PACAP+TBI. Normal saline or PACAP (1 µg/5 µL) was administered intracerebroventricularly 20 minutes before TBI. Right parietal cortical contusion was produced via a weight-dropping method. Brains were extracted 24 hours after trauma. Histological changes in brains were examined by HE staining. The numbers of CD4(+) and CD8(+) T cells in blood and the spleen were detected via flow cytometry. RESULTS: In injured brain regions, edema, hemorrhage, inflammatory cell infiltration, and swollen and degenerated neurons were observed under a light microscope, and the neurons were disorderly arrayed in the hippocampi. Compared to the sham group, average CD4(+) CD8(-) lymphocyte counts in blood and the spleen were significantly decreased in rats that received TBI+vehicle, and CD4(-) CD8(+) were increased. In rats administered PACAP prior to TBI, damage was attenuated as evidenced by significantly increased CD4(+), and decreased CD8(+), T lymphocytes in blood and the spleen. CONCLUSION: Pretreatment with PACAP may protect against TBI by influencing periphery T cellular immune function.

[Mechanistic study of nimesulide on enhancing gammadeltaT cell-mediated killing of gastric cancer cells].

AIM: To explore the effect of nimesulide on human gammadeltaT cell function. METHODS: gammadeltaT cells were cultured routinely, collected on the 9th day, and then induced with nimesulide at indicated concentrations (0.25 micromol/L, 0.5 micromol/L, 1 micromol/L, 2 micromol/L, 4 micromol/L, respectively). Twenty-four hours after induction, the supernatants were collected to detect IFN-gamma, TNF-alpha and IL-12, whereas the cells were assayed for perforin, granzyme B and NKG2D by flow cytometry (FCM) and for killing of gastric cancer cells by LDH. RESULTS: Nimesulide (1 micromol/L) caused gammadeltaT cells to express more of perforin and granzyme B (62.8% and 72.7%, respectively) than the control group (51.4% and 60.9%, respectively) (P<0.05). Likewise, nimesulide (1 micromol/L) enabled gammadeltaT cells to secret more of IFN-gamma and TNF-alpha (262.3 ng/L and 177.5 ng/L, respectively) than the control group (196.1 ng/L and 158.5 ng/L, respectively) (P<0.05). Nimesulide did not affect IL-12 secreting capability of gammadeltaT cells as compared with the control group (P>0.05). Nimesulide-stimulated gammadeltaT cells killed more of SGC-7901 and BCG-823 gastric cancer cells (73% and 70%, respectively) than the control group(54% and 53%, respectively) (P<0.05). CONCLUSION: Nimesulide made gammadeltaT cells to express more perforin and granzyme B and to secret more IFN-gamma and TNF-alpha into the supernatant, leading to higher killing rate of SGC-7901 and BCG-823 gastric cancer cells. The above data provides experimental basis on the clinical use of nimesulide to prevent and treat digestive tract tumors.

[The effect of aspirin on human's gammadeltaT cells killing digestive system tumor cell lines].

AIM: To explore the effect of aspirin on human's gammadeltaT cells killing digestive system tumor cell lines. METHODS: Use the isopentenyl pyrophosphate method to amplify human peripheral blood gammadeltaT cells in vitro. Various concentrations of aspirin were used to induce gammadeltaT cells and digestive system tumor cells SGC-7901, SW-1990, SW-480, SW-1116, LOVO cells lines, LDH assays was used to measure the cytotoxic activity of gammadeltaT cells, flow cytometry analysis the apoptosis percentage of before and after induced gammadeltaT cells and SGC-7901, SW-1990, SW-480, SW-1116, LOVO cell lines. RESULTS: gammadeltaT cells were cultivated for ten days which proliferation ratio increase from 4.21% to 70.35%. When gammadeltaT cells induced via aspirin concentrations rang from 0.4 mmol/L to 0.8 mmol/L which cytotoxic activity on the five kinds of tumor cell lines was the highest, if aspirin's concentration surpassing 3.2 mmol/L, cytotoxic activity of gammadeltaT cells present decrease tendency. SGC-7901, SW-1990, SW-480, SW-1116, LOVO cells lines were induced by different concentrations of aspirin for 24 hours, just SW-480, SW-1116 and LOVO were enhanced as far as the cytotoxic activity of gammadeltaT cells on these cell lines was concerned, other groups and control group have no notable changed. The apoptosis percentage of gammadeltaT cells(52.71%) were induced by aspirin which concentration was 3.2 mmol/L, which was strikingly higher than SGC-7901, SW-1990, SW-480, SW-1116, LOVO cells lines, (respectively 7.88%, 8.89%, 6.21%, 4.47% and 3.67%). CONCLUSION: When aspirin's concentration for clinical routine used can enhance the effect of gammadeltaT cells killing the tumor cells, if surpassing this concentration can obvious inhibited the proliferation capacity and cytotoxic activity of gammadeltaT cells and augment apoptosis ratio, however, it is not obvious for digestive tract tumor lines.

[Characterization and heavy metal adsorption properties of schwertmannite synthesized by bacterial oxidation of ferrous sulfate solutions].

An amorphous ferric hydroxysulfate named schwertmannite was synthesized by using bacterial oxidation of ferrous iron by resting A. ferrooxidans cells in acid ferrous sulfate solution (pH 2.5). The identification of the bacterially oxidized iron precipitate was carried out by SEM, XRD, FTIR and chemical analysis. Results showed that resting cells of A. ferrooxidans LX5 could completely oxidize ferrous ions to ferric ions in the FeSO4-H2O system after 2 days of incubation. The solution pH decreased from an initial 2.5 to 2.10, and about 15% of the ferrous iron was transformed into the red-brown precipitates. The subsequent characterization of the precipitates showed that the biotic, synthetic ferric hydroxysulfate was schwertmannite. Sorption edge measurement showed that the adsorption behavior of metal cations (e. g., Cu2+, Zn2+ and Cr3+) was pH-dependent. The adsorption increased with an increase in pH, and the maximum was found in a pH range of 6.0-7.0. For solution concentration of 50 mg x L(-1), the maximum adsorption efficiencies of Cu2+, Zn2+ and Cr3+ were 99.3%, 99.4% and 87.6%, respectively.

[Specific anti-leukemic cell effect mediated by dendritic cells pulsed with chronic myelogenous leukemia lysate antigen in vitro].

To investigate the specific antileukemia effect of dendritic cells (DC) pulsed with chronic myelogenous leukemic lysate antigen (CLA), dendritic cells from patients with chronic myelogenous leukemia (CML) were pulsed by CLA, and then cocultured with cytokine-induced killer (CIK) cells from CML patients (CIK + CLA-DC group). The cytotoxic activity in vitro was measured by using a lactate dehydrogenase release assay, and compared with CIK + DC, CIK and CIK + CLA groups. The results showed that under an effector-target ratio of 25:1, the cytotoxic activity of CIK + CLA-DC, CIK + DC, CIK and CIK + CLA groups against autologous CML cells was (68.8 +/- 14.2)%, (52.5 +/- 9.4)%, (20.6 +/- 7.5)% and (24.2 +/- 8.7)%, respectively. CIK + CLA-DC group displayed a strongest cytotoxic activity. When K562 and Raji cells acted as target cells and CIK as effectors, the cytotoxic activity against autologous CML cells in CIK + CLA-DC group (68.8 +/- 14.2)% was much higher than that against K562 cells (14.6 +/- 6.2)% and Raji cells (12.7 +/- 10.2)%, respectively. In conclusion, coculture of CIK cells with DC led to a significant increase in cytotoxic activity. The cytotoxicity could be further increased by DC pulse with CML cell lysate antigen, and cytotoxicity mediated by CML lysate antigen possess stronger specificity.

[Clinical observation on adoptive immunotherapy with autologous cytokine-induced killer cells for advanced malignant tumor].

BACKGROUND & OBJECTIVE: Cytokine-induced killer (CIK) cells have the characteristics of rapid proliferation, high efficiency, and broad spectrum in killing of tumor cells. However, there was few report about its clinical application on treatment for cancer patients. The current study was designed to evaluate the effect of adoptive transfer of autologous CIK cells on the patients with advanced malignant tumor. METHODS: Peripheral blood mononuclear cells of the patients with advanced malignant tumor were separated by fractionation on Ficoll-Hypaque gradient, then cultured in the medium containing IFN-gamma, IL-2, and CD3McAb for 7 days in vitro, and than the cultured auto-CIK cells were transfused back to the patients. The numbers of transferred CIK cells per patient were 5-15 x 10(9) in one course of treatment. Among these patients, 47 cases received chemotherapy, 3 cases received radiotherapy before CIK cells transfusion. The interval between chemoradiotherapy and immunotherapy was over 2 to 4 weeks. RESULTS: Among 63 patients receiving CIK cells immunotherapy, the total effective rate (PR + MR) was 44.46% (28). In the patients with increasing of CEA level in serum, 14 cases showed reduction of serum CEA and 1 cases remained increasing after the treatment with CIK cell. In the patients with increasing of AFP level in serum, similarly, 9 cases showed reduction of serum AFD and 1 case remained increasing. The absolute members of CD3, CD4, and CD8T cells increased to over 45% after being treated with CIK cells. Among treated patients, the appetite of 51 cases and performance and sleep of 32 cases got improved. Among 18 cases, 13 cases showed the pain relief. CONCLUSION: Adoptive immunotherapy of auto-CIK cells can significantly enhance cellular immune functions and improve subjective symptoms, but without side effects, so this is a safe and effective treatment for the patients with malignant tumor.