No abstract available.
Animals ; Indomethacin* ; Killer Cells, Lymphokine-Activated* ; Mice*

No abstract available.
Humans ; Killer Cells, Lymphokine-Activated* ; Killer Cells, Natural*

No abstract available.
Interferon-alpha* ; Interleukin-2* ; Killer Cells, Lymphokine-Activated*

OBJECTIVESTo evaluate the effects of K562-dendritic cell (DC) and Raji-DC fusion vaccines on the cytotoxicity of cord blood (CB) derived cytokine-induced killer/natural killer (CIK/NK) cells.

METHODSDC and CIK/NK cells were derived from CB mononuclear cells. CB-DC were fused with inactivated K562 or Raji cells by PEG to form K562 or Raji-DC fusion vaccine. The CIK/NK cells stimulated by different co-culture antigens were three groups: K562-DC or Raji-DC fusion vaccine group, inactivated K562 or Raji plus DC group, and CB-DC alone group. The cytotoxicity of CIK/NK cells stimulated by different co-culture antigens was measured by MTT test.

RESULTSAll the antigens used for stimulation could enhance the cytotoxicity of CB-CIK/NK cells, with no specificity difference. At 20:1 effector-target ratio, the cytolytic activities of K562-DC and Raji-DC fusion vaccine groups against Raji cells were (75.44 +/- 4.19)% and (81.33 +/- 4.18)% respectively (P < 0.05); and that of inactivated K562 + DC and Raji + DC group against Raji cells were (73.12 +/- 4.22)% and (80.49 +/- 4.27)%, respectively (P < 0.01). There was no significant difference in the cytotoxicity to K562 cells between the two fusion vaccine groups (P > 0.05). The cytotoxicity of CB-CIK/NK cells immunized by Raji cells was higher than that by K562 cells. In CIK/NK cells co-stimulated by the same tumor antigen, there was no significant difference in the cytotoxicity between DC fusion vaccine group and inactivated cells plus DC group to different tumor cells.

CONCLUSIONSThe cytotoxicity of CB-CIK/NK cells to tumor cells was not specific. There was no significant difference in the cytotoxic activity of CB-CIK/NK cells between the DC fusion vaccine group and inactivated cells plus DC group.

Cancer Vaccines ; immunology ; pharmacology ; Cells, Cultured ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; Fetal Blood ; cytology ; Humans ; K562 Cells ; Killer Cells, Lymphokine-Activated ; immunology ; Killer Cells, Natural ; immunology

BACKGROUND: Nitric oxide (NO), a cytotoxic molecule is produced in various tissues including tumor cells during interleukin-2 (IL-2) therapy . Lymphokine-activated killer (LAK) cells are induced during IL-2 therapy, and have cytotoxic activity against tumor cells. The current study investigated the effects of NO synthesized in target cells or exposure of target cells to NO on the sensitivity of target cells to LAK cell cytotoxicity. METHODS: Cytotoxicity was measured using 4 h chromium release assays. LAK cells which were induced by a 4 day incubation of BALB/c mouse splenocytes with IL-2 (6,000 IU/mL) were employed as effector cells. RD-995 skin tumor cells originated from a C3H/HeN mouse were employed as target cells. NO synthesis in target cells was induced by a 24 h incubation of RD-995 cells with IFN gamma (25 U/mL), TNF (50 U/mL) and IL-1 (20 U/mL). S-nitrosyl acetylpenicillamine (SNAP), an NO donor, was used to expose target cells to NO. N(G) -monomethyl-L-arginine (MLA) and carboxy-PTIO were added during cytotoxicity assays to inhibit NO synthesis, and to scavenge NO produced by target cells, respectively. RESULTS: Sensitivity of NO-producing RD-995 cells to LAK cell cytotoxicity was decreased by addition of MLA and carboxy-PTIO during cytotoxicity assays. However, the two reagents had no effect on the sensitivity of non-NO-producing RD-995 cells. Pretreatment of RD-995 target cells with SNAP increased the sensitivity in comparison with untreated cells. CONCLUSIONS: Sensitivity of target cells to LAK cell cytotoxicity is increased by target cell NO synthesis or exposure to NO. Further studies are needed to evaluate whether these in vitro results have relevance to in vivo phenomena.
Animals ; Chromium ; Humans ; Indicators and Reagents ; Interleukin-1 ; Interleukin-2 ; Killer Cells, Lymphokine-Activated* ; Lymphocytes ; Mice ; Nitric Oxide* ; Skin ; Tissue Donors

BACKGROUND: Murine IL-2-induced lymphokine-activated killers can be divided into two mutually exclusive subset:NK1.1'CD8 and NK1.1 CD8+. However, there is a strong evidence that NK cell may belong to T cell lineage. Recently novel lymphocyte subsets, present in the adult murine thymus, CD3+NK1.1'TCRap(TNK) cell is readily identifiable in fresh obtained murine adult CD4 CD8 thymocytes. MATERIAL AND METHOD: We sorted out CD4 and CD8 (double negative.' DN) cells and CD8+ cells from murine spleen and cultivated these cells with IL-2. And the surface B220, CD8, NK1. 1 and cytopasmic NK1.1 was analysed simultaneously to see whether these cells can be switched to the other subtype of cells. RESULT: Purified DN cells were switched to several subtype of cells'. CD8'B220+(LAK cells), NK1.1'B220+(LAK cells), CD8 B220, cytoplasmic NK1.1+B220 cells. Purified CD8 cells were switched to CD8+B220' LAK cells and cytoplasmic NK1.1+ CD8+ B220+ and cytoplasmic NK1.1' CD8 B220 cells. In addition, the CD8' cells originated from DN cells do not express the cytoplasmic NK1.1 in contrary to the sorted CD8 cells. CONCLUSION: Our results indicated that these will be useful models to investigating CD8 precursor potentials in populations of CD4 CD8 (doble negative) cells and relationship of NK1.1 These results also supports the hypothesis that T cells and NK cells have same ontogeny and CD8 effector functions are potentially diverse and could be exploited by various conditions that switch off host protected cytolytic response. These model offer a way to study the molecular regulation of CD8 gene expression.
Adult ; Cell Lineage ; Cytoplasm ; Gene Expression ; Humans ; Interleukin-2* ; Interleukins* ; Killer Cells, Lymphokine-Activated ; Killer Cells, Natural ; Lymphocyte Subsets ; Spleen ; T-Lymphocytes ; Thymocytes ; Thymus Gland

This study was aimed to investigate the proliferation activities and phenotype changes of DC, CIK and DC-CIK, and their cytotoxicity against hepatocarcinoma cells in co-culture of DC with CIK. Peripheral blood mononuclear cells (PBMNC) were isolated from healthy adult donors. After incubation of PBMNC for 2 hours, DCs were induced from adherent cells by some cytokines and CIKs were generated from non-adherent cells. Mature DCs were harvested after incubation for 9 days, and then were co-cultured with CIK at ratio of 1:5 for 3 days. The cytotoxicity activity against SMMC-7721 hepatocellular carcinoma cell line was detected by MTT assay. The results showed that CIK cells were able to lyse SMMC-7721 hepatocellular carcinoma cells at low ratios of effector to target. This effect was significantly enhanced by co-culture with DCs. It is concluded that CIK cells have high lytic activity against 7721 hepatocellular carcinoma cell line, which can be enhanced by co-culture with DC. DC-CIK cells are highly effective immune cells.
Carcinoma, Hepatocellular ; immunology ; pathology ; Cells, Cultured ; Coculture Techniques ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; Humans ; Killer Cells, Lymphokine-Activated ; cytology ; immunology ; Liver Neoplasms ; immunology ; pathology

The purpose was to investigate the effect of phytohemagglutinin (PHA) on proliferation and cytotoxicity of cytokine-induced killer (CIK). Peripheral blood mononuclear cells (PBMNCs) from healthy donors were divided into two groups. Cells were resuspended and maintained in complete medium containing of 10% autologous plasma. CIK cells were cultured by traditional method in group one. The other group cells were added PHA to stimulate PBMNCs for 24 hours, then cultured like incubating CIK cells. Their cytotoxicity to different target cells was evaluated by (51)Cr release assay. The results showed that the proliferation multiples of CIK and PHA-CIK cells were both high, however, the latter was much higher than CIK with significance (P < 0.05). Cells in each group cells showed high cytotoxicity. At the same high effector/target ratio PHA-CIK cells cytotoxicity was stronger than CIK cells when targets were K562 cells or acute leukemia cells (P < 0.05). In conclusion, PHA-CIK cells exhibit stronger proliferation and cytotoxicity than CIK cells, and the result provides an experimental basis for biotherapy.
Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cells, Cultured ; Coculture Techniques ; Cytotoxicity Tests, Immunologic ; Cytotoxicity, Immunologic ; drug effects ; Humans ; K562 Cells ; Killer Cells, Lymphokine-Activated ; cytology ; immunology ; Leukocytes, Mononuclear ; cytology ; immunology ; Phytohemagglutinins ; pharmacology

In order to investigate whether cytokine-induced killer (CIK) cells can induce apoptosis of bcr-abl(+) K562 cells, apoptosis of K562 cells and CEM cells induced by CIK cells, etoposide or camptothecin was detected with flow cytometry DNA assay. RT-PCR showed that K562 cells expressed the bcr-abl fusion gene, K 562 cells, K562 cells/etoposide or K562 cells/camptothecin groups showed no sub-G(1) peak. K562 cells/CIK cells group showed sub-G(1) peak (38.1%). CEM cells showed no sub-G(1) peak. CEM cells/camptothecin or CEM cells/etoposide groups showed sub-G(1) peak (23.5% or 32.3% respectively). CEM cells/CIK cells group showed sub-G(1) peak (45.4%). Etoposide or camptothecin did not induce apoptosis of K562 cells. CIK cells induce apoptosis of K562 cells. Bcr-abl fusion gene prevented apoptosis induced by etoposide or camptothecin, but did not prevent apoptosis induced by CIK cells. This property may support the observed adoptive immunologic effect of allogeneic bone marrow transplantation and donor lymphocyte transfusions of CML case relapsing after allogeneic bone marrow transplantation.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; immunology ; Camptothecin ; pharmacology ; Coculture Techniques ; Cytotoxicity, Immunologic ; Etoposide ; pharmacology ; Flow Cytometry ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; K562 Cells ; drug effects ; immunology ; metabolism ; Killer Cells, Lymphokine-Activated ; cytology ; immunology

Total RNA was extracted from human LAK cell, and a cDNA encoding mature peptide HMG-17 and its alpha helix domain was amplified by RT-PCR. The recombinant prokaryotic expression vector pGEX-1lambdaT-HMG-17 and pGEX-1lambdaT HMG-17alpha helix was constructed. Using affinity chromatography, thrombin cleaving and AU-PAGE elution, we obtained the purified HMG-17. Analyses of MIC, MEC and MBC indicated that HMG-17 and HMG-17alpha had strong antibacterial activity. MIC of the alpha-helic domain was almost the same as that of HMG17, suggesting that the alpha-helic structure would be essential for the antibacterial activity of HMG-17.
Anti-Bacterial Agents ; biosynthesis ; pharmacology ; Escherichia coli ; genetics ; metabolism ; HMGN2 Protein ; biosynthesis ; genetics ; pharmacology ; Humans ; Killer Cells, Lymphokine-Activated ; chemistry ; Peptides ; genetics ; pharmacology ; Prokaryotic Cells ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology