ABSTRACT
Background: To assess the increased cellular immunity of Peripheral Blood Mononuclear Cells (PBMC) derived LAK cells from endometriosis patients towards endometriosis cell cultures after stimulation with IL-2. Methods: This study is a quasi-experimental study of pre and post treatment using controls. Phenotype evaluation of CD3+CD4+, CD3+CD8+ and CD56+ effector cells of PBMC from endometriosis patients and controls was performed. Cytotoxicity test of PBMC from endometriosis patients and control towards Daudi, K562 cell line and endometriosis cell cultures using 51Chromium release assay was also carried out. Results: Phenotype evaluation of PBMC from endometriosis patients (n = 10) and controls (n = 6) were done prior to and after IL-2 stimulation. Before IL-2 stimulation, CD3+CD4+, CD56+ from endometriosis group (n = 10) tend to be lower than control (n=6) whereas CD3+CD8+ were higher in endometriosis group than controls. After IL-2 stimulation, CD3+ CD8+, CD56+ of PBMC from endometriosis group were significantly increased (p < 0.05). Cytotoxicity test revealed a significant increase (p < 0.05) in both PBMC’s effector cells from endometriosis and control group towards target cells, Daudi, and K562 cell lines after IL-2 stimulation. PBMC’s effector cells cytotoxicity from both endometriosis and control towards target endometriosis cell cultures were also elevated after IL-2 stimulation. Conclusion: LAK cells derived IL-2 stimulated PBMC from endometriosis patients increased cellular immunity towards endometriosis cell cultures.
Subject(s)
Endometriosis , Killer Cells, Lymphokine-ActivatedABSTRACT
Objective To identify the changes in proliferative activity and cytotoxicity of lymphokine activated killer(LAK) cells after in vitro co-culture with dendritic cells from umbilical cord blood(CBDC). Methods CBDCs were induced by using GM-CSF and IL-4,and then co-cultrued with allogenic LAK at different proportions.Proliferative activity and cytotoxicity of LAK cells were measured by MTT assay.Results The typical CBDCs were induced in vitro,and the cultured CBDCs stimulated the proliferation and cytotoxicity of LAK cells.The optimal proportion of CBDC and LAK was 1:10,at which the activated LAK showed the highest cytotoxic effect on the tumor cells.Conclusion The CBDCs can enhance the proliferation and cytotoxicity of LAK cells.
ABSTRACT
Objective:As the function of ??T cells and NK in the immunological therapy is showing more and more important,the characteristics of ?? T cells,NK and LAK were analyzed comparatively after that they were richened by isolating and incubating in vitro.Methods:The cells were collected using MACS after the cells were panned respectively with special monoclonal antibodies.Then the characteristics including proliferation,phenotype,cytotoxin and the down regulation blocked by specific antibodies were analyzed.Results:The ?? T cells can expand 600～800 times after culturing 2 weeks and the percent of CD3,CD8 and ?? expressed on the collected cells were 72.29%,58.02% and 65.98% respectively ?? T cells showed the high cytotoxin to K562(NK sensitive cell line),Raji and XG 7 cell lines(NK non sensitive cell lines).The percentage of cytotoxin reached 35.98%,52.27% and 69.08% respectively compared with ??T respectively.No obviously change of percent of ??T cytotxic ability to these target cells were observed using special MHC class I monoclonal antibody to block the ??T cell before coculturing the target cells with ?? T cells.Conclusion:All of ?? T cells,NK and LAK showed the non specific cytotoxin to tumor cells.The cytotoxic capability of ?? T cells did not be effected after blocking with MHC class I monoclonal antibody.These results demonstrated that ?? T cell could kill more kinds of tumor cells than NK and LAK.
ABSTRACT
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.
Subject(s)
Adult , Humans , Cell Lineage , Cytoplasm , Gene Expression , Interleukin-2 , Interleukins , Killer Cells, Lymphokine-Activated , Killer Cells, Natural , Lymphocyte Subsets , Spleen , T-Lymphocytes , Thymocytes , Thymus GlandABSTRACT
Some murine (YAC, P815 and SP20) and human (Molt4, Raji and HR7) tumour cell lines were (i) treated with IFN-γ for inducing enhanced expression of MHC class I antigen, or (ii) given a brief treatment with citrate buffer (pH 3.0), which resulted in denaturation of class I MHC antigens on these tumour cells. IFL-γ or acid treated tumour cells were used as unlabelled competing targets in cold target inhibition assays. The results indicated that the competing ability of acid-treated tumour cells remained unaltered, whereas IFN-γ treated tumour cells competed with significantly less efficiency. These results have been evaluated in light of the current view of NK cell development and the expression of inhibitory receptors for MHC class I molecules (IRMs), on NK cells. A modified view on NK cell heterogeneity based upon IRM expression has been proposed which reconciles several apparently discordant observations about the activity and role of NK cells. Two classes of NK cells have been proposed. Type I NK ceils have target recognition receptors which do not recognize autologous normal cells, lack IRMs, and may participate in first line of defence against transformed cells in vivo. Type II NK cells have target recognition receptors for autologous normal cells and express at least one selfreactive IRM in order to prevent auto-killing. Type II NK cells participate in killing those transformed cells which down-regulate their MHC class I expression in order to escape cytotoxic T-cell surveillance. It is also postulated that mechanism of inverse correlation of target cell MHC class I expression levels and their susceptibility to NK cells, involves interference model of missing self hypothesis for type I NK cells and inhibitory signal model of missing self hypothesis for type II NE cells. Finally, it is proposed that acid treatment of tumour cells enhances their lysis susceptibility by making them additionally susceptible to type II NK cells, rather than enhancing their killing by type I NK cells. This proposition would explain the lack of effect of acid treatment on the competing ability of tumour cells, when target cells are only lysed by type I NE cells.
ABSTRACT
The peripheral blood lymphocytes (PBLs) from 111 cancer patients were isolated and cultured respectively for 23 - 27 days in the medium mainly conditioned by IL-2 and PHA. With ~(125) I-UdR release method, sampling in random way, we examed the cytotoxicities of PBLs and lymphokine-activated-killer (LAK) cells in different culture periods in vitro. The statistic analysis on sufficient data gave the following results: 1. The cytotoxicity against K562 cells increased from 34.78 ?25% of the PBLs to 68.04 ?17.3% of the cells cultured for 8-13 days, afterward, kept about 70% to 23 - 27 days. The constitutional proportion patterns showed that the freshly isolated samples dispersed at a wide range of cytotoxicities (10 - 90%), and that most of the cultured samples ( ~ 85%) concentrated on the range of higher cytotoxicities (50 ~ 95% ) after 8-13 days. 2. The cytotoxicity against Raji cells rose from 8.9 ?8% of the fresh PBLs to 42.1 ?22% of the LAK cells at 8 - 13 days, and maintained about 35% in the following periods. The constitutional proportion patterns of the cytotoxicity against Raji illustrated that all the fresh PBL samples were below 25% of cytotoxicity, and that during the culture, one part of the samples ( ~ 30%) acquired the higher cytotoxicities (50 -90% ), but the other part of the samples ( - 40%) remained at lower cytotoxicities (below 35% ) . The mechanisms behind these phenomena are worth further investigating.
ABSTRACT
Highly active adherent LAK cells (A-LAK) with monocytes depleted by phenylalahine methyl ester (PME) were cultured from peripheral blood lymphocytes of patients with hepatocellular carcinoma (HCC). Results showed that A-LAK cells cultured in about 14 days had expanded better and faster than that of nonadherent LAK cells (NA-LAK) with their greatest expansion varied from 23 to 243 fold .A-LAK cells showed a trend of increase in TH cell subgroup and decrease in Ts cell sub-group as well as significant difference of TH/TS ratio. The IL-2R expression increased from 34.1%to 64.3%. A-LAK cells had a higher cytotoxicity (64.6%)than that of NA-LAK cells (42.8%).Further clinical application of A-LAK cells may improve biotherapeutic effect on HCC patients compared with that of NA-LAK cells .
ABSTRACT
Objective: To investigate the role and mechanism of histamine in regulation of C57BL/6 mice spleen derived LAK activity in vivo. Methods: The C57BL/6 mice carrying B16 pulmonary metastatic melanoma were treated with LAK/IL 2/histamine therapy or LAK/IL 2 therapy with the aim of evaluating both anti tumor responses in vivo. Results: It was found that the addition of histamine effectively potentiated the anti metastatic effect produced by LAK/IL 2 therapy and induced regression of NK resistant B16 pulmonary metastatic melanoma. Survival period was significantly prolonged in mice receiving LAK/IL 2/histamine as compared with LAK/IL 2 therapy alone. The effect of histamine was completely blocked by H 2 R antagonist ranitidine, and mimicked by dimaprit, a H 2 R agonist. Conclusion:Histamine, via specific activation of H 2 R, may be an important regulator of LAK cells activity; histamine and LAK/IL 2 synergistically induce more potent antitumor efficacy in vivo .
ABSTRACT
TNF gene was transfected into murine LAK cells by retrovirus. Our results showed that TNF gene-transfected LAK cells secreted TNF more than normal LAK cells and control gehe-transfected LAK cells. The in vitro growth ability and cytotoxicity of TNF gene-transfected LAK cells were augmented significantly.The cytotoxicity of ,TNF gene-transfected LAK cells was markedly inhibited by anti - TNF monoclonal antibody, indicating that the, above augmentation was mediated by TNF secreted by transfected LAK cells. Significant therapeutic effect on the ascitic liver carcinoma.-bearing mice was achieved by i.p. injection of low dosage TNF gene transfected LAK cells and IL - 2.
ABSTRACT
Although TNF alone was not capable of inducing LAK activity and could not enhance the LAK activity induced by IL-2 at optimal concentration (1000U/ml), TNF (500U/ml) was found to act synergistically with IL-2 at suboptimal concentration (10～100U/ml). This TNF/IL-2 synergistic effect was blocked by anti-TNF McAb (Z8) or anti-IL-2R? chain McAb (TU27). When TNF and IL-2/LAK cells were combined to treat tumor-bearing mice,TNF could potentiate significantly the therapeutic effect of IL-2/LAK cells in vivo.
ABSTRACT
In the present study it was observed that the reactivity of splenocytes to Con A,LPS and IL-2 was enhanced in the mice which were given Total Soyasaponin(Ts)Via mouth?We also found that the level of IL—2 in splenocytes and the activity of NK and LAK cells were obviously higher than that in the control group.
ABSTRACT
Peripheral blood mononuclear cells (PBMNC) isolated from patients with acute leukemia (AL) and from normal controls were cultured in medium containing 1000 units/ml of recombinant interleukin 2 (IL-2). Marked LAK activity was induced on the third culture day in the normal controls,, with the highest cytotoxicity appearing between day 3 and 5 whereas induction of LAK activity in the AL patients began on the 5th day of culture, with the peak level appearing at day 15, showing that the peak of LAK activity was significantly delayed in AL. LAK cells surface phenotyping tests showed that CD8+ and CD16+ positive cells began to increase significantly from day 5 and reached the highest level at week 3, whereas CD4+ subclass began to decrease on day 5 and dropped to the nadir at week 3. The proportion of CD8+ and CD16+ cells were positively cor related with LAK activity, but, that of the CD4+ cell was inversely related with the LAK activity.