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1.
Immunogenic proteins with identical molecular mass (64kDa) were purified from a syngeneic spontaneous T cell leukaemia line, designated LB3, and lymphoblast extracts both derived from BALB/c mice. The 64-kDa protein was purified by a sequence of biochemical steps from cell extracts containing protease inhibitors. The following steps were included in the purification pathway: Sephadex G-100 gel filtration, anion-exchange chromatography, concanavalin A (ConA) affinity chromatography, and preparative gel electrophoresis. The immunogenic fraction isolated in each step was subjected to the next step along the purification pathway. The immunogenicity of the separated fractions was measured by a lymph-node proliferation assay, which is indicative of delayed-type hypersensitivity. The final 64-kDa isolated protein of blast cells induced in BALB/c mice an efficient lymphnode proliferation response, which was detected in the regional lymph node after challenge with the final isolated protein of LB3 cells and vice versa. In addition to their identical molecular mass, both proteins were eluted from an anion exchange column with the same NaCl concentration (0.57 M) and both expressed affinity to the ConA-Sepharose column, suggesting that they are glycosylated. The specificity of the immunological responses induced or elicited with the various isolated proteins was also shown. The implications of these findings are discussed. 相似文献
2.
Xu G Zhang Y Zhang L Ren G Shi Y 《Biochemical and biophysical research communications》2008,375(4):666-670
Bone marrow stromal cells (BMSCs) have been shown to promote the growth and survival of a wide variety of tumors. However, in the present study, we found that BMSCs induced apoptosis of lymphoma cells in the presence of INFγ and TNF. IFNγ and TNF dramatically induced the expression of inducible nitric oxide synthase (iNOS) by BMSCs in culture, and BMSCs generated from iNOS knockout mice did not induce apoptosis of lymphoma cells in the presence of IFNγ and TNF. In addition, we found that IFNγ and TNF also increased IL-6 expression by BMSCs, and anti-IL-6 further increased the killing of tumor cells by BMSCs. Taken together, our findings indicate that BMSCs induce apoptosis of lymphoma cells in the presence of IFNγ and TNF, and that the proapoptotic effect of BMSCs is mediated by nitric oxide. Our findings suggest a possibility to harness this proapoptotic feature of BMSCs for the development of novel therapeutic strategy to eliminate tumor cells, especially tumor cells in bone marrow. 相似文献
3.
Liposomal delivery of CTL epitopes to dendritic cells 总被引:5,自引:0,他引:5
The induction of strong and long lasting T-cell response, CD4+ or CD8+, is a major requirement in the development of efficient vaccines. An important aspect involves delivery of antigens to dendritic cells (DCs) as antigen presenting cells (APCs) for the induction of potent antigen-specific CD8+ T lymphocyte (CTLs) responses. Protein or peptide-based vaccines become an attractive alternative to the use of live cell vaccines to stimulate CTL responses for the treatment of viral diseases or malignancies. However, vaccination with proteins or synthetic peptides representing discrete CTL epitopes have failed in most instances due to the inability for exogenous antigens to be properly presented to T cells via major histocompatibility complex (MHC) class I molecules. Modern vaccines, based on either synthetic or natural molecules, will be designed in order to target appropriately professional APCs and to co-deliver signals able to facilitate activation of DCs. In this review, we describe the recent findings in the development of lipid-based formulations containing a combination of these attributes able to deliver tumor- or viral-associated antigens to the cytosol of DCs. We present in vitro and pre-clinical studies reporting specific immunity to viral, parasitic infection and tumor growth. 相似文献
4.
We have shown, in a preliminary report, that macrophages can induce strand breaks in the DNA of co-cultured tumor cells (Chong et al., 1988). The present study is designed to determine if oxygen-centered species generated by the cell-free enzyme-substrate combination of hypoxanthine and xanthine oxidase can induce similar lesions and to identify the specific mediator(s). We report that co-incubation of murine mammary tumor cell lines with hypoxanthine and xanthine oxidase leads to the induction of DNA-strand breaks as determined by fluorescence analysis of DNA unwinding (FADU) assay or alkaline elution techniques. This damage is preventable by catalase which removes hydrogen peroxide but no protection is provided by agents to remove or prevent the formation of superoxide anion (superoxide dismutase), or hydroxyl radical (mannitol or the iron chelator o-phenanthroline). Likewise, cyclooxygenase or lipoxygenase inhibitors of arachidonate metabolism (indomethacin, nordihydroguaiaretic acid, caffeic acid) or bromophenacyl bromide do not alter the degree of DNA scission. Treatment with higher doses of oxygen species leads to significant toxicity as determined by evaluation of cell growth potential or colony-forming ability. Again, toxicity is prevented only by the presence of catalase. Tumor cells are able to rejoin strand breaks at lower, less toxic doses. When comparing different tumor cell subpopulations at various stages of progression, i.e., metastatic vs. nonmetastatic, for sensitivity to hydrogen peroxide-induced strand breakage, we found that at lower concentrations (less than 5μM) metastatic populations are sensitive whereas nonmetastatic populations exhibit no significant breakage. At higher concentrations of hydrogen peroxide, all lines were sensitive, suggesting that a lower threshold of sensitivity may exist for more progressed tumour cell lines. 相似文献
5.
6.
Immunotherapy of cancer could be possible in cases in which competent effector T cells can be induced. Such an approach depends on expression of tumour-specific antigens by the tumour cells and on the availability of sufficient costimulatory support for activation of cytotoxic T lymphocytes. Here, a strategy for helper T cell recruitment for induction of tumour-specific cytotoxic immune responses is presented. Allogenic MHC class II molecules were introduced into tumour cells by cell fusion. These hybrid cells, when injected into mice, induced rejection of an established tumour. The contribution of CD4-expressing helper T cells in the induction phase and of CD8-expressing T cells in the effector phase of the immune response was demonstrated. The approach described could be applicable to cases in which a suitable tumour antigen is present but not identified; it employs regulatory interactions that govern physiological immune responses and is designed to be minimally invasive. 相似文献
7.
Allogeneic gastric cancer cell-dendritic cell hybrids induce tumor antigen (carcinoembryonic antigen) specific CD8+ T cells 总被引:3,自引:0,他引:3
Matsumoto S Saito H Tsujitani S Ikeguchi M 《Cancer immunology, immunotherapy : CII》2006,55(2):131-139
The development of protocols for the ex vivo generation of dendritic cells (DCs) has led to intensive research of their potential
use in immunotherapy. Accumulating results show the efficacy of this treatment on melanomas which are highly immunogenic.
However, its efficacy remains unclear in other tumors. In this study, allogeneic gastric cancer cell–DC hybrids were used
to determine the efficacy of this type of immunotherapy in gastric cancer. Fusion cells of DC and allogeneic gastric cancer
cells were generated by polyethylene glycol (PEG) and electrofusion. These hybrids were used to induce tumor associated antigen
(TAA) specific cytotoxic T lymphocytes (CTLs). The DCs were successfully fused with the allogeneic gastric cancer cells resulting
in hybrid cells. These hybrid cells were functional as antigen-presenting cell because they induced allogeneic CD4+ T cells
proliferation. CD8+ T cells stimulated by the MKN-45-DC hybrid cells were able to kill MKN-45 when used for immunization.
The CTLs killed another gastric cancer cell line, MKN-1, as well as a melanoma cell line, 888mel, suggesting the recognition
of a shared tumor antigen. MKN-45 specific CTLs can recognize carcinoembryonic antigen (CEA), indicating that the killing
is due to tumor antigens as well as alloantigens. This approach suggests the possible use of allogeneic gastric cancer cell–DC
hybrids in DC based immunotherapy for gastric cancer treatment. 相似文献
8.
Asfour IA El-Tehewi MM Ahmed MH Abdel-Sattar MA Moustafa NN Hegab HM Fathey OM 《Biological trace element research》2009,127(3):200-210
The present study was undertaken to explore the effect of administration of high doses of sodium selenite on the apoptosis
of lymphoma cells in patients with non-Hodgkin’s lymphoma (NHL). Forty patients with newly diagnosed NHL were randomly divided
into two groups. Group I received standard chemotherapy, whereas group II received adjuvant sodium selenite 0.2 mg kg−1 day−1 for 7 days in addition to chemotherapy. Flow cytometry was used for monitoring of lymphoma cells apoptosis at the time of
diagnosis and after therapy in the two groups. Sodium selenite administration resulted in significant increase in percentage
of apoptotic lymphoma cells after therapy in group II (78.9 ± 13.3% versus 58.9 ± 18.9%, p < 0.05). In addition, patients who received sodium selenite treatment demonstrated statistically significant increase in
percentage of reduction of cervical and axillary lymphadenopathy, decrease in splenic size, and decreased percentage of bone
marrow infiltration. Also, we found a statistically significant decrease in cardiac ejection fraction (CEF) in group I and
no reduction in CEF in patients who received sodium selenite ‘group II’, denoting the cardioprotective effect of selenium.
It is concluded that sodium selenite administration at the dosage and duration chosen has synergistic effect to chemotherapy
in inducing apoptosis and, consequently, could improve clinical outcome. 相似文献
9.
M. Stoeck C. Marland-Noske M. Manasterski R. Zawatzky S. Horn V. Möbus P. Schlag V. Schirrmacher 《Cancer immunology, immunotherapy : CII》1993,37(4):240-244
In order to understand further the effects of Newcastle-disease-virus(NDV)-modified tumour vaccines we investigated the feasibility of isolating lymphocytes from the site of injection of patients undergoing postoperative active specific immunization (ASI) with autologous NDV-modified tumour cells. Delayed-type-hypersensitivity(DTH)-like reactions from five cancer patients were surgically removed, minced and the tissue particles were digested with collagenase and DNase. Lymphoid cells recovered were expanded in a highly efficient limiting-dilution analysis system optimized for T cell growth [Moretta et al. (1983) J Exp Med 157: 743] and lymphocyte microcultures (clonal probability >0.8) could be grown for up to 1 year. Analysis of the microcultures for phenotype and function showed that the majority were positive for CD4 (92%) and TCR (96%). Concanavalin-A-induced production of interleukin-2 (IL-2), IL-6, interferon and tumour necrosis factor was detected in more than 70% of the microcultures. Lectin-dependent cytotoxicity was only very rarely observed. The general characteristics of the microcultures obtained support the notion of a DTH-like reaction taking place at the site of tumour cell challenge. The possibility of in vitro expansion and cultivation of T lymphocytes from ASI vaccination sites should help to elucidate further the role of these cells in active specific immunization against autologous tumour cells.This study was supported by Dr.-Mildred-Scheel-Stiftung and the Tumorzentrum Heidelberg 相似文献
10.
Efficient uptake and rapid degradation of plasmid DNA by murine dendritic cells via a specific mechanism 总被引:3,自引:0,他引:3
Yoshinaga T Yasuda K Ogawa Y Takakura Y 《Biochemical and biophysical research communications》2002,299(3):389-394
In spite of the important roles of dendritic cells in DNA-based therapies, the cellular uptake mechanism of plasmid DNA (pDNA) in dendritic cells is poorly understood. The present study was undertaken to investigate the binding and uptake of pDNA in vitro using a murine dendritic cell line, DC2.4 cells. A significant and time-dependent cellular association of [32P]pDNA with DC2.4 cells was observed at 37 degrees C and this fell markedly at 4 degrees C. The binding and uptake of [32P]pDNA were significantly inhibited by cold pDNA, polyinosinic acid (poly[I]), dextran sulfate, or heparin, but not by polycytidylic acid (poly[C]), dextran, or EDTA, suggesting that a specific mechanism mediated by a receptor like the macrophage scavenger receptor may be involved. The TCA precipitation experiments showed that DC2.4 cells rapidly endocytosed and degraded a significant amount of [32P]pDNA at 37 degrees C and released the degradation products into the medium. The pDNA degradation was also significantly inhibited by poly[I], but not poly[C]. The rate of pDNA degradation by DC2.4 cells was significantly higher than that by macrophages. A confocal microscopic study using fluorescein-labeled pDNA confirmed the rapid internalization and degradation of pDNA by the dendritic cells. Taken together, these results indicate that pDNA is efficiently taken up and rapidly digested by the dendritic cells via a specific mechanism. These findings may suggest the important role of the dendritic cells in the innate immune system for host defense. 相似文献
11.
CD38 has been widely characterised both as an ectoenzyme and as a receptor. In the present paper, we investigated the role of CD38 as possible modulator of apoptosis. CD38-positive (CD38(+)) and negative (CD38(-)) fractions, obtained by sorting CD38(+) cells from lymphoma T (Jurkat) and lymphoma B (Raji) and by transfecting lymphoma LG14 and myeloid leukemia K562 cell lines, were used. Cellular subpopulations were exposed to different triggers (H(2)O(2), UV-B, alpha-TOS and hrTRAIL) and the extent of apoptosis was determined by Annexin V-FITC/PI assay. Our data showed that, in lymphoma cells, propensity to apoptosis was significantly linked to CD38 expression and that, remarkably, such response was independent of the nature of the trigger used. Inhibition of CD38 expression by antisense oligonucleotides treatment resulted in CD38-silenced fractions which were as prone to apoptosis as CD38(-) ones. Notably, susceptibility of K562 to apoptosis-inducing challenges was not affected by CD38 expression. 相似文献
12.
G. Heuff H. S. A. Oldenburg H. Boutkan J. J. Visser R. H. J. Beelen N. Van Rooijen C. D. Dijkstra S. Meyer 《Cancer immunology, immunotherapy : CII》1993,37(2):125-130
The evidence that Kupffer cells are capable of controlling metastatic growth in the liver in vivo is largely circumstantial. The best approach when studying natural cytotoxicity activities of Kupffer cells is to investigate the effect of Kupffer cell elimination on tumour growth. Until now it has not been possible to eliminate Kupffer cells without affecting other cell populations. We have recently developed a new method to eliminate Kupffer cells selectively: intravenous injection of liposome-encapsulated (dichloromethylene)bisphosphonate (Cl2MDP-liposomes) leads to effective elimination of all Kypffer cells, without affecting non-phagocytic cells. Wag/Rij rats were injected with Cl2MDP-liposomes. After 48 h, rats were inoculated with syngeneic CC531 colon carcinoma cells by injection in the portal system. The results show a strongly enhanced tumour growth in the liver of the Cl2MDP-liposometreated rats. In these animals, livers were almost completely replaced by tumour and had increased in weight, whereas in the control groups only a few (four to eight) small (1-mm) tumour nodules were found. These data show that selective elimination of Kupffer cells results in enhanced tumour growth in the liver, implying that Kupffer cells play a crucial role in controlling tumour growth in the liver. 相似文献
13.
Grande S Luciani AM Rosi A Palma A Giovannini C Sapora O Guidoni L Viti V 《FEBS letters》2007,581(4):637-643
1H MRS signals of glutathione and of free glutamate were examined in samples from cultured tumour cells, namely MCF-7 from mammary carcinoma and TG98 from malignant glioma, with the aim of relating signal intensities to aspects of GSH metabolism. Spectra of cells harvested at different cell densities suggest that GSH and glu signal intensities are related to cell density and proliferation and their ratio is dependent on the activity of the gamma-glutamyl cysteine synthetase. The hypothesis is confirmed by experiments performed on cells treated with buthionine sulfoximine that inhibits the enzyme activity. 相似文献
14.
Michele Caraglia Pierosandro Tagliaferri Pierpaolo Correale Geppino Genua Antonio Pinto Silvana Del Vecchio Giuseppe Esposito Angelo Raffaele Bianco 《Cancer immunology, immunotherapy : CII》1993,37(3):150-156
We report that cytosine arabinoside (Ara-C), a cytosine analogue that at low doses causes phenotypical changes on human leukemia cells in vitro and in vivo, induces growth inhibition of oropharyngeal cancer KB and lung adenocarcinoma A549 cell lines. An increase in the number of epidermal growth factor and transferrin receptors (EGFR, TrfR) is induced by Ara-C on these cells. Maximal EGFR up-regulation occurs 96 h after the beginning of Ara-C exposure while maximal TrfR up-regulation is detected 24 h later. These effects occur without changes in the affinity of EGFR and TrfR for their ligands. Two classes of EGF-binding sites with aK
d of 0.055 nM and 2.3 nM respectively, and one class of transferrin-binding sites with aK
d of about 4 nM are detected on both untreated and Ara-C-treated KB cells. [3H]Thymidine uptake is clearly stimulated on KB cells by nanomolar concentrations of EGF and transferrin, whereas in Ara-C-treated cells [3H]thymidine uptake is not increased by EGF and transferrin under conditions where maximal EGFR and TrfR up-regulation occurs. The enhanced EGF and transferrin binding is paralleled by a twofold increase of in vitro targeting of Ara-C-treated KB and A549 cells with anti-EGFR 108.1 mAb and anti-TrfR OKT9 mAb. We propose that Ara-C could provide a new approach for the improvement of the therapeutic index of anti-EGFR and anti-TrfR immunoconjugates.This work has been supported by the Italian Association for Cancer Research (A.I.R.C.) and by the National Council of Research (C.N.R.) of Italy, contract 92.02274.PF39 相似文献
15.
A. Mouras I. Negrutiu Y. Dessaux 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1987,74(2):253-260
Summary Phenotypic and genetic variations of tumour cells were analysed both quantitatively and qualitatively in clones and subclones of a crown-gall strain. Thus, growth rates, grafting tests, octopine synthesis, estimations of the T-DNA contents, modifications in the numbers, and structures of chromosomes were examined. Phenotypic variations are closely associated with genetic changes, including variation in chromosome number (which is shown to be non-specific to the tumoral state) and, above all, variation in the copy-number of T-DNA sequences per cell, and structural rearrangements of chromosomes. Such rearrangements are characterized by specific marker chromosomes in the tumour cells and they correlate with the degree of oncogenicity of the cells. 相似文献
16.
《Journal of enzyme inhibition and medicinal chemistry》2013,28(2):360-369
Carbonic anhydrase IX (CA IX) is a hypoxia-regulated enzyme, overexpressed in many types of human cancer. CA IX is involved in pH homeostasis, contributing to extracellular acidification and tumourigenesis. Acidification of the extracellular milieu can impact upon cellular uptake of chemotherapeutic drugs by favouring weak acids (e.g. melphalan), but limiting access of weak bases (e.g. doxorubicin). We investigated whether alterations of CA IX activity affected anti-cancer drug uptake and toxicity. CA inhibitor acetazolamide (AZM) enhanced doxorubicin toxicity but reduced melphalan toxicity in cell lines that highly expressed CA IX under anoxic conditions (HT29 and MDA435 CA9/18). The toxicity changes reflected modification of passive drug uptake. AZM did not alter toxicity or uptake in cells with low CA IX activity (HCT116 and MDA435 EV1). AZM lowered intracellular pH in HT29 and MDA435 CA9/18 cells under anoxic conditions. CA IX activity has chemomodulatory properties and is an attractive target for anti-cancer therapy. 相似文献
17.
A novel method for the comparative analysis of cell motility by direct viewing was developed and used in a preliminary study of sarcoma cells. Three cell lines from the RPS family of sarcomas in inbred LEW/CUB rats which differ in the incidence of spontaneous metastasis were studied. A system of four different culture conditions, designed to mimic the stress within the tumour, was created by changing and combining two variables: the pH of the medium (with 2% calf serum only), which was either physiological at 7.4 or 6.6; and the adhesiveness of the culture substratum, which was either at a standard level or decreased. It was found that changing the pH from 7.4 to 6.6 in a single step slowed the cells down, and also changed the way in which they moved, from «walking« in random directions to moving in a more directional way. Making the culture surface less adhesive sped up cell locomotion. Decreasing the adhesiveness of the culture substratum and the pH simultaneously stimulated the migration of highly metastasizing cells preferentially. Thus we have found a way to distinguish between highly metastasizing A297Nb sarcoma cells and poorly metastasizing T15 and non-metastasizing K2 sarcoma cells, a distinction that could not be made by a simple examination of the cells. It is concluded that a comparison of cell motility by directly viewing the cells under different conditions may be useful when investigating the in vitro motility properties of malignant cells. 相似文献
18.
Thomas S. Vates Garry L. Brake S. K. Majumdar Gail L. Ferguson 《In vitro cellular & developmental biology. Plant》1986,22(6):305-310
Summary Cis-diamminedichloroplatinum II (cis-DDP), an antitumor drug and the inactive trans-isomer were studied to evaluate their
effects on cell multiplication, DNA synthesis, and surface morphology of the murine erythroleukemia cells (clone 6A11A). Short-term
treatment of cells (1h) with 5 and 10μg/ml of cis-DDP resulted in a significant inhibition of cell multiplication. Continuous
treatment with cis-DDP (up to 144 h) significantly arrested cell growth at 1,5, and 10μg/ml. The cells exposed to 10 μg/ml
trans-DDP exhibited a slight decrease in cell multiplication; however, the 25-μg/ml treatments showed a modest inhibition
of cell growth. Continuous treatment with cis-DDP resulted in a concentrationdependent decrease in DNA synthesis, although
low-dose treatment (0.05 and 0.1 μg/ml), with a few exceptions, had no relative inhibitory effect. Likewise, trans-DDP treatments
decreased tritiated thymidine incorporation; however, this inhibitory effect was not as drastic as with corresponding concentrations
of cis-DDP. Scanning electron microscope studies revealed the formation of many giant cells and blebs at all short-term treatment
concentrations of cis-DDP past the 48 h interval. Continuous treatment of cis-DDP at 1 μg/ml concentration produced giant
cells with minute holes, whereas the 5 and 10 μg/ml exposure resulted in the formation of blebs and large holes and reduction
of microvilli past the 48-h treatment period. At higher concentrations the continuous treatment of cis-DDP completely destroyed
the cells. The surface morphology of trans-isomer treated cells, in most instances, resembled the corresponding untreated
control cells. 相似文献
19.
Todryk SM Eaton J Birchall L Greenhalgh R Soars D Dalgleish AG Melcher AA Pandha HS 《Cancer immunology, immunotherapy : CII》2004,53(4):323-330
Vaccination with established tumour cell lines may circumvent the problem of obtaining autologous tumour cells from patients, but may also need immunological adjuvants. Up-regulation of heat shock proteins within tumour cell vaccines has resulted in increased immunogenicity in some models, but this has yet to be demonstrated for allogeneic (MHC-disparate) cell vaccines. This was investigated here using a rat model for prostate tumour cell vaccination. Heating of tumour cells (42°C, 1 h) elicited significant increases in HSP70 expression. Vaccination with heated autologous PAIII cells elicited protection against PAIII challenge in 60% of rats >50 days compared to 0% with unheated vaccine and was associated with an increased Th1 (IFN) immune response. Heated allogeneic MLL cells elicited significant protection against PAIII challenge, in contrast to unheated cells. The principle was confirmed in two mouse models, although the allogeneic melanoma vaccine K1735 elicited the best protection when heated and administered mixed with autologous dendritic cells. Thus, while heating of vaccine cells in some models is highly beneficial, and is a means of enhancing immunogenicity without genetic modification or inclusion of potentially toxic adjuvants, additional immune enhancement may be required. 相似文献
20.
Aladin F Lautscham G Humphries E Coulson J Blake N 《Cancer immunology, immunotherapy : CII》2007,56(8):1143-1152
It is becoming increasingly apparent that the majority of tumours display defects in the MHC class I antigen processing pathway, particularly low levels of the transporters-associated with antigen processing (TAP) and tapasin. Thus, immunotherapy approaches targeting such tumours with CD8+ cytotoxic T lymphocytes (CTL) requires strategies to overcome these defects. Previously we had identified an antigen processing pathway by which cytosolically derived hydrophobic peptides could be presented in the absence of TAP. Here we show in the tapasin-negative cell line 721.220 that a number of these hydrophobic TAP-independent peptides can also be presented in a tapasin-independent manner. Yet when these experiments were extended to tumour cell lines derived from small cell lung cancer (SCLC), which we show to be tapasin deficient in addition to TAP-negative, the TAP-, tapasin-independent peptides were not presented. This lack of presentation could be rectified by pre-treatment of SCLC cells with IFNgamma. Alternatively, by directing the TAP-, tapasin-independent peptides into the endoplasmic reticulum (ER) via an ER signal sequence, these peptides were presented efficiently by SCLC cells. We infer from this data that the TAP-independent pathway for presentation of hydrophobic peptides generates a low concentration of peptide in the ER and, for tumour cells which also lack tapasin, this concentration of antigenic peptide is insufficient to load onto MHC class I molecules. Thus, for immunotherapeutic approaches to target SCLC and other tumours with defects in the MHC class I antigen processing pathway it will be important to consider strategies that address tapasin-defects. 相似文献