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  • 1
    ISSN: 1432-1211
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0851
    Keywords: Key words Newcastle disease virus (NDV) ; Bispecific antibodies ; Hemagglutinin-neuraminidase ; T cell activation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  A new type of cancer vaccine for therapeutic application in cancer patients is described. It consists of three components. (1) autologous tumor cells, (2) Newcastle Disease Virus (NDV), to be used for infection and (3) bispecific antibodies (bsAb) which attach to the viral hemagglutinin neuraminidase (HN) molecule on the infected tumor cells. A standardized procedure has been developed for generating virus infected human autologous tumor cell vaccines (ATV-NDV) which includes cell dissociation, removal of leukocytes and cell debris, gamma-irradiation and cryopreservation. Infection with the non-virulent strain NDV Ulster is performed within 30 min of co-incubation. While virus infection already increased immunogenicity of the tumor vaccine, further augmentation of T cell stimulatory capacity is achieved by attachment of specially designed bi-specific antibodies (bs HN × CD28 or bs HN × CD3).
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0851
    Keywords: Key words Tumour resistance ; Cytokine mRNA ; T cell activation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  This study examines whether a correlation may be found between Th1- or Th2-type cytokine responses and resistance or susceptibility to tumour growth. Cytokine profiles were investigated in a well-defined mouse tumour model in which the injection site and the genetic background determine the phenotype of either tumour resistance or tumour susceptibility. DBA/2-derived ESb lymphoma variant cells with high metastatic capacity were inoculated into syngeneic mice either s.c., where they grow and metastasize, or into the ear pinna (i.e.), where they do not grow because of induction of protective immunity. Alternatively, the tumour cells were injected s.c. or i.e. into allogeneic B10.D2 mice, which are resistant to the tumour although they are identical at the MHC locus. Between 1 and 10 days after tumour cell injection the spleen-derived mRNA was tested for cytokine gene expression or the spleen cells were analysed by FACScan for T cell activation. The strongest cytokine response was observed in i.e. inoculated B10.D2 mice. This was characterized by an early (days 2–3) peak of interferon γ (INF-γ), interleukin-2 (IL-2), IL-2 receptor α (IL-2Rα) and IL-4. The cytokine mRNA response of i.e. inoculated DBA/2 mice was quite similar except that no IFN-γ could be detected. In s.c. inoculated B10.D2 mice, the IL-2, IL-2Rα and IFN-γ responses were weaker than after i.e. injection while the IL-4 response was comparable. The most striking difference between these cytokine profiles from tumour-resistant mice and those of s.c. inoculated tumour-susceptible DBA/2 mice was a delay in the latter in the IL-2, IL-2Rα and IFN-γ responses and the observation that the IL-4 response was not down-regulated. The persisting IL-4 response could down-regulate a Th1-type response and thereby explain tumour susceptibility as a consequence of host conditioning.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-0851
    Keywords: Active specific immunization ; NDV-modified tumour cells ; Microcultures ; Tumour vaccines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract 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 TCRαβ (96%). Concanavalin-A-induced production of interleukin-2 (IL-2), IL-6, interferon γ and tumour necrosis factor α 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.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-0711
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1433-8726
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Thirty patients with advanced renal cell carcinoma (RCC), 23 of whom had distant metastases in at least one organ, were entered after nephrectomy into a protocol involving vaccination with Newcastle disease virus (NDV)-modified autologous tumour material, with a subsequent induction week and repetitive bi-weekly cycles of interleukin-2 (rIL-2) and interferon α2b/rIFN-α2b at a lower s. c. dose (1.5 million Cetus units m−2 day−1 every 12 h on 2 days and 3 million IU/m2 once a day on days 1, 3 and 5). The inpatient treatment was followed by a maintenance phase during which 0.3 million Cetus units/m2 rIL-2 was given s. c. every 12 h on days 1–5 and 3 million IU m−2 day−1 rIFN-α2b was administered on days 1, 3, and 5 on an outpatient basis. All but 3 patients completed the induction week and 6 weeks of outpatient therapy. No grade 3 or 4 toxicities occurred during the therapy. Therapy was discontinued for 3 patients because of rapid tumour progression. Of the 23 evaluable RCC patients, 3 exhibited a complete response and 4 displayed partial remission, 7 showed stable disease during 1–18 months (median = 5 months) of therapy, and progression was seen in 9. We conclude that vaccination with autologous tumour material combined with s. c. rIL-2 and rIFN-α2b administration can induce regressions in patients with advanced RCC and that even in non-responding patients a more favourable course of the disease can be achieved.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1573-7276
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Various murine tumour sublines which differed considerably in their in vivo metastatic capacity were tested in vitro for their ability to invade normal tissue. For this purpose we developed two quantitative tests, a Boyden chamber endothelial cell invasion assay and a brain tissue microsphere invasion assay. The invasion of [75Se]methionine-prelabelled tumour cells into the normal tissues was followed by measuring the percentage of tumour-associated label in the brain microspheres or the endothelial monolayers after 12–48 h of co-cultivation. Clear and comparable differences existed in both assays between the amount of radiolabel found in the normal tissues after a co-cultivation with the different tumour lines. In three of the four tumour lines invasiveness correlated with metastatic capacity. The fourth line, a plastic adherent variant, was highly invasive but low metastatic. The ability of tumour cells to invade normal tissue, therefore, while necessary for the generation of metastases, is not in itself sufficient. Since both assays are independent of time-consuming histological sectioning and staining and allow a quantitative determination of invasive capacity of tumour cells grown as single cell suspensions they appear well suited for experimental manipulation and for screening of anti-invasive drugs.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1573-7276
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A variant of the highly metastatic mouse lymphoma ESb was isolated from ESb suspension culture by repeated selection for adhesiveness to plastic. The adhesion variant ESb-MP was shown to have greatly reduced malignant potential and a change in organotropism. ESb-MP cells metastasized into the kidney with a high frequency (80% of mice), whereas the ESb cells only rarely infiltrated the kidney (4% of mice). Two subclones of the adherent variant (ESb-M2 A8, ESb-M2 H6) showed a reduced malignant potentialin vivo very similar to the ESb-MP cell line, but only the clone ESb-M2 A8 infiltrated the kidney. Cell motility and chemotaxis of the various tumor lines was tested in a modified Boyden chamber assay. ESb-MP cells always migrated about 45 μm into a nitrocellulose filter of 8 μm pore size during 4 h incubation, whereas ESb cells and the two subclones migrated only to a distance of 12–25 μm. There was no apparent correlation betweenin vitro motility and malignancyin vivo. The addition of several factors known to be chemoattractant for granulocytes (C5a, FMLP) or for tumor cells (FMLP, laminin, fibronectin) did not affect the migration of ESb or ESb-MP cell lines. Enhanced migration of the ESb-MP cell line was, however, observed when kidney conditioned medium (KCM) was used as attractant in the lower chamber. This was not true for ESb cells. Additionally, within the subclones of the ESb-MP cell line, there was a remarkable correlation between kidney metastasisin vivo and responsiveness to KCMin vitro. The motility stimulating effect of KCM was due to a chemotactic activity. ESb cells, which did not respond to KCM, responded to other factor(s) derived from inflammatory exudate fluid. We conclude that the stimulation of tumor cell motility by an organotropic factor was highly selective for certain tumor lines. The correlation, between responsivenessin vitro and kidney metastasisin vivo, suggests that it is not so much the random migration of tumor cells which plays a role in metastasis but the motility following selective stimulation by tissue-derived factors and chemoattractants.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1573-7276
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Peritoneal macrophages from normal DBA/2 mice were found to bind significantly more cells of a syngeneic low metastatic lymphoma line (Eb) than cells of a high metastatic variant (ESb) derived therefrom. These differences were observed in three different assays, at 4°C and at 37°C, and at various ratios of macrophages to tumor cells. Upon co-culture with normal macrophages, a tumor cytostatic effect was consistently observed with Eb but not with ESb tumor cells. Further experiments indicated that macrophages exerted their growth inhibitory effect via direct tumor cell contact. Pre-treatment of tumor cells with neuraminidase or pre-treatment of macrophages with lens culinaris lectin increased the numbers of macrophages binding Eb and ESb tumor cells. Addition of D-galactose or D-mannose at 50 mM concentration led to an increase of tumor cell binding and tumor cytostatic activity. Taken together, these results suggest (i) that carbohydrates play a role in tumor cell recognition by macrophages and (ii) that the differences observed between Eb and ESb tumor cells may be due to differences in the expression of carbohydrates. Pre-activation of the macrophages by lymphokine(s) led to a short increase in their tumor cell binding capacity. Lymphokine activation resulted in a strong but also short-lived increase of tumor cytostatic potential. This was effective against both the low and the high metastatic tumor line.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1573-7276
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Repeated administration of the hepatic lectin blocking agents D-galactose or arabinogalactan completely prevented the settling of metastatic cells of sarcoma L-1 tumor in the liver of Balb/c mice and greatly reduced the colonization process of highly metastatic ESb lymphoma cells of the liver of DBA/2 mice. Therefore, when hepatic lectins were blocked with competitive glycoconjugates, tumor cell colonization of the liver could be prevented in two different model systems.
    Type of Medium: Electronic Resource
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