Experiments were designed to assess age-related changes in the transplantability of human tumours xenografted in congenitally athymic (nu/nu) mice. It has been found that the number of progressively growing human tumour xenografts decreased significantly with increasing age of BALB/c nu/nu recipients. These findings, taken together with a previously recognized increase in the frequency of endogenous interleukin 2 (IL-2)-producing cells with age of nu/nu mice, prompted us to investigate whether administration of exogenous IL-2 to young adult nu/nu mice could change the transplantability of human tumours in the mice. Peritumoral administration of exogenous interleukin 2 to 8-week-old nu/nu mice inhibited the growth of the human tumour xenografts. In vitro activation of nu/nu splenocytes with exogenous Il-2 resulted in the generation of killer cells which have been found to be cytolytic when allowed to react with human tumour targets in 51Cr cytotoxicity assay. In addition, it has been found that the percentage of IL-2-activated Thy 1.2+ and ASGM1+ cells substantially increased with increasing age of nu/nu spleen cell donors. These findings are compatible with the hypothesis that the observed age-related decrease in takes of human tumour xenografts might be determined by the increasing level of IL-2 production and subsequent maturation of IL-2-dependent effector cells.
- MeSH
- antigeny povrchové analýza MeSH
- buněčné dělení MeSH
- buňky K aktivované lymfokiny imunologie MeSH
- cytotoxicita imunologická MeSH
- experimentální sarkom patologie MeSH
- HeLa buňky MeSH
- interleukin-2 farmakologie MeSH
- karcinom z přechodných buněk patologie MeSH
- lidé MeSH
- lymfom T-buněčný patologie MeSH
- myši nahé růst a vývoj MeSH
- myši MeSH
- nádory děložního čípku patologie MeSH
- nádory močového měchýře patologie MeSH
- průtoková cytometrie MeSH
- slezina růst a vývoj imunologie MeSH
- stárnutí fyziologie MeSH
- transplantace heterologní MeSH
- transplantace nádorů * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antigeny povrchové MeSH
- interleukin-2 MeSH
We have analyzed 14 human urothelial cell lines in two different transformation grades for the production of a tumor-associated-alpha-2-macroglobulin. It has been shown in radioimmunoprecipitation and immunoblotting tests that human urothelial cells in vitro do not synthesize the wide-spectrum proteinase inhibitor alpha-2-macroglobulin. Four of the tested cell lines were additionally tested for the expression of specific mRNA which resulted in negative findings. The significance of the absence of alpha-2-macroglobulin in the human urothelial cells is discussed.
- MeSH
- alfa-makroglobuliny biosyntéza imunologie MeSH
- invazivní růst nádoru MeSH
- králíci MeSH
- lidé MeSH
- melanom metabolismus MeSH
- nádorová transformace buněk MeSH
- nádorové buňky kultivované MeSH
- nádory močového měchýře metabolismus MeSH
- precipitinové testy MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- alfa-makroglobuliny MeSH
We have prepared a retroviral expression construct, pPS-IL-2, in which human IL-2 cDNA has been inserted into the polylinker region, and have used the retroviral vector to introduce the functional IL-2 gene into a fibroblast cell line, RAT-1. Peritumoral administration of IL-2-producing RAT-1 cells into congenitally athymic (nu/nu) mice carrying subcutaneous transplants of human carcinoma cells inhibited the growth of the human tumour xenografts.
- MeSH
- buňky NK imunologie MeSH
- experimentální nádory genetika imunologie terapie MeSH
- fibroblasty imunologie transplantace MeSH
- genetické inženýrství * MeSH
- genetické vektory MeSH
- HeLa buňky imunologie patologie MeSH
- interleukin-2 biosyntéza genetika MeSH
- lidé MeSH
- myši nahé MeSH
- myši MeSH
- Retroviridae genetika MeSH
- transformace genetická MeSH
- transplantace nádorů MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- interleukin-2 MeSH
The production of interleukin 2 (IL-2) by phytohaemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC) from 21 patients with transitional cell carcinoma of the urinary bladder (BTCC) and 16 control blood donors was measured with a solid phase enzyme immunoassay based on the dual antibody immunometric sandwich principle. PBMC from patients with invasive BTCC (grade III-IV) showed a defect in the production of IL-2. The concentration of IL-2 in the supernatants of PBMC cultures from these patients was substantially lower (0.4 +/- 0.1 U/ml) than that observed in the supernatants of PBMC cultures from patients with non-invasive BTCC, grade II (1.5 +/- 0.7 U/ml), and from tumour-free controls (1.4 +/- 0.8 U/ml). These results suggest an immune dysfunction based on quantitatively impaired IL-2 production in patients with invasive BTCC and indicate that exogenous IL-2 could be used as an immunological response modifier for the treatment of these patients.
- MeSH
- aktivace lymfocytů MeSH
- fytohemaglutininy MeSH
- interleukin-2 biosyntéza MeSH
- karcinom z přechodných buněk imunologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- lymfocyty imunologie metabolismus MeSH
- nádory močového měchýře imunologie MeSH
- senioři MeSH
- techniky in vitro MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- fytohemaglutininy MeSH
- interleukin-2 MeSH
The present study was performed to localize in the IL-2 molecule the active site responsible for interaction with the IL-2 receptor. To predict the receptor binding site on the IL-2 molecule, a computer programme based on the hypothesis that the active site will contain parts of the protein molecule having a high tendency to form a bend was utilized. The tendency to form a bend was evaluated by assessing the probability of beta-turn occurrence; the highest probability was found in the tetrapeptide Asn-Pro-Lys-Leu, occupying the positions 33-36 of the IL-2 molecule. Accordingly, the hexadecapeptide H-Cys-Nle-Gly-Ile-Asn-Asn-Tyr-Lys-Asn-Pro-Lys-Leu-Thr-Arg-Met-Leu-NH2 that spans over the predicted tetrapeptide Asn-Pro-Lys-Leu and comprises the region 27-40 from the IL-2 amino acid sequence was synthesized. This synthetic (I-16) peptide was found to selectively inhibit the IL-2-dependent uptake of 3H-TDR by CTLL cells, apparently by competing with IL-2 for the IL-2 receptor. The synthetic I-16 hexadecapeptide was conjugated to carrier (BSA) protein and used for immunization of rabbits. Resulting I-16 antibodies were capable of binding specifically to the I-16 hexadecapeptide in indirect ELISA test; they reacted substantially with IL-2-producing but not with IL-2-non-producing Jurkat cells in indirect cell membrane immunofluorescence, and inhibited activation of killer spleen cells with human recombinant IL-2 as detected by 51Cr microcytotoxicity assay. Taken together, these results suggest that at least one of the receptor contact sites of the IL-2 is localized within the N-terminal part of the molecule in the region defined by amino acids 27-40 and coded for by the exon 1 and 2.
- MeSH
- aktivace lymfocytů MeSH
- biotest MeSH
- buněčná imunita MeSH
- cytotoxicita imunologická MeSH
- ELISA MeSH
- epitopy MeSH
- fluorescenční protilátková technika MeSH
- imunologické techniky MeSH
- interleukin-2 * imunologie metabolismus MeSH
- oligopeptidy chemická syntéza imunologie metabolismus MeSH
- protilátky MeSH
- receptory imunologické metabolismus MeSH
- receptory interleukinu-2 MeSH
- vazebná místa MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- epitopy MeSH
- interleukin-2 * MeSH
- oligopeptidy MeSH
- protilátky MeSH
- receptory imunologické MeSH
- receptory interleukinu-2 MeSH
Highly purified recombinant human interleukin 2 induced cytotoxicity of lymphocytes from urinary bladder carcinoma patients and from control healthy donors when added during an 18-h 51Cr microcytotoxicity assay against bladder carcinoma (T24) target cells. Similar levels of killer cell activation were detected in mononuclear cell preparations from bladder carcinoma patients and control healthy donors; hence, no defect in the responsiveness of bladder carcinoma patients' lymphocytes to interleukin 2 could be observed. The effect of the recombinant interleukin 2 was dose-dependent. Addition of monoclonal antibody 7E9 directed against cell-type restricted antigen associated with the T24 target cells and capable of inducing antibody-dependent cellular cytotoxicity could not increase the cytotoxicity-inducing effects of interleukin 2.
- MeSH
- aktivace lymfocytů MeSH
- buněčná cytotoxicita závislá na protilátkách MeSH
- buňky NK imunologie MeSH
- cytotoxicita imunologická MeSH
- imunoterapie MeSH
- interleukin-2 imunologie MeSH
- lidé MeSH
- monoklonální protilátky MeSH
- nádory močového měchýře imunologie terapie MeSH
- techniky in vitro MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- interleukin-2 MeSH
- monoklonální protilátky MeSH
In previous communications we have demonstrated that crude rat interleukin 2 and partially purified mouse interleukin 2 were capable of inhibiting growth of transplantable, MC-induced mouse sarcomas in syngeneic recipients. Here we report that repeated peritumoral injections of highly purified human recombinant interleukin 2 can inhibit growth of these mouse sarcomas and prolong survival of tumour-bearing mice. These findings taken together and the ready availability of high doses of recombinant human interleukin 2 substantiate our proposal for initiation of clinical trials using local administration of the interleukin 2 (Bubeník et al. 1983) in selected cancer patients.
- MeSH
- experimentální sarkom chemicky indukované terapie MeSH
- imunoterapie MeSH
- inbrední kmeny myší MeSH
- interleukin-2 imunologie izolace a purifikace MeSH
- lidé MeSH
- methylcholanthren MeSH
- myši MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- interleukin-2 MeSH
- methylcholanthren MeSH
Selectivity of mouse monoclonal antibody 7E9 (IGG3) directed against human urinary bladder carcinoma cells has been examined by indirect membrane immunofluorescence, using a panel of 31 human cell lines. The 7E9 monoclonal antibody discriminated between urinary bladder carcinoma cells and normal urothelium or cells of non-urothelial origin, although occasional reactions with bladder carcinoma-unrelated cell types were observed. The 7E9 antibody was purified by protein A affinity chromatography, labeled with 131I and used for gamma scintigraphy in nude mice xenografted with human urinary bladder carcinoma T24. The 7E9 antibody was capable of locating the T24 xenografts in nude mice; it localized preferentially in the T24 tissue compared to normal mouse tissues. The T24 xenografts could not be detected by gamma scintigraphy with 131I-labelled monoclonal antibody against human mammary carcinoma cells and two other control antibodies. Likewise, the 131I-labelled 7E9 antibody was not capable of locating human mammary carcinoma xenografts in nude mice.
- MeSH
- buněčné linie MeSH
- fluorescenční protilátková technika MeSH
- karcinom z přechodných buněk diagnostické zobrazování imunologie MeSH
- lidé MeSH
- monoklonální protilátky * imunologie MeSH
- myši nahé MeSH
- myši MeSH
- nádory močového měchýře diagnostické zobrazování imunologie MeSH
- nádory prsu imunologie MeSH
- protilátky nádorové * imunologie MeSH
- radioisotopová scintigrafie MeSH
- radioizotopy jodu MeSH
- specificita protilátek MeSH
- tkáňová distribuce MeSH
- transplantace nádorů MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- monoklonální protilátky * MeSH
- protilátky nádorové * MeSH
- radioizotopy jodu MeSH
