Treatment of murine EL4 T cell lymphoma with N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer conjugates of doxorubicin (Dox) leads to complete tumor regression and to the development of therapy-dependent longlasting cancer resistance. This phenomenon occurs with two types of Dox conjugates tested, despite differences in the covalent linkage of Dox to the polymer carrier. Such a cancer resistance cannot fully express in conventional treatment with free Dox, due to substantial immunotoxicity of the treatment, which was not observed in the polymer conjugates. In this study, calreticulin (CRT) translocation and high mobility group box-1 protein (HMGB1) release was observed in EL4 cells treated with a conjugate releasing Dox by a pH-dependent manner. As a result, the treated tumor cells were engulfed by dendritic cells (DC) in vitro, and induced their expression of CD80, CD86, and MHC II maturation markers. Conjugates with Dox bound via an amide bond only increased translocation of HSPs to the membrane, which led to an elevated phagocytosis but was not sufficient to induce increase of the maturation markers on DCs in vitro. Both types of conjugates induced engulfment of the target tumor cells in vivo, that was more intense than that seen with free Dox. It means that the induction of anti-tumor immunity documented upon treatment of EL4 lymphoma with HPMA-bound Dox conjugates does not rely solely on CRT-mediated cell death, but involves multiple mechanisms.
- MeSH
- antigeny CD80 metabolismus MeSH
- antigeny CD86 metabolismus MeSH
- apoptóza účinky léků MeSH
- chemorezistence účinky léků MeSH
- dendritické buňky cytologie imunologie MeSH
- doxorubicin aplikace a dávkování analogy a deriváty chemie toxicita MeSH
- fagocytóza MeSH
- kalretikulin metabolismus MeSH
- koncentrace vodíkových iontů MeSH
- kyseliny polymethakrylové aplikace a dávkování chemie toxicita MeSH
- lymfom T-buněčný farmakoterapie imunologie MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nosiče léků chemie MeSH
- protein HMGB1 metabolismus MeSH
- proteiny teplotního šoku metabolismus MeSH
- protinádorové látky aplikace a dávkování chemie toxicita MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antigeny CD80 MeSH
- antigeny CD86 MeSH
- doxorubicin-N-(2-hydroxypropyl)methacrylamide copolymer conjugate MeSH Prohlížeč
- doxorubicin MeSH
- kalretikulin MeSH
- kyseliny polymethakrylové MeSH
- nosiče léků MeSH
- protein HMGB1 MeSH
- proteiny teplotního šoku MeSH
- protinádorové látky MeSH
The cytostatic effects of polymeric conjugates based on N-(2-hydroxypropyl)methacrylamide copolymers (PHPMA) and containing doxorubicin bound through amide and hydrazone bonds (mixed conjugates) were compared with the cytostatic effects of monoconjugates containing drug bound through an amide or hydrazone bond. One group of mixed conjugates was formed from two comonomers containing doxorubicin bound to the methacryloyl group through a spacer and an amide (DOX(AM)) or hydrazone (DOX(HYD)) bond via copolymerization with HPMA. A second group of mixed conjugates was formed from two different interconnected HPMA copolymers, one containing DOX(AM) and the other DOX(HYD), forming a high-molecular-weight branched structure. The third mixed polymeric system was a simple mixture of monoconjugates DOX(AM)-PHPMA and DOX(HYD)-PHPMA. Simultaneous treatment with all mixed forms of the polymeric derivatives of doxorubicin significantly increased antitumor efficacy after application of monoconjugates, suggesting a synergizing effect that could be used in designing new doxorubicin-containing therapeutic systems.
- MeSH
- akrylamidy chemie MeSH
- amidy chemie MeSH
- buněčné linie MeSH
- doxorubicin chemie farmakologie MeSH
- fluorescenční mikroskopie MeSH
- hydrazony chemie MeSH
- lidé MeSH
- lymfom T-buněčný farmakoterapie MeSH
- molekulární struktura MeSH
- myši inbrední C57BL MeSH
- myši nahé MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- polymery chemická syntéza chemie MeSH
- protinádorová antibiotika chemie farmakologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- akrylamidy MeSH
- amidy MeSH
- doxorubicin MeSH
- hydrazony MeSH
- N-(2-hydroxypropyl)methacrylamide MeSH Prohlížeč
- polymery MeSH
- protinádorová antibiotika MeSH
N-(2-Hydroxypropyl)methacrylamide (HPMA) copolymer carrier containing the anticancer drug doxorubicin bound either by a proteolytically degradable bond (non-targeted PK1 or targeted with alpha-CD71 mAb) or by a hydrolytically degradable bond were synthesised and tested in vivo for various biological properties. Mouse 38C13 B-cell lympoma was used as a well established and defined cell line for this study. 38C13 cells are sensitive to free doxorubicin and IC50 was very low, about 0.014 microM. PK1 showed a strongly decreased cytostatic effect, IC50 being 12.6 microM. alpha-CD71 targeted conjugate, which can be considered as an antibody-targeted form of PK1, had IC50 0.358 microM. HPMA copolymer with doxorubicin bound via a hydrolytically sensitive bond (HYD conjugate) showed a high cytostatic effect with IC50 about 0.052 microM. We demonstrated that HYD conjugate inhibited DNA synthesis and induced p21(Waf1/Cip1) protein expression (p21(Waf1/Cip1) is cyclin-dependent kinase inhibitor which blocks cell cycle progression) as quickly as free doxorubicin, whereas PK1 acted much more slowly. Similarly, apoptosis induction measured by Annexin V binding and Caspase 3 activity was detected later after incubation of cells with PK1 or alpha-CD71 targeted conjugate. Apoptosis was manifested by elevation of bax and bad mRNA levels, which was much more rapid and intense in the case of free doxorubicin and HYD conjugate. Expression of antiapoptotic genes as well as cyclin-dependent kinases was surprisingly not affected.
- MeSH
- akrylamidy chemická syntéza metabolismus farmakologie MeSH
- apoptóza účinky léků MeSH
- buněčná membrána účinky léků patologie MeSH
- buněčný cyklus genetika MeSH
- DNA antagonisté a inhibitory genetika metabolismus MeSH
- doxorubicin chemická syntéza metabolismus farmakologie MeSH
- exprese genu účinky léků genetika MeSH
- geny myc účinky léků genetika MeSH
- hydrazony chemická syntéza metabolismus farmakologie MeSH
- hydrolýza MeSH
- inhibiční koncentrace 50 MeSH
- inhibitor p21 cyklin-dependentní kinasy MeSH
- kaspasa 3 MeSH
- kaspasy škodlivé účinky účinky léků metabolismus MeSH
- ligandy * MeSH
- messenger RNA genetika MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- preklinické hodnocení léčiv metody MeSH
- proliferace buněk účinky léků MeSH
- protein Bad MeSH
- protein X asociovaný s bcl-2 MeSH
- proteiny buněčného cyklu genetika metabolismus MeSH
- protoonkogenní proteiny c-bcl-2 genetika metabolismus MeSH
- receptory transferinu účinky léků genetika MeSH
- thymidin metabolismus MeSH
- transportní proteiny genetika metabolismus MeSH
- tritium MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- akrylamidy MeSH
- Bad protein, mouse MeSH Prohlížeč
- Bax protein, mouse MeSH Prohlížeč
- Casp3 protein, mouse MeSH Prohlížeč
- Cdkn1a protein, mouse MeSH Prohlížeč
- DNA MeSH
- doxorubicin MeSH
- hydrazony MeSH
- inhibitor p21 cyklin-dependentní kinasy MeSH
- kaspasa 3 MeSH
- kaspasy MeSH
- ligandy * MeSH
- messenger RNA MeSH
- N-(2-hydroxypropyl)methacrylamide co-polymer-doxorubicin conjugate MeSH Prohlížeč
- protein Bad MeSH
- protein X asociovaný s bcl-2 MeSH
- proteiny buněčného cyklu MeSH
- protoonkogenní proteiny c-bcl-2 MeSH
- receptory transferinu MeSH
- thymidin MeSH
- transportní proteiny MeSH
- tritium MeSH
When feeding on vertebrate host ticks (ectoparasitic arthropods and potential vectors of bacterial, rickettsial, protozoal, and viral diseases) induce both innate and specific acquired host-immune reactions as part of anti-tick defenses. In a resistant host immune defense can lead to reduced tick viability, sometimes resulting in tick death. Tick responds to the host immune attack by secreting saliva containing pharmacologically active molecules and modulating host immune response. Tick saliva-effected immunomodulation at the attachment site facilitates both tick feeding and enhances the success of transmission of pathogens from tick into the host. On the other hand, host immunization with antigens from tick saliva can induce anti-tick resistance and is seen to be able to induce immunity against pathogens transmitted by ticks. Many pharmacological properties of saliva described in ticks are shared widely among other blood-feeding arthropods.
- MeSH
- arachnida jako vektory MeSH
- imunita účinky léků MeSH
- infestace klíšťaty imunologie MeSH
- interakce hostitele a parazita účinky léků imunologie MeSH
- klíšťata chemie imunologie mikrobiologie MeSH
- nemoci přenášené klíšťaty imunologie přenos MeSH
- slinné proteiny a peptidy farmakologie MeSH
- slinné žlázy fyziologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- slinné proteiny a peptidy MeSH
Tick salivary gland extract (SGE) was previously shown to inhibit murine T cell proliferation. In mice, SGE has an inhibitory effect on Th1 and a stimulatory effect on Th2 cytokine elaboration. In the present study, tick-mediated immunomodulation of human T cell proliferation and cytokine elaboration was analyzed using human peripheral blood mononuclear cells (PBMCs) stimulated with concanavalin A (Con A) or lipopolysaccharide (LPS). Using flow cytometry, tick saliva-induced changes were investigated in human mononuclear cell subpopulations. SGE from Ixodes ricinus dose-dependently inhibited human T cell proliferation. This finding supports the flow cytometry data, showing that the percentage of Con A-activated HLA-DR-CD3+ T lymphocytes and CD4+ CD8+ double-positive T cells decreased after SGE treatment. SGE significantly inhibited the in vitro production of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) secreted by Th1 lymphocytes. In contrast, the elaboration of IL-4, IL-6, and IL-10 secreted by Th2 lymphocytes was significantly stimulated by I. ricinus SGE. Similarly, the production of both IL-1alpha and IL-1beta was significantly stimulated after SGE treatment. These data indicate that the tick-induced immunomodulatory events in humans are similar to those previously described in a murine model.
- MeSH
- adjuvancia imunologická MeSH
- aktivace lymfocytů MeSH
- cytokiny biosyntéza MeSH
- klíště imunologie MeSH
- kultivované buňky MeSH
- lidé MeSH
- slinné žlázy imunologie MeSH
- T-lymfocyty cytologie imunologie MeSH
- Th2 buňky cytologie imunologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- adjuvancia imunologická MeSH
- cytokiny MeSH
Schistosome cercariae of the genus Trichobilharzia are the causative agent of swimmers' itch. In order to characterize the changes in parasites during and after the penetration of the host skin, in vitro and in vivo (in ducks and mice) transformations of T. szidati cercariae to schistosomula were performed. Ultrastructural observation revealed that cercariae possess a simple outer tegumental membrane with a thick glycocalyx. As with human schistosomes, the latter structure disappears during transformation and a new double membrane with putative protective function is formed. Our biochemical and immunological observations showed that the carbohydrate-rich glycocalyx of cercariae is readily bound by lectins and antibodies. The in vitro transformation to schistosomula can be detected by enhanced reactivity of 2 lectin probes (PNA and ConA) with the surface. The in vivo-transformed (skin and lung) schistosomula appear to have few surface ligands for the 12 lectin probes being tested. Similarly, the cercarial surface and its remnants on the in vitro-produced schistosomula is recognized by sera from immunized mice and humans with cercarial dermatitis; the tissue schistosomula fail to react with these antibodies. The loss of surface targets as a part of parasite immune evasion within the host is discussed.
- MeSH
- elektronová mikroskopie MeSH
- epitopy analýza MeSH
- fluorescenční protilátková technika nepřímá MeSH
- fluorescenční protilátková technika přímá MeSH
- interakce hostitele a parazita MeSH
- kachny MeSH
- lidé MeSH
- myši inbrední BALB C parazitologie MeSH
- myši MeSH
- protilátky helmintové analýza MeSH
- sacharidy analýza MeSH
- Schistosoma chemie růst a vývoj imunologie fyziologie ultrastruktura MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- epitopy MeSH
- protilátky helmintové MeSH
- sacharidy MeSH
- MeSH
- atmosférický tlak MeSH
- epistaxe epidemiologie etiologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- počasí * MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Československo MeSH
- Klíčová slova
- APPENDECTOMY *,
- MeSH
- apendektomie * MeSH
- lidé MeSH
- senioři MeSH
- Check Tag
- lidé MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
