BACKGROUND: The HIV (human immunodeficiency virus) population remains a global concern whose treatment is effective, though not yet optimal. Immune based therapies have thus far been disappointing and still need to be explored further. Based on published data suggesting that the functions of cytotoxic CD8+ T lymphocytes (CTL) can be improved by histamine, we investigated the effect of histamine in vitro on HIV-1 specific CD8+ T lymphocytes in HIV+ subjects. RESULTS: 60 HIV+ subjects were included in the study. We evaluated CTL function by IFNγ (interferon gamma) production (using the enzyme-linked immunospot assay (Elispot), BD Bioscience). Changes in the production of IFNγ after incubation with histamine were compared with the levels of total IgE (immunoglobulin E, measured using a Dade Behring analyzer), because histamine is endogenously released through IgE. Activation of HIV-specific CTL by histamine occurs via H2R (histamine receptors). Thus we attempted to block this activation using cimetidine (antagonist H2R). CONCLUSIONS: We found an increase in IFNγ production after the activation of HIV-1 specific CD8+ T lymphocytes by histamine (this elevation was blocked by cimetidine), furthermore, we demonstrated a negative correlation between the production of IFNγ and levels of total IgE.

• MeSH
• CD8-pozitivní T-lymfocyty imunologie   MeSH
• histamin imunologie   MeSH
• HIV infekce imunologie   MeSH
• HIV-1 imunologie   MeSH
• imunoglobulin E biosyntéza   imunologie   MeSH
• interferon gama biosyntéza   imunologie   MeSH
• lidé MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

The granzyme B-induced cell death has been traditionally viewed as a primary mechanism that is used by cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells to eliminate harmful target cells including allogeneic, virally infected and tumour cells. Granzyme B (GrB) is the most abundant serine protease which is stored in secretory granules of CTLs and NK cells. After recognition of the target cell, the engaged CTLs and NK cells vectorially secrete GrB along with other granule proteins including perforin into the immunological synapse. From this submicroscopic intercellular cleft GrB translocates into the cytoplasm of the target cell. Although several models have been proposed to explain the GrB delivery mechanism, conclusive understanding of this process remains still elusive. Once in the cytoplasm, GrB cleaves and activates, or inactivates, multiple protein substrates, resulting eventually into apoptotic demise of the target cell. This review is focused on the gene structure and expression of GrB, its biosynthesis and activation, delivery mechanisms into the target cell cytoplasm, direct proteolytic involvement in activation of several pro-apoptotic pathways, and on regulation of its activity in cancer cells. Moreover, emphasis is given to the GrB-mediated anticancer effects and future clinical applications of the GrB-based and tumour-targeted recombinant fusion constructs.

• MeSH
• aktivace enzymů MeSH
• apoptóza genetika   MeSH
• biologické modely MeSH
• granzymy genetika   metabolismus   fyziologie   MeSH
• lidé MeSH
• nádory enzymologie   genetika   metabolismus   MeSH
• regulace genové exprese enzymů MeSH
• regulace genové exprese u nádorů MeSH
• tkáňová distribuce MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

• MeSH
• cytotoxické T-lymfocyty imunologie   MeSH
• dendritické buňky imunologie   MeSH
• finanční podpora výzkumu jako téma MeSH
• imunoterapie metody   MeSH
• myši MeSH
• Papillomaviridae imunologie   MeSH
• peptidy imunologie   MeSH
• protinádorové vakcíny terapeutické užití   MeSH
• slezina imunologie   MeSH
• techniky in vitro MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• srovnávací studie MeSH

Multiple myeloma has been considered a weakly immunogenic malignancy that can cause profound defects in the immune system. An important issue for the immunotherapy of myeloma is the identification of appropriate tumor-associated antigens (TAAs). Recently, hTERT (human telomerase reverse transcriptase) was detected on a majority of human malignancies. In the studies reported here, we studied antigen-specific and HLA-A2-restricted cytotoxic activity against an ARH77 myeloma cell line in vitro. An HLA-A2-specific hTERT-derived nonapeptide ((540)ILAKFLHWL(548)) was used as a TAA. Myeloma-specific cytotoxic activity of hTERT-reactive cytotoxic lymphocytes (CTLs) was established by repeated stimulation of the CTLs via dendritic cells loaded with hTERT-derived nonapeptide. These studies were able to demonstrate that hTERT-reactive T-lymphocytes can be identified and expanded using relatively simple in vitro techniques consisting of antigen-specific stimulation, immunomagnetic sorting, and then induction of rapid expansion.

• MeSH
• aktivace lymfocytů MeSH
• antigeny nádorové imunologie   MeSH
• cytotoxické T-lymfocyty imunologie   MeSH
• dendritické buňky imunologie   MeSH
• lidé MeSH
• mnohočetný myelom imunologie   MeSH
• nádorové buněčné linie MeSH
• peptidové fragmenty genetika   imunologie   MeSH
• telomerasa genetika   imunologie   metabolismus   MeSH
• transdukce genetická MeSH
• transfekce MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

• MeSH
• cytotoxické T-lymfocyty MeSH
• MHC antigeny I. třídy MeSH

Despite novel treatment strategies, multiple myeloma (MM) remains an incurable disease with low immunogenicity and multiple immune defects. We developed an ex vivo strategy for inducing myeloma-specific cytotoxic T lymphocytes (CTLs) and demonstrate the possibility of identification and long-term in vivo monitoring of individual myeloma-specific T-cell clones using the most sensitive clonotypic assay that is able to detect low frequencies of T-cell clones (1 clonotypic cell in 10(6) cells). Ten patients with MM were examined for the presence of tumour-reactive T cells using dendritic cells loaded with autologous tumour cells. All patients had detectable myeloma-reactive T cells in vitro. Expanded myeloma-reactive T cells demonstrated specific cytotoxic effects against autologous tumour cells in vitro (median 39.6% at an effector:target ratio of 40:1). The clonality of myeloma-specific T cells was studied with a clonotypic assay, which demonstrated both oligoclonal and monoclonal populations of myeloma-specific T cells. CD8(+) CTLs were the most immunodominant myeloma-specific T-cell clones and clinical responses were closely associated with the in vivo expansion and long-term persistence of individual CD8(+) T-cell clones, usually at very low frequencies (10(-3)-10(-6)). We conclude that the clonotypic assay is the most sensitive tool for immunomonitoring of low-frequency T cells.

• MeSH
• aktivace lymfocytů imunologie   MeSH
• antigeny nádorové imunologie   MeSH
• buněčná diferenciace imunologie   MeSH
• buněčné klony imunologie   MeSH
• cytotoxicita imunologická imunologie   MeSH
• cytotoxické T-lymfocyty imunologie   MeSH
• dendritické buňky imunologie   MeSH
• hypervariabilní oblasti genetika   imunologie   MeSH
• imunodominantní epitopy imunologie   MeSH
• imunomagnetická separace metody   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mnohočetný myelom imunologie   terapie   MeSH
• molekulární sekvence - údaje MeSH
• monitorování imunologické metody   MeSH
• nádorové buňky kultivované MeSH
• následné studie MeSH
• prezentace antigenu imunologie   MeSH
• sekvence aminokyselin MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• adenylátcyklasy imunologie   MeSH
• antigeny virové genetika   imunologie   MeSH
• Bordetella pertussis enzymologie   MeSH
• CD8-pozitivní T-lymfocyty imunologie   MeSH
• cytotoxické T-lymfocyty fyziologie   MeSH
• epitopy imunologie   MeSH
• myši MeSH
• rekombinantní proteiny farmakologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH

Anti-viral and anti-tumor vaccines aim to induce cytotoxic CD8+ T cells (CTL) and antibodies. Conserved protein antigens, such as p24 from human immunodeficiency virus, represent promising component for elicitation CTLs, nevertheless with suboptimal immunogenicity, if formulated as recombinant protein. To enhance immunogenicity and CTL response, recombinant proteins may be targeted to dendritic cells (DC) for cross presentation on MHCI, where mannose receptor and/or other lectin receptors could play an important role. Here, we constructed liposomal carrier-based vaccine composed of recombinant p24 antigen bound by metallochelating linkage onto surface of nanoliposomes with surface mannans coupled by aminooxy ligation. Generated mannosylated proteonanoliposomes were analyzed by dynamic light scattering, isothermal titration, and electron microscopy. Using murine DC line MutuDC and murine bone marrow derived DC (BMDC) we evaluated their immunogenicity and immunomodulatory activity. We show that p24 mannosylated proteonanoliposomes activate DC for enhanced MHCI, MHCII and CD40, CD80, and CD86 surface expression both on MutuDC and BMDC. p24 mannosylated liposomes were internalized by MutuDC with p24 intracellular localization within 1 to 3 h. The combination of metallochelating and aminooxy ligation could be used simultaneously to generate nanoliposomal adjuvanted recombinant protein-based vaccines versatile for combination of recombinant antigens relevant for antibody and CTL elicitation.

• MeSH
• antigeny MeSH
• dendritické buňky MeSH
• HIV-1 * MeSH
• lidé MeSH
• liposomy metabolismus   MeSH
• mannany metabolismus   MeSH
• myši MeSH
• rekombinantní proteiny metabolismus   MeSH
• vakcíny proti AIDS * imunologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

U dospělých pacientů s chronickou hepatitidou C (HC), genotypem 1b budou z periferní krve v Praze a Hradci Králové vyšetřovány tyto markery: 1) Exprese aktivačních znaků CD38 a HLA-DR na cytotoxických T lymfocytech (CTL) k získání informace o aktivaci imunitního systému. 2) Stanovení FoxP3 (Treg), tlumící cytotoxické lymfocyty (CTLs), potlačující eliminaci viru HC. 3) Další cytokiny budou vyšetřovány v laboratoři v Praze metodikou FlowCytomix: IL-1b, IL-2, IL-4, IL-5, IL-6, IL-10, IL-13, IFNg, TGF-b a TNF? a 4) Imunologické markery v laboratoři v Albany (NY, USA) pomocí technologie GCSPCE (grating-coupled surface plasmon coupled emission) s cílem určení těch, které mají diagnostický a prognostický význam. Markery budou sledovány u těchto pacientů:1. Soubor 40 pacientů léčených pegylovaným interferonem alfa s ribavirinem. 2. Soubor 20 pacientů, kteří eliminovali HCV infekci bez antivirové léčby a 3. Kontrolní soubor 30 zdravých osob; The following markers will be examined in the blood of adult patients with chronic hepatitis C (HC), genotype 1b in Prague and Hradec Králove: 1) Expression of activation markers CD38 a HLA-DR on the cytotoxic T lymphocytes (CTLs) presents the information about activation of immune system. 2) Determination of FoxP3 (Treg) that suppress CTLs and viral elimination is inhibited. 3) Next cytokines will be analysed in laboratory in Prague using FlowCytomix methodology: IL-1b, IL-2, IL-4, IL-5, IL-6, IL-10, IL-13, IFNg, TGF-b and TNF? and 4) Next imunological markers will be assessed in Albany (NY,USA) by GCSPCE (grating-coupled surface plasmon coupled emission) technology with the aim to assess what markers have diagnostic and prognostic relevance. Markers will be monitored in these patients: 1) Group of 40 patients (pts) treated with PegIFN alfa with RBV. 2) 20 pts that eliminated HCV infection without antiviral treatment and 3) 30 healthy persons as control group.

• MeSH
• antigeny CD38 MeSH
• biologické markery MeSH
• chronická hepatitida C imunologie   MeSH
• cytotoxické T-lymfocyty MeSH
• genotypizační techniky MeSH
• hepatitida C - protilátky MeSH
• hepatitida C MeSH
• HLA-DR antigeny MeSH
• interferon alfa MeSH
• interleukiny analýza   MeSH
• molekulární biologie MeSH
• regulační T-lymfocyty MeSH

• Konspekt
• Biochemie. Molekulární biologie. Biofyzika
• NLK Obory
• biologie
• infekční lékařství
• virologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

• MeSH
• AIDS MeSH
• cytotoxické T-lymfocyty MeSH
• HIV-1 MeSH
• Langerhansovy buňky MeSH
• vakcíny proti AIDS MeSH