BACKGROUND: Antiviral prophylaxis against cytomegalovirus has been associated with reduced risk of allograft rejection and improved allograft survival after renal transplantation. This phenomenon might not be fully explained by preventing the indirect effects of cytomegalovirus. The effect of antiviral agents on lymphocyte function in patients treated with modern immunosuppression has not been studied to date. METHODS: Adult renal transplant recipients were assigned to 3-month prophylaxis with either valganciclovir (900 mg once daily; n=19) or valacyclovir (2 g four times daily; n=17) as part of an ongoing randomized trial. Subsets of lymphocytes, lymphocyte proliferation and/or cytokine production after in vitro mitogen stimulation were evaluated at the end of prophylaxis and 1 month after withdrawal of antiviral drugs. RESULTS: Lymphocyte proliferation was significantly decreased both after phytohemagglutinine (25% ±15% versus 32% ±18%; P=0.025) and concanavalin A stimulation (17% ±9% versus 25% ±16%; P=0.011) during valganciclovir, but not valacyclovir therapy. Moreover, a lower activated T-cell count (CD3(+)HLA-DR(+) cells) was noted in valganciclovir-treated patients (13% ±10% versus 17% ±12% of total CD3(+) T-cells; P=0.005). CONCLUSIONS: Valganciclovir suppresses lymphocyte proliferation and activation in patients after renal transplantation.
- MeSH
- acyklovir aplikace a dávkování analogy a deriváty terapeutické užití MeSH
- aktivace lymfocytů účinky léků MeSH
- antivirové látky aplikace a dávkování terapeutické užití MeSH
- cytomegalovirové infekce farmakoterapie etiologie prevence a kontrola virologie MeSH
- Cytomegalovirus účinky léků imunologie MeSH
- dospělí MeSH
- fytohemaglutininy farmakologie MeSH
- ganciklovir aplikace a dávkování analogy a deriváty terapeutické užití MeSH
- imunosupresiva aplikace a dávkování škodlivé účinky MeSH
- imunosupresivní léčba škodlivé účinky MeSH
- konkanavalin A farmakologie MeSH
- kultivované buňky MeSH
- lidé středního věku MeSH
- lidé MeSH
- počet lymfocytů MeSH
- přežívání štěpu účinky léků MeSH
- proliferace buněk účinky léků MeSH
- rejekce štěpu prevence a kontrola MeSH
- T-lymfocyty - podskupiny cytologie účinky léků imunologie MeSH
- transplantace ledvin imunologie MeSH
- valin aplikace a dávkování analogy a deriváty terapeutické užití MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
OBJECTIVES: We studied a) mitogen lectin (PHA) evoked changes of Na+/K+-ATPase activity in functionally different lymphocytes or brain cortex cells and b) quantitative relationship between PHA- evoked early enzyme activation and late lymphocyte proliferation were analyzed. MATERIALS AND METHODS: We performed biochemical analyses of Pi released from ATP by Na+/K+-ATPase activity. Lymphocyte proliferation was assayed by 3H-thymidine incorporation. RESULTS: We demonstrated PHA stimulated Na+/K+-ATPase activity of mouse spleen lymphocytes or freshly isolated brain cortex cells. Besides this, we estimated high stimulation of Na+/K+-ATPase activity and subsequent late 3H-thymidine incorporation into pig lymphocytes as both PHA dose and K+ ion concentration dependent. CONCLUSIONS: Thus, early PHA dose-dependent stimulation of Na+/K+-ATPase activity is a more general response in different animal species and functionally different cells. We measured both cell type- and PHA-dose dependent enzyme activity stimulation. We can suggest that intensity of early PHA induced Na+/K+-ATPase activation could be in relationship to subsequent elevated level of T lymphocyte proliferation. The Na+/K+-ATPase can be a part of mitogen lectin evoked signal transduction mechanisms.
- MeSH
- aktivace lymfocytů účinky léků MeSH
- buněčné kultury MeSH
- draslík farmakologie MeSH
- fytohemaglutininy farmakologie MeSH
- mitogeny farmakologie MeSH
- mozková kůra účinky léků metabolismus MeSH
- myši inbrední CBA MeSH
- myši MeSH
- prasata MeSH
- slezina účinky léků metabolismus MeSH
- sodíko-draslíková ATPasa metabolismus MeSH
- T-lymfocyty metabolismus MeSH
- thymus účinky léků metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The main aim of this study was to compare the reaction of quiescent and proliferating, i.e. phytohemagglutinin (PHA)-stimulated, human peripheral blood mononuclear cells (PBMCs) to gamma-radiation, and analyse changes of proteins related to repair of DNA damage and apoptosis, such as gammaH2A.X, p53, p53 phosphorylation at serines-15 and -392, and p21 and their dose dependence. Freshly isolated PBMCs in peripheral blood are predominantly quiescent, in G(0) phase, and with very low amounts of proteins p53 and p21. Using confocal microscopy we detected dose dependent (0.5-5 Gy) induction of foci containing gammaH2A.X (1 h after gamma-ray exposure), which are formed around radiation-induced double strand breaks of DNA. Apoptosis was detected from 24 h after irradiation by the dose of 4 Gy onwards by Annexin V binding and lamin B cleavage. Seventy two hours after irradiation 70% of CD3(+) lymphocytes were A(+). Neither increase in p53 nor its phosphorylation on serine-392 after irradiation was detected in these cells. However, massive increase in p21 (cyclin-dependent kinase inhibitor 1A) was detected after irradiation, which can be responsible for late occurrence of apoptosis in these quiescent cells. PHA-stimulation itself (72 h) caused an increase in early apoptosis (A(+)PI(-)) in comparison to non-stimulated PBMCs (38% A(+) resp. 13.4%). After PHA-stimulation also the amount of gammaH2A.X, p53, and p21 increased, but no phosphorylation of p53 on serine-392 or -15 was detected. Reaction to gamma-radiation was different in PHA-stimulated lymphocytes: the p53 pathway was activated and p53 was phosphorylated on serines-15 and -392 4 h after irradiation by the dose of 4 Gy. Phosphorylation of p53 at serine-15 increased in a dose-dependent manner in the studied dose range 0.2-7.5 Gy. Also the amount of p21 increased after irradiation. Seventy two hours after irradiation of PHA-stimulated CD3(+) T lymphocytes by the dose of 4 Gy 65% of cells were A(+).
- MeSH
- apoptóza účinky záření MeSH
- financování organizované MeSH
- fosforylace MeSH
- fytohemaglutininy farmakologie MeSH
- histony chemie metabolismus MeSH
- lidé MeSH
- lymfocyty MeSH
- nádorový supresorový protein p53 chemie metabolismus MeSH
- poškození DNA MeSH
- techniky in vitro MeSH
- vztah dávky záření a odpovědi MeSH
- záření gama MeSH
- Check Tag
- lidé MeSH
As previous studies suggested, nuclear myosin I (NMI) and actin have important roles in DNA transcription. In this study, we characterized the dynamics of these two proteins during transcriptional activation in phytohemagglutinin (PHA) stimulated human lymphocytes. The stimulation led to strong up-regulation of NMI both on the mRNA and protein level, while actin was relatively stably expressed. The intranuclear distribution of actin and NMI was evaluated using immunogold labeling. In nucleoli of resting cells, actin was localized predominantly to fibrillar centers (FCs), while NMI was located mainly to the dense fibrillar component (DFC). Upon stimulation, FCs remained the main site of actin localization, however, an accumulation of both actin and NMI in the DFC and in the granular component was observed. In the nucleoplasm of resting lymphocytes, both actin and NMI were localized mostly in condensed chromatin. Following stimulation, the majority of both proteins shifted towards the decondensed chromatin. In transcriptionally active cells, both actin and NMI colocalized with nucleoplasmic transcription sites. These results demonstrate that actin and NMI are compartmentalized in the nuclei where they can dynamically translocate depending on transcriptional activity of the cells.
- MeSH
- aktiny metabolismus MeSH
- buněčné jadérko metabolismus účinky léků ultrastruktura MeSH
- buněčné jádro metabolismus účinky léků ultrastruktura MeSH
- financování organizované MeSH
- fytohemaglutininy farmakologie MeSH
- genetická transkripce imunologie MeSH
- lidé MeSH
- lymfocyty mikrobiologie účinky záření virologie MeSH
- messenger RNA metabolismus MeSH
- myosin typu I metabolismus MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- transmisní elektronová mikroskopie MeSH
- Check Tag
- lidé MeSH
- MeSH
- buněčné jadérko genetika MeSH
- daktinomycin farmakologie MeSH
- finanční podpora výzkumu jako téma MeSH
- fytohemaglutininy farmakologie MeSH
- genetická transkripce účinky záření MeSH
- jaderné proteiny genetika chemie MeSH
- lidé MeSH
- replikace DNA účinky záření MeSH
- ribozomální DNA genetika účinky léků MeSH
- T-lymfocyty fyziologie účinky léků MeSH
- transport proteinů účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- srovnávací studie MeSH
- MeSH
- autoantigeny farmakologie MeSH
- diabetes mellitus 1. typu imunologie krev MeSH
- ELISPOT MeSH
- finanční podpora výzkumu jako téma MeSH
- fytohemaglutininy farmakologie MeSH
- interferon gama analýza MeSH
- leukocyty mononukleární imunologie MeSH
- lidé MeSH
- protilátky analýza MeSH
- tetanový toxoid farmakologie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- MeSH
- fytohemaglutininy farmakologie MeSH
- interferon alfa biosyntéza MeSH
- interferonové induktory MeSH
- interferony biosyntéza MeSH
- kinetika MeSH
- lektiny farmakologie MeSH
- lidé MeSH
- prasata MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
Cell-mediated cytotoxicity of adherent and non-adherent subpopulations of mouse lymph node cells sensitized in vitro on monolayers of syngeneic, methylcholanthrene-induced sarcoma cells was examined by inhibition of 3H-thymidine incorporation. The cell-mediated cytotoxicity of nylon wood-adherent, non-adherent and unfractionated lymph node cells was found to be equally efficient.
- MeSH
- aktivace lymfocytů účinky léků MeSH
- antigeny nádorové imunologie MeSH
- buněčná adheze * MeSH
- cytotoxické testy imunologické * MeSH
- experimentální sarkom chemicky indukované imunologie MeSH
- fytohemaglutininy farmakologie MeSH
- imunizace * MeSH
- imunoglobuliny analýza MeSH
- lymfocyty imunologie MeSH
- methylcholanthren MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nylony MeSH
- techniky in vitro MeSH
- transplantace nádorů MeSH
- vlna MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
