Highly purified human recombinant interleukin 2 induced cytotoxicity in mouse spleen cells against mouse sarcoma cells when added during the 51Cr microcytotoxicity assay. It elicited similar levels of killer cell activation as did human lymphoid (Jurkat leukaemia-derived) or mouse lymphoid (EL-4 leukaemia-derived) IL-2 preparations. The susceptibility of six MC-induced mouse sarcomas to the cytolytic effect of lymphokine-activated killer cells was compared. Five (MC11, MC13, MC14, MC15, MC16) of six mouse sarcoma cell lines examined were sensitive in vitro to the LAK cell effect, whereas one cell line (MC12) was resistant. Since the sensitive and resistant target cell lines had been induced with the same carcinogen and in mice of the same genotype, they represent a very useful model for investigation of target cell structures responsible for the sensitivity to the LAK cell effect.

• MeSH
• buňky NK účinky léků   imunologie   MeSH
• experimentální sarkom imunologie   terapie   MeSH
• imunoterapie MeSH
• inbrední kmeny myší MeSH
• interleukin-2 farmakologie   terapeutické užití   MeSH
• lidé MeSH
• myši MeSH
• rekombinantní proteiny farmakologie   terapeutické užití   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• interleukin-2 MeSH
• rekombinantní proteiny MeSH

The proliferation and development of cytotoxic T cells was investigated in human peripheral blood mononuclear cell (PBMC) cultures stimulated with an antigenic extract from Candida albicans (MPPS), or with the purified protein derivative from Mycobacterium tuberculosis (PPD), or with human recombinant interleukin 2 (rIL-2). Microbial antigen- and rIL-2-induced cytotoxic T cells were able to lyse both natural killer (NK) sensitive and resistant targets. No correlation was observed between the development of T cell cytotoxicity and interferon (IFN) production in vitro. The addition of anti-class II monoclonal antibodies at the beginning of MPPS/PPD-stimulated cultures inhibited the cell proliferation, IFN production and T cell cytotoxicity, while all these cellular activities were not inhibited by anti-class II antibodies in rIL-2-stimulated cultures. Finally, antibodies to class I determinants inhibit T cell cytotoxicity, suggesting a role of such determinants in the development of the non-adaptive immunity to microbial infections.

• MeSH
• antigeny bakteriální farmakologie   MeSH
• buněčné dělení účinky léků   MeSH
• Candida albicans MeSH
• cytotoxické T-lymfocyty cytologie   účinky léků   imunologie   MeSH
• dospělí MeSH
• HLA antigeny metabolismus   farmakologie   MeSH
• interleukin-2 farmakologie   MeSH
• lidé MeSH
• monoklonální protilátky MeSH
• Mycobacterium tuberculosis MeSH
• rekombinantní proteiny farmakologie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny bakteriální MeSH
• HLA antigeny MeSH
• interleukin-2 MeSH
• monoklonální protilátky MeSH
• rekombinantní proteiny MeSH

Regulatory T cells (Tregs) are indispensable for maintaining self-tolerance by suppressing conventional T cells. On the other hand, Tregs promote tumor growth by inhibiting anticancer immunity. In this study, we identified that Tregs increase the quorum of self-reactive CD8+ T cells required for the induction of experimental autoimmune diabetes in mice. Their major suppression mechanism is limiting available IL-2, an essential T-cell cytokine. Specifically, Tregs inhibit the formation of a previously uncharacterized subset of antigen-stimulated KLRK1+ IL-7R+ (KILR) CD8+ effector T cells, which are distinct from conventional effector CD8+ T cells. KILR CD8+ T cells show superior cell-killing abilities in vivo. The administration of agonistic IL-2 immunocomplexes phenocopies the absence of Tregs, i.e., it induces KILR CD8+ T cells, promotes autoimmunity, and enhances antitumor responses in mice. Counterparts of KILR CD8+ T cells were found in the human blood, revealing them as a potential target for immunotherapy.

As well as protecting us from invading pathogens, like bacteria or viruses, our immune system can also identify dangerous cells of our own that may cause the body harm, such as cancer cells. Once detected, a population of immune cells called cytotoxic T cells launch into action to kill the potentially harmful cell. However, sometimes the immune system makes mistakes and attacks healthy cells which it misidentifies as being dangerous, leading to autoimmune diseases. Special immune cells called T regulatory lymphocytes, or ‘Tregs’, can suppress the activity of cytotoxic T cells, preventing them from hurting the body’s own cells. While this can have a positive impact and reduce the effects of autoimmunity, Tregs can also make the immune system less responsive to cancer cells and allow tumors to grow. But how Tregs alter the behavior of cytotoxic T cells during autoimmune diseases and cancer is poorly understood. While multiple mechanisms have been proposed, none of these have been tested in living animal models of these diseases. To address this, Tsyklauri et al. studied Tregs in laboratory mice which had been modified to have autoimmune diabetes, which is when the body attacks the cells responsible for producing insulin. The experiments revealed that Tregs take up a critical signaling molecule called IL-2 which cytotoxic T cells need to survive and multiply. As a result, there is less IL-2 molecules available in the environment, inhibiting the cytotoxic T cells’ activity. Furthermore, if Tregs are absent and there is an excess of IL-2, this causes cytotoxic T cells to transition into a previously unknown subset of T cells with superior killing abilities. Tsyklauri et al. were able to replicate these findings in two different groups of laboratory mice which had been modified to have cancer. This suggests that Tregs suppress the immune response to cancer cells and prevent autoimmunity using the same mechanism. In the future, this work could help researchers to develop therapies that alter the behavior of cytotoxic T cells and/or Tregs to either counteract autoimmune diseases, or help the body fight off cancer.

• Klíčová slova
• IL-2, T cells, autoimmunity, cytotoxic, immune suppression, immunology, inflammation, mouse, regulatory T cells,
• MeSH
• CD8-pozitivní T-lymfocyty MeSH
• diabetes mellitus 1. typu * patologie   MeSH
• imunologická tolerance MeSH
• interleukin-2 MeSH
• lektinové receptory NK-buněk - podrodina K MeSH
• lidé MeSH
• myši MeSH
• receptory interleukinu-7 MeSH
• regulační T-lymfocyty * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• interleukin-2 MeSH
• KLRK1 protein, human MeSH Prohlížeč
• Klrk1 protein, mouse MeSH Prohlížeč
• lektinové receptory NK-buněk - podrodina K MeSH
• receptory interleukinu-7 MeSH

Nanomaterials, including zinc oxide nanoparticles (ZnO NPs), have a great application potential in many fields, such as medicine, the textile industry, electronics, and cosmetics. Their impact on the environment must be carefully investigated and specified due to their wide range of application. However, the amount of data on possible negative effects of ZnO NPs on plants at the cellular level are still insufficient. Thus, we focused on the effect of ZnO NPs on tobacco BY-2 cells, i.e., a widely accepted plant cell model. Adverse effects of ZnO NPs on both growth and biochemical parameters were observed. In addition, reactive oxygen and nitrogen species visualizations confirmed that ZnO NPs may induce oxidative stress. All these changes were associated with the lipid peroxidation and changes in the plasma membrane integrity, which together with endoplasmatic reticulum and mitochondrial dysfunction led to autophagy and programmed cell death. The present study demonstrates that the phytotoxic effect of ZnO NPs on the BY-2 cells is very complex and needs further investigation.

• Klíčová slova
• BY-2 cells, ZnO nanoparticles, autophagy, oxidative stress, phytotoxicity, programmed cell death,
• Publikační typ
• časopisecké články MeSH

The immunoregulatory properties of mesenchymal stem cells (MSCs) have been well documented in various models in vitro and in vivo. Furthermore, a population of regulatory B cells (Bregs) that produce relatively high concentrations of IL-10 has been recently described. To study the relationship between MSCs and Bregs, we analyzed the effects of MSCs on IL-10 production by lipopolysaccharide (LPS)-activated mouse B cells. The production of IL-10 by B cells remained preserved in the presence of MSCs and was even significantly enhanced by IFN-γ. However, the production of IL-10 was strongly suppressed in cultures containing MSCs and IFN-γ. Preincubation of MSCs, but not of B cells, with IFN-γ induced the suppression of IL-10 secretion in cultures containing MSCs and B cells. The supernatants from IFN-γ-treated MSCs had no inhibitory effect, and the suppression of IL-10 production was abrogated if the MSCs and B cells were separated in a transwell system. Analysis of the gene expression of IFN-γ- or IFN-γ and LPS-treated MSCs revealed a strong upregulation of genes for indoleamine-2,3-dioxygenase (IDO), cyclooxygenase-2 (Cox-2) and programmed cell death-ligand 1 (PD-L1). While the inhibition of IDO activity or the inclusion of the neutralization monoclonal antibody anti-PD-L1 did not abrogate the suppression, indomethacin, an inhibitor of Cox-2, completely inhibited the MSC-mediated suppression of IL-10 production. Accordingly, the production of IL-10 by B cells was inhibited by exogenous prostaglandin E2. The results thus suggest that IFN-γ-treated MSCs strongly inhibit IL-10 production by activated B cells by a mechanism requiring cell contact and involving the Cox-2 pathway.

• Klíčová slova
• B cells, Cyclooxygenase-2, IL-10 production, Immunosuppression, Mesenchymal stem cells,
• MeSH
• aktivace lymfocytů účinky léků   MeSH
• antigeny CD274 antagonisté a inhibitory   genetika   imunologie   MeSH
• B-lymfocyty cytologie   účinky léků   imunologie   MeSH
• cyklooxygenasa 2 genetika   imunologie   MeSH
• difuzní komory kultivační MeSH
• dinoproston farmakologie   MeSH
• indolamin-2,3,-dioxygenasa genetika   imunologie   MeSH
• indomethacin farmakologie   MeSH
• inhibitory cyklooxygenasy farmakologie   MeSH
• interferon gama farmakologie   MeSH
• interleukin-10 antagonisté a inhibitory   genetika   imunologie   MeSH
• kokultivační techniky MeSH
• kultivační média speciální farmakologie   MeSH
• lipopolysacharidy farmakologie   MeSH
• mezenchymální kmenové buňky cytologie   účinky léků   imunologie   MeSH
• mezibuněčná komunikace imunologie   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• neutralizující protilátky farmakologie   MeSH
• primární buněčná kultura MeSH
• regulace genové exprese MeSH
• signální transdukce MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny CD274 MeSH
• Cd274 protein, mouse MeSH Prohlížeč
• cyklooxygenasa 2 MeSH
• dinoproston MeSH
• IL10 protein, mouse MeSH Prohlížeč
• indolamin-2,3,-dioxygenasa MeSH
• indomethacin MeSH
• inhibitory cyklooxygenasy MeSH
• interferon gama MeSH
• interleukin-10 MeSH
• kultivační média speciální MeSH
• lipopolysacharidy MeSH
• neutralizující protilátky MeSH
• Ptgs2 protein, mouse MeSH Prohlížeč

It has been previously found that local administration of X63-m-IL-2 cells transformed by interleukin 2 (IL-2) cDNA and constitutively producing large quantities of IL-2 mediated regressions of murine plasmacytomas and 3-methyl-cholanthrene-induced sarcomas transplanted in syngeneic mice. Here we show that killer cells generated by cultivation of spleen cells in supernatants from X63-m-IL-2 cultures (LAK) or by co-cultivation of murine splenocytes with X63-m-IL-2 cells were cytolytic for natural killer (NK)-sensitive as well as NK-resistant target cells, including the IL-2-producing X63-m-IL-2 cells. Spleen cells cultured in X63-m-IL-2 supernatants or co-cultivated with X63-m-IL-2 cells yielded predominantly Thy 1.2+, CD3+, LFA-1+ lymphocytes. The in vitro results suggest that the LAK cells generated due to the IL-2 production by genetically engineered cells probably help to terminate the treatment by killing the IL-2 producers.

• MeSH
• aktivace lymfocytů * MeSH
• buněčné linie MeSH
• buňky K aktivované lymfokiny imunologie   MeSH
• cytotoxicita imunologická * MeSH
• DNA genetika   MeSH
• imunofenotypizace MeSH
• interleukin-2 biosyntéza   genetika   MeSH
• kultivované buňky MeSH
• myši MeSH
• průtoková cytometrie MeSH
• slezina imunologie   MeSH
• transfekce * MeSH
• transformované buněčné linie MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA MeSH
• interleukin-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

The mechanisms of obesity and type 2 diabetes (T2D)-associated impaired fracture healing are poorly studied. In a murine model of T2D reflecting both hyperinsulinemia induced by high-fat diet and insulinopenia induced by treatment with streptozotocin, we examined bone healing in a tibia cortical bone defect. A delayed bone healing was observed during hyperinsulinemia as newly formed bone was reduced by -28.4 ± 7.7% and was associated with accumulation of marrow adipocytes at the defect site +124.06 ± 38.71%, and increased density of SCA1+ (+74.99 ± 29.19%) but not Runx2+ osteoprogenitor cells. We also observed increased in reactive oxygen species production (+101.82 ± 33.05%), senescence gene signature (≈106.66 ± 34.03%), and LAMIN B1- senescent cell density (+225.18 ± 43.15%), suggesting accelerated senescence phenotype. During insulinopenia, a more pronounced delayed bone healing was observed with decreased newly formed bone to -34.9 ± 6.2% which was inversely correlated with glucose levels (R2 = 0.48, P < .004) and callus adipose tissue area (R2 = .3711, P < .01). Finally, to investigate the relevance to human physiology, we observed that sera from obese and T2D subjects had disease state-specific inhibitory effects on osteoblast-related gene signatures in human bone marrow stromal cells which resulted in inhibition of osteoblast and enhanced adipocyte differentiation. Our data demonstrate that T2D exerts negative effects on bone healing through inhibition of osteoblast differentiation of skeletal stem cells and induction of accelerated bone senescence and that the hyperglycemia per se and not just insulin levels is detrimental for bone healing.

• Klíčová slova
• bone healing, insulin-resistance, insulinopenia, senescence, type 2 diabetes,
• MeSH
• diabetes mellitus 2. typu * komplikace   MeSH
• fraktury kostí * MeSH
• hojení fraktur MeSH
• hyperinzulinismus * MeSH
• kmenové buňky MeSH
• kostní svalek MeSH
• lidé MeSH
• myši MeSH
• obezita komplikace   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND & AIMS: Patients with decompensated cirrhosis suffer from recurrent infections and inadequate responses to prophylactic vaccinations. However, many patients present with hypergammaglobulinemia (HGG), indicating a sustained ability to generate antibody responses. As follicular T helper (Tfh) cells are central facilitators of humoral immunity, we hypothesized that Tfh cell responses may be altered in advanced liver disease and we aimed to identify the mechanisms underlying any such alterations. METHODS: Tfh, regulatory T (Treg) cells, B cells, circulating cytokines and immunoglobulins were analyzed in cohorts of patients with compensated (n = 37) and decompensated cirrhosis (n = 82) and in non-cirrhotic controls (n = 45). Intrahepatic T cells were analyzed in 8 decompensated patients. The influence of IL-2 on Tfh cell function was evaluated in vitro, including Tfh cell cloning and T cell-B cell co-cultures with clones and primary tonsil-derived Tfh cells. RESULTS: Tfh cell frequencies were reduced in patients with decompensated cirrhosis, with phenotypic signatures indicative of increased IL-2 signaling. Soluble IL-2 receptor (sCD25) was elevated in these patients and CD4 T cells were more responsive to IL-2 signaling, as characterized by STAT5 phosphorylation. IL-2 exposure in vitro diminished the Tfh phenotype and resulted in impaired Tfh helper function in co-culture experiments with naïve B cells. Tfh cells were barely detectable in cirrhotic livers. IL-2 signatures on Tfh cells in decompensated patients correlated with immunoglobulin levels, which were found to be associated with improved survival. CONCLUSIONS: Tfh cell impairment represents a previously underestimated feature of cirrhosis-associated immune dysfunction that is driven by IL-2. The presence of HGG in decompensated patients predicts an intact Tfh cell compartment and is associated with a favorable outcome. LAY SUMMARY: Patients with advanced cirrhosis often fail to generate protective immunity after prophylactic vaccinations and suffer from recurring infections that are associated with high mortality. Follicular T helper (Tfh) cells are specialized CD4 T cells that enable the emergence of antibody responses against microbial pathogens. This report demonstrates that Tfh cells are impaired in patients with advanced cirrhosis due to interleukin-2 signaling, a cytokine that is known to impair the generation of Tfh cells.

• Klíčová slova
• CD4 T cells, Cellular immunity, Hepatic decompensation, Immunosuppression,
• MeSH
• B-lymfocyty imunologie   MeSH
• dospělí MeSH
• folikulární pomocné T-buňky imunologie   MeSH
• hypergamaglobulinemie komplikace   MeSH
• imunoglobulin G krev   MeSH
• interleukin-2 krev   MeSH
• jaterní cirhóza krev   komplikace   imunologie   MeSH
• kohortové studie MeSH
• kokultivační techniky MeSH
• kultivované buňky MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorové supresorové proteiny metabolismus   MeSH
• prognóza MeSH
• průřezové studie MeSH
• regulační T-lymfocyty imunologie   MeSH
• senioři MeSH
• signální transdukce imunologie   MeSH
• transkripční faktor STAT5 metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• IL2 protein, human MeSH Prohlížeč
• imunoglobulin G MeSH
• interleukin-2 MeSH
• nádorové supresorové proteiny MeSH
• STAT5A protein, human MeSH Prohlížeč
• transkripční faktor STAT5 MeSH

Fusarium-derived mycotoxin deoxynivalenol (DON) usually induces diarrhea, vomiting and gastrointestinal inflammation. We studied the cytotoxic effect of DON on porcine small intestinal epithelium using the intestinal porcine epithelial cell line IPEC-J2. We screened out differentially expressed genes (DEGs) using RNA-seq and identified 320 upregulated genes and 160 downregulated genes. The enrichment pathways of these DEGs focused on immune-related pathways. DON induced proinflammatory gene expression, including cytokines, chemokines and other inflammation-related genes. DON increased IL1A, IL6 and TNF-α release and DON activated the phosphorylation of extracellular signal-regulated kinase-1 and-2 (ERK1/2), JUN N-terminal kinase (JNK) and p38 MAPK. A p38 inhibitor attenuated DON-induced IL6, TNF-α, CXCL2, CXCL8, IL12A, IL1A, CCL20, CCL4 and IL15 production, while an ERK1/2 inhibitor had only a small inhibitory effect on IL15 and IL6. An inhibitor of p38 MAPK decreased the release of IL1A, IL6 and TNF-α and an inhibitor of ERK1/2 partly attenuated protein levels of IL6. These data demonstrate that DON induces proinflammatory factor production in IPEC-J2 cells by activating p38 and ERK1/2.

• Klíčová slova
• IPEC-J2 cells, MAPKs, RNA-seq, deoxynivalenol, inflammation,
• MeSH
• buněčné linie MeSH
• epitelové buňky účinky léků   imunologie   metabolismus   MeSH
• interleukin-1 genetika   MeSH
• interleukin-6 genetika   MeSH
• MAP kinasový signální systém účinky léků   genetika   imunologie   MeSH
• mitogenem aktivované proteinkinasy p38 metabolismus   MeSH
• prasata MeSH
• střevní sliznice účinky léků   imunologie   metabolismus   MeSH
• TNF-alfa genetika   MeSH
• transkriptom účinky léků   MeSH
• trichotheceny toxicita   MeSH
• viabilita buněk účinky léků   MeSH
• zánět MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• deoxynivalenol MeSH Prohlížeč
• interleukin-1 MeSH
• interleukin-6 MeSH
• mitogenem aktivované proteinkinasy p38 MeSH
• TNF-alfa MeSH
• trichotheceny MeSH