SKP1-CUL1-F-box protein (SCF) ubiquitin ligases are versatile protein complexes that mediate the ubiquitination of protein substrates. The direct substrate recognition relies on a large family of F-box-domain-containing subunits. One of these substrate receptors is FBXO38, which is encoded by a gene found mutated in families with early-onset distal motor neuronopathy. SCFFBXO38 ubiquitin ligase controls the stability of ZXDB, a nuclear factor associated with the centromeric chromatin protein CENP-B. Loss of FBXO38 in mice results in growth retardation and defects in spermatogenesis characterized by deregulation of the Sertoli cell transcription program and compromised centromere integrity. Moreover, it was reported that SCFFBXO38 mediates the degradation of PD-1, a key immune-checkpoint inhibitor in T cells. Here, we have re-addressed the link between SCFFBXO38 and PD-1 proteolysis. Our data do not support the notion that SCFFBXO38 directly or indirectly controls the abundance and stability of PD-1 in T cells.
- MeSH
- antigeny CD279 * metabolismus genetika MeSH
- F-box proteiny * metabolismus genetika MeSH
- lidé MeSH
- myši MeSH
- proteinligasy komplexu SCF metabolismus genetika MeSH
- proteolýza MeSH
- T-lymfocyty metabolismus MeSH
- ubikvitinace MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
PURPOSE: Common variable immunodeficiency (CVID) is the most frequent symptomatic primary immunodeficiency, with heterogeneous clinical presentation. Our goal was to analyze CD8 T cell homeostasis in patients with infection only CVID, compared to those additionally affected by dysregulatory and autoimmune phenomena. METHODS: We used flow and mass cytometry evaluation of peripheral blood of 40 patients with CVID and 17 healthy donors. RESULTS: CD8 T cells are skewed in patients with CVID, with loss of naïve and increase of effector memory stages, expansion of cell clusters with high functional exhaustion scores, and a highly activated population of cells with immunoregulatory features, producing IL-10. These findings correlate to clinically widely used B cell-based EURO classification. Features of exhaustion, including loss of CD127 and CD28, and expression of TIGIT and PD-1 in CD8 T cells are strongly associated with interstitial lung disease and autoimmune cytopenias, whereas CD8 T cell activation with elevated HLA-DR and CD38 expression predict non-infectious diarrhea. CONCLUSION: We demonstrate features of advanced differentiation, exhaustion, activation, and immunoregulatory capabilities within CD8 T cells of CVID patients. Assessment of CD8 T cell phenotype may allow risk assessment of CVID patients and provide new insights into CVID pathogenesis, including a better understanding of mechanisms underlying T cell exhaustion and regulation.
- MeSH
- analýza přežití MeSH
- antigeny CD279 genetika účinky léků MeSH
- doba přežití bez progrese choroby MeSH
- ipilimumab farmakologie terapeutické užití MeSH
- karcinom z renálních buněk * farmakoterapie patologie MeSH
- lidé MeSH
- nivolumab MeSH
- protinádorové látky imunologicky aktivní terapeutické užití MeSH
- protokoly protinádorové kombinované chemoterapie MeSH
- výsledek terapie MeSH
- Check Tag
- lidé MeSH
PURPOSE: The aim of this study was to investigate the potential association and combined prognostic significance of the frequency of primary cilia (PC), programmed cell death protein-1 receptor (PD1) and CD8+ tumor infiltrating lymphocytes (TIL) in patients with clear cell renal cancer (ccRCC). METHODS: The frequency of PC, PD1 expression and the frequency of intratumoral CD8+ TIL were evaluated in 104 ccRCC patients. RESULTS: The median frequency of PC was 0.003. The expression of PD1+ cells were <5% in 52 patients, 5-25% in 34 patients and 26-50% in 13 patients and >50% in 5 patients. Intratumoral CD8+ TIL were evaluable in all patients: negative in 1 patient, <25% in 63, 26-50% in 29 and >50% in 11 patients. Overall survival (OS) according to the frequency of PC was significantly shorter in patients with higher frequency (≥0.002) than in patients with lower frequency (<0.002) (p<0.001). Median OS was significantly shorter in patients with higher (25%) CD8+ TIL and higher (>25%) PD1+ expression than in patients with lower (<25%) expression (4.6 vs. 97. years, p=0.006 and 2.9 vs. 8.9 years, p=0.006, respectively). CONCLUSIONS: The present study provides the first data on the potential association and combined prognostic significance of frequency of PC, PD1+ cells and CD8+ TIL in patients with clear cell renal cancer.
- MeSH
- antigeny CD279 genetika MeSH
- CD8-pozitivní T-lymfocyty patologie MeSH
- cilie genetika patologie MeSH
- dospělí MeSH
- Kaplanův-Meierův odhad MeSH
- karcinom z renálních buněk genetika patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- přežití bez známek nemoci MeSH
- prognóza * MeSH
- regulace genové exprese u nádorů MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- tumor infiltrující lymfocyty patologie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Klíčová slova
- KEYTRUDA (pembrolizumab),
- MeSH
- antigeny CD279 genetika účinky léků MeSH
- humanizované monoklonální protilátky aplikace a dávkování MeSH
- lidé MeSH
- nemalobuněčný karcinom plic * diagnostické zobrazování farmakoterapie patologie MeSH
- nežádoucí účinky léčiv epidemiologie etiologie MeSH
- regulace genové exprese u nádorů účinky léků MeSH
- senioři MeSH
- stupeň nádoru MeSH
- Check Tag
- lidé MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
Adoptive cell therapy (ACT) is becoming a prominent alternative therapeutic treatment for cancer patients relapsing on traditional therapies. In parallel, antibodies targeting immune checkpoint molecules, such as cytotoxic-T-lymphocyte-associated antigen 4 (CTLA-4) and cell death protein 1 pathway (PD-1), are rapidly being approved for multiple cancer types, including as first line therapy for PD-L1-expressing non-small-cell lung cancer. The combination of ACT and checkpoint blockade could substantially boost the efficacy of ACT. In this study, we generated a novel self-delivering small interfering RNA (siRNA) (sdRNA) that knocked down PD-1 expression on healthy donor T cells as well as patient-derived tumor-infiltrating lymphocytes (TIL). We have developed an alternative chemical modification of RNA backbone for improved stability and increased efficacy. Our results show that T cells treated with sdRNA specific for PD-1 had increased interferon γ (IFN-γ) secreting capacity and that this modality of gene expression interference could be utilized in our rapid expansion protocol for production of TIL for therapy. TIL expanded in the presence of PD-1-specific sdRNA performed with increased functionality against autologous tumor as compared to control TIL. This method of introducing RNAi into T cells to modify the expression of proteins could easily be adopted into any ACT protocol and will lead to the exploration of new combination therapies.
- MeSH
- antigeny CD279 genetika metabolismus MeSH
- buněčná a tkáňová terapie metody MeSH
- HeLa buňky MeSH
- imunoterapie adoptivní metody MeSH
- interferon gama genetika metabolismus MeSH
- lidé MeSH
- melanom imunologie metabolismus terapie MeSH
- nádory plic imunologie metabolismus terapie MeSH
- průtoková cytometrie MeSH
- RNA interference fyziologie MeSH
- T-lymfocyty metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
Purpose Hodgkin Reed-Sternberg (HRS) cells evade antitumor immunity by multiple means, including gains of 9p24.1/ CD274(PD-L1)/ PDCD1LG2(PD-L2) and perturbed antigen presentation. Programmed death 1 (PD-1) receptor blockade is active in classic Hodgkin lymphoma (cHL) despite reported deficiencies of major histocompatibility complex (MHC) class I expression on HRS cells. Herein, we assess bases of sensitivity to PD-1 blockade in patients with relapsed/refractory cHL who were treated with nivolumab (anti-PD-1) in the CheckMate 205 trial. Methods HRS cells from archival tumor biopsies were evaluated for 9p24.1 alterations by fluorescence in situ hybridization and for expression of PD ligand 1 (PD-L1) and the antigen presentation pathway components-β2-microglobulin, MHC class I, and MHC class II-by immunohistochemistry. These parameters were correlated with clinical responses and progression-free survival (PFS) after PD-1 blockade. Results Patients with higher-level 9p24.1 copy gain and increased PD-L1 expression on HRS cells had superior PFS. HRS cell expression of β2-microglobulin/MHC class I was not predictive for complete remission or PFS after nivolumab therapy. In contrast, HRS cell expression of MHC class II was predictive for complete remission. In patients with a > 12-month interval between myeloablative autologous stem-cell transplantation and nivolumab therapy, HRS cell expression of MHC class II was associated with prolonged PFS. Conclusion Genetically driven PD-L1 expression and MHC class II positivity on HRS cells are potential predictors of favorable outcome after PD-1 blockade. In cHL, clinical responses to nivolumab were not dependent on HRS cell expression of MHC class I.
- MeSH
- antigeny CD274 antagonisté a inhibitory biosyntéza genetika imunologie MeSH
- antigeny CD279 antagonisté a inhibitory biosyntéza genetika imunologie MeSH
- beta-2-mikroglobulin biosyntéza genetika imunologie MeSH
- buňky Reedové-Sternberga účinky léků imunologie patologie MeSH
- doba přežití bez progrese choroby MeSH
- Hodgkinova nemoc farmakoterapie genetika imunologie patologie MeSH
- kohortové studie MeSH
- lidé MeSH
- lidské chromozomy, pár 9 MeSH
- MHC antigeny II. třídy biosyntéza genetika imunologie MeSH
- nivolumab terapeutické užití MeSH
- prediktivní hodnota testů MeSH
- prezentace antigenu MeSH
- protinádorové látky imunologicky aktivní terapeutické užití MeSH
- výsledek terapie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- klinické zkoušky, fáze II MeSH
- multicentrická studie MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
Mesenchymal stem cells (MSCs) represent a population of cells which have the ability to regulate reactivity of T and B lymphocytes by multiple mechanisms. The immunoregulatory activities of MSCs are strictly influenced by the cytokine environment. Here we show that two functionally distinct cytokines, interleukin-4 (IL-4) and interferon-γ (IFN-γ), significantly potentiate the ability of MSCs to inhibit IL-10 production by activated regulatory B cells (Bregs). However, MSCs in the presence of IL-4 or IFN-γ inhibit the IL-10 production by different mechanisms. Preincubation of MSCs with IFN-γ led to the suppression, but pretreatment with IL-4 of neither MSCs nor B cells resulted in the suppression of IL-10 production. The search for candidate regulatory molecules expressed in cytokine-treated MSCs revealed different patterns of the gene expression. Pretreatment of MSCs with IFN-γ, but not with IL-4, induced expression of indoleamine-2,3-dioxygenase, cyclooxygenase-2 and programmed cell death-ligand 1. To identify the molecule(s) responsible for the suppression of IL-10 production, we used specific inhibitors of the putative regulatory molecules. We found that indomethacine, an inhibitor of cyclooxygenase-2 (Cox-2) activity, completely abrogated the inhibition of IL-10 production in cultures containing MSCs and IFN-γ, but had no effect on the suppression in cell cultures containing MSCs and IL-4. The results show that MSCs can inhibit the response of B cells to one stimulus by different mechanisms in dependence on the cytokine environment and thus support the idea of the complexity of immunoregulatory action of MSCs.
- MeSH
- aktivace lymfocytů účinky léků imunologie MeSH
- antigeny CD279 genetika imunologie metabolismus MeSH
- buněčné mikroprostředí účinky léků imunologie MeSH
- cyklooxygenasa 2 genetika imunologie metabolismus MeSH
- cytokiny imunologie metabolismus farmakologie MeSH
- ELISA MeSH
- exprese genu účinky léků genetika imunologie MeSH
- indolamin-2,3,-dioxygenasa genetika imunologie metabolismus MeSH
- interferon gama farmakologie MeSH
- interleukin-10 imunologie metabolismus MeSH
- interleukin-4 farmakologie MeSH
- interleukin-6 genetika imunologie metabolismus MeSH
- kokultivační techniky MeSH
- kultivované buňky MeSH
- mezenchymální kmenové buňky účinky léků imunologie metabolismus MeSH
- myši MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- regulační B-lymfocyty účinky léků imunologie metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
