Plasmacytoid dendritic cells (pDCs) are a minority subset of dendritic cells that despite their tiny quantity play an important role in the immune system, especially in antiviral immunity. They are known mostly as the major producers of type I IFN, which they secrete upon stimulation of endosomal Toll-like receptors 7 and 9 with viral RNA and DNA. However, the functionality of pDCs is more complex, as they were shown to be also involved in autoimmunity, inflammation, and cancer. In the context of the tumor microenvironment, pDCs mostly show substantial functional defects and thus contribute to establishing immunosuppressive micromilieu. Indeed, tumor-infiltrating pDCs were shown to be predominantly pro-tumorigenic, with reduced ability to produce IFNα and capacity to prime regulatory T cells via the ICOS/ICOS-L pathway. Here we describe in detail a method to assess the functional capacity of pDCs upon exposure to tumor-derived cell culture supernatants. The same technique can be implemented with minimal variations to test any soluble factor's impact on pDC phenotype and function.

• MeSH
• dendritické buňky * imunologie   metabolismus   MeSH
• lidé MeSH
• nádorové mikroprostředí imunologie   MeSH
• nádory * imunologie   patologie   metabolismus   MeSH
• průtoková cytometrie metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Intratumoral tertiary lymphoid structures (TLSs) have been associated with improved outcome in various cohorts of patients with cancer, reflecting their contribution to the development of tumor-targeting immunity. Here, we demonstrate that high-grade serous ovarian carcinoma (HGSOC) contains distinct immune aggregates with varying degrees of organization and maturation. Specifically, mature TLSs (mTLS) as forming only in 16% of HGSOCs with relatively elevated tumor mutational burden (TMB) are associated with an increased intratumoral density of CD8+ effector T (TEFF) cells and TIM3+PD1+, hence poorly immune checkpoint inhibitor (ICI)-sensitive, CD8+ T cells. Conversely, CD8+ T cells from immunologically hot tumors like non-small cell lung carcinoma (NSCLC) are enriched in ICI-responsive TCF1+ PD1+ T cells. Spatial B-cell profiling identifies patterns of in situ maturation and differentiation associated with mTLSs. Moreover, B-cell depletion promotes signs of a dysfunctional CD8+ T cell compartment among tumor-infiltrating lymphocytes from freshly isolated HGSOC and NSCLC biopsies. Taken together, our data demonstrate that - at odds with NSCLC - HGSOC is associated with a low density of follicular helper T cells and thus develops a limited number of mTLS that might be insufficient to preserve a ICI-sensitive TCF1+PD1+ CD8+ T cell phenotype. These findings point to key quantitative and qualitative differences between mTLSs in ICI-responsive vs ICI-irresponsive neoplasms that may guide the development of alternative immunotherapies for patients with HGSOC.

• MeSH
• CD8-pozitivní T-lymfocyty MeSH
• ektopické lymfoidní struktury * MeSH
• fenotyp MeSH
• lidé MeSH
• nádorové mikroprostředí MeSH
• nádory plic * MeSH
• nádory vaječníků * patologie   MeSH
• nemalobuněčný karcinom plic * MeSH
• tumor infiltrující lymfocyty MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

The profile of the antitumor immune response is an important factor determining patient clinical outcome. However, the influence of the tissue contexture on the composition of the tumor microenvironments of virally induced tumors is not clearly understood. Therefore, we analyzed the immune landscape of two HPV-associated malignancies: oropharyngeal squamous cell carcinoma (OPSCC) and squamous cell carcinoma of uterine cervix (CESC). We employed multiplex immunohistochemistry and immunofluorescence to evaluate the density and spatial distribution of immune cells in retrospective cohorts of OPSCC and CESC patients. This approach was complemented by transcriptomic analysis of purified primary tumor cells and in silico analysis of publicly available RNA sequencing data. Transcriptomic analysis showed similar immune profiles in OPSCC and CESC samples. Interestingly, immunostaining of OPSCC tissues revealed high densities of immune cells in both tumor stroma and tumor epithelium, whereas CESC samples were mainly characterized by the lack of immune cells in the tumor epithelium. However, in contrast to other immune cell populations, polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) were abundant in both segments of CESC samples and CESC-derived tumor cells expressed markedly higher levels of the PMN-MDSC chemoattractants CXCL1, CXCL5, and CXCL6 than OPSCC tumor cells. Taken together, despite their having the same etiologic agent, the immune infiltration pattern significantly differs between OPSCC and CESC, with a noticeable shift toward prominent MDSC infiltration in the latter. Our data thus present a rationale for a diverse approach to targeted therapy in patients with HPV-associated tumors of different tissue origins.

• Publikační typ
• časopisecké články MeSH

Imunoterapie je slibnou léčebnou metodou, která si rychle získala své místo v léčbě mnoha malignit. Předpoklad jejího účinku u nádorů hlavy a krku vychází z vysoké mutační zátěže těchto nádorů. Klinické studie fáze I byly v souladu s preklinickými předpoklady. Následné studie fáze II a III potvrdily účinnost inhibitorů kontrolních bodů nivolumabu (OPDIVO) v druhé linii paliativní léčby a v případě pembrolizumabu v první linii léčby recidivujícího a metastazujícího karcinomu hlavy a krku. V České republice bylo použití pemobrolizumabu schváleno v roce 2014. V jasně definovaných indikacích je nyní léčba pembrolizumabem (Keytruda) hrazena i z veřejného zdravotního pojištění. V České republice však stále zůstává zlatým standardem v první linii léčby karcinomu hlavy a krku chirurgická léčba následovaná adjuvantní onkologickou léčbou. Nicméně přístup je vždy individuální a nejlepší léčebná modalita se volí na základě rozsahu primárního onemocnění a celkového stavu a preferencí pacienta.

Immunotherapy is a promising therapeutic modality that has rapidly gained its place in the treatment of many malignancies. The assumption of its effect in head and neck tumors stems from the high mutational load of these tumors. Phase I clinical trials were consistent with preclinical assumptions. Subsequent phase II and III trials confirmed the efficacy of the check-point inhibitors nivolumab (OPDIVO) in second-line palliation, and in the case of pembrolizumab, in first-line treatment of recurrent and metastatic head and neck cancer. In the Czech Republic, the use of pemobrolizumab was approved in 2014. In clearly defined indications, pembrolizumab therapy (Keytruda) is now also covered by public health insurance. However, in the Czech Republic, surgical treatment followed by adjuvant oncological treatment still remains the gold standard in the first-line treatment of head and neck cancer. Nevertheless, the approach is always individual and the best treatment modality is chosen based on the extent of the primary disease and the overall condition and preferences of the patient.

• MeSH
• humanizované monoklonální protilátky farmakologie   škodlivé účinky   terapeutické užití   MeSH
• imunoterapie * metody   škodlivé účinky   trendy   MeSH
• inhibitory kontrolních bodů farmakologie   škodlivé účinky   terapeutické užití   MeSH
• klinická studie jako téma MeSH
• lidé MeSH
• nádory hlavy a krku * epidemiologie   farmakoterapie   terapie   MeSH
• nivolumab farmakologie   škodlivé účinky   terapeutické užití   MeSH
• protinádorové látky imunologicky aktivní farmakologie   škodlivé účinky   terapeutické užití   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH

Organic inclusions in lime binders provide useful samples for radiocarbon dating of historical objects. Two Czech castles Týřov and Pyšolec from Late Middle Ages were explored, and tens of charcoals were found in their walls. The radiocarbon content of the charcoals was measured with accelerator mass spectrometry. The dating results showed that none of the charcoals were younger than the known historical ages (Týřov: 1260 - 1270, Pyšolec: 1300 - 1340), but some were considerably older. Two charcoals from Pyšolec castle dated to Palaeolithic, likely originating from fluvial sediments added as an aggregate to the mortar. When excluding these two charcoals, the others indicated most likely dates being 50-100 y older than the building dates of the castles. This systemic effect corresponds to the age of wood used for lime burning and shall be accounted for when dating mortars using charcoals.

• MeSH
• dřevěné a živočišné uhlí * MeSH
• dřevo MeSH
• radioaktivní datování * metody   MeSH
• Publikační typ
• časopisecké články MeSH

Karcinomy hlavy a krku tvoří prognosticky nepříznivou skupinu onemocnění zatíženou jak vysokou mortalitou, tak i morbiditou způsobenou klasickými léčebnými postupy, jako je chirurgie, radioterapie a chemoterapie. Tyto nádory však patří mezi onemocnění s početným zastoupením imunitních buněk, které je ještě umocněno u tumorů indukovaných HPV, a jsou tak velmi perspektivním cílem imunoterapeutických postupů. Imunoterapie za využití blokátorů kontrolních bodů imunitního systému se v posledních letech stala jedním z pilířů onkologické terapie, kterou lze aplikovat již v 1. linii léčby u rekurentního a metastatického karcinomu hlavy a krku. Na tuto léčbu ale překvapivě reaguje pouze malá část pacientů. Podrobná analýza imunitního infiltrátu v nádorovém mikroprostředí je pak jedním z klíčů ke zlepšení léčebných výsledků a k efektivnější stratifikaci pacientů, kteří mohou z aplikace imunoterapie a zmírnění klasické terapie profitovat.

Head and neck cancer is prognostically unfavourable group of diseases burdened by high mortality and morbidity following classic treatment regimens, such as surgery, radiotherapy and chemotherapy. Nevertheless, these tumors are highly infiltrated by immune cells, especially in case of HPV induced tumors, and constitute a perspective target for immune-based treatment. Immunotherapy with checkpoint inhibitors has become one of the basic pillars of cancer therapy, that is applicable as a first line treatment in recurrent and metastatic head and neck cancer. However, only a small fraction of patients is responsive to the therapy. In-depth analysis of immune cells in the tumor microenvironment could be the key element for improvement of current treatment regimens and effective stratification of patients that could benefit from immunotherapy and de-intensification of classic therapeutic protocols.

• Klíčová slova
• tumor-infiltrující leukocyty,
• MeSH
• imunoterapie MeSH
• infekce papilomavirem MeSH
• lidé MeSH
• nádorové mikroprostředí MeSH
• nádory hlavy a krku * imunologie   terapie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH

Plasmacytoid dendritic cells (pDCs) are the most potent type I interferon-producing cells and play an important role in antiviral immunity. Tumor-infiltrating pDCs were shown to be predominantly pro-tumorigenic, with reduced ability to produce interferon alpha (IFNα) and confirmed capacity to prime regulatory T cells (Tregs) by the ICOS/ICOS-L pathway. Because a significant number of HNSCCs are induced by human papillomaviruses and show markedly different immune profiles than non-virally induced tumors, we compared the phenotype and functional capacity of HNSCC-infiltrating pDCs to the HPV status of the tumor. We observed a reduced capacity of pDCs to produce IFNα upon toll-like receptor activation in HPV-negative samples and a rather uncompromised functionality in HPV-associated tumors. Additionally, supernatants from non-virally induced but not HPV-associated tumor cell suspensions significantly inhibited IFNα production by peripheral blood-derived pDCs. We identified IL-10 and TNFα as the soluble pDC-suppressive factors with the highest variability between HPV-negative and HPV-positive tumor-derived supernatants. Additionally, we observed a positive correlation of tumor-infiltrating pDCs with Tregs in HPV-negative samples but not in virally induced tumors. Overall, our study indicates that the immunosuppressive cytokine milieu rich in IL-10 and TNFα in HPV-negative but not in HPV-positive HNSCC significantly affects the functional capacity of tumor-infiltrating pDCs, and such dysfunctional pDCs may further support the immunosuppressive tumor microenvironment by promoting the expansion of Tregs in the tumor tissue.

• MeSH
• biologické markery MeSH
• cytokiny metabolismus   MeSH
• dendritické buňky imunologie   metabolismus   patologie   MeSH
• dlaždicobuněčné karcinomy hlavy a krku etiologie   metabolismus   patologie   MeSH
• exprese genu MeSH
• infekce papilomavirem komplikace   virologie   MeSH
• interferon alfa imunologie   metabolismus   MeSH
• lidé MeSH
• náchylnost k nemoci MeSH
• nádorové mikroprostředí * imunologie   MeSH
• regulační T-lymfocyty imunologie   metabolismus   MeSH
• studie případů a kontrol MeSH
• T-lymfocyty - podskupiny imunologie   metabolismus   MeSH
• virová transformace buněk MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Head and neck squamous cell carcinoma (HNSCC) is a highly heterogeneous disease that affects more than 800,000 patients worldwide each year. The variability of HNSCC is associated with differences in the carcinogenesis processes that are caused by two major etiological agents, namely, alcohol/tobacco, and human papillomavirus (HPV). Compared to non-virally induced carcinomas, the oropharyngeal tumors associated with HPV infection show markedly better clinical outcomes and are characterized by an immunologically "hot" landscape with high levels of tumor-infiltrating lymphocytes. However, the standard of care remains the same for both HPV-positive and HPV-negative HNSCC. Surprisingly, treatment de-escalation trials have not shown any clinical benefit in patients with HPV-positive tumors to date, most likely due to insufficient patient stratification. The in-depth analysis of the immune response, which places an emphasis on tumor-infiltrating immune cells, is a widely accepted prognostic tool that might significantly improve both the stratification of HNSCC patients in de-escalation trials and the development of novel immunotherapeutic approaches.

• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

BACKGROUND: Standard treatment of oropharyngeal squamous cell carcinoma (OPSCC) is associated with high morbidity, whereas immunotherapeutic approaches using PD-1:PD-L1 checkpoint blockade only show moderate response rates in OPSCC patients. Therefore, a better stratification of patients and the development of novel therapeutic protocols are crucially needed. The importance of tumor-infiltrating B cells (TIL-Bs) in shaping antitumor immunity remains unclear; therefore, we analyzed frequency, phenotype, prognostic value and possible roles of TIL-Bs in OPSCC. METHODS: We utilized transcriptomic analysis of immune response-related genes in 18 OPSCC samples with respect to human papillomavirus (HPV) status. The density and localization of CD20+, CD8+ and DC-LAMP+ cells were subsequently analyzed in 72 tissue sections of primary OPSCC samples in relation to patients' prognosis. The immunohistochemical approach was supplemented by flow cytometry-based analysis of phenotype and functionality of TIL-Bs in freshly resected primary OPSCC tissues. RESULTS: We observed significantly higher expression of B cell-related genes and higher densities of CD20+ B cells in HPV-associated OPSCC samples. Interestingly, CD20+ TIL-Bs and CD8+ T cells formed non-organized aggregates with interacting cells within the tumor tissue. The densities of both intraepithelial CD20+ B cells and B cell/CD8+ T cell interactions showed prognostic significance, which surpassed HPV positivity and CD8+ TIL density in stratification of OPSCC patients. High density of TIL-Bs was associated with an activated B cell phenotype, high CXCL9 production and high levels of tumor-infiltrating CD8+ T cells. Importantly, the abundance of direct B cell/CD8+ T cell interactions positively correlated with the frequency of HPV16-specific CD8+ T cells, whereas the absence of B cells in tumor-derived cell cultures markedly reduced CD8+ T cell survival. CONCLUSIONS: Our results indicate that high abundance of TIL-Bs and high density of direct B cell/CD8+ T cell interactions can predict patients with excellent prognosis, who would benefit from less invasive treatment. We propose that in extensively infiltrated tumors, TIL-Bs might recruit CD8+ T cells via CXCL9 and due to a highly activated phenotype contribute by secondary costimulation to the maintenance of CD8+ T cells in the tumor microenvironment.

• MeSH
• adjuvantní chemoradioterapie MeSH
• aktivace lymfocytů MeSH
• B-lymfocyty imunologie   MeSH
• CD8-pozitivní T-lymfocyty imunologie   MeSH
• dlaždicobuněčné karcinomy hlavy a krku imunologie   mortalita   terapie   virologie   MeSH
• dospělí MeSH
• infekce papilomavirem imunologie   mortalita   terapie   virologie   MeSH
• Kaplanův-Meierův odhad MeSH
• kohortové studie MeSH
• krční disekce MeSH
• lidé středního věku MeSH
• lidé MeSH
• mezibuněčná komunikace imunologie   MeSH
• nádorové mikroprostředí imunologie   MeSH
• nádory orofaryngu imunologie   mortalita   terapie   virologie   MeSH
• orofarynx patologie   chirurgie   MeSH
• Papillomaviridae imunologie   izolace a purifikace   MeSH
• prognóza MeSH
• progrese nemoci MeSH
• senioři MeSH
• tumor infiltrující lymfocyty imunologie   MeSH
• výběr pacientů MeSH
• výsledek terapie 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

PURPOSE: In multiple oncological settings, expression of the coinhibitory ligand PD-L1 by malignant cells and tumor infiltration by immune cells expressing coinhibitory receptors such as PD-1, CTLA4, LAG-3, or TIM-3 conveys prognostic or predictive information. Conversely, the impact of these features of the tumor microenvironment on disease outcome among high-grade serous carcinoma (HGSC) patients remains controversial. EXPERIMENTAL DESIGN: We harnessed a retrospective cohort of 80 chemotherapy-naïve HGSC patients to investigate PD-L1 expression and tumor infiltration by CD8+ T cells, CD20+ B cells, DC-LAMP+ dendritic cells as well as by PD-1+, CTLA4+, LAG-3+, and TIM-3+ cells in relation with prognosis and function orientation of the tumor microenvironment. IHC data were complemented with transcriptomic and functional studies on a second prospective cohort of freshly resected HGSC samples. In silico analysis of publicly available RNA expression data from 308 HGSC samples was used as a confirmatory approach. RESULTS: High levels of PD-L1 and high densities of PD-1+ cells in the microenvironment of HGSCs were strongly associated with an immune contexture characterized by a robust TH1 polarization and cytotoxic orientation that enabled superior clinical benefits. Moreover, PD-1+TIM-3+CD8+ T cells presented all features of functional exhaustion and correlated with poor disease outcome. However, although PD-L1 levels and tumor infiltration by TIM-3+ cells improved patient stratification based on the intratumoral abundance of CD8+ T cells, the amount of PD-1+ cells failed to do so. CONCLUSIONS: Our data indicate that PD-L1 and TIM-3 constitute prognostically relevant biomarkers of active and suppressed immune responses against HGSC, respectively.

• MeSH
• antigen CTLA-4 imunologie   metabolismus   MeSH
• antigeny CD274 imunologie   metabolismus   MeSH
• buněčný receptor 2 viru hepatitidy A imunologie   metabolismus   MeSH
• CD8-pozitivní T-lymfocyty imunologie   MeSH
• dospělí MeSH
• epiteliální ovariální karcinom imunologie   metabolismus   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• membránové glykoproteiny asociované s lyzozomy imunologie   metabolismus   MeSH
• míra přežití MeSH
• nádorové biomarkery imunologie   metabolismus   MeSH
• nádorové proteiny imunologie   metabolismus   MeSH
• prognóza MeSH
• regulace genové exprese u nádorů * MeSH
• retrospektivní studie MeSH
• senioři MeSH
• serózní cystadenokarcinom imunologie   metabolismus   patologie   MeSH
• tumor infiltrující lymfocyty imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH