| 1. |
- Hartana, Ciputra Adijaya, et al.
(författare)
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Urothelial bladder cancer may suppress perforin expression in CD8+ T cells by an ICAM-1/TGFβ2 mediated pathway
- 2018
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Ingår i: PLOS ONE. - : Public Library Science. - 1932-6203. ; 13:7
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Tidskriftsartikel (refereegranskat)abstract
- The immune system plays a significant role in urothelial bladder cancer (UBC) progression, with CD8+ T cells being capable to directly kill tumor cells using perforin and granzymes. However, tumors avoid immune recognition by escape mechanisms. In this study, we aim to demonstrate tumor immune escape mechanisms that suppress CD8+ T cells cytotoxicity. 42 patients diagnosed with UBC were recruited. CD8+ T cells from peripheral blood (PB), sentinel nodes (SN), and tumor were analyzed in steady state and in vitro-stimulated conditions by flow cytometry, RT-qPCR, and ELISA. Mass spectrometry (MS) was used for identification of proteins from UBC cell line culture supernatants. Perforin was surprisingly found to be low in CD8+ T cells from SN, marked by 1.8-fold decrease of PRF1 expression, with maintained expression of granzyme B. The majority of perforin-deficient CD8+ T cells are effector memory T (TEM) cells with exhausted Tc2 cell phenotype, judged by the presence of PD-1 and GATA-3. Consequently, perforin-deficient CD8+ T cells from SN are low in T-bet expression. Supernatant from muscle invasive UBC induces perforin deficiency, a mechanism identified by MS where ICAM-1 and TGFβ2 signaling were causatively validated to decrease perforin expression in vitro. Thus, we demonstrate a novel tumor escape suppressing perforin expression in CD8+ T cells mediated by ICAM-1 and TGFβ2, which can be targeted in combination for cancer immunotherapy.
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| 2. |
- Winerdal, Malin E, et al.
(författare)
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FOXP3 and survival in urinary bladder cancer
- 2011
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Ingår i: BJU International. - 1464-4096 .- 1464-410X. ; 108:10, s. 1672-1678
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Tidskriftsartikel (refereegranskat)abstract
- OBJECTIVE:To investigate the possible impact of FOXP3 expression in T-cells, as well as in tumour cells, on long-term survival in patients with urinary bladder cancer (UBC) invading muscle.PATIENTS AND METHODS:In a retrospective study, tumour specimens from 37 patients cystectomized for T1-T4 UBC during 1999-2002 at the Karolinska University Hospital were examined by immunohistochemistry for tumour expression and/or infiltration of immune cells expressing FOXP3 as well as CD3. The results obtained were correlated with clinicopathological parameters, where the primary and secondary outcomes investigated were overall survival and progression-free survival, respectively.RESULTS:Infiltration of CD3(+) and FOXP3(+) lymphocytes (≥3 cells per high-power field) were both correlated with better survival, and this relationship persisted throughout the whole study period (all P < 0.05). Patients with FOXP3(+) tumour cells had decreased long-term survival compared to those patients with FOXP3(-) tumours (P < 0.05). Despite a limited amount of patient material, the results of the present study indicate that FOXP3 expression, in both lymphocytes and tumour cells, is an important prognostic factor in UBC.CONCLUSIONS:FOXP3 expression in UBC cells is associated with decreased long-term survival and thus may be a novel negative prognostic factor in UBC invading muscle. By contrast, the presence of FOXP3(+) tumour-infiltrating lymphocytes was correlated with a positive prognosis. Because FOXP3 is up-regulated upon activation in human T-cells, FOXP3 may serve more as an activation marker than as a regulatory T-cell indicator in this case. These results support the need for larger prospective studies aiming to confirm the results obtained and to examine the underlying mechanisms in detail.
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| 3. |
- Winerdal, Malin E., et al.
(författare)
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Urinary Bladder Cancer Tregs Suppress MMP2 and Potentially Regulate Invasiveness
- 2018
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Ingår i: CANCER IMMUNOLOGY RESEARCH. - : American Association for Cancer Research (AACR). - 2326-6066 .- 2326-6074. ; 6:5, s. 528-538
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Tidskriftsartikel (refereegranskat)abstract
- Regulatory T cells (Treg) have long been considered one-sided suppressors of antitumor immune responses and hence associated with poor patient outcome in cancer. However, evidence is mounting of a paradoxical positive prognostic effect of Tregs on certain malignancies, including urinary bladder cancer (UBC). This discrepancy has partly been attributed to the shear misidentification of Tregs, but also to the inflammatory profile of the tumor. Our aim was to determine whether tumor-infiltrating Forkhead box P3+ (FOXP3+) cells confer a stable Treg phenotype and to investigate putative beneficial Treg functions, focusing on tumor-promoting inflammatory pathways in UBC. Patients (n = 52) with suspected UBC were prospectively included. We show, by using a broad range of analytical approaches, that tumor-infiltrating CD4+FOXP3+ T cells in UBC phenotypically, functionally, and epigenetically represent a true Treg population. At the invasive front of UBC tumors, we found an inverse relationship between Treg frequency and expression of matrix metalloproteinase 2 (MMP2), a key proinvasive factor induced by tumor-promoting inflammation. Correspondingly, a significant, dose-dependent Treg-mediated downregulation of MMP2 protein and mRNA expression was observed in both macrophages and UBC cells. Also, we found that Treg frequency specifically at the invasive front positively correlated with survival. Thus, we identify Treg-mediated suppression of MMP2 in the tumor microenvironment as a mechanism explaining the paradoxical positive prognostic impact of tumor-infiltrating Tregs in UBC.
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| 4. |
- Lindstrand, Anna, et al.
(författare)
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Genome sequencing is a sensitive first-line test to diagnose individuals with intellectual disability
- 2022
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Ingår i: Genetics in Medicine. - : ELSEVIER SCIENCE INC. - 1098-3600 .- 1530-0366. ; 24:11, s. 2296-2307
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Tidskriftsartikel (refereegranskat)abstract
- Purpose: Individuals with intellectual disability (ID) and/or neurodevelopment disorders (NDDs) are currently investigated with several different approaches in clinical genetic diagnostics. Methods: We compared the results from 3 diagnostic pipelines in patients with ID/NDD: genome sequencing (GS) first (N = 100), GS as a secondary test (N = 129), or chromosomal microarray (CMA) with or without FMR1 analysis (N = 421). Results: The diagnostic yield was 35% (GS -first), 26% (GS as a secondary test), and 11% (CMA/FMR1). Notably, the age of diagnosis was delayed by 1 year when GS was performed as a secondary test and the cost per diagnosed individual was 36% lower with GS first than with CMA/FMR1. Furthermore, 91% of those with a negative result after CMA/FMR1 analysis (338 individuals) have not yet been referred for additional genetic testing and remain undiagnosed. Conclusion: Our findings strongly suggest that genome analysis outperforms other testing strategies and should replace traditional CMA and FMR1 analysis as a first-line genetic test in individuals with ID/NDD. GS is a sensitive, time-and cost-effective method that results in a confirmed molecular diagnosis in 35% of all referred patients. (c) 2022 The Authors. Published by Elsevier Inc. on behalf of American College of Medical Genetics and Genomics. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
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| 5. |
- Winerdal, Max
(författare)
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Inflammation, caffeine and adenosine in neonatal hypoxic ischemic brain injury
- 2014
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Background: Brain injury during the neonatal period has potentially lifelong consequences for a child. Perinatal infections and inflammation can induce preterm birth and unfavorable cognitive development, Thus inflammation has received enthusiastic interest for potential therapeutic approaches seeking to protect the newborn brain. Experimental evidence demonstrates that inflammation induces brain injury succeeding the initial insult. A key cytokine in brain injury is the tumor necrosis factor (TNF-α), with reported detrimental cytotoxic effects on selected neuronal populations. Nonetheless, important functions of TNF-α in cerebral homeostasis and development have also been described. Caffeine is used against apneas of prematurity, with noticeable protection against cognitive delay and cerebral palsy. The main effects of caffeine at clinically relevant doses are mediated through inhibition of adenosine receptors. Adenosine is formed from adenosine triphosphate, the main transporter of chemical energy in the cell, which is readily cleaved to adenosine upon extracellular release or extensive leakage from injured necrotic cells. Hence, adenosine signaling is tightly interrelated with local energy levels and cell injury. In addition, adenosine modulates inflammatory responses in profound ways. Methods: In mouse models of premature excitotoxic lesions and full term hypoxic ischemic brain injury we investigated blockade of TNF-α with and without interleukin IL-1 or lipopolysaccharide (LPS) induced systemic inflammation. Furthermore, in the hypoxic ischemic model we developed a flow cytometry based method to investigate temporal distribution of brain infiltrating and splenetic immune cells and their activation. To analyze the data in an unbiased way, we next adapted a data driven gating methodology. Moreover, we used principal component analysis to discriminate between experimentally entangled variables. Utilizing these techniques, we explored the effect of genetic inactivation of adenosine A1 and A2A receptors in the hypoxic ischemic model. We also tested the unselective, competitive adenosine receptor antagonist caffeine and assessed the effect on outcome and immune activation. Results: Blockade of TNF-α protected the brain against excitotoxic lesions in the presence but not absence of systemic inflammation. No protection was observed in the full term hypoxic ischemic model. Persistent lymphocyte activation was found three months after the lesion. Moreover, spleenocytes harvested five months after neonatal brain damage proliferated when stimulated with brain homogenate in contrast to sham operated counterparts. Adenosine A1 receptor deficient mice acquired significantly larger infarcts and associated adverse behavioral outcome compared to wild type. There were specific alterations in the immune responses induced after brain injury, including impaired cytotoxic function and dysregulation of regulatory B-lymphocytes. Adenosine A2A receptor knockout mice developed increased atrophy compared to wild type after hypoxic ischemia, an effect accompanied by functional deficits in behavioral tests. Furthermore, a compensated functional insufficiency was estimated in the regulatory T- lymphocyte compartment in combination with a seemingly inadequate number of myeloid derived suppressor like cells, accompanied by a reversed, increased response in innate antigen presenting cells in the knockout. Finally, we report neuroprotective properties of 5 mg/kg caffeine given directly after neonatal brain injury. Discussion: TNF-α blockade could potentially protect against preterm excitotoxic brain injury. Only patients with concurrent systemic inflammation would potentially benefit. Moreover, concern about adverse effects exists, why TNF-α blockade for neonatal brain injury is likely not clinically applicable in the near future. Persistent long term cerebral adaptive immune activation, preceded by systemic immune activation in spleen was discovered. Remarkably, spleenocytes from animals subjected to brain injury responded to brain antigen five months after brain damage, whereas spleenocytes from uninjured did not, suggesting formation of immunological memory that might affect long term outcome and provoke autoimmunity later in life. To avoid bias from manual gating of flow cytometry data we developed a data driven approach adapted for brain infiltrating immune cells. Furthermore, we deployed principal component analysis to verify biological relevance in the pattern of immune activation and to discriminate between genotype and injury size effects, since they are experimentally inseparable. Thus we could predict genotype and whether they acquired brain injury or not, from the flow cytometric immune activation pattern alone. Adenosine A1 receptor deficient mice display signs of regulatory B-lymphocyte dysfunction that imply a novel adenosinergic mechanism of B-lymphocyte regulation. In addition, these animals displayed signs of altered cellular cytotoxic immunity. Thus considerable effects on immune activation were present in the A1 receptor knockouts compared to wild type, adding another mechanism linked to worse outcome after hypoxic ischemic brain injury in these animals. Deletion of the adenosine A2A receptor similarly causes worse outcome, however, the alteration of the immune response is completely different. Fundamental changes were observed in regulatory populations like monocyte derived suppressor like cells and regulatory T-lymphocytes. Extensive activation of cytotoxic populations in the adenosine A2A receptor knockout links insufficient regulatory immune function with adverse behavioral and morphological outcome. We also propose a novel hypothesis that short term blockade of adenosine A2A receptors offers neuroprotection whereas long term blockade is detrimental by immunological mechanisms. Thus we tested the tentative therapeutic potential of caffeine, an unselective competitive antagonist of adenosine receptors. Caffeine 5mg/kg given directly after the insult resulted in reduced injury size after neonatal hypoxic ischemia. Since caffeine is a relatively well studied substance with negligible adverse long term effect in technically sound studies absent of significant bias, this approach has a clear clinical relevance. Are we ready for a clinical trial?
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| 6. |
- Zirakzadeh, A. Ali, et al.
(författare)
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Tumour-associated B cells in urothelial urinary bladder cancer
- 2020
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Ingår i: Scandinavian Journal of Immunology. - : Wiley-Blackwell. - 0300-9475 .- 1365-3083. ; 91:2
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Tidskriftsartikel (refereegranskat)abstract
- Tumour infiltrating B cells and CD38+ plasma cells have been correlated with survival in different malignancies but their role in urinary bladder cancer is unclear. IL-10 is a multifunctional cytokine with both anti-inflammatory and immunostimulatory properties, that can be released by regulatory B cells (Bregs). We have stained paraffin-embedded tumour sections from 31 patients with invasive urothelial urinary bladder cancer with respect to CD20+ B cells, CD38+ cells, IL-10-expressing cells, IgG, C1q and C3a and analysed the impact of these markers on survival. Interestingly, we observe tumour-associated CD20+ B cells forming follicle-like structures in tumours of some patients. We demonstrate that follicle-like structures, tumour-associated CD38+ cells, IL-10 produced by non-B cells, tumour infiltrating IgG and activation of the complement system, may associate to longer survival of urinary bladder cancer patients. IL-10 expression by tumour-associated Bregs may instead negatively affect prognosis. More research is needed to fully understand the role of B cells and IL-10 in urinary bladder cancer.
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