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Urban, Constantin F.ORCID iD iconorcid.org/0000-0003-1438-1134
Alternative names
Publications (10 of 60) Show all publications
Khamzeh, A., Rudin, A. D., Venkatakrishnan, V., Stylianou, M., Sanchez Klose, F. P., Urban, C. F., . . . Christenson, K. (2024). High levels of short-chain fatty acids secreted by Candida albicans hyphae induce neutrophil chemotaxis via free fatty acid receptor 2. Journal of Leukocyte Biology, 115(3), 536-546
Open this publication in new window or tab >>High levels of short-chain fatty acids secreted by Candida albicans hyphae induce neutrophil chemotaxis via free fatty acid receptor 2
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2024 (English)In: Journal of Leukocyte Biology, ISSN 0741-5400, E-ISSN 1938-3673, Vol. 115, no 3, p. 536-546Article in journal (Refereed) Published
Abstract [en]

Candida albicans belongs to our commensal mucosal flora and in immune-competent individuals in the absence of epithelial damage, this fungus is well tolerated and controlled by our immune defense. However, C. albicans is an opportunistic microorganism that can cause different forms of infections, ranging from superficial to life-threatening systemic infections. C. albicans is polymorphic and switches between different phenotypes (e.g. from yeast form to hyphal form). C. albicans hyphae are invasive and can grow into tissues to eventually reach circulation. During fungal infections, neutrophils in particular play a critical role for the defense, but how neutrophils are directed toward the invasive forms of fungi is less well understood. We set out to investigate possible neutrophil chemoattractants released by C. albicans into culture supernatants. We found that cell-free culture supernatants from the hyphal form of C. albicans induced both neutrophil chemotaxis and concomitant intracellular calcium transients. Size separation and hydrophobic sorting of supernatants indicated small hydrophilic factors as responsible for the activity. Further analysis showed that the culture supernatants contained high levels of short-chain fatty acids with higher levels from hyphae as compared to yeast. Short-chain fatty acids are known neutrophil chemoattractants acting via the neutrophil free fatty acid receptor 2. In line with this, the calcium signaling in neutrophils induced by hyphae culture supernatants was blocked by a free fatty acid receptor 2 antagonist and potently increased in the presence of a positive allosteric modulator. Our data imply that short-chain fatty acids may act as a recruitment signal whereby neutrophils can detect C. albicans hyphae.

Place, publisher, year, edition, pages
Oxford University Press, 2024
Keywords
acetate, GPR43, granulocyte, infection, inflammation
National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:umu:diva-221856 (URN)10.1093/jleuko/qiad146 (DOI)37992073 (PubMedID)2-s2.0-85186083719 (Scopus ID)
Funder
Swedish Research Council, 2019-01123Swedish Research Council, 2022-00561
Available from: 2024-03-12 Created: 2024-03-12 Last updated: 2024-03-12Bibliographically approved
Unger, L., Skoluda, S., Backman, E., Amulic, B., Ponce-Garcia, F. M., Etiaba, C. N. .., . . . Urban, C. F. (2023). Candida albicans induces neutrophil extracellular traps and leucotoxic hypercitrullination via candidalysin. EMBO Reports, 24(11), Article ID e57571.
Open this publication in new window or tab >>Candida albicans induces neutrophil extracellular traps and leucotoxic hypercitrullination via candidalysin
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2023 (English)In: EMBO Reports, ISSN 1469-221X, E-ISSN 1469-3178, Vol. 24, no 11, article id e57571Article in journal (Refereed) Published
Abstract [en]

The peptide toxin candidalysin, secreted by Candida albicans hyphae, promotes stimulation of neutrophil extracellular traps (NETs). However, candidalysin alone triggers a distinct mechanism for NET-like structures (NLS), which are more compact and less fibrous than canonical NETs. Candidalysin activates NADPH oxidase and calcium influx, with both processes contributing to morphological changes in neutrophils resulting in NLS formation. NLS are induced by leucotoxic hypercitrullination, which is governed by calcium-induced protein arginine deaminase 4 activation and initiation of intracellular signalling events in a dose- and time-dependent manner. However, activation of signalling by candidalysin does not suffice to trigger downstream events essential for NET formation, as demonstrated by lack of lamin A/C phosphorylation, an event required for activation of cyclin-dependent kinases that are crucial for NET release. Candidalysin-triggered NLS demonstrate anti-Candida activity, which is resistant to nuclease treatment and dependent on the deprivation of Zn2+. This study reveals that C. albicans hyphae releasing candidalysin concurrently trigger canonical NETs and NLS, which together form a fibrous sticky network that entangles C. albicans hyphae and efficiently inhibits their growth.

Place, publisher, year, edition, pages
John Wiley & Sons, 2023
Keywords
chronic granulomatous disease, fungal immunology, histone citrullination, polymorphonuclear leucocytes, reactive oxygen species
National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:umu:diva-215387 (URN)10.15252/embr.202357571 (DOI)001081020500001 ()37795769 (PubMedID)2-s2.0-85173538219 (Scopus ID)
Funder
Swedish Research Council, VR-MH2018-05909Swedish Research Council, VR-MH2020-01764Swedish Research Council, VR-MH2022-00850The Kempe Foundations, CK-2033,U16Wellcome trust, 214229_Z_18_ZGerman Research Foundation (DFG), 390713860Swedish Research Council, 2019-01123Swedish Heart Lung Foundation, 2019-01123Stiftelsen Konung Gustaf V:s 80-årsfondRegion Västra Götaland, TUAGBG-917531
Available from: 2023-10-30 Created: 2023-10-30 Last updated: 2024-01-04Bibliographically approved
Silao, F. G., Jiang, T., Bereczky-Veress, B., Kühbacher, A., Ryman, K., Uwamahoro, N., . . . Ljungdahl, P. O. (2023). Proline catabolism is a key factor facilitating Candida albicans pathogenicity. PLoS Pathogens, 19(11 NOVEMBER), Article ID e1011677.
Open this publication in new window or tab >>Proline catabolism is a key factor facilitating Candida albicans pathogenicity
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2023 (English)In: PLoS Pathogens, ISSN 1553-7366, E-ISSN 1553-7374, Vol. 19, no 11 NOVEMBER, article id e1011677Article in journal (Refereed) Published
Abstract [en]

Candida albicans, the primary etiology of human mycoses, is well-adapted to catabolize proline to obtain energy to initiate morphological switching (yeast to hyphal) and for growth. We report that put1-/- and put2-/- strains, carrying defective Proline UTilization genes, display remarkable proline sensitivity with put2-/- mutants being hypersensitive due to the accumulation of the toxic intermediate pyrroline-5-carboxylate (P5C), which inhibits mitochondrial respiration. The put1-/- and put2-/- mutations attenuate virulence in Drosophila and murine candidemia models and decrease survival in human neutrophils and whole blood. Using intravital 2-photon microscopy and label-free non-linear imaging, we visualized the initial stages of C. albicans cells infecting a kidney in real-time, directly deep in the tissue of a living mouse, and observed morphological switching of wildtype but not of put2-/- cells. Multiple members of the Candida species complex, including C. auris, are capable of using proline as a sole energy source. Our results indicate that a tailored proline metabolic network tuned to the mammalian host environment is a key feature of opportunistic fungal pathogens.

Place, publisher, year, edition, pages
Public Library of Science (PLoS), 2023
National Category
Microbiology in the medical area Microbiology
Identifiers
urn:nbn:se:umu:diva-216679 (URN)10.1371/journal.ppat.1011677 (DOI)2-s2.0-85175854519 (Scopus ID)
Available from: 2023-11-16 Created: 2023-11-16 Last updated: 2023-11-16Bibliographically approved
Shchukarev, A., Backman, E., Watts, S., Salentinig, S., Urban, C. F. & Ramstedt, M. (2021). Applying Cryo-X-ray Photoelectron Spectroscopy to Study the Surface Chemical Composition of Fungi and Viruses. Frontiers in Chemistry, 9, Article ID 666853.
Open this publication in new window or tab >>Applying Cryo-X-ray Photoelectron Spectroscopy to Study the Surface Chemical Composition of Fungi and Viruses
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2021 (English)In: Frontiers in Chemistry, E-ISSN 2296-2646, Vol. 9, article id 666853Article in journal (Refereed) Published
Abstract [en]

Interaction between microorganisms and their surroundings are generally mediated via the cell wall or cell envelope. An understanding of the overall chemical composition of these surface layers may give clues on how these interactions occur and suggest mechanisms to manipulate them. This knowledge is key, for instance, in research aiming to reduce colonization of medical devices and device-related infections from different types of microorganisms. In this context, X-ray photoelectron spectroscopy (XPS) is a powerful technique as its analysis depth below 10 nm enables studies of the outermost surface structures of microorganism. Of specific interest for the study of biological systems is cryogenic XPS (cryo-XPS). This technique allows studies of intact fast-frozen hydrated samples without the need for pre-treatment procedures that may cause the cell structure to collapse or change due to the loss of water. Previously, cryo-XPS has been applied to study bacterial and algal surfaces with respect to their composition of lipids, polysaccharides and peptide (protein and/or peptidoglycan). This contribution focuses onto two other groups of microorganisms with widely different architecture and modes of life, namely fungi and viruses. It evaluates to what extent existing models for data treatment of XPS spectra can be applied to understand the chemical composition of their very different surface layers. XPS data from model organisms as well as reference substances representing specific building blocks of their surface were collected and are presented. These results aims to guide future analysis of the surface chemical composition of biological systems.

Place, publisher, year, edition, pages
Frontiers Media S.A., 2021
Keywords
cryo-XPS, virus, fungi, reference data, bacteriophage, surface chemistry, cell wall
National Category
Other Chemistry Topics Biophysics Physical Chemistry Microbiology Other Medical Biotechnology
Identifiers
urn:nbn:se:umu:diva-183667 (URN)10.3389/fchem.2021.666853 (DOI)000660083200001 ()34124001 (PubMedID)2-s2.0-85107576942 (Scopus ID)
Funder
The Kempe Foundations, JCK-1720
Available from: 2021-05-28 Created: 2021-05-28 Last updated: 2023-03-24Bibliographically approved
Shankar, M., Uwamahoro, N., Backman, E., Holmberg, S., Niemiec, M. J., Roth, J., . . . Urban, C. F. (2021). Immune Resolution Dilemma: Host Antimicrobial Factor S100A8/A9 Modulates Inflammatory Collateral Tissue Damage During Disseminated Fungal Peritonitis. Frontiers in Immunology, 12, Article ID 553911.
Open this publication in new window or tab >>Immune Resolution Dilemma: Host Antimicrobial Factor S100A8/A9 Modulates Inflammatory Collateral Tissue Damage During Disseminated Fungal Peritonitis
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2021 (English)In: Frontiers in Immunology, E-ISSN 1664-3224, Vol. 12, article id 553911Article in journal (Refereed) Published
Abstract [en]

Intra-abdominal infection (peritonitis) is a leading cause of severe disease in surgical intensive care units, as over 70% of patients diagnosed with peritonitis develop septic shock. A critical role of the immune system is to return to homeostasis after combating infection. S100A8/A9 (calprotectin) is an antimicrobial and pro-inflammatory protein complex used as a biomarker for diagnosis of numerous inflammatory disorders. Here we describe the role of S100A8/A9 in inflammatory collateral tissue damage (ICTD). Using a mouse model of disseminated intra-abdominal candidiasis (IAC) in wild-type and S100A8/A9-deficient mice in the presence or absence of S100A9 inhibitor paquinimod, the role of S100A8/A9 during ICTD and fungal clearance were investigated. S100A8/A9-deficient mice developed less ICTD than wild-type mice. Restoration of S100A8/A9 in knockout mice by injection of recombinant protein resulted in increased ICTD and fungal clearance comparable to wild-type levels. Treatment with paquinimod abolished ICTD and S100A9-deficient mice showed increased survival compared to wild-type littermates. The data indicates that S100A8/A9 controls ICTD levels and antimicrobial activity during IAC and that targeting of S100A8/A9 could serve as promising adjunct therapy against this challenging disease.

Place, publisher, year, edition, pages
Frontiers Media S.A., 2021
Keywords
Candida albicans, host-pathogen interactions, host-targeted agents, inflammation, peritonitis, S100A8/A9 complex, sepsis
National Category
Immunology in the medical area
Identifiers
urn:nbn:se:umu:diva-181798 (URN)10.3389/fimmu.2021.553911 (DOI)000627778800001 ()2-s2.0-85102439343 (Scopus ID)
Funder
The Kempe Foundations, SMK-1453Swedish Research Council, VR-M 2017-01681Swedish Research Council, 2014-02281
Available from: 2021-04-01 Created: 2021-04-01 Last updated: 2024-01-17Bibliographically approved
Poloni, J. A., Garcia, C. D., Rotta, L. N. & Urban, C. F. (2021). Neutrophils phagocytosing fungal hyphae in urinary sediment. Jornal Brasileiro de Nefrologia, 43(3), 431-433
Open this publication in new window or tab >>Neutrophils phagocytosing fungal hyphae in urinary sediment
2021 (English)In: Jornal Brasileiro de Nefrologia, ISSN 0101-2800, E-ISSN 2175-8239, Vol. 43, no 3, p. 431-433Article in journal (Refereed) Published
Abstract [en]

The Phagocytosis of fungal structures by neutrophils is a well-documented function of these immune cells. However, neutrophil phagocytosis of hyphal structures in the urine sediment is not usually observed during routine sample evaluation. This is a case of hyphal phagocytosis by neutrophils in the urine of a kidney allograft recipient patient.

Abstract [pt]

A fagocitose de estruturas fúngicas por neutrófilos é uma função bem documentada destas células imunes. No entanto, a fagocitose de hifas por neutrófilos no sedimento urinário não é normalmente observada durante avaliação de rotina de amostras. Este é um caso de fagocitose de hifas por neutrófilos na urina de um paciente receptor de aloenxerto renal.

Place, publisher, year, edition, pages
FapUNIFESP (SciELO), 2021
Keywords
Phagocytosis, Hyphae, Urinalysis, Neutrophils, Fagocitose, Hifas, Urinálise, Neutrófilos
National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:umu:diva-188493 (URN)10.1590/2175-8239-jbn-2019-0245 (DOI)000852619700020 ()33350430 (PubMedID)2-s2.0-85116172663 (Scopus ID)
Available from: 2021-10-11 Created: 2021-10-11 Last updated: 2023-09-05Bibliographically approved
De Samber, B., De Rycke, R., De Bruyne, M., Kienhuis, M., Sandblad, L., Bohic, S., . . . Vincze, L. (2020). Effect of sample preparation techniques upon single cell chemical imaging: A practical comparison between synchrotron radiation based X-ray fluorescence (SR-XRF) and Nanoscopic Secondary Ion Mass Spectrometry (nano-SIMS). Analytica Chimica Acta, 1106, 22-32
Open this publication in new window or tab >>Effect of sample preparation techniques upon single cell chemical imaging: A practical comparison between synchrotron radiation based X-ray fluorescence (SR-XRF) and Nanoscopic Secondary Ion Mass Spectrometry (nano-SIMS)
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2020 (English)In: Analytica Chimica Acta, ISSN 0003-2670, E-ISSN 1873-4324, Vol. 1106, p. 22-32Article in journal (Refereed) Published
Abstract [en]

Analytical capabilities of Nanoscopic Secondary Ion Mass Spectrometry (nano-SIMS) and Synchrotron Radiation based X-ray Fluorescence (SR nano-XRF) techniques were compared for nanochemical imaging of polymorphonuclear human neutrophils (PMNs). PMNs were high pressure frozen (HPF), cryosubstituted, embedded in Spurr's resin and cut in thin sections (500 nm and 2 mu m for both techniques resp.) Nano-SIMS enabled nanoscale mapping of isotopes of C, N, O, P and S, while SR based nano-XRF enabled trace level imaging of metals like Ca, Mn, Fe, Ni, Cu and Zn at a resolution of approx. 50 nm. The obtained elemental distributions were compared with those of whole, cryofrozen PMNs measured at the newly developed ID16A nano-imaging beamline at the European Synchrotron Radiation Facility (ESRF) in Grenoble, France. Similarities were observed for elements more tightly bound to the cell structure such as phosphorus and sulphur, while differences for mobile ions such as chlorine and potassium were more pronounced. Due to the observed elemental redistribution of mobile ions such as potassium and chlorine, elemental analysis of high pressure frozen (HPF), cryo-substituted and imbedded cells should be interpreted critically. Although decreasing analytical sensitivity occurs due to the presence of ice, analysis of cryofrozen cells - close to their native state - remains the golden standard. In general, we found nanoscale secondary ion mass spectrometry (nano-SIMS) and synchrotron radiation based nanoscopic X-ray fluorescence (SR nano-XRF) to be two supplementary alternatives for nanochemical imaging of single cells at the nanoscale. 

Place, publisher, year, edition, pages
Elsevier, 2020
Keywords
Synchrotron radiation, X-ray fluorescence, XRF, Nano-SIMS, Cell imaging, Sample preparation
National Category
Analytical Chemistry
Identifiers
urn:nbn:se:umu:diva-169366 (URN)10.1016/j.aca.2020.01.054 (DOI)000519110500002 ()32145852 (PubMedID)2-s2.0-85079038777 (Scopus ID)
Available from: 2020-04-07 Created: 2020-04-07 Last updated: 2023-03-24Bibliographically approved
Urban, C. F. & Backman, E. (2020). Eradicating, retaining, balancing, swarming, shuttling and dumping: a myriad of tasks for neutrophils during fungal infection. Current Opinion in Microbiology, 58, 106-115
Open this publication in new window or tab >>Eradicating, retaining, balancing, swarming, shuttling and dumping: a myriad of tasks for neutrophils during fungal infection
2020 (English)In: Current Opinion in Microbiology, ISSN 1369-5274, E-ISSN 1879-0364, Vol. 58, p. 106-115Article in journal (Refereed) Published
Abstract [en]

Opportunistic, invasive mycoses in immunocompromised patients remain challenging for health care with unacceptably high levels of morbidity and mortality. Neutrophils are essential in host protection against invasive mycoses. Upon development of acute infection, neutrophils are recruited from circulation to the infected tissue, where they exert a considerable variety of effector functions with the ultimate task to eradicate invading microbes. Effector functions include recognition, phagocytosis and intracellular killing of microorganisms via oxidative and non-oxidative mechanisms, excretion of antimicrobial factors from intracellular storages (degranulation), release of neutrophil extracellular traps (NETs) and of extracellular vesicles (EVs), as well as generation of cytokines and chemokines to modulate immune responses. Herein, we describe recent findings which further our understanding of the roles of neutrophils during opportunistic fungal infections which could serve as starting point for the development of immune-targeted interventions to improve clinical management of affected individuals.

Place, publisher, year, edition, pages
Elsevier, 2020
National Category
Immunology in the medical area
Identifiers
urn:nbn:se:umu:diva-176945 (URN)10.1016/j.mib.2020.09.011 (DOI)000601245600016 ()33091824 (PubMedID)2-s2.0-85092790416 (Scopus ID)
Funder
Swedish Research Council, 2017-01681
Available from: 2020-11-20 Created: 2020-11-20 Last updated: 2023-03-23Bibliographically approved
Lopes, J. P., Stylianou, M., Backman, E., Holmberg, S., Ekoff, M., Nilsson, G. & Urban, C. F. (2019). Cryptococcus neoformans Induces MCP-1 Release and Delays the Death of Human Mast Cells. Frontiers in Cellular and Infection Microbiology, 9, Article ID 289.
Open this publication in new window or tab >>Cryptococcus neoformans Induces MCP-1 Release and Delays the Death of Human Mast Cells
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2019 (English)In: Frontiers in Cellular and Infection Microbiology, E-ISSN 2235-2988, Vol. 9, article id 289Article in journal (Refereed) Published
Abstract [en]

Cryptococcosis, caused by the basidiomycete Cryptococcus neoformans, is a life-threatening disease affecting approximately one million people per year worldwide. Infection can occur when C. neoformans cells are inhaled by immunocompromised people. In order to establish infection, the yeast must bypass recognition and clearance by immune cells guarding the tissue. Using in vitro infections, we characterized the role of mast cells (MCs) in cryptococcosis. We found that MCs recognize C. neoformans and release inflammatory mediators such as tryptase and cytokines. From the latter group MCs released mainly CCL-2/MCP-1, a strong chemoattractant for monocytic cells. We demonstrated that supernatants of infected MCs recruit monocytes but not neutrophils. During infection with C. neoformans, MCs have a limited ability to kill the yeast depending on the serotype. C. neoformans, in turn, modulates the lifespan of MCs both, by presence of its polysaccharide capsule and by secreting soluble modulators. Taken together, MCs might have important contributions to fungal clearance during early stages of cryptocococis where these cells regulate recruitment of monocytes to mucosal tissues.

Place, publisher, year, edition, pages
Frontiers Media S.A., 2019
Keywords
innate immunity, Cryptococcus neoformans, mast cells, monocytes, monocyte chemoattractant protein 1/CCL-2, fungi
National Category
Immunology in the medical area Microbiology in the medical area
Identifiers
urn:nbn:se:umu:diva-162849 (URN)10.3389/fcimb.2019.00289 (DOI)000480550400001 ()31456952 (PubMedID)2-s2.0-85071629810 (Scopus ID)
Available from: 2019-09-06 Created: 2019-09-06 Last updated: 2023-03-24Bibliographically approved
Singel, K. L., Grzankowski, K. S., Khan, A. N., Grimm, M. J., D'Auria, A. C., Morrell, K., . . . Segal, B. H. (2019). Mitochondrial DNA in the tumour microenvironment activates neutrophils and is associated with worse outcomes in patients with advanced epithelial ovarian cancer. British Journal of Cancer, 120(2), 207-217
Open this publication in new window or tab >>Mitochondrial DNA in the tumour microenvironment activates neutrophils and is associated with worse outcomes in patients with advanced epithelial ovarian cancer
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2019 (English)In: British Journal of Cancer, ISSN 0007-0920, E-ISSN 1532-1827, Vol. 120, no 2, p. 207-217Article in journal (Refereed) Published
Abstract [en]

BACKGROUND: Advanced cancer causes necrosis and releases damage-associated molecular patterns (DAMPs). Mitochondrial DAMPs activate neutrophils, including generation of neutrophil extracellular traps (NETs), which are injurious, thrombogenic, and implicated in metastasis. We hypothesised that extracellular mitochondrial DNA (mtDNA) in ascites from patients with epithelial ovarian cancer (EOC) would correlate with worse outcomes.

METHODS: Banked ascites supernatants from patients with newly diagnosed advanced EOC were analysed for mtDNA, neutrophil elastase, and activation of healthy donor neutrophils and platelets. TCGA was mined for expression of SELP and ELANE.

RESULTS: The highest quartile of ascites mtDNA correlated with reduced progression-free survival (PFS) and a higher likelihood of disease progression within 12-months following primary surgery (n = 68, log-rank, p = 0.0178). NETs were detected in resected tumours. Ascites supernatants chemoattracted neutrophils, induced NETs, and activated platelets. Ascites exposure rendered neutrophils suppressive, based on abrogation of ex vivo stimulated T cell proliferation. Increased SELP mRNA expression correlated with worse overall survival (n = 302, Cox model, p = 0.02).

CONCLUSION: In this single-centre retrospective analysis, ascites mtDNA correlated with worse PFS in advanced EOC. Mitochondrial and other DAMPs in ascites may activate neutrophil and platelet responses that facilitate metastasis and obstruct anti-tumour immunity. These pathways are potential prognostic markers and therapeutic targets.

Place, publisher, year, edition, pages
Nature Publishing Group, 2019
National Category
Cell and Molecular Biology Cancer and Oncology
Research subject
Infectious Diseases; cell research; Immunology
Identifiers
urn:nbn:se:umu:diva-156192 (URN)10.1038/s41416-018-0339-8 (DOI)000456328200009 ()30518816 (PubMedID)2-s2.0-85058029797 (Scopus ID)
Funder
NIH (National Institute of Health), R01CA188900NIH (National Institute of Health), T32CA108456NIH (National Institute of Health), T32CA085183NIH (National Institute of Health), K01LM012100
Available from: 2019-02-07 Created: 2019-02-07 Last updated: 2023-03-23Bibliographically approved
Projects
Opportunistic mycoses and innate immune mechanisms of recognition, eliminitation and tolerance [2011-02393_VR]; Umeå UniversityIdentificattion of innate immune mechanisms in opportunistic mycoses and exploitation as antifungal therpaies [2014-02281_VR]; Umeå UniversityExploiting innate immune mechanisms to develop new treatment for invasive mycoses [2017-01681_VR]; Umeå UniversityNew strategies for therapies against mycoses [2018-05909_VR]; Umeå University
Organisations
Identifiers
ORCID iD: ORCID iD iconorcid.org/0000-0003-1438-1134

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