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  • 1.
    Beyer, Sarah
    et al.
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Kimani, Martha
    Chemical and Optical Sensing Division, Bundesanstalt für Materialforschung und -prüfung (BAM), Richard-Willstätter Straße 11, 12489 Berlin, Germany.
    Zhang, Yuecheng
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Verhassel, Alejandra
    Institute of Biomedicine, University of Turku, 20520 Turku, Finland; FICAN West Cancer Centre, Turku University Hospital, 20520 Turku, Finland.
    Sternbæk, Louise
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces. Phase Holographic Imaging AB, SE-223 63 Lund, Sweden.
    Wang, Tianyan
    Department of Molecular Biology, Umeå University, SE-901 87 Umeå, Sweden.
    Persson, Jenny L.
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces. Department of Molecular Biology, Umeå University, SE-901 87 Umeå, Sweden.
    Härkönen, Pirkko
    Institute of Biomedicine, University of Turku, 20520 Turku, Finland; FICAN West Cancer Centre, Turku University Hospital, 20520 Turku, Finland.
    Johansson, Emil
    Department of Chemistry, Umeå University, SE-901 87 Umeå, Sweden; Umeå Centre for Microbial Research, Umeå University, SE-901 87 Umeå, Sweden.
    Caraballo, Remi
    Department of Chemistry, Umeå University, SE-901 87 Umeå, Sweden; Umeå Centre for Microbial Research, Umeå University, SE-901 87 Umeå, Sweden.
    Elofsson, Mikael
    Department of Chemistry, Umeå University, SE-901 87 Umeå, Sweden; Umeå Centre for Microbial Research, Umeå University, SE-901 87 Umeå, Sweden.
    Gawlitza, Kornelia
    Chemical and Optical Sensing Division, Bundesanstalt für Materialforschung und -prüfung (BAM), Richard-Willstätter Straße 11, 12489 Berlin, Germany.
    Rurack, Knut
    Chemical and Optical Sensing Division, Bundesanstalt für Materialforschung und -prüfung (BAM), Richard-Willstätter Straße 11, 12489 Berlin, Germany.
    Ohlsson, Lars
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    El-Schich, Zahra
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Gjörloff Wingren, Anette
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Stollenwerk, Maria M
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Fluorescent Molecularly Imprinted Polymer Layers against Sialic Acid on Silica-Coated Polystyrene Cores-Assessment of the Binding Behavior to Cancer Cells.2022In: Cancers, ISSN 2072-6694, Vol. 14, no 8, article id 1875Article in journal (Refereed)
    Abstract [en]

    Sialic acid (SA) is a monosaccharide usually linked to the terminus of glycan chains on the cell surface. It plays a crucial role in many biological processes, and hypersialylation is a common feature in cancer. Lectins are widely used to analyze the cell surface expression of SA. However, these protein molecules are usually expensive and easily denatured, which calls for the development of alternative glycan-specific receptors and cell imaging technologies. In this study, SA-imprinted fluorescent core-shell molecularly imprinted polymer particles (SA-MIPs) were employed to recognize SA on the cell surface of cancer cell lines. The SA-MIPs improved suspensibility and scattering properties compared with previously used core-shell SA-MIPs. Although SA-imprinting was performed using SA without preference for the α2,3- and α2,6-SA forms, we screened the cancer cell lines analyzed using the lectins Maackia Amurensis Lectin I (MAL I, α2,3-SA) and Sambucus Nigra Lectin (SNA, α2,6-SA). Our results show that the selected cancer cell lines in this study presented a varied binding behavior with the SA-MIPs. The binding pattern of the lectins was also demonstrated. Moreover, two different pentavalent SA conjugates were used to inhibit the binding of the SA-MIPs to breast, skin, and lung cancer cell lines, demonstrating the specificity of the SA-MIPs in both flow cytometry and confocal fluorescence microscopy. We concluded that the synthesized SA-MIPs might be a powerful future tool in the diagnostic analysis of various cancer cells.

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  • 2.
    Dao Nyesiga, Gillian
    et al.
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces. Idogen AB, Lund, Sweden.
    Pool, Lieneke
    Idogen AB, Lund, Sweden.
    Englezou, Pavlos C
    Idogen AB, Lund, Sweden.
    Hylander, Terese
    Idogen AB, Lund, Sweden.
    Ohlsson, Lars
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Appelgren, Daniel
    Department of Health, Medicine and Caring Sciences, Faculty of Medicine and Health Sciences, Linköping University, Linköping, Sweden.
    Sundstedt, Anette
    Idogen AB, Lund, Sweden.
    Tillerkvist, Kristina
    Idogen AB, Lund, Sweden.
    Romedahl, Hanne R
    Idogen AB, Lund, Sweden.
    Wigren, Maria
    Idogen AB, Lund, Sweden.
    Tolerogenic dendritic cells generated in vitro using a novel protocol mimicking mucosal tolerance mechanisms represent a potential therapeutic cell platform for induction of immune tolerance.2023In: Frontiers in Immunology, E-ISSN 1664-3224, Vol. 14, article id 1045183Article in journal (Refereed)
    Abstract [en]

    Dendritic cells (DCs) are mediators between innate and adaptive immunity and vital in initiating and modulating antigen-specific immune responses. The most important site for induction of tolerance is the gut mucosa, where TGF-β, retinoic acid, and aryl hydrocarbon receptors collaborate in DCs to induce a tolerogenic phenotype. To mimic this, a novel combination of compounds – the synthetic aryl hydrocarbon receptor (AhR) agonist IGN-512 together with TGF-β and retinoic acid – was developed to create a platform technology for induction of tolerogenic DCs intended for treatment of several conditions caused by unwanted immune activation. These in vitro-generated cells, designated ItolDCs, are phenotypically characterized by their low expression of co-stimulatory and activating molecules along with high expression of tolerance-associated markers such as ILT3, CD103, and LAP, and a weak pro-inflammatory cytokine profile. When co-cultured with T cells and/or B cells, ItolDC-cultures contain higher frequencies of CD25+Foxp3+ regulatory T cells (Tregs), CD49b+LAG3+ ‘type 1 regulatory (Tr1) T cells, and IL-10-producing B cells and are less T cell stimulatory compared to cultures with matured DCs. Factor VIII (FVIII) and tetanus toxoid (TT) were used as model antigens to study ItolDC antigen-loading. ItolDCs can take up FVIII, process, and present FVIII peptides on HLA-DR. By loading both ItolDCs and mDCs with TT, antigen-specific T cell proliferation was observed. Cryo-preserved ItolDCs showed a stable tolerogenic phenotype that was maintained after stimulation with LPS, CD40L, or a pro-inflammatory cocktail. Moreover, exposure to other immune cells did not negatively impact ItolDCs’ expression of tolerogenic markers. In summary, a novel protocol was developed supporting the generation of a stable population of human DCs in vitro that exhibited a tolerogenic phenotype with an ability to increase proportions of induced regulatory T and B cells in mixed cultures. This protocol has the potential to constitute the base of a tolDC platform for inducing antigen-specific tolerance in disorders caused by undesired antigen-specific immune cell activation.

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  • 3.
    El-Schich, Zahra
    et al.
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Zhang, Yuecheng
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Feith, Marek
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Department of Physiology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
    Beyer, Sarah
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Sternbæk, Louise
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces. Phase Holographic Imaging AB, Lund, Sweden.
    Ohlsson, Lars
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Stollenwerk, Maria Magdalena
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Gjörloff Wingren, Anette
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Molecularly imprinted polymers in biological applications.2020In: BioTechniques, ISSN 0736-6205, E-ISSN 1940-9818, Vol. 69, no 6Article in journal (Refereed)
    Abstract [en]

    Molecularly imprinted polymers (MIPs) are currently widely used and further developed for biological applications. The MIP synthesis procedure is a key process, and a wide variety of protocols exist. The templates that are used for imprinting vary from the smallest glycosylated glycan structures or even amino acids to whole proteins or bacteria. The low cost, quick preparation, stability and reproducibility have been highlighted as advantages of MIPs. The biological applications utilizing MIPs discussed here include enzyme-linked assays, sensors, in vivo applications, drug delivery, cancer diagnostics and more. Indeed, there are numerous examples of how MIPs can be used as recognition elements similar to natural antibodies

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  • 4.
    Falk, Magnus
    et al.
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Psotta, Carolin
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Cirovic, Stefan
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Ohlsson, Lars
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Shleev, Sergey
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Electronic Tongue for Direct Assessment of SARS-CoV-2-Free and Infected Human Saliva-A Feasibility Study2023In: Biosensors, ISSN 2079-6374, Vol. 13, no 7, article id 717Article in journal (Refereed)
    Abstract [en]

    An electronic tongue is a powerful analytical instrument based on an array of non-selective chemical sensors with a partial specificity for data gathering and advanced pattern recognition methods for data analysis. Connecting electronic tongues with electrochemical techniques for data collection has led to various applications, mostly within sensing for food quality and environmental monitoring, but also in biomedical research for the analyses of different bioanalytes in human physiological fluids. In this paper, an electronic tongue consisting of six electrodes (viz., gold, platinum, palladium, titanium, iridium, and glassy carbon) was designed and tested in authentic (undiluted, unpretreated) human saliva samples from eight volunteers, collected before and during the COVID-19 pandemic. Investigations of 11 samples using differential pulse voltammetry and a principal component analysis allowed us to distinguish between SARS-CoV-2-free and infected authentic human saliva. This work, as a proof-of-principle demonstration, provides a new perspective for the use of electronic tongues in the field of enzyme-free electrochemical biosensing, highlighting their potential for future applications in non-invasive biomedical analyses.

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  • 5.
    Fernström, Johan
    et al.
    Department of Clinical Sciences Lund, Psychiatry, Faculty of Medicine, Lund University, Lund, Sweden; Office for Psychiatry and Habilitation, Psychiatric Clinic Lund, Region Skåne, Sweden..
    Ohlsson, Lars
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Asp, Marie
    Department of Clinical Sciences Lund, Psychiatry, Faculty of Medicine, Lund University, Lund, Sweden; Office for Psychiatry and Habilitation, Psychiatric Clinic Lund, Region Skåne, Sweden..
    Lavant, Eva
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Holck, Amanda
    Department of Clinical Sciences Lund, Psychiatry, Faculty of Medicine, Lund University, Lund, Sweden; Office for Psychiatry and Habilitation, Psychiatric Clinic Lund, Region Skåne, Sweden..
    Grudet, Cécile
    Department of Clinical Sciences Lund, Psychiatry, Faculty of Medicine, Lund University, Lund, Sweden..
    Westrin, Åsa
    Department of Clinical Sciences Lund, Psychiatry, Faculty of Medicine, Lund University, Lund, Sweden.; Office for Psychiatry and Habilitation, Psychiatry Research Skåne, Region Skåne, Sweden..
    Lindqvist, Daniel
    Department of Clinical Sciences Lund, Psychiatry, Faculty of Medicine, Lund University, Lund, Sweden.; Office for Psychiatry and Habilitation, Psychiatry Research Skåne, Region Skåne, Sweden..
    Plasma circulating cell-free mitochondrial DNA in depressive disorders2021In: PLOS ONE, E-ISSN 1932-6203, Vol. 16, no 11, article id e0259591Article in journal (Refereed)
    Abstract [en]

    BACKGROUND: Plasma circulating cell-free mitochondrial DNA (ccf-mtDNA) is an immunogenic molecule and a novel biomarker of psychiatric disorders. Some previous studies reported increased levels of ccf-mtDNA in unmedicated depression and recent suicide attempters, while other studies found unchanged or decreased ccf-mtDNA levels in depression. Inconsistent findings across studies may be explained by small sample sizes and between-study variations in somatic and psychiatric co-morbidity or medication status.

    METHODS: We measured plasma ccf-mtDNA in a cohort of 281 patients with depressive disorders and 49 healthy controls. Ninety-three percent of all patients were treated with one or several psychotropic medications. Thirty-six percent had a personality disorder, 13% bipolar disorder. All analyses involving ccf-mtDNA were a priori adjusted for age and sex.

    RESULTS: Mean levels in ccf-mtDNA were significantly different between patients with a current depressive episode (n = 236), remitted depressive episode (n = 45) and healthy controls (n = 49) (f = 8.3, p<0.001). Post-hoc tests revealed that both patients with current (p<0.001) and remitted (p = 0.002) depression had lower ccf-mtDNA compared to controls. Within the depressed group there was a positive correlation between ccf-mtDNA and "inflammatory depression symptoms" (r = 0.15, p = 0.02). We also found that treatment with mood stabilizers lamotrigine, valproic acid or lithium was associated with lower ccf-mtDNA (f = 8.1, p = 0.005).

    DISCUSSION: Decreased plasma ccf-mtDNA in difficult-to-treat depression may be partly explained by concurrent psychotropic medications and co-morbidity. Our findings suggest that ccf-mtDNA may be differentially regulated in different subtypes of depression, and this hypothesis should be pursued in future studies.

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  • 6.
    Gustafsson, Anna
    et al.
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Prgomet, Zdenka
    Malmö University, Faculty of Odontology (OD).
    Jankovskaja, Skaidre
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Ruzgas, Tautgirdas
    Malmö University, Biofilms Research Center for Biointerfaces. Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Engblom, Johan
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Ohlsson, Lars
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Gjörloff Wingren, Anette
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Effect of IFN-γ on the kynurenine/tryptophan ratio in monolayer-cultured keratinocytes and a 3D reconstructed human epidermis model2020In: Journal of dermatological science (Amsterdam), ISSN 0923-1811, E-ISSN 1873-569X, Vol. 99, no 3, p. 177-184, article id S0923-1811(20)30234-6Article in journal (Refereed)
    Abstract [en]

    BACKGROUND: Interferon-gamma (IFN-γ) represents a potent inducer for keratinocyte inflammatory and immune activation in vitro. Since tryptophan (trp) conversion to kynurenine (kyn) is involved in inflammation, the topical kyn/trp ratio may serve as a biomarker of skin inflammation. However, the trp metabolism in keratinocytes exposed to IFN-γ is not yet fully understood.

    OBJECTIVE: The aim of this study was to establish a human epidermis model in order to quantify cytokine and kyn/trp secretion from IFN-γ stimulated cells and tissues. Moreover, to compare the cell response of 2D-cultured keratinocytes and the 3D epidermis model.

    METHODS: Polycarbonate filters were used on which primary keratinocytes could attach and stratify in order to form the typical layers of reconstructed human epidermis (RHE). After IFN-γ treatment, secretion of kyn/trp was measured by high performance liquid chromatography. Gene and protein expression of indoleamine 2,3-dioxygenase 1 (IDO) was analyzed with real-time PCR and immunohistochemistry. The secretion of cytokines was quantified with ELISA.

    RESULTS: Trp catabolism to kyn was significantly increased (P < 0.01) in the 2D culture in response to IFN-γ treatment. Before kyn secretion, IDO was strongly upregulated (P < 0.001). IFN-γ treatment also increased the secretion of IL-6 and IL-8 from the keratinocytes. In the RHE, IDO was upregulated by IFN-γ, and kyn secretion could be detected. Interestingly, IDO expression was only present in the basal cells of the RHE.

    CONCLUSION: Our results suggest that IFN-γ acts as an inducer of trp degradation preferentially in undifferentiated keratinocytes, indicated by the IDO expression in the basal layer of the RHE.

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  • 7.
    Gustafsson, Anna
    et al.
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Ventorp, Filip
    Department of Clinical Sciences, Division of Psychiatry, Lund University, Lund, Sweden.
    Wisen, Anita G. M.
    Department of Health Sciences, Division of Physiotherapy, Lund University, Lund, Sweden.
    Ohlsson, Lars
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Ljunggren, Lennart
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Westrin, Åsa
    Department of Clinical Sciences, Division of Psychiatry, Lund University, Lund, Sweden.
    Effects of Acute Exercise on Circulating Soluble Form of the Urokinase Receptor in Patients With Major Depressive Disorder2017In: Biomarker Insights, E-ISSN 1177-2719, Vol. 12, p. 1-6Article in journal (Refereed)
    Abstract [en]

    Inflammation has been proposed to play a role in the generation of depressive symptoms. Previously, we demonstrated that patients with major depressive disorder (MDD) have increased plasma levels of the soluble form of the urokinase receptor (suPAR), a marker for low-grade inflammation. The aim of this study was to test the hypothesis that acute exercise would induce inflammatory response characterized by increased suPAR and elucidate whether patients with MDD display altered levels of suPAR in response to acute exercise. A total of 17 patients with MDD and 17 controls were subjected to an exercise challenge. Plasma suPAR (P-suPAR) was analyzed before, during, and after exercise. There was a significantly higher baseline P-suPAR in the patients with MDD, and the dynamic changes of P-suPAR during the exercise were significantly lower in the patients with MDD, compared with the controls. This study supports the hypothesis that an activation of systemic inflammatory processes, measured as elevated P-suPAR, is involved in the pathophysiology of depression. The study concludes that P-suPAR is influenced by acute exercise, most likely due to release from activated neutrophils.

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  • 8.
    Lindqvist, D.
    et al.
    Department of Clinical Sciences Lund, Psychiatry, Lund University, Lund, Sweden; Department of Psychiatry, School of Medicine, University of California San Francisco, School of Medicine, San Francisco, CA, USA; Psychiatric Clinic, Lund, Division of Psychiatry, Lund, Sweden.
    Fernström, J.
    Department of Clinical Sciences Lund, Psychiatry, Lund University, Lund, Sweden; Psychiatric Clinic, Lund, Division of Psychiatry, Lund, Sweden.
    Grudet, C.
    Department of Clinical Sciences Lund, Psychiatry, Lund University, Lund, Sweden.
    Ljunggren, Lennart
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Träskman-Bendz, L.
    Department of Clinical Sciences Lund, Psychiatry, Lund University, Lund, Sweden.
    Ohlsson, Lars
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Westrin, Åsa
    Department of Clinical Sciences Lund, Psychiatry, Lund University, Lund, Sweden; Psychiatric Clinic, Lund, Division of Psychiatry, Lund, Sweden.
    Increased plasma levels of circulating cell-free mitochondrial DNA in suicide attempters: associations with HPA-axis hyperactivity2016In: Translational Psychiatry, E-ISSN 2158-3188, Vol. 6, no 12, article id e971Article in journal (Refereed)
    Abstract [en]

    Preclinical data suggest that chronic stress may cause cellular damage and mitochondrial dysfunction, potentially leading to the release of mitochondrial DNA (mtDNA) into the bloodstream. Major depressive disorder has been associated with an increased amount of mtDNA in leukocytes from saliva samples and blood; however, no previous studies have measured plasma levels of free-circulating mtDNA in a clinical psychiatric sample. In this study, free circulating mtDNA was quantified in plasma samples from 37 suicide attempters, who had undergone a dexamethasone suppression test (DST), and 37 healthy controls. We hypothesized that free circulating mtDNA would be elevated in the suicide attempters and would be associated with hypothalamic-pituitary-adrenal (HPA)-axis hyperactivity. Suicide attempters had significantly higher plasma levels of free-circulating mtDNA compared with healthy controls at different time points (pre- and post-DST; all P-values < 2.98E - 12, Cohen's d ranging from 2.55 to 4.01). Pre-DST plasma levels of mtDNA were positively correlated with post-DST cortisol levels (rho = 0.49, P < 0.003). Suicide attempters may have elevated plasma levels of free-circulating mtDNA, which are related to impaired HPA-axis negative feedback. This peripheral index is consistent with an increased cellular or mitochondrial damage. The specific cells and tissues contributing to plasma levels of free-circulating mtDNA are not known, as is the specificity of this finding for suicide attempters. Future studies are needed in order to better understand the relevance of increased free-circulating mtDNA in relation to the pathophysiology underlying suicidal behavior and depression.

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  • 9.
    Lindqvist, Daniel
    et al.
    UCSF, San Francisco, CA USA; Lund Univ, Lund, Sweden.
    Fernström, Johan
    Lund Univ, Lund, Sweden.
    Grudet, Cécile
    Lund Univ, Lund, Sweden.
    Ljunggren, Lennart
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Träskman-Bendz, Lil
    Lund Univ, Lund, Sweden.
    Ohlsson, Lars
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Westrin, Åsa
    Lund Univ, Lund, Sweden.
    Increased Plasma Levels of Circulating Cell-Free Mitochondrial DNS in Suicide Attempters - Associations with HPA-Axis Hyperactivity2017In: Biological Psychiatry, ISSN 0006-3223, E-ISSN 1873-2402, Vol. 81, no 10, p. S228-S228, article id 565Article in journal (Other academic)
    Abstract [en]

    Background: Preclinical data suggest that chronic stress may cause cellular damage and mitochondrial dysfunction, potentially leading to the release of mitochondrial DNA (mtDNA) into the bloodstream. Major Depressive Disorder has been associated with an increased amount of mtDNA in leukocytes from saliva samples and blood, but no previous studies have measured plasma levels of free-circulating mtDNA in a clinical psychiatric sample.Methods: In this study, free circulating mtDNA was quantified in plasma samples from 37 suicide attempters, who had undergone a dexamethasone suppression test (DST), and 37 healthy controls. We hypothesized that free circulating mtDNA would be elevated in the suicide attempters and associated with hypothalamic pituitary adrenal (HPA)-axis hyperactivity. Results: Suicide attempters had significantly higher plasma levels of free-circulating mtDNA compared to healthy controls at different time points (pre- and post-DST) (all p-values ,2.98E-12, Cohen’s d ranging from 2.55-4.01). Pre-DST plasma levels of mtDNA were positively correlated with postDST cortisol levels (rho50.49, p,0.003).Conclusions: Suicide attempters may have elevated plasma levels of free-circulating mtDNA, which are related to impaired HPA-axis negative feedback. This peripheral index is consistent with increased cellular or mitochondrial damage. The specific cells and tissues contributing to plasma levels of free-circulating mtDNA are not known, as is the specificity of this finding for suicide attempters. Future studies are needed in order to better understand the relevance of increased freecirculating mtDNA in relation to the pathophysiology underlying suicidal behavior and depression.

  • 10.
    Lindqvist, Daniel
    et al.
    Department of Clinical Sciences Lund, Psychiatry, Faculty of Medicine, Lund University, Lund, Sweden; Office for Psychiatry and Habilitation, Psychiatry Research Skåne, Lund, Sweden.
    Furmark, Tomas
    Department of Psychology, Uppsala University, Uppsala, Sweden.
    Lavebratt, Catharina
    Department of Molecular Medicine and Surgery, Karolinska Institutet, and Center for Molecular Medicine, Karolinska University Hospital, Stockholm, Sweden.
    Ohlsson, Lars
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Månsson, Kristoffer N T
    Center for Psychiatry Research, Department of Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden.
    Plasma circulating cell-free mitochondrial DNA in social anxiety disorder2022In: Psychoneuroendocrinology, ISSN 0306-4530, E-ISSN 1873-3360, Vol. 148, article id 106001Article in journal (Refereed)
    Abstract [en]

    OBJECTIVE: To investigate plasma levels of circulating cell-free mitochondrial DNA (ccf-mtDNA) in patients with social anxiety disorder (SAD) and healthy controls (HC).

    METHODS: In this study, 88 participants (46 patients with SAD and 42 HCs) were enrolled and both ccf-mtDNA and peripheral blood mononuclear cells (PBMC) mtDNA copy number (mtDNA-cn) were measured at up to three times per individual (9-11 weeks apart). SAD patients also received cognitive behavioral therapy (CBT) between the second and third time-point.

    RESULTS: SAD patients had significantly lower ccf-mtDNA compared to HCs at all time points, but ccf-mtDNA did not change significantly after CBT, and was not associated with severity of anxiety symptoms. Plasma ccf-mtDNA did not significantly correlate with PBMC mtDNA-cn in patients.

    CONCLUSION: This is the first report of lower ccf-mtDNA in patients with an anxiety disorder. Our findings could reflect a more chronic illness course in SAD patients with prolonged periods of psychological stress leading to decreased levels of ccf-mtDNA, but future longitudinal studies are needed to confirm or refute this hypothesis.

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  • 11.
    Lindqvist, Daniel
    et al.
    Faculty of Medicine, Department of Clinical Sciences, Psychiatry, Lund University, Lund, Sweden; Department of Psychiatry, University of California San Francisco (UCSF) School of Medicine, San Francisco, CA, USA; Psychiatric Clinic, Lund, Division of Psychiatry, Lund, Sweden.
    Wolkowitz, Owen M.
    Department of Psychiatry, University of California San Francisco (UCSF) School of Medicine, San Francisco, CA, USA.
    Picard, Martin
    Division of Behavioral Medicine, Department of Psychiatry, Columbia University Medical Center, New York, NY, USA; Department of Neurology and Columbia Translational Neuroscience Initiative, Columbia University Medical Center, New York, NY, USA; Columbia Aging Center, Columbia University Medical Center, New York, NY, USA.
    Ohlsson, Lars
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Bersani, Francesco S.
    Department of Psychiatry, University of California San Francisco (UCSF) School of Medicine, San Francisco, CA, USA; Department of Neurology and Psychiatry, Sapienza University of Rome, Rome, Italy.
    Fernström, Johan
    Faculty of Medicine, Department of Clinical Sciences, Psychiatry, Lund University, Lund, Sweden; Psychiatric Clinic, Lund, Division of Psychiatry, Lund, Sweden.
    Westrin, Åsa
    Faculty of Medicine, Department of Clinical Sciences, Psychiatry, Lund University, Lund, Sweden; Psychiatric Clinic, Lund, Division of Psychiatry, Lund, Sweden.
    Hough, Christina M.
    Department of Psychiatry, University of California San Francisco (UCSF) School of Medicine, San Francisco, CA, USA; Department of Psychology, University of California, Los Angeles (UCLA), Los Angeles, CA, USA.
    Lin, Jue
    Department of Biochemistry and Biophysics, University of California San Francisco (UCSF) School of Medicine, San Francisco, CA, USA.
    Reus, Victor I.
    Department of Psychiatry, University of California San Francisco (UCSF) School of Medicine, San Francisco, CA, USA.
    Epel, Elissa S.
    Department of Psychiatry, University of California San Francisco (UCSF) School of Medicine, San Francisco, CA, USA.
    Mellon, Synthia H.
    Department of OB/GYN and Reproductive Sciences, University of California San Francisco (UCSF) School of Medicine, San Francisco, CA, USA.
    Circulating cell-free mitochondrial DNA, but not leukocyte mitochondrial DNA copy number, is elevated in major depressive disorder2018In: Neuropsychopharmacology, ISSN 0893-133X, E-ISSN 1740-634X, Vol. 43, no 7, p. 1557-1564Article in journal (Refereed)
    Abstract [en]

    Major depressive disorder (MDD) has been linked to mitochondrial defects, which could manifest in mitochondrial DNA (mtDNA) polymorphisms or mutations. Additionally, copy number of mtDNA (mtDNA-cn) can be quantified in peripheral blood mononuclear cells (PBMC)s, indirectly reflecting cellular energetics, or in the circulating cell-free mtDNA (ccf-mtDNA) levels, which may reflect a fraction of the mitochondrial genome released during cellular stress. Few studies have examined ccf-mtDNA in MDD, and no studies have tested its relationship with intracellular mtDNA-cn or with antidepressant treatment response. Here, mtDNA levels were quantified in parallel from: (i) PBMCs and (ii) cell-free plasma of 50 unmedicated MDD subjects and 55 controls, in parallel with PBMC telomere length (TL) and antioxidant enzyme glutathione peroxidase (GpX) activity. MtDNA measures were repeated in 19 MDD subjects after 8 weeks of open-label SSRI treatment. In analyses adjusted for age, sex, BMI, and smoking, MDD subjects had significantly elevated levels of ccf-mtDNA (F = 20.6, p = 0.00002). PBMC mtDNA-cn did not differ between groups (p > 0.4). In preliminary analyses, we found that changes in ccf-mtDNA with SSRI treatment differed between SSRI responders and non-responders (F = 6.47, p = 0.02), with the non-responders showing an increase in ccf-mtDNA and responders not changing. Baseline ccf-mtDNA was positively correlated with GpX (r = 0.32, p = 0.001), and PBMC mtDNA correlated positively with PBMC TL (r = 0.38, p = 0.0001). These data suggest that plasma ccf-mtDNA and PBMC mtDNA-cn reflect different cellular processes and that the former may be more reflective of certain aspects of MDD pathophysiology and of the response to SSRI antidepressants.

  • 12.
    Lindqvist, Daniel
    et al.
    Univ Calif San Francisco, San Francisco, CA 94143 USA; Lund Univ, Lund, Sweden.
    Wolkowitz, Owen
    Univ Calif San Francisco, San Francisco, CA 94143 USA.
    Picard, Martin
    Columbia Univ, New York, NY 10027 USA.
    Ohlsson, Lars
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Bersani, Francesco Saverio
    Sapienza Univ Rome, Rome, Italy.
    Fernström, Johan
    Lund Univ, Lund, Sweden.
    Westrin, Åsa
    Lund Univ, Lund, Sweden.
    Hough, Christina
    Univ Calif San Francisco, San Francisco, CA 94143 USA.
    Lin, Jue
    Univ Calif San Francisco, San Francisco, CA 94143 USA.
    Grudet, Cecile
    Lund Univ, Lund, Sweden.
    Ljunggren, Lennart
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Traskman-Bendz, Lil
    Lund Univ, Lund, Sweden.
    Reus, Victor
    Univ Calif San Francisco, San Francisco, CA 94143 USA.
    Epel, Elissa
    Univ Calif San Francisco, San Francisco, CA 94143 USA.
    Mellon, Synthia H.
    Univ Calif San Francisco, San Francisco, CA 94143 USA.
    Circulating Cell-Free Mitochondrial DNA - a Novel Marker of Mitochondrial Stress Associated With Suicidality and Major Depressive Disorder2018In: Biological Psychiatry, ISSN 0006-3223, E-ISSN 1873-2402, Vol. 83, no 9, suppl. 1, p. S25-S26, article id 62Article in journal (Other academic)
    Abstract [en]

    Background: Mitochondrial DNA copy number (mtDNA-cn), which represents the number of mitochondrial genomes per cell, can be quantified in peripheral blood mononuclear cells (PBMC) and is thought to reflect variations in mitochondrial biogenesis. Additionally, mtDNA may be released at low levels into the circulation from mitochondria under cellular stress, resulting in circulating cell-free mtDNA (ccf-mtDNA) detectable in plasma. The source or physiological significance of ccf-mtDNA in psychiatric illness is unknown but may reflect cell damage, cell death, or bioenergetic compromise. Methods: We enrolled suicide attempters (across diagnoses), non-suicidal subjects with Major Depressive Disorder (MDD), and healthy controls (all medication-free) in two independent cohorts (n=110 & n=74). MtDNA was quantified in cell-free plasma and in PBMCs. Results: Ccf-mtDNA was elevated in suicide attempters and in non-suicidal MDD subjects, compared to healthy controls. These group effects were very large (Cohen’s d ranging from 0.9 to 4.0, all p<0.00001). Ccf-mtDNA and cellular PBMC mtDNA-cn were not significantly correlated with each other (r=0.02, p=0.87), suggesting they reflect different processes. Ccf-mtDNA correlated with post-dexamethasone cortisol (r=0.5, p<0.001), suggesting that HPA-axis hyperactivity may be associated with cellular damage and release of ccf-mtDNA into the blood. Ccf-mtDNA also directly correlated with the antioxidant enzyme glutathione peroxidase (r=0.32, p=0.001), possibly reflecting a compensatory attempt to upregulate antioxidant defence mechanisms due to cellular stress. Conclusions: Ccf-mtDNA may represent a novel marker of cellular stress, which is increased in certain psychiatric conditions. These results call for replication in larger cohorts and in longitudinal studies.

  • 13.
    Ohlsson, Lars
    et al.
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Exley, Christopher
    Darabi, Anna
    Sandén, Emma
    Siesjö, Peter
    Eriksson, Håkan
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Aluminium based adjuvants and their effects on mitochondria and lysosomes of phagocytosing cells2013In: Journal of Inorganic Biochemistry, ISSN 0162-0134, E-ISSN 1873-3344, Vol. 128, p. 229-236Article in journal (Refereed)
    Abstract [en]

    Aluminium oxyhydroxide, Al(OH)3 is one of few compounds approved as an adjuvant in human vaccines. However, the mechanism behind its immune stimulating properties is still poorly understood. In vitro co- culture of an aluminium adjuvant and the human monocytic cell line THP-1 resulted in reduced cell proliferation. Inhibition occurred at concentrations of adjuvant several times lower than would be found at the injection site using a vaccine formulation containing an aluminium adjuvant. Based on evaluation of the mitochondrial membrane potential, THP-1 cells showed no mitochondrial rupture after co-culture with the aluminium adju- vant, instead an increase in mitochondrial activity was seen. The THP-1 cells are phagocytosing cells and after co-culture with the aluminium adjuvant the phagosomal pathway was obstructed. Primary or early phagosomes mature into phagolysosomes with an internal pH of 4.5 – 5 and carry a wide variety of hydrolysing enzymes. Co-culture with the aluminium adjuvant yielded a reduced level of acidic vesicles and cathepsin L activity, a proteolytic enzyme of the phagolysosomes, was almost completely inhibited. THP-1 cells are an appropriate in vitro model in order to investigate the mechanism behind the induction of a phagocytosing antigen presenting cell into an inflammatory cell by aluminium adjuvants. Much information will be gained by investigating the phagosomal pathway and what occurs inside the phagosomes and to elucidate the ultimate fate of phagocytosed aluminium particles.

  • 14.
    Ohlsson, Lars
    et al.
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Gustafsson, Anna
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Lavant, Eva
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Suneson, K
    Faculty of Medicine, Department of Clinical Sciences Lund, Psychiatry, Lund University, Lund, Sweden.
    Brundin, L
    Center for Neurodegenerative Science, Van Andel Research Institute, Grand Rapids, MI, USA.
    Westrin, Å
    Faculty of Medicine, Department of Clinical Sciences Lund, Psychiatry, Lund University, Lund, Sweden.
    Ljunggren, Lennart
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Lindqvist, D
    Faculty of Medicine, Department of Clinical Sciences Lund, Psychiatry, Lund University, Lund, Sweden.
    Leaky gut biomarkers in depression and suicidal behavior.2019In: Acta Psychiatrica Scandinavica, ISSN 0001-690X, E-ISSN 1600-0447, Vol. 139, no 2, p. 185-193Article in journal (Refereed)
    Abstract [en]

    OBJECTIVE: Inflammation is associated with major depressive disorder (MDD) and suicidal behavior. According to the 'leaky gut hypothesis', increased intestinal permeability may contribute to this relationship via bacterial translocation across enterocytes. We measured plasma levels of gut permeability markers, in patients with a recent suicide attempt (rSA), MDD subjects with no history of a suicide attempt (nsMDD), and healthy controls (HC), and related these markers to symptom severity and inflammation. METHOD: We enrolled rSA (n = 54), nsMDD (n = 13), and HC (n = 17). Zonulin, intestinal fatty acid binding protein (I-FABP), soluble CD14, and interleukin-6 (IL-6) were quantified in plasma. Montgomery-Asberg Depression Rating Scale (MADRS) and Suicide Assessment Scale (SUAS) were used for symptom assessments. RESULTS: The rSA group displayed higher I-FABP and lower zonulin levels compared with both the nsMDD and the HC groups (all P < 0.001). IL-6 correlated positively with I-FABP (r = 0.24, P < 0.05) and negatively with zonulin (r = -0.25, P < 0.05). In all subjects, I-FABP levels correlated positively with MADRS (r = 0.25, P < 0.05) and SUAS scores (r = 0.38, P < 0.001), and the latter correlation was significant also in the nsMDD group (r = 0.60, P < 0.05). CONCLUSION: The 'leaky gut hypothesis' may improve our understanding of the link between inflammation and suicidal behavior. These findings should be considered preliminary until replicated in larger cohorts.

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  • 15.
    Ohlsson, Lars
    et al.
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Hall, Anna
    Department of Clinical Chemistry, SUS, Region Skane, Malmö, Sweden.
    Lindahl, Hanne
    Department of Clinical Pathology, SUS, Region Skane, Malmö, Sweden.
    Danielsson, Ravi
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Gustafsson, Anna
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Lavant, Eva
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Ljunggren, Lennart
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Increased level of circulating cell-free mitochondrial DNA due to a single bout of strenuous physical exercise2020In: European Journal of Applied Physiology, ISSN 1439-6319, E-ISSN 1439-6327, Vol. 120, p. 897-905Article in journal (Refereed)
    Abstract [en]

    Purpose Physical exercise is reported to affect the immune response in various ways. Thus, the levels of pro-inflammatory cytokines as well as the abundance of circulating leukocytes are changed. In this study, the occurence of circulating cell-free mitochondrial DNA (cfmtDNA) and nuclear DNA (nDNA) was investigated in connection with a single bout of strenuous physical exercise. Methods Healthy volunteers performed a controlled ergo-spirometry cycle test and venous blood samples were taken at different time-points to analyze the concentration of blood components before, during and after the test. The number of circulating leukocytes was measured, as well as secretion of the soluble urokinase activator receptor (suPAR). Results Cf-mtDNA significantly increased during exercise, compared to baseline values and after 30 and 90 min of rest. Circulating leukocytes increased during exercise, but returned to baseline levels afterwards. Surface expression of the urokinase plasminogen activating receptor (uPAR) on neutrophils decreased significantly during exercise. The concentration of suPAR tended to increase during exercise but only significantly after 90 min of rest. Conclusion Increased concentration of cf-mtDNA indicates that cell damage takes place during high intensity training. Hypoxia and tissue damage are likely causes of cf-mtDNA from muscle cells. The levels of cf-mtDNA remain high during the initial rest, due to the decreasing numbers of leukocytes normally clearing the plasma from cf-mtDNA. The increased levels of suPAR further emphasize that strenuous physical exercise causes a reaction similar to inflammation. Further studies are needed to detect the source of increased cf-mtDNA and the corresponding increase of suPAR liberation.

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  • 16.
    Pankratov, Dmitry
    et al.
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Engineering Enzymology, A.N. Bach Institute of Biochemistry, Moscow, 119 071, Russian Federation; Kurchatov NBICS Centre, National Research Centre Kurchatov Institute, Moscow, 123182, Russian Federation.
    Ohlsson, Lars
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Gudmundsson, Petri
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Halak, Sanela
    Medical Imaging and Physiology, Skåne University Hospital, Malmö, 205 06, Sweden.
    Ljunggren, Lennart
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Blum, Zoltan
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Shleev, Sergey
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Engineering Enzymology, A.N. Bach Institute of Biochemistry, Moscow, 119 071, Russian Federation; Kurchatov NBICS Centre, National Research Centre Kurchatov Institute, Moscow, 123182, Russian Federation.
    Ex vivo electric power generation in human blood using an enzymatic fuel cell in a vein replica2016In: RSC Advances, E-ISSN 2046-2069, Vol. 6, no 74, p. 70215-70220Article in journal (Refereed)
    Abstract [en]

    Here we report an enzymic fuel cell in a vein replica that generates sustained electricity, enough to power an e-​ink display, in an authentic human blood stream. We also detail a simple and safe approach for fuel cell evaluation under homeostatic conditions. Our results demonstrate proof-​of-​principle operation of a biocompatible and safe biodevice that could be implanted in superficial human veins, which we anticipate to be a starting point for more sophisticated investigations of personal sources of electricity.

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  • 17.
    Seghatoleslami, Sepideh
    et al.
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Ohlsson, Lars
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Hamberg, Kristina
    Malmö högskola, Faculty of Odontology (OD).
    Carlsson, Peter
    Malmö högskola, Faculty of Odontology (OD).
    Ericson, Dan
    Malmö högskola, Faculty of Odontology (OD).
    Ljunggren, Lennart
    Malmö högskola, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
    Quantitative detection of Streptococcus mutans from saliva using FTATM elute cards and real-time polymerase chain reaction2013In: American Journal of Molecular Biology, ISSN 2161-6620, E-ISSN 2161-6663, Vol. 3, no 3, p. 148-152Article in journal (Refereed)
    Abstract [en]

    Dental caries is a localized, transmissible, pathological infection process that ends up in the destruction of hard dental tissue. Numerous reports have shown the close relationship between salivary levels of Streptococcus mutans and dental caries. As S. mutans, is considered to be the principle etiological agent of dental caries, the development of a quick and convenient method for detection and quantification of these bacteria from patient saliva samples would simplify diagnosis and treatment. The purpose of this study was to compare a new means of quantifying bacteria using FTATM Elute cards and Real-Time Polymerase Chain Reaction to a conventional culture-based assay using oral S. mutans as a model sample. A total of 60 different saliva samples were investigated. The results show a significant negative correlation between the two methods, with a correlation coefficient of −0.577 (Spearman’s Correlation) and p < 0.01. The method demonstrates a high sensitivity, specificity and reliable quantitative results, covering a large range of bacterial concentrations.

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  • 18.
    Tassidis, Helena
    et al.
    Department of Natural Science, Kristianstad University, Kristianstad, Sweden.
    Jankovskaja, Skaidre
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Awad, Kassem
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Ohlsson, Lars
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Gjörloff Wingren, Anette
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Gustafsson, Anna
    Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
    Investigation of tryptophan to kynurenine degradation in response to interferon-γ in melanoma cell lines2024In: Biochemistry and Biophysics Reports, ISSN 2405-5808, Vol. 37, article id 101612Article in journal (Refereed)
    Abstract [en]

    Background and aim: Melanoma is a fatal form of skin cancer that carries a grave prognosis if the cancer cells spread and form metastases. The Kynurenine (Kyn) pathway is activated by the enzyme indoleamine 2,3-dioxygenase 1 (IDO-1) and has been shown to have a role in tumour progression. We have previously shown that interferon-γ (IFN-γ) acts as an inducer of tryptophan (Trp) degradation to Kyn in keratinocytes of the basal layer in a 3D epidermis model. Before extending our reconstructed human epidermis model to not only contain keratinocytes but also fibroblasts and melanocytes/melanoma cells, we have in this study set out to investigate possible differences between primary adult melanocytes and six melanoma cell lines regarding the expression of the immune checkpoint inhibitors IDO-1 and programmed death ligand 1 (PD-L1) together with Kyn production.

    Methods: The melanocytes and melanoma cells were stimulated with 1–20 ng/ml of IFN-γ and the levels of Trp to Kyn degradation were monitored with high-performance liquid chromatography (HPLC). To analyze the viability of the cell types after IFN-γ treatment, an MTT assay was performed. mRNA quantity of IDO-1, PD-L1 and IFN-γ receptor (IFN-GR1) was analyzed with qPCR.

    Results: After 24 h, only the metastatic cell line WM-266-4 was affected by all concentrations of IFN-γ, whereas at 48 h, the higher IFN-γ concentrations gave a more pronounced effect on the viability in all cell types. Trp was detected at various levels in the culture medium from all cell types before and after IFN-γ treatment. The degradation to Kyn was detected in primary melanocytes, Mel Juso, and Mel Ho cell lines after 24 h of treatment and low levels of IFN-γ. However, the higher concentration of IFN-γ, 20 ng/ml, induced Kyn to various degrees in all cell types after 24 h. The change in mRNA quantity of IDO-1 and PD-L1 was similar in all cell types.

    Conclusion: To conclude, no significant difference in upregulation of the immune checkpoint inhibitors PD-L1 and IDO-1 was seen between primary tumour and metastatic melanoma. IFN-γ stimulation of melanocytes and different stages of melanoma cell lines resulted in an increased Kyn/Trp ratio in the more aggressive melanoma cells when a high concentration was used (20 ng/ml) but when a lower concentration of IFN-γ (5 ng/ml) was used an increased Kyn/Trp ratio were detected in media from primary melanocytes and early-stage melanoma.

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