| 1. |
- Gunaltay, Sezin, 1986-, et al.
(författare)
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Clinical and immunologic effects of faecal microbiota transplantation in a patient with collagenous colitis
- 2017
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Ingår i: World Journal of Gastroenterology. - : Baishideng Publishing Group Co. - 1007-9327 .- 2219-2840. ; 23:7, s. 1319-1324
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Tidskriftsartikel (refereegranskat)abstract
- One to six percent of patients with microscopic colitis are refractory to medical treatment. The effect of faecal microbiota transplantation (FMT) in active collagenous colitis (CC) has, to the best of our knowledge, never been reported before. Here, we report the effect of repeated FMT in a patient with CC. The patient presented with severe symptoms including profuse diarrhea and profound weight loss. Although she responded to budesonide in the beginning, she became gradually refractory to medical treatment, and was therefore treated with FMT. The patient remained in remission for 11 mo after the third faecal transplantation. The immunomodulatory effect of the therapy was evaluated using flow cytometry, which showed alterations in the profile of intraepithelial and lamina propria lymphocyte subsets after the second transplantation. Our observations indicate that FMT can have an effect in CC, which support the hypothesis that luminal factors, influencing the intestinal microbiota, are involved in the pathogenesis of CC.
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| 2. |
- Günaltay, Sezin, 1986-, et al.
(författare)
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Differential expression of interleukin-1/Toll-like receptor signaling regulators in microscopic and ulcerative colitis
- 2014
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Ingår i: World Journal of Gastroenterology. - : WJG Press. - 1007-9327 .- 2219-2840. ; 20:34, s. 12249-12259
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Tidskriftsartikel (refereegranskat)abstract
- AIM: To investigate Toll-like receptor (TLR) signaling regulators in microscopic and ulcerative colitis patients.METHODS: Total RNA and microRNA were isolated from fresh frozen colonic biopsies of non-inflamed controls and patients with active or in-remission collagenous colitis (CC), lymphocytic colitis (LC), or ulcerative colitis (UC). We compared expressions of interleukin-1 receptor-associated kinase (IRAK)-2, IRAK-M, interleukin (IL)-37, microRNA (miR)-146a, miR-155, and miR-21 using quantitative real time reverse transcription polymerase chain reaction.RESULTS: IRAK-M expression was increased in LC patients with active disease in histopathological remission (LC-HR; P = 0.02) and UC patients (P = 0.01), but no differences in IRAK-2 expression were detected compared to controls. miR-146a, -155 and -21 expressions were increased in LC-HR (P = 0.04, 0.07, and 0.004) and UC (P = 0.02, 0.04 and 0.03) patients. miR-146a and miR-21 expressions were significantly enhanced in UC patients compared to UC remission (UC-R; P = 0.01 and 0.04). Likewise, active CC patients showed significantly increased expression of miR-155 (P = 0.003) and miR-21 (P = 0.006). IL-37 expression was decreased in both CC (P = 0.03) and LC (P = 0.04) patients with a similar trend in UC patients but not statistically significant, whilst it was increased in UC-R patients compared to controls (P = 0.02) and active UC (P = 0.001).CONCLUSION: The identification of differentially expressed miRNAs, IL-37, and IRAK-M suggests different pathophysiologic mechanisms in various disease stages in LC, CC, and UC. (C) 2014 Baishideng Publishing Group Inc. All rights reserved.
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| 3. |
- Günaltay, Sezin, 1986-
(författare)
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Dysregulated mucosal immune responses in microscopic colitis patients
- 2016
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Microscopic colitis (MC), comprising collagenous colitis (CC) and lymphocytic colitis (LC) is a common cause of chronic watery diarrhea. The diagnosis relies on typical histopathological changes observed upon microscopic examination. The studies in this thesis investigated innate and adaptive immune responses in the colonic mucosa of MC patients, also comparing patients with active disease (CC and LC) and histopathologically in remission (CC/LC-HR). We first analyzed expression of interleukin-1/Toll-like receptor (IL-1/TLR) signaling regulators in MC patients (Paper I). Our results showed enhanced IRAK-M, microRNA-146a, -155 and -21 expressions, whereas IL-37 gene expression was reduced in CC and LC patients as compared to non-inflamed controls. These results suggest different pathophysiological mechanisms in MC patients. The mixed inflammatory cell infiltrations seen in the lamina propria of MC patients might be a result of dysregulated expression of chemotactic mediators. In Paper II, we showed that MC patients display mainly an increased expression of chemokines and chemokine receptors in active disease as compared to noninflamed controls. In Paper III, we examined if the decreased IL-37 expression seen in Paper I could mediate the upregulation of chemokines seen in Paper II. We showed that a relatively small reduction in the ability of epithelial cells to produce IL-37 results in mainly increased chemokine expressions in a pattern similar to the findings in Paper II. In order to understand the nature of infiltrating T cells commonly observed in MC patients, we analyzed the T cell receptor (TCR) β chains in colonic biopsies of MC patients (Paper IV). Our results showed significant differences in TCRβ repertoire, which suggests selectively expanded T cell clones in active MC and histopathologically in remission patients. Altogether, these results i) increase the knowledge of MC pathogenesis by showing changes in TLR signaling regulators, enhanced chemokine and their receptor expressions involved in a mixed immune cell infiltrations and selectively expanded T cell clones in CC and LC patients, as well as in histopathological remission ii) might potentially increase the possibility of more target-specific therapies based on IL-37 induction, chemokines or chemokine receptor inhibitions, or hindering T cell infiltration according to TCR clonality.
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| 4. |
- Gunaltay, Sezin, 1986-, et al.
(författare)
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Enhanced levels of chemokines and their receptors in the colon of microscopic colitis patients indicate mixed immune cell recruitment
- 2015
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Ingår i: Mediators of Inflammation. - : Hindawi Limited. - 0962-9351 .- 1466-1861.
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Tidskriftsartikel (refereegranskat)abstract
- Microscopic colitis (MC), comprising collagenous colitis (CC) and lymphocytic colitis (LC), is a common cause of chronic diarrhea. Various immune cell infiltrations in the epithelium and lamina propria are seen in MC immunopathology. We compared gene and protein expressions of different immune cell attracting chemokines and their receptors in colon biopsies from MC patients in active disease or histopathological remission (CC/LC-HR) with controls, using qRT-PCR and Luminex, respectively. CC and LC patients with active disease demonstrated a mixed chemokine profile with significantly enhanced gene and/or protein expressions of the chemokines CCL2, CCL3, CCL4, CCL5, CCL7, CCL22, CXCL8, CXCL9, CXCL10, CXCL11, and CX(3)CL1 and the receptors CCR2, CCR3, CCR4, CXCR1, CXCR2, and CX(3)CR1. Enhanced chemokine/chemokine receptor gene and protein levels in LC-HR patients were similar to LC patients, whereas CC-HR patients demonstrated almost normalized levels. These findings expand the current understanding of the involvement of various immune cells in MC immunopathology and endorse chemokines as potential diagnostic markers as well as therapeutic candidates. Moreover, this study further supports the hypothesis that CC and LC are two different entities due to differences in their immunoregulatory responses.
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| 5. |
- Gunaltay, Sezin, 1986-, et al.
(författare)
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Oligoclonal T-cell Receptor Repertoire in Colonic Biopsies of Patients with Microscopic Colitis and Ulcerative Colitis
- 2017
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Ingår i: Inflammatory Bowel Diseases. - : Lippincott Williams & Wilkins. - 1078-0998 .- 1536-4844. ; 23:6, s. 932-945
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Tidskriftsartikel (refereegranskat)abstract
- Background: Microscopic colitis (MC), comprising collagenous colitis (CC) and lymphocytic colitis (LC), is a type of variation of inflammatory bowel diseases. Local T-cell infiltration in the mucosa plays a major role in MC immunopathology.Methods: To understand diversity and clonality of infiltrating T cells, we analyzed the T-cell receptor beta (TCR beta) chains in colonic biopsies of MC, ulcerative colitis (UC), and their remission counterparts (CC/LC-HR [histological remission] or UC-R [remission]) compared with patients with non-inflamed colons using next-generation sequencing.Results: Compared with controls and patients with CC, patients with LC had significantly lower diversity with significantly lower evenness and richness in TCRVb-Jb gene segments. Similarly, patients with LC-HR had lower diversity because of significantly lower TCRVb-Jb clone richness. Patients with UC and UC-R showed significantly higher diversity and richness. Univariate and multivariate analyses were performed to identify TCRVb-Jb gene segments differentiating disease types from controls or their remission counterparts. Patients with LC were discriminated from controls by 12 clones and from patients with CC by 8 clones. Neither univariate nor multivariate analyses showed significance for patients with CC or CC-HR compared with controls. Patients with UC and UC-R had 16 and 14 discriminating clones, respectively, compared with controls.Conclusions: Altogether, patients with MC and UC showed an oligoclonal TCRb distribution. TCRVb-Jb clone types and their diversity were distinctive between patients with CC and LC, as well as for patients with UC, suggesting different pathophysiological mechanisms according to disease type and stage. This study suggests that CC and LC are different entities because of differences in immunoregulatory responses, as mirrored by their T-cell repertoire.
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| 6. |
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| 7. |
- Günaltay, Sezin, 1986-, et al.
(författare)
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Reduced IL-37 Production Increases Spontaneous Chemokine Expressions in Colon Epithelial Cells
- 2017
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Ingår i: Digestive Diseases and Sciences. - : Springer. - 0163-2116 .- 1573-2568. ; 62:5, s. 1204-1215
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Tidskriftsartikel (refereegranskat)abstract
- Background and Aim: Microscopic colitis, comprising collagenous colitis and lymphocytic colitis, is a common cause of chronic diarrhea. Previously, we showed enhanced chemokine productions in microscopic colitis patients, indicating dysregulated immune cell chemotaxis in the immunopathogenesis. We also showed decreased mRNA of IL-37, mainly regarded as an anti-inflammatory cytokine, in the colonic mucosa of these patients, potentially an important factor for the chronicity of the colitis. Our aim in this study was to understand the possible role of IL-37 in chemokine production using a cell line model.Methods: A colon epithelial cell line, T84, was stimulated with the TLR5 ligand flagellin. IL-37 protein production was reduced 20% using the CRISPR/Cas9 system, and the changes in chemokine mRNA and protein expressions were compared to cells transfected with empty plasmid.Results: The 20% reduction in IL-37 protein levels spontaneously increased CCL5, CXCL8, CXCL10, and CXCL11 mRNA and protein expressions. CCL2 mRNA and protein levels were enhanced upon TLR5 stimulation. CCL3, CCL20, and CX3CL1 mRNA expressions were increased either spontaneously or following TLR5 stimulation, whereas CCL4 and CCL22 mRNA expressions were significantly decreased.Conclusions: Even a minor decrease in the ability of colon epithelial cells to produce IL-37 results in altered chemokine expression, mainly an increase in the production of several chemokines. Our results indicate that a decreased IL-37 expression by colon epithelial cells may be an important factor for increasing the recruitment of immune cells and subsequently developing microscopic colitis.
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| 8. |
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| 9. |
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| 10. |
- Jayaprakash, Kartheyaene, 1983-, et al.
(författare)
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PKC, ERK/p38 MAP kinases and NF-B targeted signalling play a role in the expression and release of IL-1β and CXCL8 in Porphyromonas gingivalis-infected THP1 cells
- 2017
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Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS). - : John Wiley & Sons. - 0903-4641 .- 1600-0463. ; 125:7, s. 623-633
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Tidskriftsartikel (refereegranskat)abstract
- Porphyromonas gingivalis is a keystone pathogen in periodontitis and is gaining importance in cardiovascular pathogenesis. Protease-activated receptors (PARs), toll-like receptors (TLRs) and nucleotide-binding oligomerization domain (NOD) on monocytes recognize the structural components on P. gingivalis, inducing inflammatory intermediates. Here, we elucidate the modulation of PARs, TLRs, NODs, and the role of MAPK and NF-B in IL-1 and CXCL8 release. THP1 cells were stimulated with P. gingivalis wild-type W50 and its isogenic gingipain mutants: Rgp mutant E8 and Kgp mutant K1A. We observed modulation of PARs, TLRs, NOD, IL-1 and CXCL8 expression by P. gingivalis. Gingipains hydrolyse IL-1 and CXCL8, which is more evident for IL-1 accumulation at 24 h. Inhibition of PKC (protein kinase C), p38 and ERK (extracellular signal-regulated kinases) partially reduced P. gingivalis-induced IL-1 at 6 h, whereas PKC and ERK reduced CXCL8 at both 6 and 24 h. Following NF-B inhibition, P. gingivalis-induced IL-1 and CXCL8 were completely suppressed to basal levels. Overall, TLRs, PARs and NOD possibly act in synergy with PKC, MAPK ERK/p38 and NF-B in P. gingivalis-induced IL-1 and CXCL8 release from THP1 cells. These pro-inflammatory cytokines could affect leucocytes in circulation and exacerbate other vascular inflammatory conditions such as atherosclerosis.
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| 11. |
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| 12. |
- Wallenhammar, Amélie, 1986-, et al.
(författare)
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Developing a novel strategy to determine new Tick-borne encephalitis foci
- 2018
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Ingår i: 15th Smögen Summer Symposium on Virology, Smögen, Sweden, August 23-25, 2018..
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Konferensbidrag (refereegranskat)abstract
- TBEV is the most important viral tick-borne zoonosis in Europe, and infection may lead to severeCNS disease, including encephalitis and myelitis. Climate changes have increased the tickdistribution in Sweden, increasing the risk areas of Tick-borne encephalitis (TBE) at severalregions including the Örebro county. The TBE virus (TBEV) is usually transmitted to humans viatick bites, however oral transmission through consumption of non-pasteurized dairy products havealso been described. Both TBEV and antibodies against the viral proteins have been detected inmilk of goats, sheep and cattle. Since the prevalence of TBEV in the tick population is low there is a need for new and robust surveillance techniques identifying novel risk areas of TBEV at earlystages.In this study we have developed a strategy for identifying new TBEV foci. We have collected raw milk and colostrum samples from sheep and goats in the Örebro region. The milk samples were analyzed for the presence of TBEV antibodies by ELISA. In addition, the ELISA results were further verified by an in-house Western Blot assay where milk samples were used asprimary antibody to detect the Envelope-protein of TBEV. This method has so far revealed two novel TBEV foci within the Örebro county. Questing ticks and ticks feeding on sheep have been collected at areas of TBEV positive milk. The ticks are currently being analyzed for TBEV by PCR. The specific Örebro strains will be isolated from TBEV positive ticks and the viral genomeswill be further characterized using established next-generation sequencing technique.
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| 13. |
- Wallenhammar, Amélie, 1986-, et al.
(författare)
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Revealing new tick-borne encephalitis foci by screening antibodies in sheep milk
- 2019
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Ingår i: 16th Smögen Symposium on Virology, August 22-24, 2019.
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Konferensbidrag (refereegranskat)abstract
- Climate changes have increased the tick-distribution in Sweden, and the prevalence of ticks has been predicted to increase towards the northern parts of the country, increasing the risk of tick-borne zoonosis in new regions. Tick-borne encephalitis (TBE) is the most important viral tick-borne zoonosis in Sweden as well as in Europe. TBE virus (TBEV) infection often leads to severe CNS disease, including encephalitis and severe myelitis, which may lead to paralysis and respiratory failure in humans. TBEV and antibodies against TBEV are excreted in milk of goats, sheep and cattle and the virus can be ingested orally by consumption of non-pasteurized dairy products. Since the prevalence of TBEV in the tick population is low there is a need for new and robust surveillance techniques identifying new risk areas of TBEV at early stages.In this study we have developed a novel strategy for identifying new TBEV foci. We have collected raw milk and colostrum samples from sheep and goats in Örebro County, Sweden. The milk samples were analyzed for the presence of TBEV antibodies by ELISA, and validated by an in-house Western Blot assay where milk samples were used as primary antibody to detect purified TBEV E-protein. By monitoring TBEV antibodies in milk we have found three novel foci in the Örebro County which also overlap with the plausible place of infection of registered human TBE cases reported during 2009-2018. Furthermore, the stability of TBEV in milk and raw milk was studied at different temperatures. Our data indicates that keeping unpasteurized milk at 4 ˚C will preserve the infectivity of TBEV for several days. Ticks have also been collected from areas with TBEV positive milk. We aim to extract total RNA from the sampled ticks, followed by TBEV detection by nested-PCR and next-generation sequencing.Here we present a novel technique to reveal risk areas of TBE in Sweden, which is robust, reliable, and non-invasive and can accordingly be used to map TBEV “hotspots”. In the TBE foci, more than 50 % of the tested animals were antibody positive, and TBEV infectivity in refrigerated milk was preserved, stressing the importance of pasteurization before consumption.
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| 14. |
- Wallenhammar, Amélie, 1986-, et al.
(författare)
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Revealing new tick-borne encephalitis virus foci by screening antibodies in sheep milk
- 2020
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Ingår i: Parasites & Vectors. - : BioMed Central. - 1756-3305. ; 13:1
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Tick distribution in Sweden has increased in recent years, with the prevalence of ticks predicted to spread towards the northern parts of the country, thus increasing the risk of tick-borne zoonoses in new regions. Tick-borne encephalitis (TBE) is the most significant viral tick-borne zoonotic disease in Europe. The disease is caused by TBE virus (TBEV) infection which often leads to severe encephalitis and myelitis in humans. TBEV is usually transmitted to humans via tick bites; however, the virus can also be excreted in the milk of goats, sheep and cattle and infection may then occur via consumption of unpasteurised dairy products. Virus prevalence in questing ticks is an unreliable indicator of TBE infection risk as viral RNA is rarely detected even in large sample sizes collected at TBE-endemic areas. Hence, there is a need for robust surveillance techniques to identify emerging TBEV risk areas at early stages.METHODS: Milk and colostrum samples were collected from sheep and goats in Örebro County, Sweden. The milk samples were analysed for the presence of TBEV antibodies by ELISA and validated by western blot in which milk samples were used to detect over-expressed TBEV E-protein in crude cell extracts. Neutralising titers were determined by focus reduction neutralisation test (FRNT). The stability of TBEV in milk and colostrum was studied at different temperatures.RESULTS: In this study we have developed a novel strategy to identify new TBEV foci. By monitoring TBEV antibodies in milk, we have identified three previously unknown foci in Örebro County which also overlap with areas of TBE infection reported during 2009-2018. In addition, our data indicates that keeping unpasteurised milk at 4 °C will preserve the infectivity of TBEV for several days.CONCLUSIONS: Altogether, we report a non-invasive surveillance technique for revealing risk areas for TBE in Sweden, by detecting TBEV antibodies in sheep milk. This approach is robust and reliable and can accordingly be used to map TBEV "hotspots". TBEV infectivity in refrigerated milk was preserved, emphasising the importance of pasteurisation (i.e. 72 °C for 15 s) prior to consumption.
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