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| 3. |
- Fryk, Emanuel, et al.
(författare)
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Galectin-1 correlates with inflammatory markers and T regulatory cells in children with type 1 diabetes and/or celiac disease
- 2024
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Ingår i: Clinical and Experimental Immunology. - : Oxford University Press. - 1365-2249 .- 0009-9104. ; 215:3, s. 240-250
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Tidskriftsartikel (refereegranskat)abstract
- Type 1 diabetes (T1D) and celiac disease (CeD) are common autoimmune diseases in children where the pathophysiology is not fully characterized. The autoimmune process involves a complex scenario of both inflammatory and regulatory features. Galectin-1 (GAL-1) has a wide range of biological activities e.g. interaction with immune cells. We examined the relationship between GAL-1 and soluble immune markers and T-cell subsets in a cohort of children with T1D and/or CeD relative to healthy children. GAL-1, together with several soluble immune markers [e.g. interleukins (IL)], tumor necrosis factor (TNF), acute phase proteins, and matrix metalloproteinases (MMP) were measured in sera from children with T1D and/or CeD by fluorochrome (Luminex) technique using children without these diseases as a reference. Subgroups of T cells, including T-regulatory (Treg) cells, were analysed by flow cytometry. Association between GAL-1, pro-inflammatory markers, and Treg cells differed depending on which illness combination was present. In children with both T1D and CeD, GAL-1 correlated positively with pro-inflammatory markers (IL-1 beta, IL-6, and TNF-alpha). Composite scores increased the strength of correlation between GAL-1 and pro-inflammatory markers, Th1-associated interferon (IFN)-gamma, and T1D-associated visfatin. Contrary, in children diagnosed with exclusively T1D, GAL-1 was positively correlated to CD25hi and CD25hiCD101+ Treg cells. For children with only CeD, no association between GAL-1 and other immune markers was observed. In conclusion, the association observed between GAL-1, soluble immune markers, and Treg cells may indicate a role for GAL-1 in the pathophysiology of T1D and, to some extent, also in CeD. Type 1 diabetes (T1D) and celiac disease (CeD) are common autoimmune diseases in children where pathophysiology is not fully characterized. The autoimmune process involves a complex scenario of both inflammatory and regulatory features. Galectin-1 (GAL-1) has a wide range of biological activities e.g. interaction with immune cells and the association observed between GAL-1, soluble immune markers, and T regulatory (Treg) cells may indicate a role for GAL-1 in the pathophysiology of children with T1D and, to some extent, also in children with CeD. Graphical Abstract
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| 7. |
- Ross, Owen A., et al.
(författare)
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Mitochondrial DNA damage in lymphocytes : a role in immunosenescence?
- 2002
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Ingår i: Experimental Gerontology. - : Elsevier. - 0531-5565 .- 1873-6815. ; 37:2-3, s. 329-340
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Tidskriftsartikel (refereegranskat)abstract
- An age-related increase of DNA damage/mutation has been previously reported in human lymphocytes. The high copy number and mutation rate make the mtDNA genome an ideal candidate for assessing damage and to act as a potential biomarker of ageing. In the present study, two assays were developed to evaluate the level of mtDNA4977 and the accumulation of point mutations with age. A competitive polymerase chain reaction (PCR) methodology incorporating three primers was used to detect and quantify the levels of mtDNA4977 and a novel heteroduplex reference strand conformational analysis (RSCA) technique was used to analyse the accumulation of point mutations. The assays were applied to an in vitro model of T cell ageing and ex vivo DNA samples from an elderly cohort of subjects and a younger control group. The mtDNA4977 was detected in all the DNA samples examined but only a very low concentration was observed and no age-related increase or accumulation was observed. No accumulation of point mutations was identified using RSCA within the T cell clones as they were aged or the ex vivo lymphocytes from the elderly cohort. A higher level of variation was observed within the ex vivo DNA samples, verifying the high resolution of RSCA and its ability to identify different mtDNA species, although no correlation with age was observed. The low level of mtDNA damage observed with respect to the ex vivo lymphocyte DNA samples within this study may be due in part to the high turnover of blood cells/mtDNA, which may inhibit the accumulation of genetically abnormal mtDNA that may play a role in immunosenescence. A similar explanation may also apply to the in vitro model of T cell ageing if the vast majority of the cells are replicating rather than entering senescence.
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| 9. |
- Tompa, Andrea
(författare)
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Pinpointing biomarkers of importance for children with combined type 1 diabetes and celiac disease
- 2021
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Type 1 diabetes (T1D) and celiac disease are both characterized by an autoimmune feature. The diseases also share the same risk genes, and thereby patients have an increased risk of developing the other disease subsequently. The pattern of peripheral T and B cell subsets and soluble immune markers (cytokines, chemokines, acute phase proteins, adipocytokines and matrix metalloproteinases) are not yet well characterized in children with a combination of the common pediatric immunological disorders, T1D and celiac disease. To better understand the complex pathophysiology, it is important to gain a deeper knowledge of alterations present in the peripheral immune profile in children with these autoimmune diseases. Pinpointing biomarkers, e.g. peripheral immune markers, can hopefully contribute to the improvement of prognosis, diagnosis, and disease management. Flow cytometry is useful for studying different immune cells, but several pre-analytical factors may affect the outcome. In order to generate reliable results, it is important to evaluate the impact of different pre-analytical factors that possibly can lead to in vitro alterations of the immune cells.Aim: The overall aim of this thesis was to increase our knowledge of peripheral immune marker patterns in children with a combined diagnosis of T1D and celiac disease, with a focus on T and B cell subsets and soluble immune markers; by immunological methods evaluated and adapted for this purpose.Methods: This thesis comprises methodological and cross-sectional studies. The methodological studies are based on whole blood collected from sixty blood donors to examine the impact of pre-analytical factors (anticoagulant, sample handling time, isolation and cryopreservation) that may affect the immune cells (Study I, II). The cross-sectional studies include blood samples collected from a total of 103 participants (children with T1D and/or celiac disease or no diagnosis at all). The pattern of peripheral B (Study II) and T (Study III) cell subsets were examined by flow cytometry. Nearly thirty soluble immune markers were quantified in serum by Luminex technology (Study IV).Results: Peripheral lymphocytes were stable in whole blood samples up to 24 hours, regardless of the anticoagulant. Generally, T and B cell subsets were not affected by isolation and cryopreservation. Children with combined T1D and celiac disease had higher percentages of terminally differentiated memory T helper cells, lower percentages of effector memory T cytotoxic cells and lower expression of suppressive immune markers on regulatory T cells compared with the other study groups. Further, children with combined T1D and celiac disease had a higher percentage of memory B and lower percentages of naive B cells than children with either T1D or celiac disease. Contrary, children with single diagnoses had an inverted naive/memory B cell pattern compared to children with combined diagnoses. Several of the "classical" (cytokines, chemokines), as well as "non-classical" (acute phase proteins, adipocytokines, matrix metalloproteinases) immune markers, were lower in children with combined diagnoses compared to the other study groups.Conclusions: Based on our results, we conclude that whole blood samples stored up to 24 hours are feasible for flow cytometric analysis of lymphocyte subsets, regardless of the type of anticoagulant. Further, isolated and cryopreserved immune cells are feasible for flow cytometric analysis of T and B cell subsets. Impairment in the T and B cells mediated immune regulation in children with combined T1D and celiac disease seems to be clearly divergent from those seen in children with exclusively one of these two autoimmune diseases. Children with combined T1D and celiac disease appear to have a suppressed immune profile, including "classical" and "non-classical" immune markers. The methodological studies provide deeper knowledge of how reliable results can be obtained in studies of peripheral immune cells, e.g., in children with autoimmune diseases. The knowledge obtained by this thesis also brings a better understanding of the pattern of peripheral immune markers in T1D and/or celiac disease. This could potentially contribute to promoting the improvement of prognosis, diagnosis, and disease management.
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| 10. |
- Tompa, Andrea, 1969, et al.
(författare)
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Shift in the B cell subsets between children with type 1 diabetes and/or celiac disease
- 2024
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Ingår i: Clinical and Experimental Immunology. - : Oxford University Press. - 1365-2249 .- 0009-9104. ; 216:1, s. 36-44
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Tidskriftsartikel (refereegranskat)abstract
- Our purpose was to characterize the pattern of B cell subsets in children with a combined diagnosis of type 1 diabetes (T1D) and celiac disease (C) since children with single or double diagnosis of these autoimmune diseases may differ in peripheral B cell subset phenotype patterns. B cells were analyzed with flow cytometry for the expression of differentiation/maturation markers to identify transitional, naive, and memory B cells. Transitional (CD24(hi)CD38(hi)CD19(+)) and memory Bregs (mBregs; CD24(hi)CD27(+)CD19(+), CD1d(+)CD27(+)CD19(+), and CD5(+)CD1d(+)CD19(+)) were classified as B cells with regulatory capacity. Children with a combined diagnosis of T1D and C showed a pattern of diminished peripheral B cell subsets. The B cells compartment in children with combined diagnosis had higher percentages of memory B subsets and Bregs, including activated subsets, compared to children with either T1D or C. Children with combined diagnosis had a lower percentage of naive B cells (CD27(-)CD19(+); IgD(+)CD19(+)) and an increased percentage of memory B cells (CD27(+)CD19(+); IgD(-)CD19(+)). A similar alteration was seen among the CD39(+) expressing naive and memory B cells. Memory Bregs (CD1d(+)CD27(+)CD19(+)) were more frequent, contrary to the lower percentage of CD5(+) transitional Bregs in children with a combined diagnosis. In children with either T1D or C, the peripheral B cell compartment was dominated by naive cells. Differences in the pattern of heterogeneous peripheral B cell repertoire subsets reflect a shifting in the B cell compartment between children with T1D and/or C. This is an immunological challenge of impact on the pathophysiology of these autoimmune diseases. A larger compartment of naive B and Breg subsets, including activated subsets, and smaller compartment of the memory B and Breg subsets are seen in children with isolated type 1 diabetes (T1D) or celiac disease (C). In children with the combined diagnosis, a shifting of the naive B subset compartment toward a more mature/differentiated B cell compartment was observed, indicating another type of disturbance of the B cell mediated immune regulation compared to what is present in the context of only one isolated autoimmune disease. Differences in the pattern of heterogeneous peripheral B cell repertoire subsets reflect a shifting in the B cell compartment between children with T1D and/or C.
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| 12. |
- Tompa, Andrea, et al.
(författare)
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Subsets of CD4+, CD8+, and CD25hi Lymphocytes Are in General Not Influenced by Isolation and Long-Term Cryopreservation
- 2018
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Ingår i: Journal of Immunology. - : American Association of Immunologists. - 0022-1767 .- 1550-6606. ; 201:6, s. 1799-1809
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Tidskriftsartikel (refereegranskat)abstract
- Several key factors can affect the outcome of immunological studies; isolation/cryopreservation can possibly alter T, B, NK, and T-regulatory (Treg) cell marker expression patterns. Blood samples from 50 blood donors supplemented with Na-heparin or K2EDTA were handled within 4 and 24 h after blood sampling. PBMC were isolated with different density gradients. Flow cytometric analysis of intracellular and extracellular CD markers was performed on blood samples freshly isolated PBMC, and PBMC was thawed 6 and 12 mo post-cryopreservation for the purpose of identifying B, NK, Th, T-cytotoxic, and Treg cells. No differences were observed in the percentages for CD3+, CD3+CD4+, CD3+CD8+, CD19+, or CD56+CD16+ cells within 24 h of sampling regardless of which supplement or isolation techniques were used. Differentiated (diff) CD4+ cells were in general less affected by isolation and cryopreservation than diff CD8+ cells. Terminally diff effector CD4+ and CD8+ cells were not affected by either isolation of lymphocytes or cryopreservation. In contrast, naive and early-diff effector memory CD4+ and CD8+ cells were affected by isolation and cryopreservation. The percentages of Treg cells defined as CD4+CD25hi expressing CD101 or CD129, CD4+CD25hiCD127−, and CD4+CD25hiCD127−FOXP3+, respectively, remained stable after isolation and cryopreservation. Subsets expressing CD127, with or without FOXP3, were not affected by isolation/cryopreservation. Subsets expressing CD39, contrary to CD45RA, on CD4+CD25+CD127− cells with or without FOXP3 were not affected by either isolation or cryopreservation. In conclusion, subsets of CD4+, CD8+, and CD25hi lymphocytes are in general not influenced by isolation and long-term cryopreservation.
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| 13. |
- Tompa, Andrea, et al.
(författare)
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Suppressed immune profile in children with combined type 1 diabetes and celiac disease
- 2020
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Ingår i: Clinical and Experimental Immunology. - : John Wiley & Sons. - 0009-9104 .- 1365-2249. ; 201:3, s. 244-257
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Tidskriftsartikel (refereegranskat)abstract
- Children diagnosed with a combination of type 1 diabetes (T1D) and celiac disease (CD) show a dysregulated T helper (Th) 1/Th17 response. Besides the cellular involvement, several soluble immune markers are involved in the autoimmune process of both T1D and CD. Only few studies have examined the peripheral pattern of different cytokines, chemokines, and acute phase proteins (APP) in children with combined T1D and CD. To our knowledge no studies have evaluated the serum levels of adipocytokines and matrix metalloproteinases (MMPs) in this context. The purpose of the present study was to acquire more knowledge and to gain deeper understanding on peripheral immunoregulatory milieu in children with both T1D and CD. The study included children diagnosed with both T1D and CD (n=18), children with T1D (n=27) or CD (n=16), and reference children (n=42). Sera were collected and analysis of twenty-eight immune markers (cytokines, chemokines, APPs, adipocytokines and MMPs) was performed with Luminex technique. The major findings showed that children with double diagnosis had lower serum levels of interleukin (IL)-22, monocyte chemoattractant protein (MIP)-1α, monocyte chemoattractant protein (MCP)-1, procalcitonin, fibrinogen, visfatin and matrix metalloproteinase (MMP)-2. These results indicate a suppressed immune profile in children with combined T1D and CD, including Th17 cytokines, chemokines, acute phase proteins, adipocytokines and MMPs. We conclude that besides cytokines and chemokines, other immune markers, e.g. APPs, adipocytokines and MMPs, are of importance for further investigations to elucidate the heterogeneous immune processes present in patients diagnosed with T1D in combination with CD.
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| 14. |
- Tompa, Andrea, et al.
(författare)
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Suppressed immune profile in children with type 1 diabetes in combination with celiac disease
- 2019
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Konferensbidrag (refereegranskat)abstract
- Introduction: Cytokines, chemokines, acute phase proteins (APP), adipocytokines and matrix metalloproteinases (MMP) are involved in different pathophysiological processes of inflammatory character. The role of the different immune markers and the peripheral immunoregulatory milieu in children diagnosed with type 1 diabetes (T1D) in combination with celiac disease (CD) is not fully understood and is not well studied. The purpose of the present study was therefore to acquire more knowledge and to gain deeper understanding on peripheral immunoregulatory milieu in children with T1D and/or CD.Methods: The study included children diagnosed with T1D in combination with CD (n=18), children with T1D (n=27) or CD (n=16), and reference children (n=42).Blood samples were collected, and serum stored in -80°C until analysis, avoiding multiple freeze-thaw cycles. The inflammatory cyto/chemokines (IL-1β, -5, -6, -8, -9, -10, -13, -15, -17A, -22, -25, -33, IFN-γ, TNF-α, G-CSF, MCP-1, MIP-1α, MIP-1β), diabetes related immune markers (visfatin, resistin), APP (procalcitonin (PTC), ferritin, tissue protein activator, fibrinogen, serum amyloid A) and matrix metalloproteinases (MMP-1, -2, -3) were analyzed with Luminex technique using Bio-Plex assays. Hierarchical cluster analysis was used to identify similarities/differences in immune profiles between children with double diagnosis and children with single diagnosis and reference children. Mann-Whitney U test was used for comparison of the different diagnosis groups within the clusters and whole cohort, respectively.Results: The largest cluster included 75% of the participants and the diagnose distribution in the cluster were very similar to the distribution in the whole study cohort. The remaining 25% were divided in two smaller clusters representing 15.5% and 6.5% respectively. The major finding of this study showed that children with double diagnosis had (1) lower serum levels of IL-22, MCP-1, PCT, visfatin and MMP-2 compared to children with T1D; and (2) lower serum levels of the APC associated chemokine MIP-1α compared to reference children, observed in the main cluster. Most of these observations were also seen in the whole cohort. Conclusion: Our observations indicate decreased serum levels of IL-22, MIP-1α, MCP-1, PCT, visfatin and MMP-2 in children diagnosed with T1D in combination with CD. These results indicate a suppressed immune profile including Th17 cytokines, chemokines, acute phase proteins, diabetes-related and matrix metalloproteinase immune markers. Functional studies of the involved immune cells (CD4+ Treg, CD8+ Treg, NK-cells and dendritic cells) could contribute to elucidate the heterogeneous immunological processes in children with more than one autoimmune disease.
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| 15. |
- Åkesson, Karin, et al.
(författare)
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Low expression of CD39(+)/CD45RA(+) on regulatory T cells (T-reg) cells in type 1 diabetic children in contrast to high expression of CD101(+)/CD129(+) on T-reg cells in children with coeliac disease
- 2015
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Ingår i: Clinical and Experimental Immunology. - : Wiley: 12 months. - 0009-9104 .- 1365-2249. ; 180:1, s. 70-82
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Tidskriftsartikel (refereegranskat)abstract
- Type 1 diabetes (T1D) and coeliac disease are both characterized by an autoimmune feature. As T1D and coeliac disease share the same risk genes, patients risk subsequently developing the other disease. This study aimed to investigate the expression of T helper (Th), T cytotoxic (Tc) and regulatory T cells (T-reg) in T1D and/or coeliac disease children in comparison to healthy children. Subgroups of T cells (Th:CD4(+) or Tc:CD8(+)); naive (CD27(+)CD28(+)CD45RA(+)CCR7(+)), central memory (CD27(+)CD28(+)CD45RA(-)CCR7(+)), effector memory (early differentiated; CD27(+)CD28(+)CD45RA(-)CCR7(-) and late differentiated; CD27(-)CD28(-)CD45RA(-)CCR7(-)), terminally differentiated effector cells (TEMRA; CD27(-)CD28(-)CD45RA(+)CCR7(-)) and T-reg (CD4(+)CD25(+)FOXP3(+)CD127(-)) cells, and their expression of CD39, CD45RA, CD101 and CD129, were studied by flow cytometry in T1D and/or coeliac disease children or without any of these diseases (reference group). Children diagnosed with both T1D and coeliac disease showed a higher percentage of TEMRA CD4(+) cells (Pless than005), but lower percentages of both early and late effector memory CD8(+) cells (Pless than005) compared to references. Children with exclusively T1D had lower median fluorescence intensity (MFI) of forkhead box protein 3 (FoxP3) (Pless than005) and also a lower percentage of CD39(+) and CD45RA(+) within the T-reg population (CD4(+)CD25(+)FOXP3(+)CD127(-)) (Pless than005). Children with exclusively coeliac disease had a higher MFI of CD101 (Pless than001), as well as a higher percentage of CD129(+) (Pless than005), in the CD4(+)CD25(hi) lymphocyte population, compared to references. In conclusion, children with combined T1D and coeliac disease have a higher percentage of differentiated CD4(+) cells compared to CD8(+) cells. T1D children show signs of low CD39(+)/CD45RA(+) T-reg cells that may indicate loss of suppressive function. Conversely, children with coeliac disease show signs of CD101(+)/CD129(+) T-reg cells that may indicate suppressor activity.
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