Hypomorphic IL2RG mutations may lead to milder phenotypes than X-SCID, named variably as atypical... more Hypomorphic IL2RG mutations may lead to milder phenotypes than X-SCID, named variably as atypical X-SCID or X-CID. We report an 11-year-old boy with a novel c. 172C>T;p.(Pro58Ser) mutation in IL2RG, presenting with atypical X-SCID phenotype. We also review the growing number of hypomorphic IL2RG mutations causing atypical X-SCID. We studied the patient’s clinical phenotype, B, T, NK, and dendritic cell phenotypes, IL2RG and CD25 cell surface expression, and IL-2 target gene expression, STAT tyrosine phosphorylation, PBMC proliferation, and blast formation in response to IL-2 stimulation, as well as protein-protein interactions of the mutated IL2RG by BioID proximity labeling. The patient suffered from recurrent upper and lower respiratory tract infections, bronchiectasis, and reactive arthritis. His total lymphocyte counts have remained normal despite skewed T and B cells subpopulations, with very low numbers of plasmacytoid dendritic cells. Surface expression of IL2RG was reduce...
Gut microbiota participates in diverse metabolic and homeostatic functions related to health and ... more Gut microbiota participates in diverse metabolic and homeostatic functions related to health and well-being. Its composition varies between individuals, and depends on factors related to host and microbial communities, which need to adapt to utilize various nutrients present in gut environment. We profiled fecal microbiota in 63 healthy adult individuals using metaproteomics, and focused on microbial CAZy (carbohydrate-active) enzymes involved in glycan foraging. We identified two distinct CAZy profiles, one with many Bacteroides-derived CAZy in more than one-third of subjects (n = 25), and it associated with high abundance of Bacteroides in most subjects. In a smaller subset of donors (n = 8) with dietary parameters similar to others, microbiota showed intense expression of Prevotella-derived CAZy including exo-beta-(1,4)-xylanase, xylan-1,4-beta-xylosidase, alpha-l-arabinofuranosidase and several other CAZy belonging to glycosyl hydrolase families involved in digestion of complex ...
The onset and development of type 1 diabetes (T1D) occurs in genetically predisposed individuals,... more The onset and development of type 1 diabetes (T1D) occurs in genetically predisposed individuals, and is attributed to autoimmune destruction of pancreatic beta-cells involving a multitude of immune mechanisms. Defects in immune regulation may play a central role in T1D, involving impaired function and communication of both myeloid and lymphoid cells of the innate and adaptive immune compartments. Dendritic cells and regulatory T (Treg) cells are part of this network, which seem to be hampered in their quest to control and regulate tissue-destructive autoimmunity. Recent studies have shown that in vivo activated CD16- blood monocytes exhibiting proinflammatory features are present in diabetic subjects. These monocytes may govern T cell-mediated immune responses towards the development of tissue-destructive Th1 and Th17 subtypes, and give rise to inflammatory macrophages in tissues. Differential effects of cytokines IFN-gamma and IL-4 in the development of inflammatory macrophages, a...
Abstract: Fas ligation induces apoptosis of acti-vated T cells via the caspase cascade but can al... more Abstract: Fas ligation induces apoptosis of acti-vated T cells via the caspase cascade but can also mediate costimulatory signals to naı̈ve T cells at the time of activation. We have previously shown that Fas ligation of naı̈ve CD4 T cells activated by den-dritic cells induces death or accelerates their pro-liferation and increases interferon- (IFN-) pro-duction. To understand this costimulation, we in-vestigated the roles of caspases and nuclear factor (NF)-B in survival and proliferation of responding T cells. Fas ligation increased caspase-3 and-8 activities during T cell activation, irrespective of cell fate. The accelerated proliferation induced by Fas ligation could be reduced by selective inhibi-tion of both caspases. It is interesting that inhibi-tion of NF-B simultaneously with Fas ligation in-hibited the increased IFN- production and caused uniform death of all responding T cells. Thus, Fas-mediated costimulation of naı̈ve CD4 T cells is driven by active caspases, and NF-B...
Inflammation is an acute manifestation of the activation of the immune defense. The immunological... more Inflammation is an acute manifestation of the activation of the immune defense. The immunological regulation of an inflammatory reaction is disturbed in many diseases, and inflammation is also involved in numerous conditions that are often considered non-inflammatory such as atherosclerosis, ischemic tissue injury, obesity and Alzheimer's disease. Especially recent progress in the research on the innate immune system has improved our understanding on the manifold cause-effect relationships in the regulation of the immune reactions and opened up new possibilities to manage inflammation.
Macrophages play an important role in innate immune reactions, and peritoneal macrophages guard t... more Macrophages play an important role in innate immune reactions, and peritoneal macrophages guard the sterility of this compartment mainly against microbial threat from the gut. Type-1 Diabetes (T1D) is an autoimmune disease in which gut microbiota and gut immune system appear to play a role. We have observed increased permeability and free radical production of gut epithelium in nonobese diabetic (NOD) mice. Impaired barrier function could lead to bacterial endotoxin leakage to peritoneal cavity. To seek for extraintestinal consequences of impaired barrier function, we characterized peritoneal lavage cells from young NOD mice after weaning. We detected a rapid increase in the number of macrophages in 1-2 weeks from weaning in NOD mice compared to C57BL/6 and BALB/c mice, and to NOD mice fed an antidiabetogenic diet (ProSobee), which improves gut barrier function. Upon LPS stimulation ex vivo, macrophages produced low amounts of TNF-α and showed reduced expression of CD40. Expression ...
Dendritic cells have turned out to be important component in the regulation of immune responses. ... more Dendritic cells have turned out to be important component in the regulation of immune responses. In addition to various external structures they recognize the body's own intracellular structures and utilize them to construct information about threats affecting the well-being of organs and tissues. Dendritic cells are able to direct immune responses in a manner that among other things opens new dimensions to the prevention and management of autoimmune diseases, allergies, cancer and atherosclerosis. Vaccines directed to dendritic cells and modification of dendritic cells in vitro are becoming a part of the novel, targeted immunotherapy.
The reported incidence of pertussis in European countries varies considerably. We aimed to study ... more The reported incidence of pertussis in European countries varies considerably. We aimed to study specific Bordetella pertussis seroprevalence in Europe by measuring serum IgG antibody levels to pertussis toxin (anti‐PT IgG). Fourteen national laboratories participated in this study including Belgium, Denmark, Finland, Greece, Hungary, Italy, Lithuania, Malta, Norway, Poland, Portugal, Romania, Spain, and Sweden. Each country collected approximately 250 samples (N = 7903) from the age groups 20–29 years (N = 3976) and 30–39 years (N = 3927) during 2010–2013. Samples were anonymous residual sera from diagnostic laboratories and were analyzed at the national laboratories by a Swedish reference method, a commercial ELISA kit, or were sent to Sweden for analysis. The median anti‐PT IgG concentrations ranged from 4 to 13.6 IU/mL. The proportion of samples with anti‐PT IgG ≥100 IU/mL, indicating a recent infection ranged from 0.2% (Hungary) to 5.7% (Portugal). The highest proportion of sera with anti‐PT IgG levels between 50 and <100 IU/mL, indicating an infection within the last few years, was found in Portugal (12.3%) and Italy (13.9%). This study shows that the circulation of B. pertussis is quite extensive in adults, aged 20–39 years, despite well‐established vaccination programs in Europe.
Metagenomic approaches focus on taxonomy or gene annotation but lack power in defining functional... more Metagenomic approaches focus on taxonomy or gene annotation but lack power in defining functionality of gut microbiota. Therefore, metaproteomics approaches have been introduced to overcome this limitation. However, the common metaproteomics approach uses data-dependent acquisition mass spectrometry, which is known to have limited reproducibility and dynamic range when analyzing samples with complex microbial composition. In this work, we provide a proof-of-concept for data-independent acquisition (DIA) metaproteomics. To this end, we analyze metaproteomes using DIA mass spectrometry and introduce an open-source data analysis software package diatools, which enables accurate and consistent quantification of DIA metaproteomics data. We demonstrate the feasibility of our approach in gut microbiota metaproteomics using laboratory assembled microbial mixtures as well as human fecal samples.
Dendritic cells (DCs) are first in line to sense invading microbes and to deliver signals to othe... more Dendritic cells (DCs) are first in line to sense invading microbes and to deliver signals to other immune cells. Plasmacytoid DCs (pDC) produce high amounts of type I interferons (IFNs) but also regulate immune responses. Using the Clec4C (BDCA2)‐diphtheria toxin receptor mouse model allowing conditional pDC depletion, we identified an essential role for pDCs in regulating intestinal inflammation locally in the gut. In pDC‐depleted mice, Citrobacter rodentium infection led to enhanced activation of conventional DCs and induction of IFN‐γ‐producing Th1‐cells in colon‐draining lymph nodes, while induction of Foxp3+/CD25+ Treg and IL‐17‐producing Th17 cells was impaired. Concomitantly, F4/80+ macrophages accumulated into the colon lamina propria in excess, and levels of Il‐1β and Tnf transcripts increased and Foxp3+ Treg were fewer. Our results indicate that pDCs control inflammation in the gut during C. rodentium infection and that they have an important immune regulatory role in colon‐draining lymph nodes.
Metaproteomics is an emerging research area which aims to reveal the functionality of microbial c... more Metaproteomics is an emerging research area which aims to reveal the functionality of microbial communities – unlike the increasingly popular metagenomics providing insights only on the functional potential. So far, the common approach in metaproteomics has been data-dependent acquisition mass spectrometry (DDA). However, DDA is known to have limited reproducibility and dynamic range with samples of complex microbial composition. To overcome these limitations, we introduce here a novel approach utilizing data-independent acquisition (DIA) mass spectrometry, which has not been applied in metaproteomics of complex samples before. For robust analysis of the data, we introduce an open-source software package diatools, which is freely available at Docker Hub and runs on various operating systems. Our highly reproducible results on laboratory-assembled microbial mixtures and human fecal samples support the utility of our approach for functional characterization of complex microbiota. Henc...
Intestinal microbiota is implicated in the pathogenesis of autoimmune type 1 diabetes in humans a... more Intestinal microbiota is implicated in the pathogenesis of autoimmune type 1 diabetes in humans and in non-obese diabetic (NOD) mice, but evidence on its causality and on the role of individual microbiota members is limited. We investigated if different diabetes incidence in two NOD colonies was due to microbiota differences and aimed to identify individual microbiota members with potential significance. We profiled intestinal microbiota between two NOD mouse colonies showing high or low diabetes incidence by 16S ribosomal RNA gene sequencing and colonised the high-incidence colony with the microbiota of the low-incidence colony. Based on unaltered incidence, we identified a few taxa which were not effectively transferred and thereafter, transferred experimentally one of these to test its potential significance. Although the high-incidence colony adopted most microbial taxa present in the low-incidence colony, diabetes incidence remained unaltered. Among the few taxa which were not ...
Journal of immunology (Baltimore, Md. : 1950), Jun 1, 2016
Dendritic cells (DCs) bear the main responsibility for initiation of adaptive immune responses ne... more Dendritic cells (DCs) bear the main responsibility for initiation of adaptive immune responses necessary for antimicrobial immunity. In the small intestine, afferent lymphatics convey Ags and microbial signals to mesenteric lymph nodes (LNs) to induce adaptive immune responses against microbes and food Ags derived from the small intestine. Whether the large intestine is covered by the same lymphatic system or represents its own lymphoid compartment has not been studied until very recently. We identified three small mesenteric LNs, distinct from small intestinal LNs, which drain lymph specifically from the colon, and studied DC responses to the attaching and effacing pathogen Citrobacter rodentium in these. Transcriptional profiling of conventional (CD11c(high)CD103(high)) DC and plasmacytoid (plasmacytoid DC Ag-1(high)B220(+)CD11c(int)) DC (pDC) populations during steady-state conditions revealed activity of distinct sets of genes in these two DC subsets, both in small intestinal an...
Hypomorphic IL2RG mutations may lead to milder phenotypes than X-SCID, named variably as atypical... more Hypomorphic IL2RG mutations may lead to milder phenotypes than X-SCID, named variably as atypical X-SCID or X-CID. We report an 11-year-old boy with a novel c. 172C>T;p.(Pro58Ser) mutation in IL2RG, presenting with atypical X-SCID phenotype. We also review the growing number of hypomorphic IL2RG mutations causing atypical X-SCID. We studied the patient’s clinical phenotype, B, T, NK, and dendritic cell phenotypes, IL2RG and CD25 cell surface expression, and IL-2 target gene expression, STAT tyrosine phosphorylation, PBMC proliferation, and blast formation in response to IL-2 stimulation, as well as protein-protein interactions of the mutated IL2RG by BioID proximity labeling. The patient suffered from recurrent upper and lower respiratory tract infections, bronchiectasis, and reactive arthritis. His total lymphocyte counts have remained normal despite skewed T and B cells subpopulations, with very low numbers of plasmacytoid dendritic cells. Surface expression of IL2RG was reduce...
Gut microbiota participates in diverse metabolic and homeostatic functions related to health and ... more Gut microbiota participates in diverse metabolic and homeostatic functions related to health and well-being. Its composition varies between individuals, and depends on factors related to host and microbial communities, which need to adapt to utilize various nutrients present in gut environment. We profiled fecal microbiota in 63 healthy adult individuals using metaproteomics, and focused on microbial CAZy (carbohydrate-active) enzymes involved in glycan foraging. We identified two distinct CAZy profiles, one with many Bacteroides-derived CAZy in more than one-third of subjects (n = 25), and it associated with high abundance of Bacteroides in most subjects. In a smaller subset of donors (n = 8) with dietary parameters similar to others, microbiota showed intense expression of Prevotella-derived CAZy including exo-beta-(1,4)-xylanase, xylan-1,4-beta-xylosidase, alpha-l-arabinofuranosidase and several other CAZy belonging to glycosyl hydrolase families involved in digestion of complex ...
The onset and development of type 1 diabetes (T1D) occurs in genetically predisposed individuals,... more The onset and development of type 1 diabetes (T1D) occurs in genetically predisposed individuals, and is attributed to autoimmune destruction of pancreatic beta-cells involving a multitude of immune mechanisms. Defects in immune regulation may play a central role in T1D, involving impaired function and communication of both myeloid and lymphoid cells of the innate and adaptive immune compartments. Dendritic cells and regulatory T (Treg) cells are part of this network, which seem to be hampered in their quest to control and regulate tissue-destructive autoimmunity. Recent studies have shown that in vivo activated CD16- blood monocytes exhibiting proinflammatory features are present in diabetic subjects. These monocytes may govern T cell-mediated immune responses towards the development of tissue-destructive Th1 and Th17 subtypes, and give rise to inflammatory macrophages in tissues. Differential effects of cytokines IFN-gamma and IL-4 in the development of inflammatory macrophages, a...
Abstract: Fas ligation induces apoptosis of acti-vated T cells via the caspase cascade but can al... more Abstract: Fas ligation induces apoptosis of acti-vated T cells via the caspase cascade but can also mediate costimulatory signals to naı̈ve T cells at the time of activation. We have previously shown that Fas ligation of naı̈ve CD4 T cells activated by den-dritic cells induces death or accelerates their pro-liferation and increases interferon- (IFN-) pro-duction. To understand this costimulation, we in-vestigated the roles of caspases and nuclear factor (NF)-B in survival and proliferation of responding T cells. Fas ligation increased caspase-3 and-8 activities during T cell activation, irrespective of cell fate. The accelerated proliferation induced by Fas ligation could be reduced by selective inhibi-tion of both caspases. It is interesting that inhibi-tion of NF-B simultaneously with Fas ligation in-hibited the increased IFN- production and caused uniform death of all responding T cells. Thus, Fas-mediated costimulation of naı̈ve CD4 T cells is driven by active caspases, and NF-B...
Inflammation is an acute manifestation of the activation of the immune defense. The immunological... more Inflammation is an acute manifestation of the activation of the immune defense. The immunological regulation of an inflammatory reaction is disturbed in many diseases, and inflammation is also involved in numerous conditions that are often considered non-inflammatory such as atherosclerosis, ischemic tissue injury, obesity and Alzheimer's disease. Especially recent progress in the research on the innate immune system has improved our understanding on the manifold cause-effect relationships in the regulation of the immune reactions and opened up new possibilities to manage inflammation.
Macrophages play an important role in innate immune reactions, and peritoneal macrophages guard t... more Macrophages play an important role in innate immune reactions, and peritoneal macrophages guard the sterility of this compartment mainly against microbial threat from the gut. Type-1 Diabetes (T1D) is an autoimmune disease in which gut microbiota and gut immune system appear to play a role. We have observed increased permeability and free radical production of gut epithelium in nonobese diabetic (NOD) mice. Impaired barrier function could lead to bacterial endotoxin leakage to peritoneal cavity. To seek for extraintestinal consequences of impaired barrier function, we characterized peritoneal lavage cells from young NOD mice after weaning. We detected a rapid increase in the number of macrophages in 1-2 weeks from weaning in NOD mice compared to C57BL/6 and BALB/c mice, and to NOD mice fed an antidiabetogenic diet (ProSobee), which improves gut barrier function. Upon LPS stimulation ex vivo, macrophages produced low amounts of TNF-α and showed reduced expression of CD40. Expression ...
Dendritic cells have turned out to be important component in the regulation of immune responses. ... more Dendritic cells have turned out to be important component in the regulation of immune responses. In addition to various external structures they recognize the body's own intracellular structures and utilize them to construct information about threats affecting the well-being of organs and tissues. Dendritic cells are able to direct immune responses in a manner that among other things opens new dimensions to the prevention and management of autoimmune diseases, allergies, cancer and atherosclerosis. Vaccines directed to dendritic cells and modification of dendritic cells in vitro are becoming a part of the novel, targeted immunotherapy.
The reported incidence of pertussis in European countries varies considerably. We aimed to study ... more The reported incidence of pertussis in European countries varies considerably. We aimed to study specific Bordetella pertussis seroprevalence in Europe by measuring serum IgG antibody levels to pertussis toxin (anti‐PT IgG). Fourteen national laboratories participated in this study including Belgium, Denmark, Finland, Greece, Hungary, Italy, Lithuania, Malta, Norway, Poland, Portugal, Romania, Spain, and Sweden. Each country collected approximately 250 samples (N = 7903) from the age groups 20–29 years (N = 3976) and 30–39 years (N = 3927) during 2010–2013. Samples were anonymous residual sera from diagnostic laboratories and were analyzed at the national laboratories by a Swedish reference method, a commercial ELISA kit, or were sent to Sweden for analysis. The median anti‐PT IgG concentrations ranged from 4 to 13.6 IU/mL. The proportion of samples with anti‐PT IgG ≥100 IU/mL, indicating a recent infection ranged from 0.2% (Hungary) to 5.7% (Portugal). The highest proportion of sera with anti‐PT IgG levels between 50 and <100 IU/mL, indicating an infection within the last few years, was found in Portugal (12.3%) and Italy (13.9%). This study shows that the circulation of B. pertussis is quite extensive in adults, aged 20–39 years, despite well‐established vaccination programs in Europe.
Metagenomic approaches focus on taxonomy or gene annotation but lack power in defining functional... more Metagenomic approaches focus on taxonomy or gene annotation but lack power in defining functionality of gut microbiota. Therefore, metaproteomics approaches have been introduced to overcome this limitation. However, the common metaproteomics approach uses data-dependent acquisition mass spectrometry, which is known to have limited reproducibility and dynamic range when analyzing samples with complex microbial composition. In this work, we provide a proof-of-concept for data-independent acquisition (DIA) metaproteomics. To this end, we analyze metaproteomes using DIA mass spectrometry and introduce an open-source data analysis software package diatools, which enables accurate and consistent quantification of DIA metaproteomics data. We demonstrate the feasibility of our approach in gut microbiota metaproteomics using laboratory assembled microbial mixtures as well as human fecal samples.
Dendritic cells (DCs) are first in line to sense invading microbes and to deliver signals to othe... more Dendritic cells (DCs) are first in line to sense invading microbes and to deliver signals to other immune cells. Plasmacytoid DCs (pDC) produce high amounts of type I interferons (IFNs) but also regulate immune responses. Using the Clec4C (BDCA2)‐diphtheria toxin receptor mouse model allowing conditional pDC depletion, we identified an essential role for pDCs in regulating intestinal inflammation locally in the gut. In pDC‐depleted mice, Citrobacter rodentium infection led to enhanced activation of conventional DCs and induction of IFN‐γ‐producing Th1‐cells in colon‐draining lymph nodes, while induction of Foxp3+/CD25+ Treg and IL‐17‐producing Th17 cells was impaired. Concomitantly, F4/80+ macrophages accumulated into the colon lamina propria in excess, and levels of Il‐1β and Tnf transcripts increased and Foxp3+ Treg were fewer. Our results indicate that pDCs control inflammation in the gut during C. rodentium infection and that they have an important immune regulatory role in colon‐draining lymph nodes.
Metaproteomics is an emerging research area which aims to reveal the functionality of microbial c... more Metaproteomics is an emerging research area which aims to reveal the functionality of microbial communities – unlike the increasingly popular metagenomics providing insights only on the functional potential. So far, the common approach in metaproteomics has been data-dependent acquisition mass spectrometry (DDA). However, DDA is known to have limited reproducibility and dynamic range with samples of complex microbial composition. To overcome these limitations, we introduce here a novel approach utilizing data-independent acquisition (DIA) mass spectrometry, which has not been applied in metaproteomics of complex samples before. For robust analysis of the data, we introduce an open-source software package diatools, which is freely available at Docker Hub and runs on various operating systems. Our highly reproducible results on laboratory-assembled microbial mixtures and human fecal samples support the utility of our approach for functional characterization of complex microbiota. Henc...
Intestinal microbiota is implicated in the pathogenesis of autoimmune type 1 diabetes in humans a... more Intestinal microbiota is implicated in the pathogenesis of autoimmune type 1 diabetes in humans and in non-obese diabetic (NOD) mice, but evidence on its causality and on the role of individual microbiota members is limited. We investigated if different diabetes incidence in two NOD colonies was due to microbiota differences and aimed to identify individual microbiota members with potential significance. We profiled intestinal microbiota between two NOD mouse colonies showing high or low diabetes incidence by 16S ribosomal RNA gene sequencing and colonised the high-incidence colony with the microbiota of the low-incidence colony. Based on unaltered incidence, we identified a few taxa which were not effectively transferred and thereafter, transferred experimentally one of these to test its potential significance. Although the high-incidence colony adopted most microbial taxa present in the low-incidence colony, diabetes incidence remained unaltered. Among the few taxa which were not ...
Journal of immunology (Baltimore, Md. : 1950), Jun 1, 2016
Dendritic cells (DCs) bear the main responsibility for initiation of adaptive immune responses ne... more Dendritic cells (DCs) bear the main responsibility for initiation of adaptive immune responses necessary for antimicrobial immunity. In the small intestine, afferent lymphatics convey Ags and microbial signals to mesenteric lymph nodes (LNs) to induce adaptive immune responses against microbes and food Ags derived from the small intestine. Whether the large intestine is covered by the same lymphatic system or represents its own lymphoid compartment has not been studied until very recently. We identified three small mesenteric LNs, distinct from small intestinal LNs, which drain lymph specifically from the colon, and studied DC responses to the attaching and effacing pathogen Citrobacter rodentium in these. Transcriptional profiling of conventional (CD11c(high)CD103(high)) DC and plasmacytoid (plasmacytoid DC Ag-1(high)B220(+)CD11c(int)) DC (pDC) populations during steady-state conditions revealed activity of distinct sets of genes in these two DC subsets, both in small intestinal an...
Uploads