| 1. |
- Arabpour, Mohammad, et al.
(författare)
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ADP-ribosylating adjuvant reveals plasticity in cDC1 cells that drive mucosal Th17 cell development and protection against influenza virus infection
- 2022
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Ingår i: Mucosal Immunology. - : Elsevier BV. - 1933-0219. ; 15:4, s. 745-761
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Tidskriftsartikel (refereegranskat)abstract
- Migratory dendritic cells expressing CD103 are the targets for mucosal vaccines. These belong to either of two lineage-restricted subsets, cDC1 or cDC2 cells, which have been linked to priming of functionally distinct CD4 T cells. However, recent studies have identified plasticity in cDC2 cells with overlapping functions with cDC1 cells, while the converse has not been reported. We genetically engineered a vaccine adjuvant platform that targeted the cholera toxin A1 (CTA1) ADP-ribosylating enzyme to CD103(+) cDC1 and cDC2 cells using a single-chain antibody (scFv) to CD103. Unexpectedly, intranasal immunization with the CTA1-svFcCD103 adjuvant modified cDC1 cells to effectively prime Th17 cells, a function previously limited to cDC2 cells. In fact, cDC2 cells were dispensible, while cDC1 cells, lacking in Batf3-/- mice, were critical. Following intranasal immunizations isolated cDC1 cells from mLN exclusively promoted Rorgt(+) T cells and IL-17, IL-21, and IL-22 production. Strong CD8 T cell responses through antigen cross presentation by cDC1 cells were also observed. Single-cell RNAseq analysis revealed upregulation of Th17-promoting gene signatures in sorted cDC1 cells. Gene expression in isolated cDC2 cells was largely unaffected. Our finding represents a major shift of paradigm as we have documented functional plasticity in cDC1 cells.
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| 2. |
- Gribonika, Inta, et al.
(författare)
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Peyer's patch TH17 cells are dispensable for gut IgA responses to oral immunization.
- 2022
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Ingår i: Science immunology. - : American Association for the Advancement of Science (AAAS). - 2470-9468. ; 7:73
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Tidskriftsartikel (refereegranskat)abstract
- T helper 17 (TH17) cells located at the Peyer's patch (PP) inductive site and at the lamina propria effector site of the intestinal immune system are responsive to both pathogenic and commensal bacteria. Their plasticity to convert into follicular helper T (TFH) cells has been proposed to be central to gut immunoglobulin A (IgA) responses. Here, we used an IL-17A fate reporter mouse and an MHC-II tetramer to analyze antigen-specific CD4+ T cell subsets and isolate them for single-cell RNA sequencing after oral immunization with cholera toxin and ovalbumin. We found a TFH-dominated response with only rare antigen-specific TH17 cells (<8%) in the PP. A clonotypic analysis provided little support that clonotypes were shared between TFH and TH17 cells, arguing against TH17 plasticity as a major contributor to TFH differentiation. Two mouse models of TH17 deficiency confirmed that gut IgA responses to oral immunization do not require TH17 cells, with CD4CreRorcfl/fl mice exhibiting normal germinal centers in PP and unperturbed total IgA production in the intestine.
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| 3. |
- Hansson, Charlotta, et al.
(författare)
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Tr1 cell-mediated protection against autoimmune disease by intranasal administration of a fusion protein targeting cDC1 cells.
- 2023
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Ingår i: Mucosal immunology. - 1935-3456. ; 16:4, s. 486-498
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Tidskriftsartikel (refereegranskat)abstract
- Curative therapies against autoimmune diseases are lacking. Indeed, most of the currently available treatments are only targeting symptoms. We have developed a novel strategy for a therapeutic vaccine against autoimmune diseases based on intranasal administration of a fusion protein tolerogen, which consists of a mutant, enzymatically inactive, cholera toxin A1 (CTA1)-subunit genetically fused to disease-relevant high-affinity peptides and a dimer of D-fragments from protein A (DD). The CTA1 R7K mutant - myelin oligodendrocyte glycoprotein (MOG), or proteolipid protein (PLP) - DD (CTA1R7K-MOG/PLP-DD) fusion proteins effectively reduced clinical symptoms in the experimental autoimmune encephalitis model of multiple sclerosis. The treatment induced Tr1 cells, in the draining lymph node, which produced interleukin (IL)-10 and suppressed effector clusters of differentiation 4+ T-cell responses. This effect was dependent on IL-27 signaling because treatment was ineffective in bone marrow chimeras lacking IL-27Ra within their hematopoietic compartment. Single-cell RNA sequencing of dendritic cells in draining lymph nodes demonstrated distinct gene transcriptional changes of classic dendritic cells 1, including enhanced lipid metabolic pathways, induced by the tolerogenic fusion protein. Thus, our results with the tolerogenic fusion protein demonstrate the possibility to vaccinate and protect against disease progression by reinstating tolerance in multiple sclerosis and other autoimmune diseases.
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| 4. |
- Komban, Rathan, et al.
(författare)
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Activated Peyer's patch B cells sample antigen directly from M cells in the subepithelial dome.
- 2019
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Ingår i: Nature communications. - : Springer Science and Business Media LLC. - 2041-1723. ; 10:1
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Tidskriftsartikel (refereegranskat)abstract
- The germinal center (GC) reaction in Peyer's patches (PP) requires continuousaccess to antigens, but how this is achieved is not known. Here we show that activated antigen-specific CCR6+CCR1+GL7- B cells make close contact with M cells in the subepithelial dome (SED). Using in situ photoactivation analysis of antigen-specific SED B cells, we find migration of cells towards the GC. Following antigen injection into ligated intestinal loops containing PPs, 40% of antigen-specific SED B cells bind antigen within 2h, whereas unspecifc cells do not, indicating B cell-receptor involvment. Antigen-loading is not observed in M cell-deficient mice, but is unperturbed in mice depleted of classical dendritic cells (DC). Thus, we report a M cell-B cell antigen-specific transporting pathway in PP that is independent of DC. We propose that this antigen transporting pathway has a critical role in gut IgA responses, and should be taken into account when developing mucosal vaccines.
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| 5. |
- Lebrero-Fernandez, Cristina, et al.
(författare)
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Altered expression of Butyrophilin (BTN) and BTN-like (BTNL) genes in intestinal inflammation and colon cancer
- 2016
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Ingår i: Immunity Inflammation and Disease. - : Wiley. - 2050-4527. ; 4:2, s. 191-200
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Tidskriftsartikel (refereegranskat)abstract
- Several Butyrophilin (BTN) and Btn-like (BTNL) molecules control T lymphocyte responses, and are genetically associated with inflammatory disorders and cancer. In this study, we present a comprehensive expression analysis of human and murine BTN and BTNL genes in conditions associated with intestinal inflammation and cancer. Using real-time PCR, expression of human BTN and BTNL genes was analyzed in samples from patients with ulcerative colitis, irritable bowel syndrome, and colon tumors. Expression of murine Btn and Btnl genes was examined in mouse models of spontaneous colitis (Muc2(-/-)) and intestinal tumorigenesis (Apc(Min/+)). Our analysis indicates a strong association of several of the human genes with ulcerative colitis and colon cancer; while especially BTN1A1, BTN2A2, BTN3A3, and BTNL8 were significantly altered in inflammation, colonic tumors exhibited significantly decreased levels of BTNL2, BTNL3, BTNL8, and BTNL9 as compared to unaffected tissue. Colonic inflammation in Muc2(-/-) mice significantly down-regulated the expression of particularly Btnl1, Btnl4, and Btnl6 mRNA, and intestinal polyps derived from Apc(Min/+) mice displayed altered levels of Btn1a1, Btn2a2, and Btnl1 transcripts. Thus, our data present an association of BTN and BTNL genes with intestinal inflammation and cancer and represent a valuable resource for further studies of this gene family.
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| 6. |
- Lebrero-Fernandez, Cristina
(författare)
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Butyrophilin- and Butyrophilin-like genes and their role in epithelial cell-intraepithelial T lymphocyte cross-talk
- 2016
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- More than 50% of our immune system is located in the gut. The intestinal epithelium, which forms an interface between the organism and the environment, harbors intraepithelial lymphocytes (IELs) that comprise a mixture of conventional αβ T cells and unconventional αβ- and γδ T cells. IELs play important roles in regulation of gut epithelial integrity and in recognition of stressed and infected epithelial cells, and thus, are critical effector components of mucosal immunity. However, the understanding of the IEL function and their interaction with the neighboring epithelial cells is still limited. The aim of this thesis was to investigate how the Butyrophilin (Btn) and Butyrophilin-like (Btnl) molecules are involved in the epithelial cell – IEL cross-talk and hence, to characterize their role in regulating local T cell mediated immune responses in the intestinal mucosa. Btn and Btnl proteins have over the past decade emerged as novel regulators of T cell functions both in periphery and locally in the tissue, and have been shown to be genetically associated with various inflammatory and proliferative disorders. We have reported the ability of intestinal epithelial cell (iEC)-specific Btnl proteins to induce IEL activation and proliferation in conditions without exogenous stimulation, which may contribute to the upkeep of the intestinal IEL pool. We have furthermore identified novel intestinal epithelial cell expressed Btnl- heteromeric protein complexes, and demonstrated that one of them, the Btnl1-Btnl6 heteromeric complex, specifically enhances the expansion of intestinal IELs bearing the Vγ7Vδ4 receptor in vitro. We have additionally explored how iEC-specific Btnl proteins are regulated in the neonatal murine small intestine and found that Btnl- protein expression is delayed in the ontogeny and that the expression of the Btnl genes is regulated on post-transcriptional level. Our data demonstrate that the proteins are not detectable in the small intestinal epithelium of mice before 3 weeks of age, and that the appearance of Btnl1 and Btnl6 proteins correlates with the expansion of intestinal Vγ7Vδ4 IELs, further adding strength to our in vitro results. Since γδ IELs are essential for the maintenance of the homeostasis in the gut, our findings suggest that Btnl proteins have implications in the intestinal immune response. To increase the understanding of the Btn and Btnl molecules’ role in intestinal disorders, we have characterized the expression of human and mouse Btn and Btnl genes in colonic inflammation and intestinal tumors. Our results show an altered expression of the BTN and BTNL genes in these diseases and indicate an association between Btn and Btnl genes and ulcerative colitis and colon cancer. In summary, this thesis work has demonstrated that iEC-specific Btnl proteins can regulate the function of intestinal intraepithelial lymphocytes in the gut, and that Btn and Btnl genes are associated with bowel pathology. Nonetheless, further studies are necessary to identify the complete immunomodulatory implication of the Btn and Btnl family members in healthy and inflamed/infected gut mucosa.
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| 7. |
- Lebrero-Fernandez, Cristina, et al.
(författare)
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Murine Butyrophilin-Like 1 and BtnI6 Form Heteromeric Complexes in Small Intestinal Epithelial Cells and Promote Proliferation of Local T Lymphocytes
- 2016
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Ingår i: Frontiers in Immunology. - : Frontiers Media SA. - 1664-3224. ; 7:1
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Tidskriftsartikel (refereegranskat)abstract
- To date, few molecular conduits mediating the cross talk between intestinal epithelial cells and intraepithelial lymphocytes (IELs) have been described. We recently showed that butyrophilin-like (BtnI) 1 can attenuate the epithelial response to activated IELs, resulting in reduced production of proinflammatory mediators, such as IL-6 and CXCL1. We here report that like BtnI1, murine BtnI6 expression is primarily confined to the intestinal epithelium. Although BtnI1 can exist in a cell surface-expressed homomeric form, we found that it additionally forms heteromeric complexes with BtnI6, and that the engagement of BtnI1 is a prerequisite for surface expression of BtnI6 on intestinal epithelial cells. In an IEL-epithelial cell coculture system, enforced epithelial cell expression of BtnI1 significantly enhanced the proliferation of IELs in the absence of exogenous activation. The effect on proliferation was dependent on the presence of IL-2 or IL-15 and restricted to IELs upregulating CD25. In the gamma delta T-cell subset, the BtnI1-BtnI6 complex, but not BtnI1, specifically elevated the proliferation of IELs bearing the V gamma 7V delta 4 receptor. Thus, our results show that murine epithelial cell-specific BtnI proteins can form intrafamily heterocomplexes and suggest that the interaction between BtnI proteins and IELs regulates the expansion of IELs in the intestinal mucosa.
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| 8. |
- Lebrero-Fernandez, Cristina, et al.
(författare)
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The ontogeny of Butyrophilin-like (Btnl) 1 and Btnl6 in murine small intestine
- 2016
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Ingår i: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 6
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Tidskriftsartikel (refereegranskat)abstract
- Murine Butyrophilin-like (Btnl) 1 and Btnl6 are primarily restricted to intestinal epithelium where they regulate the function of intraepithelial T lymphocytes. We recently demonstrated that Btnl1 and Btnl6 can form an intra-family heterocomplex and that the Btnl1-Btnl6 complex selectively expands V gamma 7V delta 4 TCR IELs. To define the regulation of Btnl expression in the small intestine during ontogeny we examined the presence of Btnl1 and Btnl6 in the small bowel of newborn to 4-week-old mice. Although RNA expression of Btnl1 and Btnl6 was detected in the small intestine at day 0, Btnl1 and Btnl6 protein expression was substantially delayed and was not detectable in the intestinal epithelium until the mice reached 2-3 weeks of age. The markedly elevated Btnl protein level at week 3 coincided with a significant increase of gamma delta TCR IELs, particularly those bearing the V gamma 7V delta 4 receptor. This was not dependent on gut microbial colonization as mice housed in germ-free conditions had normal Btnl protein levels. Taken together, our data show that the expression of Btnl1 and Btnl6 is delayed in the murine neonatal gut and that the appearance of the Btnl1 and Btnl6 proteins in the intestinal mucosa associates with the expansion of V gamma 7V delta 4 TCR IELs.
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| 9. |
- Lycke, Nils Y, 1954, et al.
(författare)
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ADP-ribosylating enterotoxins as vaccine adjuvants
- 2018
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Ingår i: Current opinion in pharmacology. - : Elsevier BV. - 1471-4892 .- 1471-4973. ; 41, s. 42-51
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Tidskriftsartikel (refereegranskat)abstract
- Most infections are caused by pathogens that access the body at mucosal sites. Hence, development of mucosal vaccines to prevent local infection or invasion of pathogens appears highly warranted, especially since only mucosal immunization will stimulate strong local IgA responses and tissue resident memory CD4 and CD8 T cells. The most significant obstacle to developing such vaccines is the lack of approved adjuvants that can effectively and safely enhance relevant mucosal and systemic immune responses. The most potent mucosal adjuvants known today are the adenosine diphosphate (ADP)-ribosylating bacterial enterotoxins cholera toxin (CT) and Escherichia coli heat-labile toxins (LTs). Unfortunately, these molecules are also very toxic, which precludes their clinical use. However, much effort has been devoted to developing derivatives of these enterotoxins with low or no toxicity and retained adjuvant activity. Although it is fair to say that we know more about how these toxins affect the immune system than ever before, we still lack a detailed understanding of how and why these toxins are effective adjuvants. In the present review, we provide a state-of-the-art overview of the mechanism of action of the holotoxins and the strategies used for improving the toxin-based adjuvants.
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| 10. |
- Mendoza, Patricia, et al.
(författare)
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DamID transcriptional profiling identifies the Snail/Scratch transcription factor Kahuli as an Alk target in the Drosophila visceral mesoderm
- 2021
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Ingår i: Development. - : The Company of Biologists. - 0950-1991 .- 1477-9129. ; 148:23
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Tidskriftsartikel (refereegranskat)abstract
- Development of the Drosophila visceral muscle depends on Anaplastic Lymphoma Kinase (Alk) receptor tyrosine kinase (RTK) signaling, which specifies founder cells (FCs) in the circular visceral mesoderm (VM). Although Alk activation by its ligand Jelly Belly (Jeb) is well characterized, few target molecules have been identified. Here, we used targeted DamID (TaDa) to identify Alk targets in embryos overexpressing Jeb versus embryos with abrogated Alk activity, revealing differentially expressed genes, including the Snail/Scratch family transcription factor Kahuli (Kah). We confirmed Kah mRNA and protein expression in the VM, and identified midgut constriction defects in Kah mutants similar to those of pointed (pnt). ChIP and RNA-Seq data analysis defined a Kah target-binding site similar to that of Snail, and identified a set of common target genes putatively regulated by Kah and Pnt during midgut constriction. Taken together, we report a rich dataset of Alk-responsive loci in the embryonic VM and functionally characterize the role of Kah in the regulation of embryonic midgut morphogenesis.
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