| 1. |
- Huberle, Alexander, et al.
(författare)
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Advanced Intercross Line Mapping Suggests That Ncf1 (Ean6) Regulates Severity in an Animal Model of Guillain-Barre Syndrome
- 2009
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Ingår i: Journal of Immunology. - : The American Association of Immunologists. - 1550-6606 .- 0022-1767. ; 182:7, s. 4432-4438
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Tidskriftsartikel (refereegranskat)abstract
- We here present the first genetic fine mapping of experimental autoimmune neuritis (EAN), the animal model of Guillain-Barre syndrome, in a rat advanced intercross line. We identified and refined a total of five quantitative trait loci on rat chromosomes 4, 10, and 12 (RNO4, RNO10, RNO12), showing linkage to splenic IFN-gamma secretion and disease severity. All quantitative trait loci were shared with other models of complex inflammatory diseases. The quantitative trait locus showing strongest linkage to clinical disease was Ean6 and spans 4.3 Mb on RNO12, harboring the neutrophil cytosolic factor 1 (Ncf1) among other genes. Polymorphisms in Ncf1, a member of the NADPH oxidase complex, have been associated with disease regulation in experimental arthritis and encephalomyelitis. We therefore tested the Ncf1 pathway by treating rats with a NADPH oxidase complex activator and ameliorated EAN compared the oil-treated control group. By proving the therapeutic effect of stimulating the NADPH oxidase complex, our data strongly suggest the first identification of a gene regulating peripheral nervous system inflammation. Taken together with previous reports, our findings suggest a general role of Ncf1 and oxidative burst in pathogenesis of experimental autoimmune animal models. The Journal of Immunology, 2009, 182: 4432-4438.
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| 2. |
- Lobell, Anna, et al.
(författare)
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Presence of CpG DNA and the local cytokine milieu determine the efficacy of suppressive DNA vaccination in experimental autoimmune encephalomyelitis
- 1999
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Ingår i: Journal of Immunology. - 0022-1767 .- 1550-6606. ; 163:9, s. 4754-4762
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Tidskriftsartikel (refereegranskat)abstract
- We here study the adjuvant properties of immunostimulatory DNA sequences (ISS) and coinjected cytokine-coding cDNA in suppressive vaccination with DNA encoding an autoantigenic peptide, myelin basic protein peptide 68-85, against Lewis rat experimental autoimmune encephalomyelitis (EAE). EAE is an autoaggressive, T1-mediated disease of the CNS. ISS are unmethylated CpG motifs found in bacterial DNA, which can induce production of type 1 cytokines in vertebrates through the innate immune system. Because ISS in the plasmid backbone are necessary for efficient DNA vaccination, we studied the effect of one such ISS, the 5'-AACGTT-3' motif, in our system. Treatment with a DNA vaccine encoding myelin basic protein peptide 68-85 and containing three ISS of 5'-AACGTT-3' sequence suppressed clinical signs of EAE, while a corresponding DNA vaccine without such ISS had no effect. We further observed reduced proliferative T cell responses in rats treated with the ISS-containing DNA vaccine, compared with controls. We also studied the possible impact of coinjection of plasmid DNA encoding rat cytokines IL-4, IL-10, GM-CSF, and TNF-alpha with the ISS-containing DNA vaccine. Coinjection of IL-4-, IL-10-, or TNF-alpha-coding cDNA inhibited the suppressive effect of the DNA vaccine on EAE, whereas GM-CSF-coding cDNA had no effect. Coinjection of cytokine-coding cDNA with the ISS-deficient DNA vaccine failed to alter clinical signs of EAE. We conclude that the presence of ISS and induction of a local T1 cytokine milieu is decisive for specific protective DNA vaccination in EAE.
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| 3. |
- Lobell, Anna, et al.
(författare)
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Protective DNA vaccination against organ-specific autoimmunity is highly specific and discriminates between single amino acid substitutions in the peptide autoantigen
- 2000
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Ingår i: Proceedings of the National Academy of Sciences of the United States of America. - : Proceedings of the National Academy of Sciences. - 0027-8424 .- 1091-6490. ; 97:4, s. 1689-1694
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Tidskriftsartikel (övrigt vetenskapligt/konstnärligt)abstract
- DNA vaccines that encode encephalitogenic sequences in tandem can protect from subsequent experimental autoimmune encephalomyelitis induced with the corresponding peptide. The mechanism for this protection and, in particular, if it is specific for the amino acid sequence encoding the vaccine are not known. We show here that a single amino acid exchange in position 79 from serine (nonself) to threonine (self) in myelin basic protein peptide MBP68-85, which is a major encephalitogenic determinant for Lewis rats, dramatically alters the protection. Moreover, vaccines encoding the encephalitogenic sequence MBP68-85 do not protect against the second encephalitogenic sequence MBP89-101 in Lewis rats and vice versa. Thus, protective immunity conferred by DNA vaccination exquisitely discriminates between peptide target autoantigens. No bystander suppression was observed. The exact underlying mechanisms remain elusive because no simple correlation between impact on ex vivo responses and protection against disease were noted.
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| 4. |
- Lobell, Anna, et al.
(författare)
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Suppressive DNA vaccination in myelin oligodendrocyte glycoprotein peptide-induced experimental autoimmune encephalomyelitis involves a T1-biased immune response
- 2003
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Ingår i: Journal of Immunology. - : The American Association of Immunologists. - 0022-1767 .- 1550-6606. ; 170:4, s. 1806-1813
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Tidskriftsartikel (övrigt vetenskapligt/konstnärligt)abstract
- Vaccination with DNA encoding a myelin basic protein peptide suppresses Lewis rat experimental autoimmune encephalomyelitis (EAE) induced with the same peptide. Additional myelin proteins, such as myelin oligodendrocyte glycoprotein (MOG), may be important in multiple sclerosis. Here we demonstrate that DNA vaccination also suppresses MOG peptide-induced EAE. MOG(91-108) is encephalitogenic in DA rats and MHC-congenic LEW.1AV1 (RT1(av1)) and LEW.1N (RT1(n)) rats. We examined the effects of DNA vaccines encoding MOG(91-108) in tandem, with or without targeting of the hybrid gene product to IgG. In all investigated rat strains DNA vaccination suppressed clinical signs of EAE. There was no requirement for targeting the gene product to IgG, but T1-promoting CpG DNA motifs in the plasmid backbone of the construct were necessary for efficient DNA vaccination, similar to the case in DNA vaccination in myelin basic protein-induced EAE. We failed to detect any effects on ex vivo MOG-peptide-induced IFN-gamma, TNF-alpha, IL-6, IL-4, IL-10, and brain-derived neurotropic factor expression in splenocytes or CNS-derived lymphocytes. In CNS-derived lymphocytes, Fas ligand expression was down-regulated in DNA-vaccinated rats compared with controls. However, MOG-specific IgG2b responses were enhanced after DNA vaccination. The enhanced IgG2b responses together with the requirement for CpG DNA motifs in the vaccine suggest a protective mechanism involving induction of a T1-biased immune response.
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| 5. |
- Lobell, Anna, et al.
(författare)
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Vaccination with DNA encoding an immunodominant myelin basic protein peptide targeted to Fc of immunoglobulin G suppresses experimental autoimmune encephalomyelitis
- 1998
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Ingår i: Journal of Experimental Medicine. - : Rockefeller University Press. - 0022-1007 .- 1540-9538. ; 187:9, s. 1543-1548
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Tidskriftsartikel (refereegranskat)abstract
- We explore here if vaccination with DNA encoding an autoantigenic peptide can suppress autoimmune disease. For this purpose we used experimental autoimmune encephalomyelitis (EAE), which is an autoaggressive disease in the central nervous system and an animal model for multiple sclerosis. Lewis rats were vaccinated with DNA encoding an encephalitogenic T cell epitope, guinea pig myelin basic protein peptide 68-85 (MBP68-85), before induction of EAE with MBP68-85 in complete Freund's adjuvant. Compared to vaccination with a control DNA construct, the vaccination suppressed clinical and histopathological signs of EAE, and reduced the interferon gamma production after challenge with MBP68-85. Targeting of the gene product to Fc of IgG was essential for this effect. There were no signs of a Th2 cytokine bias. Our data suggest that DNA vaccines encoding autoantigenic peptides may be useful tools in controlling autoimmune disease.
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| 6. |
- Petzold, Axel, et al.
(författare)
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Neurofilament ELISA validation
- 2010
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Ingår i: JOURNAL OF IMMUNOLOGICAL METHODS. - 0022-1759. ; 352:1-2, s. 23-31
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Tidskriftsartikel (refereegranskat)
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| 7. |
- Petzold, Axel, et al.
(författare)
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Neurofilament ELISA validation
- 2010
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Ingår i: JIM - Journal of Immunological Methods. - : Elsevier BV. - 0022-1759 .- 1872-7905. ; 352:1-2, s. 23-31
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Neurofilament proteins (Nf) are highly specific biomarkers for neuronal death and axonal degeneration. As these markers become more widely used, an inter-laboratory validation study is required to identify assay criteria for high quality performance. METHODS: The UmanDiagnostics NF-light (R)enzyme-linked immunoabsorbent assays (ELISA) for the neurofilament light chain (NfL, 68kDa) was used to test the intra-assay and inter-laboratory coefficient of variation (CV) between 35 laboratories worldwide on 15 cerebrospinal fluid (CSF) samples. Critical factors, such as sample transport and storage, analytical delays, reaction temperature and time, the laboratories' accuracy and preparation of standards were documented and used for the statistical analyses. RESULTS: The intra-laboratory CV averaged 3.3% and the inter-laboratory CV 59%. The results from the test laboratories correlated with those from the reference laboratory (R=0.60, p<0.0001). Correcting for critical factors improved the strength of the correlation. Differences in the accuracy of standard preparation were identified as the most critical factor. Correcting for the error introduced by variation in the protein standards improved the correlation to R=0.98, p<0.0001 with an averaged inter-laboratory CV of 14%. The corrected overall inter-rater agreement was subtantial (0.6) according to Fleiss' multi-rater kappa and Gwet's AC1 statistics. CONCLUSION: This multi-center validation study identified the lack of preparation of accurate and consistent protein standards as the main reason for a poor inter-laboratory CV. This issue is also relevant to other protein biomarkers based on this type of assay and will need to be solved in order to achieve an acceptable level of analytical accuracy. The raw data of this study is available online.
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| 8. |
- Wahlstrom, Jan, et al.
(författare)
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Autoimmune T cell responses to antigenic peptides presented by bronchoalveolar lavage cell HLA-DR molecules in sarcoidosis
- 2009
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Ingår i: CLINICAL IMMUNOLOGY. - : Elsevier BV. - 1521-6616 .- 1521-7035. ; 133:3, s. 353-363
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Tidskriftsartikel (refereegranskat)abstract
- The etiology of sarcoidosis remains unknown. Recently, by mass spectrometric sequencing of peptides eluted from HLA-DR molecules of bronchoalveolar lavage (BAL) cells from DRB1*0301(pos) patients, we identified potential self-antigens in sarcoidosis. The aim of the present study was to investigate the capacity of selected peptides to stimulate tung and blood T cells of sarcoidosis patients using an interferon-gamma ELISPOT assay. In peripheral blood, there were strong T cell responses to a peptide derived from the cytoskeletal protein vimentin in 6 out of 11 DRBI*0301(pos) patients with active disease but not in patients with other HLA types. BAL T cell responses against peptides derived from ATP synthase or from lysyl-tRNA synthetase were detected in DRB1*0301(pos) as welt as DRB1*0301(neg) patients. By using antigenic peptides presented in vivo in the lungs of sarcoidosis patients, we have identified blood and lung T cell autoimmune responses that may help sustain the inflammation in this disease.
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| 9. |
- Wahlström, Jan, et al.
(författare)
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Identification of HLA-DR-bound peptides presented by human bronchoalveolar lavage cells in sarcoidosis
- 2007
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Ingår i: Journal of Clinical Investigation. - 0021-9738 .- 1558-8238. ; 117:11, s. 3576-3582
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Tidskriftsartikel (refereegranskat)abstract
- Sarcoidosis is an inflammatory disease of unknown etiology, most commonly affecting the lungs. Activated CD4(+) T cells accumulate in the lungs of individuals with sarcoidosis and are considered to be of central importance for inflammation. We have previously shown that Scandinavian sarcoidosis patients expressing the HLADR allele DRB1*0301 are characterized by large accumulations in the lungs of CD4(+) T cells expressing the TCR AV2S3 gene segment. This association afforded us a unique opportunity to identify a sarcoidosis-specific antigen recognized by AV2S3(+) T cells. To identify candidates for the postulated sarcoidosis-specific antigen, lung cells from 16 HLA-DRB1*0301(pos) patients were obtained by bronchoalveolar lavage. HLA-DR molecules were affinity purified and bound peptides acid eluted. Subsequently, peptides were separated by reversed-phase HPLC and analyzed by liquid chromatography-mass spectrometry. We identified 78 amino acid sequences from self proteins presented in the lungs of sarcoidosis patients, some of which were well-known autoantigens such as vimentin and ATP synthase. For the first time, to our knowledge, we have identified HLA-bound peptides presented in vivo during an inflammatory condition. This approach can be extended to characterize HLA-bound peptides in various autoimmune settings.
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| 10. |
- Weissert, Robert
(författare)
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Immunogenetics and treatment of experimental autoimmune encephalomyelitis
- 1999
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Experimental autoimmune encephalomyelities (EAE) is a model of multiple sclerosis (MS). This experimental central nervous system (CNS) disease can be induced in rats by immunization with components of the CNS myelin sheath. EAE actively induced with myelin-basic-protein (MBP) is characterized by T cell infiltration of the CNS and typically leads to a monophasic disease course in Lewis (LEW) rats. We showed that in LEW rats MBP and MBP peptide immunizations result in preferential recruitment of TCRBV8S2+ T cells. This preferential T cell receptor (TCR) recruitment has its source in a multistep molecular mimicry event and is strictly dependent on the MHC and heterologous antigen. We used this model to assess the therapeutic potential of vaccination with naked DNA. Vaccination with DNA encoding the dominant T cell epitope MBPGP63-88 can suppress disease. Myelin oligodendrocyte glycoprotein (MOG) is a minor component of the myelin sheath. Unlike MBP which is hidden in the myelin sheath, MOG is exposed on its surface and is therefore easy accessible for antibody binding. Active immunization with the extracellular part of MOG (rMOG aa1-125) leads to different CNS specific immune effector mechanisms compared to MBP. While immunization with MBP results in purely inflammatory, monophasic disease, use of MOG cae result in a demyelinating chronic disease. We analyzed the influence of the major histocompatibility complex (MHC) haplotype on disease, pathology of CNS lesions and immune responses in this new model. The MHC haplotype does not exert an all or none effect on disease susceptibility but determines the degree of disease susceptibility, recruitment of MOG-specific immunocompetent cells, clinical course and CNS pathology in a hierarchical and allele-specific manner. The major regulatory region mapped to the MHC class II genes, but also MHC class I and class III or genes further downstream influenced disease. Moreover, non-MHC genes could overcome MHC susceptibility effects. To understand the molecular basis for the observed strong MHC influences we systematically measured immunogenicity, encephalitogenicity and MHC class II peptide binding in different rat MHC haplotypes. Using overlapping synthetic peptides covering MOG 1-125 we found that immunodominance of T cell determinants was purely MHC class II dependent. Surprisingly, immunogenicity correlated only partly with encephalitogenicity. No disease ensued in resistant haplotypes despite the presence of immunogenic epitopes. In susceptible haplotypes the region between MOG 91-114 contained the main encephalitogenic determinants. Beside a T cell determinant MOG 91-108 also contains a linear B cell determinant, suggesting a role for B cell mediated immunity in encephalitogenicity. Binding patterns and the degree of promiscuity in peptide binding varied greatly between different haplotypes, with susceptible haplotypes showing higher promiscuity. MHC haplotype regulated peptide specific production of IFN-[gamma] did not correlate to MOG peptide encephalitogenicity. Such T cell responses can instead be argued to modulate disease, perhaps by influencing antigen presentation after induction of MHC expression in the periphery or in the CNS. In summary these studies demonstrate in two EAE models the dependence of MHC gene products, background genes and antigen on the clinical outcome, lesion development and immune responses. There is a high complexity of MHC driven T- and B cell determinant-selection, TCR shaping and tuning. Depending on the antigen, great differences in the molecular pathways that lead to disease manifestation come into action.
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