TY  - THES
A1  - Wunsch, Marie
T1  - Das enterische Nervensystem als mögliche Zielstruktur der Autoimmunreaktion in der Multiplen Sklerose
T1  - The enteric nervous system is a potential autoimmune target in multiple sclerosis
N2  - Bei der Multiplen Sklerose (MS) handelt es sich um eine Autoimmunerkrankung des zentralen Nervensystems (ZNS). Abhängig von der betroffenen ZNS-Region kann es zu vielfältigen Symptomen kommen. Neben neurologischen Symptomen verursacht durch ZNS-Läsionen leidet ein Großteil der MS-Patienten auch unter gastrointestinalen Funktionsstörungen. Diese gastrointestinalen Symptome wurden bisher eher auf Läsionen im Rückenmark zurückgeführt und nicht direkt in Verbindung mit der autoimmunen Ätiologie der Erkrankung gebracht.
In dieser Studie wurde das enterische Nervensystem (ENS) in einem B-Zell- und Antikörper-abhängigen Mausmodell der MS untersucht. Dafür wurde der Autoimmunprozess durch Immunisierung mit MP4, einem Fusionsprotein aus dem Myelin-Basischen-Protein (MBP) und dem Proteolipid-Protein (PLP), ausgelöst. Das ZNS und ENS wurden in den unterschiedlichen Erkrankungsstadien immunhistochemisch und elektronenmikroskopisch analysiert. Neben der Immunpathologie des ZNS konnte dabei eine Degeneration des ENS schon vor dem Einsetzen der ersten neurologischen Defizite nachgewiesen werden. Die ENS-Pathologie war antikörper-mediiert und ging einher mit einer verringerten gastrointestinalen Motilität sowie mit einer Gliose und Neurodegeneration des ENS.
Mithilfe von Immunpräzipitation und Massenspektrometrie konnten im ENS vier mögliche Zielstrukturen des Autoimmunprozesses identifiziert werden, was auf sog. epitope spreading hindeutet. Auch im Plasma von MS-Patienten konnten Antikörper gegen drei dieser Antigene nachgewiesen werden. Des Weiteren zeigten sich in Kolon-Resektaten von MS-Patienten erste Ansätze einer Neurodegeneration und Gliose des ENS. 
In dieser Studie wurde zum ersten Mal ein direkter Zusammenhang zwischen der Autoimmunreaktion gegen das ZNS und einer simultanen Reaktion gegen das ENS gezeigt. Dies kann einen Paradigmenwechsel im Verständnis der Immunpathogenese der MS anstoßen und neue therapeutische und diagnostische Ansätze initiieren.
N2  - Multiple sclerosis (MS) is a chronic autoimmune disease, in which the immune system attacks the central nervous system (CNS). Clinical symptoms and the course of the disease can vary among patients, depending on the region of the brain primarily affected. Besides the neurological symptoms caused by CNS lesions, a great amount of MS patients display functional gastrointestinal impairments. Gastrointestinal symptoms were previously explained by the presence of spinal cord lesions rather than being linked to the autoimmune pathomechanisms of the disease. 
Here, the enteric nervous system (ENS) was studied in a B cell- and antibody-dependent mouse model of MS, in which the myelin basic protein (MBP) - proteolipid protein (PLP) fusion protein MP4 was used to initiate the autoimmune attack. Immunohistochemistry and electron microscopy were performed at different stages of the disease. We noted that in addition to the immune pathology in the CNS itself, the ENS also showed signs of degeneration. ENS degeneration was evident prior to the manifestation of CNS lesions and to the onset of neurological deficits in mice. ENS pathology was antibody-mediated and accompanied by impaired gastrointestinal motility, ENS gliosis and neurodegeneration. Using immunoprecipitation and mass spectrometry, four autoimmune targets expressed by enteric glia and/or neurons could be identified suggesting that epitope spreading to antigens of the ENS had occurred. MS patients displayed plasma antibodies against three of the ENS autoantigens. Studying human colon resectates provided preliminary evidence for gliosis and neurodegeneration of the ENS in MS patients, which was absent in non-MS controls.
Overall, this study establishes a pathomechanistic link between the well-established autoimmune attack on the CNS and a simultaneous attack on the ENS that has not been described so far. These findings can initiate a paradigm shift in the current understanding of the pathomechanism of MS with diagnostic and therapeutic implications.
KW  - Multiple Sklerose
KW  - Autoantikörper
KW  - Enterisches Nervensystem
KW  - Experimentelle Autoimmune Enzephalomyelitis
KW  - Darmwandnervensystem
KW  - Autoimmunität
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-175888
ER  - 
TY  - JOUR
A1  - Wunsch, Marie
A1  - Hohmann, Christopher
A1  - Milles, Bianca
A1  - Rostermund, Christina
A1  - Lehmann, Paul V.
A1  - Schroeter, Michael
A1  - Bayas, Antonios
A1  - Ulzheimer, Jochen
A1  - Mäurer, Mathias
A1  - Ergün, Süleyman
A1  - Kuerten, Stefanie
T1  - The Correlation between the Virus- and Brain Antigen-Specific B Cell Response in the Blood of Patients with Multiple Sclerosis
JF  - Viruses
N2  - There is a largely divergent body of literature regarding the relationship between Epstein-Barr virus (EBV) infection and brain inflammation in multiple sclerosis (MS). Here, we tested MS patients during relapse (n = 11) and in remission (n = 19) in addition to n = 22 healthy controls to study the correlation between the EBV- and brain-specific B cell response in the blood by enzyme-linked immunospot (ELISPOT) and enzyme-linked immunosorbent assay (ELISA). Cytomegalovirus (CMV) was used as a control antigen tested in n = 16 MS patients during relapse and in n = 35 patients in remission. Over the course of the study, n = 16 patients were untreated, while n = 33 patients received immunomodulatory therapy. The data show that there was a moderate correlation between the frequencies of EBV- and brain-reactive B cells in MS patients in remission. In addition we could detect a correlation between the B cell response to EBV and disease activity. There was no evidence of an EBV reactivation. Interestingly, there was also a correlation between the frequencies of CMV- and brain-specific B cells in MS patients experiencing an acute relapse and an elevated B cell response to CMV was associated with higher disease activity. The trend remained when excluding seronegative subjects but was non-significant. These data underline that viral infections might impact the immunopathology of MS, but the exact link between the two entities remains subject of controversy.
KW  - B cells
KW  - CMV
KW  - EBV
KW  - ELISPOT
KW  - MS
Y1  - 2016
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-146946
VL  - 8
IS  - 4
ER  - 
TY  - JOUR
A1  - Rovituso, Damiano M.
A1  - Scheffler, Laura
A1  - Wunsch, Marie
A1  - Kleinschnitz, Christoph
A1  - Dörck, Sebastian
A1  - Ulzheimer, Jochen
A1  - Bayas, Antonios
A1  - Steinman, Lawrence
A1  - Ergün, Süleyman
A1  - Kuerten, Stefanie
T1  - CEACAM1 mediates B cell aggregation in central nervous system autoimmunity
JF  - Scientific Reports
N2  - B cell aggregates in the central nervous system (CNS) have been associated with rapid disease progression in patients with multiple sclerosis (MS). Here we demonstrate a key role of carcinoembryogenic antigen-related cell adhesion molecule1 (CEACAM1) in B cell aggregate formation in MS patients and a B cell-dependent mouse model of MS. CEACAM1 expression was increased on peripheral blood B cells and CEACAM1\(^+\) B cells were present in brain infiltrates of MS patients. Administration of the anti-CEACAM1 antibody T84.1 was efficient in blocking aggregation of B cells derived from MS patients. Along these lines, application of the monoclonal anti-CEACAM1 antibody mCC1 was able to inhibit CNS B cell aggregate formation and significantly attenuated established MS-like disease in mice in the absence of any adverse effects. CEACAM1 was co-expressed with the regulator molecule T cell immunoglobulin and mucin domain −3 (TIM-3) on B cells, a novel molecule that has recently been described to induce anergy in T cells. Interestingly, elevated coexpression on B cells coincided with an autoreactive T helper cell phenotype in MS patients. Overall, these data identify CEACAM1 as a clinically highly interesting target in MS pathogenesis and open new therapeutic avenues for the treatment of the disease.
KW  - autoimmunity
KW  - multiple sclerosis
KW  - neuroimmunology
Y1  - 2016
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-147690
VL  - 6
ER  - 
TY  - JOUR
A1  - Wunsch, Marie
A1  - Caspell, Richard
A1  - Kuerten, Stefanie
A1  - Lehmann, Paul V.
A1  - Sundararaman, Srividya
T1  - Serial measurements of apoptotic cell numbers provide better acceptance criterion for PBMC quality than a single measurement prior to the T cell assay
JF  - Cells
N2  - As soon as Peripheral Blood Mononuclear Cells (PBMC) are isolated from whole blood, some cells begin dying. The rate of apoptotic cell death is increased when PBMC are shipped, cryopreserved, or stored under suboptimal conditions. Apoptotic cells secrete cytokines that suppress inflammation while promoting phagocytosis. Increased numbers of apoptotic cells in PBMC may modulate T cell functions in antigen-triggered T cell assays. We assessed the effect of apoptotic bystander cells on a T cell ELISPOT assay by selectively inducing B cell apoptosis using α-CD20 mAbs. The presence of large numbers of apoptotic B cells did not affect T cell functionality. In contrast, when PBMC were stored under unfavorable conditions, leading to damage and apoptosis in the T cells as well as bystander cells, T cell functionality was greatly impaired. We observed that measuring the number of apoptotic cells before plating the PBMC into an ELISPOT assay did not reflect the extent of PBMC injury, but measuring apoptotic cell frequencies at the end of the assay did. Our data suggest that measuring the numbers of apoptotic cells prior to and post T cell assays may provide more stringent PBMC quality acceptance criteria than measurements done only prior to the start of the assay.
KW  - T cell assay
KW  - apoptosis
KW  - acceptance
KW  - viability
KW  - ELISPOT
Y1  - 2015
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-150213
VL  - 4
IS  - 1
ER  - 
TY  - JOUR
A1  - Wunsch, Marie
A1  - Zhang, Wenji
A1  - Hanson, Jodi
A1  - Caspell, Richard
A1  - Karulin, Alexey Y.
A1  - Recks, Mascha S.
A1  - Kuerten, Stefanie
A1  - Sundararaman, Srividya
A1  - Lehmann, Paul V.
T1  - Characterization of the HCMV-Specific CD4 T Cell Responses that Are Associated with Protective Immunity
JF  - Viruses
N2  - Most humans become infected with human cytomegalovirus (HCMV). Typically, the immune system controls the infection, but the virus persists and can reactivate in states of immunodeficiency. While substantial information is available on the contribution of CD8 T cells and antibodies to anti-HCMV immunity, studies of the T\(_{H}\)1, T\(_{H}\)2, and T\(_{H}\)17 subsets have been limited by the low frequency of HCMV-specific CD4 T cells in peripheral blood mononuclear cell (PBMC). Using the enzyme-linked Immunospot\(^{®}\) assay (ELISPOT) that excels in low frequency measurements, we have established these in a sizable cohort of healthy HCMV controllers. Cytokine recall responses were seen in all seropositive donors. Specifically, interferon (IFN)-\({\gamma}\) and/or interleukin (IL)-17 were seen in isolation or with IL-4 in all test subjects. IL-4 recall did not occur in isolation. While the ratios of T\(_{H}\)1, T\(_{H}\)2, and T\(_{H}\)17 cells exhibited substantial variations between different individuals these ratios and the frequencies were relatively stable when tested in samples drawn up to five years apart. IFN-\({\gamma}\) and IL-2 co-expressing polyfunctional cells were seen in most subjects. Around half of the HCMV-specific CD4 cells were in a reversible state of exhaustion. The data provided here established the T\(_{H}\)1, T\(_{H}\)2, and T\(_{H}\)17 characteristic of the CD4 cells that convey immune protection for successful immune surveillance against which reactivity can be compared when the immune surveillance of HCMV fails.
KW  - memory cells
KW  - hcv infection
KW  - signature
KW  - Enzyme-Linked Immunospot assay (ELISPOT)
KW  - cytokine secretion kinetics
KW  - chronic viral infection
KW  - HCMV infection
KW  - CD4 T cells
KW  - exhaustion
KW  - activation
KW  - human cytomegalovirus (HCMV)
KW  - B cells
KW  - cytomegalovirus
KW  - elispot
Y1  - 2015
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-151462
VL  - 7
SP  - 4414
EP  - 4437
ER  - 
TY  - JOUR
A1  - Bail, Kathrin
A1  - Notz, Quirin
A1  - Rovituso, Damiano M.
A1  - Schampel, Andrea
A1  - Wunsch, Marie
A1  - Koeniger, Tobias
A1  - Schropp, Verena
A1  - Bharti, Richa
A1  - Scholz, Claus-Juergen
A1  - Foerstner, Konrad U.
A1  - Kleinschnitz, Christoph
A1  - Kuerten, Stefanie
T1  - Differential effects of FTY720 on the B cell compartment in a mouse model of multiple sclerosis.
JF  - Journal of Neuroinflammation
N2  - Background:
MP4-induced experimental autoimmune encephalomyelitis (EAE) is a mouse model of multiple sclerosis (MS), which enables targeted research on B cells, currently much discussed protagonists in MS pathogenesis. Here, we used this model to study the impact of the S1P1 receptor modulator FTY720 (fingolimod) on the autoreactive B cell and antibody response both in the periphery and the central nervous system (CNS).
Methods:
MP4-immunized mice were treated orally with FTY720 for 30 days at the peak of disease or 50 days after EAE onset. The subsequent disease course was monitored and the MP4-specific B cell/antibody response was measured by ELISPOT and ELISA. RNA sequencing was performed to determine any effects on B cell-relevant gene expression. S1P\(_{1}\) receptor expression by peripheral T and B cells, B cell subset distribution in the spleen and B cell infiltration into the CNS were studied by flow cytometry. The formation of B cell aggregates and of tertiary lymphoid organs (TLOs) was evaluated by histology and immunohistochemistry. Potential direct effects of FTY720 on B cell aggregation were studied in vitro.
Results:
FTY720 significantly attenuated clinical EAE when treatment was initiated at the peak of EAE. While there was a significant reduction in the number of T cells in the blood after FTY720 treatment, B cells were only slightly diminished. Yet, there was evidence for the modulation of B cell receptor-mediated signaling upon FTY720 treatment. In addition, we detected a significant increase in the percentage of B220\(^{+}\) B cells in the spleen both in acute and chronic EAE. Whereas acute treatment completely abrogated B cell aggregate formation in the CNS, the numbers of infiltrating B cells and plasma cells were comparable between vehicle- and FTY720-treated mice. In addition, there was no effect on already developed aggregates in chronic EAE. In vitro B cell aggregation assays suggested the absence of a direct effect of FTY720 on B cell aggregation. However, FTY720 impacted the evolution of B cell aggregates into TLOs.
Conclusions:
The data suggest differential effects of FTY720 on the B cell compartment in MP4-induced EAE.
KW  - B cells
KW  - EAE
KW  - FTY720
KW  - fingolimod
KW  - multiple sclerosis
KW  - TLO
Y1  - 2017
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-157869
VL  - 14
IS  - 148
ER  -