19 research outputs found
Entschlüsselung der Immunpathogenese des Pemphigus mit neuen Forschungsansätzen: detaillierte Analyse der systemischen und lokalen T-Zell-Reaktionen
No full textThe immune system functions as a sophisticated collaboration between the innate and
adaptive immunity capable of distinguishing self from non-self. A pivotal role in
governing adaptive immune responses is attributed to CD4+ T cells. With specificity for
a particular antigen, they facilitate the establishment of protective immunity targeted at
specific pathogens, but their actions can also inadvertently give rise to autoimmunity. One
such example is pemphigus, a rare autoimmune disease affecting skin and mucous
membranes. Pemphigus manifests as blister formations and erosions and is characterized
by the loss of immune tolerance to desmosomal cadherins, particularly desmoglein
(Dsg)1 and Dsg3. Autoreactive T cells provide help to B cells and thus play a central role
in driving the production of pathogenic IgG autoantibodies against these cadherins.
However, due to their low frequencies, detection of autoreactive T cells in pemphigus has
been challenging. Traditional methods like tritiated thymidine incorporation (3H-TdR) or
enzyme-linked immunospot (ELISpot) analysis have inherent limitations, including the
inability to distinguish specific cell populations or the need for an ab initio presumption
of the significance of certain cytokines. Newer experimental approaches, focused on
direct characterization and quantification of these T cells, are based on flow cytometry.
Some make use of MHC multimers others of surface activation markers. We aimed at
establishing a method of detecting autoreactive CD4+ T cells utilizing the transient
upregulation of CD154 (CD40L) upon stimulation with relevant antigens or
immunodominant epitopes, respectively. The CD154-method of antigen-specific T cell
detection is sensitive, time-efficient, and withal allows for specific phenotypic and
functional characterization of T cells when using a comprehensive panel of
immunological parameters. Herewith, we analyzed Dsg3-specific CD4+CD154+ T cells
from peripheral blood of patients with pemphigus vulgaris (PV) in comparison to healthy
control (HC), where the expression of CD154 on ex vivo expanded CD4+ T cells
positively correlated with Dsg3 titers in pemphigus patients. We could show clonal
expansion of CD4+CD154+ T cells in response to both, the full protein Dsg3 as well as
to various epitope-specific peptides, with P2 (extracellular domain 2 of Dsg3) appearing
to be particularly immunomodulatory in PV patients. Moreover, upregulation of IL-17
and IL-21 was seen to associate with antigen-specific activation in active stages of the
disease, further supporting the concept that immune activity in PV relies, at least partially,
on Dsg3-reactive Th17/Tfh17 cell subsets. Contrary to foregoing belief that in pemphigus
the subtype of T cells responsible for supporting autoreactive B cells is IL-4- and IL-5-producing Th2 cells. With newly discovered T cell populations such as Th9 cells, Th22
cells and many others, the inherent complexity of the pathomechanisms of pemphigus
will likely become even more evident, with different subsets presumably dominating at
different stages of the disease contributing to the complex interplay of immune
dysregulation in pemphigus.
Pemphigus is an autoimmune disease mediated by systemic immune responses, however,
the inflammation is localized in the peripheral effector tissues, namely mucosa and the
skin. More and more studies shed light on the importance of local tissue-resident
immunity in pemphigus. Yet, owing to hampered peripheral tissues’ accessibility and
difficulty in extracting the resident lymphocytes, blood remains a major source of
research material. Hence, we aimed at establishing a rapid easy-to-use protocol to isolate
a sufficient number of viable immune cells from small skin biopsies that can be directly
used for a deeper characterization such as comprehensive phenotyping and functional
studies of T cells. Since it is not yet clear what role local immunity plays in the disease's
pathogenesis and how important the balance among different T cell subsets in pemphigus
is, the need for further research to unravel its complexities is inherent. These newly
established methods, CD154 for detection of Dsg-reactive T cells and rapid isolation of
lymphocytes from skin and mucosa, aid to expedite the process and allow for closer
inspection of pemphigus’ pathogenesis. Moreover, the presented comprehensive
immunophenotyping of PV patients underlines the significance of Th17/Tfh17 cells, T
cell dysregulation, enhanced B cell activation, and elevated pro-inflammatory cytokines,
which collectively contribute to the autoimmune response observed in pemphigus. Thus,
these new findings not only provide deeper insights into the pathomechanism but may
also reveal improved treatment options for pemphigus patients in the future.Das Immunsystem funktioniert als ein ausgeklügeltes Zusammenspiel zwischen der
angeborenen und der adaptiven Immunität, das in der Lage ist, zwischen selbst und fremd
zu unterscheiden. Eine zentrale Rolle bei der Steuerung der adaptiven Immunreaktionen
wird den CD4+ T-Zellen zugeschrieben. Mit ihrer Spezifität für ein bestimmtes Antigen
erleichtern sie den Aufbau einer schützenden Immunität, die sich gegen bestimmte
Krankheitserreger richtet, aber ihre Aktivität kann auch unbeabsichtigt zu Autoimmunität
führen. Ein solches Beispiel ist Pemphigus, eine seltene Autoimmunerkrankung, die Haut
und Schleimhäute betrifft. Pemphigus äußert sich durch Blasenbildung sowie Erosionen
und ist durch den Verlust der Immuntoleranz gegenüber desmosomalen Cadherinen,
insbesondere Desmoglein (Dsg)1 und Dsg3, gekennzeichnet. Autoreaktive T-Zellen
unterstützen B-Zellen und spielen dadurch eine zentrale Rolle bei der Produktion
pathogener IgG-Autoantikörper gegen diese Cadherine. Aufgrund ihrer geringen
Häufigkeit war der Nachweis autoreaktiver T-Zellen bei Pemphigus jedoch eine
Herausforderung. Herkömmliche Methoden wie Zellproliferationsassays auf Grundlage
des Einbaus des Radioisotops [3H]-Thymidin (3H-TdR) in die zelluläre DNA oder die
ELISpot-Analyse weisen inhärente Grenzen auf, darunter die Unfähigkeit, zwischen
verschiedenen Zellpopulationen zu unterscheiden oder von vornherein die wesentlichere
Bedeutung bestimmter Zytokine vorauszusetzen. Neuere experimentelle Ansätze, die sich
auf die direkte Charakterisierung und Quantifizierung dieser T-Zellen konzentrieren,
basieren auf der Durchflusszytometrie unter Verwendung von bspw. MHC-Multimeren
oder Oberflächenaktivierungsmarkern. Unser Ziel war es, eine Methode zum Nachweis
autoreaktiver CD4+ T-Zellen zu entwickeln, die eine vorübergehende Hochregulierung
von CD154 (CD40L) nach Stimulation mit relevanten Antigenen oder immundominanten
Epitopen nutzt. Die CD154-Methode der antigenspezifischen T-Zell-Detektion ist
sensitiv, zeiteffizient und ermöglicht zudem eine spezifische phänotypische und
funktionelle Charakterisierung von T-Zellen mittels umfassender Panels
immunologischer Parameter. Hiermit analysierten wir Dsg3-spezifische CD4+CD154+
T-Zellen aus dem peripheren Blut von Patienten mit Pemphigus vulgaris (PV) und
gesunden Kontrollprobanden (HC), wobei die Expression von CD154 auf ex vivo
expandierten CD4+ T-Zellen positiv mit den Dsg3-Titern bei Pemphigus-Patienten
korrelierte. Wir konnten eine klonale Expansion von CD4+CD154+ T-Zellen als
Reaktion sowohl auf das Vollprotein Dsg3 als auch auf verschiedene epitopspezifische
Peptide zeigen, wobei P2 (extrazelluläre Domäne 2 von Dsg3) bei PV-Patienten besonders immunmodulatorisch zu sein scheint. Darüber hinaus wurde eine
Hochregulierung von IL-17 und IL-21 in Verbindung mit antigenspezifischer
Aktivierung in aktiven Stadien der Krankheit festgestellt, womit das derzeitige Konzept
unterstützt wird, dass die Immunaktivität bei PV zumindest teilweise auf Dsg3-reaktiven
Th17/Tfh17-Zelluntergruppen beruht - entgegen der bisherigen Annahme, dass bei
Pemphigus der Subtyp an T-Zellen, der für die Unterstützung autoreaktiver B-Zellen
verantwortlich ist, IL-4- und IL-5-produzierende Th2-Zellen sind. Mit den neu entdeckten
T-Zell-Populationen wie Th9-Zellen, Th22-Zellen und vielen anderen wird die inhärente
Komplexität des Pathomechanismus von Pemphigus wahrscheinlich noch deutlicher
werden, wobei verschiedene Untergruppen vermutlich in unterschiedlichen Stadien der
Krankheit dominieren und zum komplexen Zusammenspiel der Immundysregulation bei
Pemphigus beitragen.
Pemphigus ist eine Autoimmunerkrankung, die durch systemische Immunreaktionen
vermittelt wird, wobei die Entzündung jedoch in den peripheren Effektorgeweben,
nämlich der Schleimhaut und der Haut, lokalisiert ist. Immer mehr Studien beleuchten
die Bedeutung der lokalen, gewebeassoziierten Immunität bei Pemphigus, doch aufgrund
der eigeschränkten Zugänglichkeit peripherer Gewebe und der Schwierigkeit
Lymphozyten aus diesen Geweben zu extrahieren, bleibt Blut eine wichtige Quelle für
Forschungsmaterial. Daher haben wir uns zum Ziel gesetzt, ein schnelles, einfach zu
handhabendes Protokoll zur Isolierung einer ausreichenden Anzahl vitaler Immunzellen
aus kleinen Hautbiopsien zu entwickeln, die direkt für eine tiefergehende
Charakterisierung, wie z. B. eine umfassende Phänotypisierung und funktionelle Studien
von T-Zellen, verwendet werden können. Da noch nicht klar ist, welche Rolle die lokale
Immunität bei der Krankheitsentstehung spielt und welche Gleichgewichtung zwischen
den verschiedenen T-Zell-Untergruppen bei Pemphigus zukommt, ist weitere Forschung
notwendig, um die Komplexität der Krankheit zu entschlüsseln. Diese neu etablierten
Methoden, d.h. die Detektion von CD154 zum Nachweis Dsg-reaktiver T-Zellen und die
schnelle Isolierung von Lymphozyten aus Haut und Schleimhaut, tragen allerdings dazu
bei, den Prozess zu beschleunigen und ermöglichen eine genauere Untersuchung der
Pemphigus-Pathogenese. Darüber hinaus unterstreicht die vorgestellte umfassende
Immunphänotypisierung von PV-Patienten die besondere Bedeutung der Th17/Tfh17-
Zellen, der Dysregulation von T-Zellen, der verstärkten Aktivierung von B-Zellen und
von proinflammatorischen Zytokinen, die gemeinsam zur Autoimmunreaktion bei
Pemphigus beitragen. Diese neuen Erkenntnisse liefern nicht nur ein tieferes Verständnis der Pathomechanismen, sondern könnten auch verbesserte zukünftige
Behandlungsmöglichkeiten für Pemphigus-Patienten aufzeigen
Polymorphisms in mirna-binding sites of nucleotide excision repair genes and colorectal cancer risk
No full textReduced DNA repair capacity and DNA damage accumulation may lead to cancer development. Regulation of and coordination between genes involved in DNA repair pathways is fundamental for maintaining genome stability, and post-transcriptional gene regulation by microRNAs (miRNAs) may therefore be of particular relevance. In this context, the presence of single nucleotide polymorphisms (SNPs) within the 3'untranslated regions of target DNA repair genes could alter the binding with specific miRNAs, modulating gene expression and ultimately affecting cancer susceptibility.In this study, we investigated the role of genetic variations in miRNA-binding sites of nucleotide excision repair (NER) genes in association with colorectal cancer (CRC) risk. From 28 NER genes, we screened among SNPs residing in their 3'untranslated regions and simultaneously located in miRNA-binding sites, with an in silico approach. Through the calculation of different binding free energy according to both alleles of identified SNPs, and with global binding free energies median providing a threshold, we selected nine NER gene variants. We tested those SNPs in 1098 colorectal cancer cases and 1469 healthy controls from the Czech Republic.Rs7356 in RPA2 and rs4596 in GTF2H1 were associated with colorectal cancer risk. After stratification for tumor location, the association of both SNPs was significant only for rectal cancer (rs7356: OR 1.52, 95% CI 1.02-2.26, P = 0.04 and rs4596: OR 0.69, 95% CI 0.50-0.94, P = 0.02; results not adjusted for multiple testing).Variation in miRNA target binding sites in the 3' untranslated region of NER genes may be important for modulating colorectal cancer risk, with a different relevance according to tumor location. © The Author 2012. Published by Oxford University Press. All rights reserved
Политика КНР по продвижению китайской литературы за рубежом в XXI веке
No full textДанная выпускная квалификационная работа посвящена изучению политики продвижения китайской литературы за рубежом в XXI веке. В процессе работы автором был проведён анализ проектов по продвижению китайской литературы за рубеж, таких как «План КНР по продвижению литературы за рубеж» и «Книжный аромат Шёлкового пути». Выявлены идейные и жанровые особенности продвигаемой литературы, а также установлены основные регионы распространения китайской литературы.
В ходе исследования автор приходит к выводу о том, что продвижение китайской литературы за рубеж является инструментом «мягкой» силы КНР, а также используется КНР для развития международных связей.This graduate paper is devoted to the PRC’s policy of promotion Chinese literature abroad in the XXI century. Within the framework of the research, the author analyzed projects for the promotion of Chinese literature abroad, such as the "Plan of the PRC to promote literature abroad" and "Book flavor of the Silk Road". The study reveals the ideological and genre features of the promoted as well as the main regions of the circulation of Chinese literature.
In the course of the research, the author comes to the conclusion that the promotion of Chinese literature abroad is an instrument of the "soft" power of the PRC. It is also used by the PRC as a way to develop international contacts
How to Achieve Sustainable Efficiency with Teleworkers: Leadership Model in Telework
No full textAbstractGlobalization, increasing competition, changes in demographic structure of population, and development of information and communication technologies (ICT) are the factors that have posed new challenges for organizations in recent years. The newly fledged conditions have vital impact on production methods and procedures as well as work arrangements in organizations. Currently, organizations cope with various times in which work is executed as well as various places where it is executed. Thanks to ICT an employee can work at the client's place, at home, in a means of transport or in a café. All these situations constitute the work arrangement in which the worker is considered to work ‘from distance’, i.e. as a teleworker.The presented approach refers to the flexible work arrangement where an organization's priority lies with its endeavour to become and stay flexible in terms of time and place of performance. Under these conditions the implementation of new ways of work arrangements presents new challenges for leadership. To accomplish this basic function of management when employees are not present at workplace or even are not available during working hours and face-to-face communication is impossible poses many difficulties.The paper focuses on comparison and identification of the differences between traditional work arrangements and telework, highlights weak points in leadership of teleworkers, and proposes a leadership model for teleworkers’ induction
Соглашение о государственно-частном партнерстве
No full textНесмотря на принятие Федерального закона "О государственно-частном партнерстве, муниципально-частном партнерстве в Российской Федерации и внесении изменений в отдельные законодательные акты Российской Федерации" от 13.07.2015 N 224-ФЗ, законодательство в указанной сфере продолжает развиваться, а судебная и административная практика находятся на этапе становления. Большое количество теоретических и практических проблем вызывают необходимость дополнительного исследования и развития указанной темы. В настоящей работе автором дана попытка дать собственную оценку проблемным ситуациям и предложить варианты дальнейшего развития законодательства.
На основе проведенного исследования автор пришел к ряду выводов и предложений, основными из которых являются выводы о гражданско-правовой природе соглашения о публично-частном партнерстве, о самостоятельности явления публично-частного партнерства, о недопустимости применения к публично-частному партнерству законодательства о приватизации, а также рекомендации установить множественность на стороне публичных партнеров, расширить перечень лиц, уполномоченных действовать в качестве публичных партнеров, расширить перечень требований к частным партнерам, гарантирующих надлежащее исполнение соглашения, оставить открытым перечень мер поддержки частного партнера и дать возможность частному партнеру участвовать в их выборе и другие.Despite adoption of the Federal law "About public-private partnership, municipal-private partnership in the Russian Federation and modification of legal acts of the Russian Federation" of 13.07.2015 N 224, the legislation in this sphere continues to develop, and judicial and administrative practice are forming now. A large number of theoretical and practical problems cause the need for further research and development of this topic. In this paper, the author attempts to give his own assessment of the problem situations and propose options for further development of the legislation.
The research led to the following findings, the main of which are the conclusions about the civil nature of the agreement on public-private partnership, the independence of this phenomenon, the inadmissibility of the application of privatization legislation to public-private partnership. The author also gives recommendations to establish plurality on the side of public partners, to expand the list of persons authorized to act as public partners, to expand the list of requirements for private partners, to leave open the list of measures to support the private partner and to allow the private partner to participate in their selection and others
Сравнительный анализ концепций биополитики в политической философии М. Фуко, Дж. Агамбена и А. Негри
No full textВыпускная квалификационная работа на тему: «Сравнительный анализ концепций биополитики в политической философии М. Фуко, Дж. Агамбена и А. Негри».
Автор: Сигарев Иван Сергеевич, студент 4 курса факультета политологии СПбГУ.
Работа состоит из введения, двух глав, заключения и списка использованных источников.
Во введении раскрывается актуальность выбранной темы, определяются объект и предмет исследования, цель работы и исследовательские задачи. Кроме того, введение содержит информацию о методологии и методах, применявшихся в процессе создания работы, а также включает в себя краткий обзор литературы, характеризующей степень разработанности темы исследования.
Первая глава носит теоретический характер, состоит из двух параграфов. В первом параграфе рассматриваются исторические предпосылки и теоретико-методологические основания концепта «биополитика». Помимо этого, в этой части работы особое внимание уделено анализу концепции биополитики Г.Челлена. Второй параграф первой главы посвящен изучению концептов биополитики и биовласти в контексте политической философии М.Фуко.
Вторая глава данной работы посвящена анализу концепций биополитики в постфукианский период развития политической философии. Глава состоит из 3 параграфов, в первом из которых приводятся взгляды Дж.Агамбена на концепцию биополитики. Второй параграф посвящён анализу онтологизации биополитики в работах А.Негри. В третьем параграфе проведён сравнительный анализ биополитических концепций М.Фуко, Дж.Агамбена и А.Негри.
В заключении обобщается проделанная работа, формулируются выводы о сущности концепта биополитики и проблемам использования данного концепта в контексте современных социально-политических проблем.
Работа содержит 80 страниц машинописного текста. Для написания было использовано 106 библиографических источников.The final thesis on the topic: «The comparative analysis of the biopolitic’s concepts in the political philosophy of M. Foucault, J. Agamben and A. Negri.»
Author: Sigarev Ivan Sergeyevich, 4th year student of the Faculty of Political Science, St. Petersburg State University.
The diploma consists of introduction, two chapters, conclusion and bibliographical list.
The introduction allows to understand relevance of the chosen topic, define the object and subject of research, the purpose of this work and research tasks. In addition, the introduction contains information about the methodology of this work and includes a brief review of the literature characterizing the degree of research topic’s development.
The first chapter is theoretical in nature and includes two paragraphs. The first one of them discusses the historical premises and theoretical and methodological foundations of the concept of "biopolitics". In addition, special attention is paid to the analysis of the concept of G. Chellen's biopolitics in this part of the work. The second paragraph of the first chapter is devoted to the study of the concepts of biopolitics and biopower in the context of M. Foucault's political philosophy.
The second chapter of this work is devoted to the analysis of the concepts of biopolitics in the post-Foucault period of political philosophy development. The chapter consists of 3 paragraphs. The first paragraph contains J. Agamben’s views on the concept of biopolitics. The second paragraph is devoted to the analysis of the ontologization of biopolitics in the A. Negri’s works. The third paragraph includes the comparative analysis of the biopolitical concepts of M. Foucault, J. Agamben and A. Negri.
The conclusion includes summary of the work, formulates conclusions about the essence of the concept of biopolitics and the problems of its implementation in the context of modern socio-political problems.
The work contains 80 pages of typewritten text. In the process of writing 106 sources were used
Detection of rare autoreactive T cell subsets in patients with pemphigus vulgaris
No full textAnalysis of T lymphocyte proliferation and activation after antigenic or mitogenic stimulation is a vital parameter used in the diagnosis of various immuno-deficiencies and during the monitoring of treatment responses. Most applied techniques are based on the incorporation of tritiated thymidine (3H-TdR) or ELISPOT analysis, both rely on rather time-consuming/-intensive ex vivo protocols or encompass inherent drawbacks such as the inability to distinguish specific cell populations (3H-TdR, ELISPOT) or focus on a single cytokine (ELISPOT). Here we aimed at characterizing the rapid expression of intracellular CD154 (CD40L) as a marker for rare antigen-specific CD4+ T cells in pemphigus vulgaris (PV). Upon stimulation with human desmoglein (Dsg) 3, the major autoantigen in PV, the expression of CD154 was significantly increased in PV patients compared to healthy controls (HC) and correlated with anti-Dsg3 IgG titers. Patients with active disease showed higher numbers of Dsg3-reactive CD4+ T cells in CXCR5+ T follicular helper cells. In remittent PV and HC, CXCR5+CD4+ T cells remained largely unaffected by Dsg3. IL-17 and IL-21 expression were significantly induced only in CD154+CD4+ T cells from PV patients, lending themselves as potential novel treatment targets. Additionally, stimulation with immunodominant Dsg3-derived epitopes strongly induced a CD4+ T cell response via CD40-CD154 interaction similar to the human Dsg3 protein. We here established a rapid ex vivo assay allowing the detection of Dsg3-reactive CD4+ T cells from activated systemically available PBMCs, which further supports the crucial concept of antigen-specific T cells in the pathogenesis of PV.Gefördert durch den Open-Access-Publikationsfonds der UB Marburg
DataSheet_1_Detection of rare autoreactive T cell subsets in patients with pemphigus vulgaris.docx
No full textAnalysis of T lymphocyte proliferation and activation after antigenic or mitogenic stimulation is a vital parameter used in the diagnosis of various immuno-deficiencies and during the monitoring of treatment responses. Most applied techniques are based on the incorporation of tritiated thymidine (3H-TdR) or ELISPOT analysis, both rely on rather time-consuming/-intensive ex vivo protocols or encompass inherent drawbacks such as the inability to distinguish specific cell populations (3H-TdR, ELISPOT) or focus on a single cytokine (ELISPOT). Here we aimed at characterizing the rapid expression of intracellular CD154 (CD40L) as a marker for rare antigen-specific CD4+ T cells in pemphigus vulgaris (PV). Upon stimulation with human desmoglein (Dsg) 3, the major autoantigen in PV, the expression of CD154 was significantly increased in PV patients compared to healthy controls (HC) and correlated with anti-Dsg3 IgG titers. Patients with active disease showed higher numbers of Dsg3-reactive CD4+ T cells in CXCR5+ T follicular helper cells. In remittent PV and HC, CXCR5+CD4+ T cells remained largely unaffected by Dsg3. IL-17 and IL-21 expression were significantly induced only in CD154+CD4+ T cells from PV patients, lending themselves as potential novel treatment targets. Additionally, stimulation with immunodominant Dsg3-derived epitopes strongly induced a CD4+ T cell response via CD40-CD154 interaction similar to the human Dsg3 protein. We here established a rapid ex vivo assay allowing the detection of Dsg3-reactive CD4+ T cells from activated systemically available PBMCs, which further supports the crucial concept of antigen-specific T cells in the pathogenesis of PV.</p
Molecular characteristics of mismatch repair genes in sporadic colorectal tumors in Czech patients
No full text
