Regulatory T cells (Tregs) are vital for maintaining a balanced immune response and their dysfunction is oftenassociated with auto-immune disorders. We have previously shown that antigen-loaded... Show moreRegulatory T cells (Tregs) are vital for maintaining a balanced immune response and their dysfunction is oftenassociated with auto-immune disorders. We have previously shown that antigen-loaded anionic liposomescomposed of phosphatidylcholine (PC) and phosphatidylglycerol (PG) and cholesterol can induce strong antigenspecificTreg responses. We hypothesized that altering the rigidity of these liposomes while maintaining theirsize and surface charge would affect their capability of inducing Treg responses. The rigidity of liposomes isaffected in part by the length and saturation of carbon chains of the phospholipids in the bilayer, and in part bythe presence of cholesterol. We used atomic force microscopy (AFM) to measure the rigidity of anionic OVA323-containing liposomes composed of different types of PC and PG, with or without cholesterol, in a molar ratio of4:1(:2) distearoyl (DS)PC:DSPG (Young's modulus (YM) 3611 ± 1271 kPa), DSPC:DSPG:CHOL(1498 ± 531 kPa), DSPC:dipalmitoyl (DP)PG:CHOL (1208 ± 538), DPPC:DPPG:CHOL (1195 ± 348 kPa),DSPC:dioleoyl (DO)PG:CHOL (825 ± 307 kPa), DOPC:DOPG:CHOL (911 ± 447 kPa), and DOPC:DOPG(494 ± 365 kPa). Next, we assessed if rigidity affects the association of liposomes to bone marrow-deriveddendritic cells (BMDCs) in vitro. Aside from DOPC:DOPG liposomes, we observed a positive correlation betweenliposomal rigidity and cellular association. Finally, we show that rigidity positively correlates with Treg responsesin vitro in murine DCs and in vivo in mice. Our findings underline the suitability of AFM to measureliposome rigidity and the importance of this parameter when designing liposomes as a vaccine delivery system. Show less
Benne, N.; Duijn, J. van; Vigario, F.L.; Leboux, R.J.T.; Veelen, P. van; Kuiper, J.; ... ; Stutter, B. 2018
Regulatory T cells (Tregs) are vital for maintaining a balanced immune response and their dysfunction is oftenassociated with auto-immune disorders. We have previously shown that antigen-loaded... Show moreRegulatory T cells (Tregs) are vital for maintaining a balanced immune response and their dysfunction is oftenassociated with auto-immune disorders. We have previously shown that antigen-loaded anionic liposomescomposed of phosphatidylcholine (PC) and phosphatidylglycerol (PG) and cholesterol can induce strong antigenspecificTreg responses. We hypothesized that altering the rigidity of these liposomes while maintaining theirsize and surface charge would affect their capability of inducing Treg responses. The rigidity of liposomes isaffected in part by the length and saturation of carbon chains of the phospholipids in the bilayer, and in part bythe presence of cholesterol. We used atomic force microscopy (AFM) to measure the rigidity of anionic OVA323-containing liposomes composed of different types of PC and PG, with or without cholesterol, in a molar ratio of4:1(:2) distearoyl (DS)PC:DSPG (Young's modulus (YM) 3611 ± 1271 kPa), DSPC:DSPG:CHOL(1498 ± 531 kPa), DSPC:dipalmitoyl (DP)PG:CHOL (1208 ± 538), DPPC:DPPG:CHOL (1195 ± 348 kPa),DSPC:dioleoyl (DO)PG:CHOL (825 ± 307 kPa), DOPC:DOPG:CHOL (911 ± 447 kPa), and DOPC:DOPG(494 ± 365 kPa). Next, we assessed if rigidity affects the association of liposomes to bone marrow-deriveddendritic cells (BMDCs) in vitro. Aside from DOPC:DOPG liposomes, we observed a positive correlation betweenliposomal rigidity and cellular association. Finally, we show that rigidity positively correlates with Treg responsesin vitro in murine DCs and in vivo in mice. Our findings underline the suitability of AFM to measureliposome rigidity and the importance of this parameter when designing liposomes as a vaccine delivery system. Show less
Benne, N.; Leboux, R.J.T.; Glandrup, M.; Duijn, J. van; Lozano Vigario, F.; Neustrup, M.A.; ... ; Slütter, B. 2019
Regulatory T cells (Tregs) are vital for maintaining a balanced immune response and their dysfunction is oftenassociated with auto-immune disorders. We have previously shown that antigen-loaded... Show moreRegulatory T cells (Tregs) are vital for maintaining a balanced immune response and their dysfunction is oftenassociated with auto-immune disorders. We have previously shown that antigen-loaded anionic liposomescomposed of phosphatidylcholine (PC) and phosphatidylglycerol (PG) and cholesterol can induce strong antigenspecificTreg responses. We hypothesized that altering the rigidity of these liposomes while maintaining theirsize and surface charge would affect their capability of inducing Treg responses. The rigidity of liposomes isaffected in part by the length and saturation of carbon chains of the phospholipids in the bilayer, and in part bythe presence of cholesterol. We used atomic force microscopy (AFM) to measure the rigidity of anionic OVA323-containing liposomes composed of different types of PC and PG, with or without cholesterol, in a molar ratio of4:1(:2) distearoyl (DS)PC:DSPG (Young's modulus (YM) 3611 ± 1271 kPa), DSPC:DSPG:CHOL(1498 ± 531 kPa), DSPC:dipalmitoyl (DP)PG:CHOL (1208 ± 538), DPPC:DPPG:CHOL (1195 ± 348 kPa),DSPC:dioleoyl (DO)PG:CHOL (825 ± 307 kPa), DOPC:DOPG:CHOL (911 ± 447 kPa), and DOPC:DOPG(494 ± 365 kPa). Next, we assessed if rigidity affects the association of liposomes to bone marrow-deriveddendritic cells (BMDCs) in vitro. Aside from DOPC:DOPG liposomes, we observed a positive correlation betweenliposomal rigidity and cellular association. Finally, we show that rigidity positively correlates with Treg responsesin vitro in murine DCs and in vivo in mice. Our findings underline the suitability of AFM to measureliposome rigidity and the importance of this parameter when designing liposomes as a vaccine delivery system. Show less
Benne, N.; Leboux, R.J.T.; Glandrup, M.; Duijn, J. van; Lozano, Vigario F.; Neustrup, M.A.; ... ; Slütter, B. 2019
Regulatory T cells (Tregs) are vital for maintaining a balanced immune response and their dysfunction is often associated with auto-immune disorders. We have previously shown that antigen-loaded... Show moreRegulatory T cells (Tregs) are vital for maintaining a balanced immune response and their dysfunction is often associated with auto-immune disorders. We have previously shown that antigen-loaded anionic liposomes composed of phosphatidylcholine (PC) and phosphatidylglycerol (PG) and cholesterol can induce strong antigen-specific Treg responses. We hypothesized that altering the rigidity of these liposomes while maintaining their size and surface charge would affect their capability of inducing Treg responses. The rigidity of liposomes is affected in part by the length and saturation of carbon chains of the phospholipids in the bilayer, and in part by the presence of cholesterol. We used atomic force microscopy (AFM) to measure the rigidity of anionic OVA323-containing liposomes composed of different types of PC and PG, with or without cholesterol, in a molar ratio of 4:1(:2) distearoyl (DS)PC:DSPG (Young's modulus (YM) 3611 ± 1271 kPa), DSPC:DSPG:CHOL (1498 ± 531 kPa), DSPC:dipalmitoyl (DP)PG:CHOL (1208 ± 538), DPPC:DPPG:CHOL (1195 ± 348 kPa), DSPC:dioleoyl (DO)PG:CHOL (825 ± 307 kPa), DOPC:DOPG:CHOL (911 ± 447 kPa), and DOPC:DOPG (494 ± 365 kPa). Next, we assessed if rigidity affects the association of liposomes to bone marrow-derived dendritic cells (BMDCs) in vitro. Aside from DOPC:DOPG liposomes, we observed a positive correlation between liposomal rigidity and cellular association. Finally, we show that rigidity positively correlates with Treg responses in vitro in murine DCs and in vivo in mice. Our findings underline the suitability of AFM to measure liposome rigidity and the importance of this parameter when designing liposomes as a vaccine delivery system. Show less
This thesis focuses on using liposomes in two different treatment strategies; vaccination (or immunotherapy) and delivery of a small molecule, and in two different disease models; cancer and... Show moreThis thesis focuses on using liposomes in two different treatment strategies; vaccination (or immunotherapy) and delivery of a small molecule, and in two different disease models; cancer and atherosclerosis. For each of these treatment strategies, the liposomal formulation was tailored to obtain the desired therapeutic effect. Chapter 2 reviews some of the most important physicochemical properties (size, shape, and rigidity) that determine the immunological effects of liposomes and other nanoparticles. In chapter 3 we present a detailed study on the effect of the rigidity of anionic liposomes, as measured by atomic force microscopy, on antigen-specific regulatory T-cell (Treg) responses. In chapter 4, we show that our optimized anionic liposomes can induce potent antigen-specific Treg responses, and can be used to delay atherosclerosis progression in a mouse model. Chapter 5 also focuses on liposomal treatment of atherosclerosis, but here targeted liposomes were prepared to successfully deliver a small molecule to foam cells in atherosclerotic plaques. In Chapter 6, we used cationic liposomes in combination with an adjuvant for cancer immunotherapy in mice. Finally, we summarize the overall findings in chapter 7 and discuss perspectives of using liposomes for vaccination and targeted drug delivery. Show less
This study concerns three central aspects of multiculturalism, which are closely related to fundamental debates on citizenship and are described as the principal dimensions of multicultural... Show moreThis study concerns three central aspects of multiculturalism, which are closely related to fundamental debates on citizenship and are described as the principal dimensions of multicultural citizenship. The first of these dimensions relates to national belonging: the recognition that citizens of various ethnic and cultural backgrounds belong to the national group. The second dimension bears upon social equality, on grounds such as ethnic origin, religion, gender and sexual orientation. The third dimension is the most defining aspect of multiculturalism: the recognition of cultural distinctiveness of the various groups in society, which is considered necessary (by proponents of multiculturalism) to achieve citizen equality. This study explores views in Dutch society on these potentially divisive issues. Essentially, this study provides insight into the question to which extent society attributes the quality ‘Dutch’ to Dutch citizens from various ethnic and cultural backgrounds. Show less
The studies described in this thesis focus on the feasibility of using carcinoembryonic antigen (CEA) as a target for immunotherapy of colorectal cancer and on the balance between anti-tumor... Show moreThe studies described in this thesis focus on the feasibility of using carcinoembryonic antigen (CEA) as a target for immunotherapy of colorectal cancer and on the balance between anti-tumor immunity and autoimmune pathology. The potential of CEA as a target antigen for immunotherapy of cancer is conceivably restricted by the fact that CEA is expressed in several abundant and vital tissues, including intestine and stomach, and is even routinely found in the serum of healthy individuals. We demonstrate that the CEA-specific CD4+ T-cell repertoire in CEA-tg mice is severely limited compared to wild-type mice and that this CD4+ T-cell tolerance for CEA was induced by the thymus. In addition we showed that CEA was expressed in medullary thymic epithelial cells (mTEC) in both mice and human beings. The latter suggests that the CEA-specific T-cell repertoire may also be tolerized in people. In further studies we have focussed on possibilities to overcome tolerance of CEA by reconstituting the T-cell repertoire of CEA-tg mice by adoptive transfer of the T-cell repertoire of CEA-immunized wild-type mice into tumor-bearing CEA-tg mice. Adoptive transfer in combination with immune modulation can result in efficient eradication of CEA-positive tumors. However, in order to prevent hazardous CEA-specific autoimmune reactions, the choice of the right immune modulation protocol is critical. Show less
Bovenkamp, F.S. van de; Dijkstra, D.J.; Kooten, C. van; Gelderman, K.A.; Trouw, L.A. 2021
C1q is the recognition molecule of the classical pathway of the complement system. By binding to its targets, such as antigen-bound immunoglobulins or C-reactive protein, C1q contributes to the... Show moreC1q is the recognition molecule of the classical pathway of the complement system. By binding to its targets, such as antigen-bound immunoglobulins or C-reactive protein, C1q contributes to the innate defense against infections. However, C1q also plays several other roles beyond its traditional role in complement activation. Circulating levels of C1q are determined in routine diagnostics as biomarker in several diseases. Decreased C1q levels are present in several autoimmune conditions. The decreased levels reflect the consumption of C1q by complement activation and serves as a biomarker for disease activity. In contrast, increased C1q levels are present in infectious and inflammatory diseases and may serve as a diagnostic biomarker. The increased levels of C1q are still incompletely understood but are suggested to modulate the adaptive immune response as C1q is known to impact on the maturation status of antigen-presenting cells and C1q impacts directly on T cells leading to decreased T-cell activity in high C1q conditions. In this review, we provide a comprehensive overview of the current literature on circulating levels of C1q in health and disease, and discuss how C1q can both protect against infections as well as maintain tolerance by regulating adaptive immunity. Show less
Pregnancy can be seen as an immunologic paradox. Even though the fetus expresses paternally inherited alloantigens it is protected from rejection by a proper regulation of the maternal immune... Show morePregnancy can be seen as an immunologic paradox. Even though the fetus expresses paternally inherited alloantigens it is protected from rejection by a proper regulation of the maternal immune system. With the studies described in this thesis, we want to get more insight in the immunologic mechanisms that play a role in pregnancy. The results of this research can help to identify underlying etiologies in patients with unexplained pregnancy complications, such as recurrent miscarriage. Identifying these causes is important for providing answers and taking away anxiety in these couples, and eventually for the development of effective therapies. Furthermore, elucidating the mechanism leading to survival or rejection of the fetal allograft is not only essential for our understanding of processes leading to normal and abnormal pregnancies, but may also result in important concepts in the field of transplantation and autoimmunity. Show less
Dendritic cells (DCs) are antigen-presenting cells (APCs) which play a key role in the regulation of immune responses. DCs are often referred to as __professional__ APCs, since their primary... Show moreDendritic cells (DCs) are antigen-presenting cells (APCs) which play a key role in the regulation of immune responses. DCs are often referred to as __professional__ APCs, since their primary function is to present antigens from pathogens or malignant cells. Consequently, there is a great deal of interest in how DCs might be exploited as a form of immunotherapy e.g. to induce immunity to cancers. However, DCs are also thought to play an important role in directing regulatory immune responses to innocuous antigens, which are targeted in autoimmune disease or during transplantation. Soluble factors secreted by DCs are crucial mediators in determining this balance between the immunogenic and regulatory arms of the immune system. One such group of factors is cytokines and one family which is gaining increasing attention is the IL-12 family. It is composed of four members; two are immunogenic and their expression has been very well characterised in DCs. The other two are regulatory, but relatively little is known about their regulation and expression in DC populations. In this thesis we aim to give a comprehensive overview of the expression and regulation of IL-12 family members in human DCs, with a particularly emphasis on IL-12, IL-27 and IL-35. Show less
Dixon, K.O.; Kooij, S.W. van der; Vignali, D.A.A.; Kooten, C. van 2015
Caspar Coolhaes was a controversial, tolerant, Spiritualist, Reformed preacher in Leiden in the late sixteenth-century. He has been called "the forerunner of Arminius and the Remonstrants."... Show moreCaspar Coolhaes was a controversial, tolerant, Spiritualist, Reformed preacher in Leiden in the late sixteenth-century. He has been called "the forerunner of Arminius and the Remonstrants." Coolhaes criticised the stricter Calvinist clergy of his day and, in, fact, found all confessions wanting. This dissertation presents an updated biographical sketch, which expands and collates what is known of Coolhaes' life. Then, it defines his ecclesiology from a detailed study of his own works, asking the question: "If Coolhaes could have designed a church for the new Dutch Republic, what would it have looked like?" Specifically, areas which concerned Coolhaes the most are laid out: his views of the relationship between secular and ecclesiastical government, his requirements for good preachers, and his concepts of diversity and Christian freedom in both the visible and invisible church. Show less
In discussing extremism, the key questions are: to whom is a duty owed and what are the limits of intolerance that are to be tolerated? Answering these questions requires examining limits and... Show moreIn discussing extremism, the key questions are: to whom is a duty owed and what are the limits of intolerance that are to be tolerated? Answering these questions requires examining limits and rights; analyzing them in the context of extremism is the ‘core’ of this book. While freedom of speech and freedom of religion are vital to democracies, the freedoms are not unlimited. Where to draw the line between permissible and impermissible is complicated. Religious and secular extremists pose dangers to society and individuals alike; the question I will seek to answer to what extent should, and does, society protect itself against a readily identifiable threat. Whether society chooses to ‘see’ that threat is essential to the discussion; examining why the threat is minimized, at best, and ignored, at worst, is a classic example of history repeating itself. The extremist, in addition to taking the law into his own hands, unequivocally rejects restrictions and limitations imposed by society intended to preserve civil and social order. As discussed in chapter one, I define extremism as a powerful combination of violence and ideology that must necessarily always be “correct” in the mind of its believers. For those believers their ideology is invariably “the truth” and must be defended at all costs. Show less
Holborough-Kerkvliet, M.D.; Kroos, S.; Wetering, R. van de; Toes, R.E.M. 2023
Autoimmune diseases are heterogeneous pathologies characterized by a breakdown of immunological tolerance to self, resulting in a chronic and aberrant immune response to self-antigens. The scope... Show moreAutoimmune diseases are heterogeneous pathologies characterized by a breakdown of immunological tolerance to self, resulting in a chronic and aberrant immune response to self-antigens. The scope and extent of affected tissues can vary greatly per autoimmune disease and can involve multiple organs and tissue types. The patho-genesis of most autoimmune diseases remains unknown but it is widely accepted that a complex interplay be-tween (autoreactive) B and T cells in the context of breached immunological tolerance drives autoimmune pathology. The importance of B cells in autoimmune disease is exemplified by the successful use of B cell tar-geting therapies in the clinic. For example, Rituximab, a depleting anti-CD20 antibody, has shown favorable results in reducing the signs and symptoms of multiple autoimmune diseases, including Rheumatoid Arthritis, Anti-Neutrophil Cytoplasmic Antibody associated vasculitis and Multiple Sclerosis. However, Rituximab depletes the entire B cell repertoire, leaving patients susceptible to (latent) infections. Therefore, multiple ways to target autoreactive cells in an antigen-specific manner are currently under investigation. In this review, we will lay out the current state of antigen-specific B cell inhibiting or depleting therapies in the context of autoimmune diseases. Show less
Multiple Sclerosis is a degenerative disease of the central nervous system (CNS), involving autoimmunity against myelin, resulting in demyelination and paralysis. Antigen-specific immunotherapy may... Show moreMultiple Sclerosis is a degenerative disease of the central nervous system (CNS), involving autoimmunity against myelin, resulting in demyelination and paralysis. Antigen-specific immunotherapy may reduce this pathological autoimmunity, without disturbing normal immune function. This could be achieved by targeting of myelin antigens towards C-type lectin receptors (CLR) that recognize carbohydrate structures and are expressed on immune cells, because targeting of CLR under steady state conditions can suppress immunity in an antigen-specific way. In vitro studies showed that mannosylation of peptides results in internalization via the mannose receptor. In this study we observed, that immunization with mannosylated peptide does not induce disease in EAE, a model for Multiple Sclerosis. Instead, antigen-specific tolerance was induced; CNS inflammation was absent and DTH responses were impaired. Using transfer of TCR transgenic T cells in vivo we visualized that immunization with mannosylated peptide enhanced antigen presentation and induced vigorous expansion of T cells. However, T cells showed reduced blast formation and did not transfer EAE, despite normal production of inflammatory cytokines and chemokines. Lymphocytes accumulated in the lymph node of tolerized mice, which was counteracted by injection of Pertussis Toxin. Established EAE or ongoing DTH responses were ameliorated after mannosylated peptide treatment. In conclusion, mannosylated myelin peptide induced tolerance to EAE due to incomplete differentiation of encephalitogenic T cells and can be used to treat ongoing autoimmunity. Therefore, mannosylated antigens may represent a novel therapeutic approach for antigen-specific modulation of autoreactive T cells in vivo. Show less
Atherosclerosis is the main pathology behind most cardiovascular diseases. It is a chronic inflammatory disease characterized by the formation of lipid-rich plaques in arteries. Atherosclerotic... Show moreAtherosclerosis is the main pathology behind most cardiovascular diseases. It is a chronic inflammatory disease characterized by the formation of lipid-rich plaques in arteries. Atherosclerotic plaques are initiated by the deposition of cholesterol-rich LDL particles in the arterial walls leading to the activation of innate and adaptive immune responses. Current treatments focus on the reduction of LDL blood levels using statins, however the critical components of inflammation and autoimmunity have been mostly ignored as therapeutic targets. The restoration of immune tolerance towards atherosclerosis-relevant antigens can arrest lesion development as shown in pre-clinical models. In this review, we evaluate the clinical development of similar strategies for the treatment of inflammatory and autoimmune diseases like rheumatoid arthritis, type 1 diabetes or multiple sclerosis and analyse the potential of tolerogenic vaccines for atherosclerosis and the challenges that need to be overcome to bring this therapy to patients. Show less
Nagy, N.A.; Lozano Vigario, F.; Sparrius, R.; Capel, T.M.M. van; Ree, R. van; Tas, S.W.; ... ; Jong, E.C. de 2023
