In this thesis we identified TCRs for the treatment of B-cell and plasma cell malignancies. In our approach we identified TCRs specific to peptides from B-cell lineage antigens, that are expressed... Show moreIn this thesis we identified TCRs for the treatment of B-cell and plasma cell malignancies. In our approach we identified TCRs specific to peptides from B-cell lineage antigens, that are expressed in B-cell malignancies, presented in various HLA alleles. These epitopes, being self-peptides, are not immunogenic in an HLA-matched setting. To overcome this, we isolated T-cell clones across HLA-barriers using donors lacking the target HLA-alleles. We used peptide-HLA multimers for isolation, followed by comprehensive potency and safety tests to select T cells with precise antigen recognition. The TCRs of selected T-cell clones were sequenced, inserted into retroviral vectors, and introduced into third-party T cells. These T cells were then evaluated for their specificity and ability to target and eliminate B-cell malignancies expressing the antigen in vitro and in in vivo xenograft models. Throughout the thesis we described multiple TCRs with potential clinical applicability for B-cell and plasma cell malignancies. In chapter 2, TCRs are described that target peptides from FCRL5 in HLA-A1, VPREB3 in HLA-A24, and BOB1 in HLA-B35. In chapter 3, TCRs target peptides from the Jchain presented in HLA-A1, -A3, -A11 and -A24. In chapter 4, TCRs target peptides from IgA in HLA-B7 and IgG in HLA-A2. Show less
Background: The new types of mRNA-containing lipid nanoparticle vaccines BNT162b2 and mRNA-1273 and the adenovirus-based vaccine AZD1222 were developed against SARS-CoV-2 and code for its spike (S)... Show moreBackground: The new types of mRNA-containing lipid nanoparticle vaccines BNT162b2 and mRNA-1273 and the adenovirus-based vaccine AZD1222 were developed against SARS-CoV-2 and code for its spike (S) protein. Several studies have investigated short-term antibody (Ab) responses after vaccination. Objective: However, the impact of these new vaccine formats with unclear effects on the long-term Ab response - including isotype, subclass, and their type of Fc glycosylation - is less explored. Methods: Here, we analyzed anti-S Ab responses in blood serum and the saliva of SARS-CoV-2 naive and non-hospitalized pre-infected subjects upon two vaccinations with different mRNA- and adenovirus-based vaccine combinations up to day 270. Results: We show that the initially high mRNA vaccine-induced blood and salivary anti-S IgG levels, particularly IgG1, markedly decrease over time and approach the lower levels induced with the adenovirus-based vaccine. All three vaccines induced, contrary to the short-term anti-S IgG1 response with high sialylation and galactosylation levels, a long-term anti-S IgG1 response that was characterized by low sialylation and galactosylation with the latter being even below the corresponding total IgG1 galactosylation level. Instead, the mRNA, but not the adenovirus-based vaccines induced long-term IgG4 responses - the IgG subclass with inhibitory effector functions. Furthermore, salivary anti-S IgA levels were lower and decreased faster in naive as compared to pre-infected vaccinees. Predictively, age correlated with lower long-term anti-S IgG titers for the mRNA vaccines. Furthermore, higher total IgG1 galactosylation, sialylation, and bisection levels correlated with higher long-term anti-S IgG1 sialylation, galactosylation, and bisection levels, respectively, for all vaccine combinations. Conclusion: In summary, the study suggests a comparable "adjuvant" potential of the newly developed vaccines on the anti-S IgG Fc glycosylation, as reflected in relatively low long-term anti-S IgG1 galactosylation levels generated by the long-lived plasma cell pool, whose induction might be driven by a recently described T-H1-driven B cell response for all three vaccines. Instead, repeated immunization of naive individuals with the mRNA vaccines increased the proportion of the IgG4 subclass over time which might influence the long-term Ab effector functions. Taken together, these data shed light on these novel vaccine formats and might have potential implications for their long-term efficacy. Show less
Stang, M.B. le; Gleeson, P.J.; Daha, M.R.; Monteiro, R.C.; Kooten, C. van 2021
IgA Nephropathy (IgAN) is the main cause of primary glomerulonephritis, globally. This disease is associated with a wide range of clinical presentations, variable prognosis and a spectrum of... Show moreIgA Nephropathy (IgAN) is the main cause of primary glomerulonephritis, globally. This disease is associated with a wide range of clinical presentations, variable prognosis and a spectrum of histological findings. More than fifty years after its first description, this heterogeneity continues to complicate efforts to understand the pathogenesis. Nevertheless, involvement of the complement system in IgAN was identified early on. Dysfunction of the immunoglobulin A (IgA) system, the principal offender in this disease, including modification of isoforms and glycoforms of IgA1, the nature of immune complexes and autoantibodies to galactose deficient IgA1 might all be responsible for complement activation in IgAN. However, the specific mechanisms engaging complement are still under examination. Research in this domain should allow for identification of patients that may benefit from complement-targeted therapy, in the foreseeable future. Show less
Grosserichter-Wagener, C.; Franco-Gallego, A.; Ahmadi, F.; Moncada-Velez, M.; Dalm, V.A.S.H.; Rojas, J.L.; ... ; Zelm, M.C. van 2020
Objective Selective IgA deficiency (sIgAD) is the most common primary immunodeficiency in Western countries. Patients can suffer from recurrent infections and autoimmune diseases because of a... Show moreObjective Selective IgA deficiency (sIgAD) is the most common primary immunodeficiency in Western countries. Patients can suffer from recurrent infections and autoimmune diseases because of a largely unknown aetiology. To increase insights into the pathophysiology of the disease, we studied memory B and T cells and cytokine concentrations in peripheral blood.Methods We analysed 30 sIgAD patients (12 children, 18 adults) through detailed phenotyping of peripheral B-cell, CD8(+) T-cell and CD4(+) T-cell subsets, sequence analysis of IGA and IGG transcripts, in vitro B-cell activation and blood cytokine measurements.Results All patients had significantly decreased numbers of T-cell-dependent (TD; CD27(+)) and T-cell-independent (TI; CD27(-)) IgA memory B cells and increased CD21(low) B-cell numbers. IgM(+)IgD(-) memory B cells were decreased in children and normal in adult patients. IGA and IGG transcripts contained normal SHM levels. In sIgAD children, IGA transcripts more frequently used IGA2 than controls (58.5% vs. 25.1%), but not in adult patients. B-cell activation after in vitro stimulation was normal. However, adult sIgAD patients exhibited increased blood levels of TGF-beta 1, BAFF and APRIL, whereas they had decreased Th1 and Th17 cell numbers.Conclusion Impaired IgA memory formation in sIgAD patients is not due to a B-cell activation defect. Instead, decreased Th1 and Th17 cell numbers and high blood levels of BAFF, APRIL and TGF-beta 1 might reflect disturbed regulation of IgA responses in vivo.These insights into B-cell extrinsic immune defects suggest the need for a broader immunological focus on genomics and functional analyses to unravel the pathogenesis of sIgAD. Show less
Jonker, E.F.F.; Uijlings, M.A.C.; Visser, L.G.; Soonawala, D. 2019
Background: The evidence for recommendations regarding vaccination in solid organ transplant recipients is sparse. There is little data comparing vaccine responses between groups on different... Show moreBackground: The evidence for recommendations regarding vaccination in solid organ transplant recipients is sparse. There is little data comparing vaccine responses between groups on different immunosuppressive drugs. This study was conducted to evaluate the antibody response to Dukoral oral cholera vaccine in renal transplant recipients (RTR). Methods: In a single-center non-randomized controlled clinical trial, healthy volunteers (n = 21) and renal transplant recipients (n = 30) were vaccinated with the oral whole cell/recombinant B subunit cholera vaccine Dukoral (Valneva Inc., Vienna, Austria). The RTR were stratified according to their maintenance immunosuppressive therapy: either prednisone and a calcineurin inhibitor (cyclosporine A or tacrolimus; P/CNI group; n = 15) or prednisone and mycophenolate (P/MMF group; n = 15). All volunteers ingested Dukoral at baseline and at day 14. Serum samples were drawn at day 0 and day 21. The primary outcome was seroconversion, defined as either a 3-fold IgA serum titer increase in anti-cholera toxin B antibodies and/or a 4-fold rise in the serum vibriocidal titer. Results: Follow-up was complete. Seroconversion after vaccination was 57% (standard error, SE 9%) in RTR and 81% (SE 9%) in healthy controls (Relative Risk, RR 0.70; 95% CI 0.48–1.02). When stratified according to maintenance immunosuppression, the seroconversion rate was 67% (SE 12%) in the P/CNI group (RR compared with controls 0.82; 95% CI 0.55–1.25) and 47% (SE 13%) in the P/MMF group (RR compared with controls 0.58; 95% CI 0.32–1.03). Conclusion: Adverse events were mild to moderate and transient. The response to Dukoral was weaker and the seroconversion rate was lower in renal transplant recipients than in healthy controls. In particular, those using mycophenolate had a poor response. Nevertheless, more than half of the transplant recipients seroconverted. Therefore oral vaccines should not be discarded as a potential tool for protection of solid organ transplant recipients. Show less
The worldwide resurgence of whooping cough (pertussis), even in highly vaccinated populations, demands improved pertussis vaccines. In this thesis a systems vaccinology approach is applied to... Show moreThe worldwide resurgence of whooping cough (pertussis), even in highly vaccinated populations, demands improved pertussis vaccines. In this thesis a systems vaccinology approach is applied to deepen knowledge of the immune responses evoked by different pertussis vaccines and compare this with a Bordetella pertussis infection since the latter induces robust protection. Infection-induced responses in mice conferred sterilizing protection that is caused by systemic immunity but more importantly by mucosal IgA, T-helper (Th)1/Th17 responses, and ‘trained’ innate immune cells in the lungs. An experimental outer membrane vesicle vaccine (omvPV) was compared with the two licensed vaccines, acellular vaccine (aPV), whole-cell vaccine (wPV) as well as a B. pertussis infection. OmvPV evoked a different immunoproteomic profile with respect to antibody levels, antigen specificity, and subclass distribution. Furthermore, omvPV confers equal protection in mice as wPV, but with a lower inflammatory response. In this thesis it is also shown that the immunization route is critical. Although subcutaneous omvPV immunization is effective, pulmonary administration lead to superior protection, comparable to infection-induced immunity and included hallmarks of protection such as pulmonary Th17 cells and mucosal IgA. The molecular and cellular signatures described in this thesis may have an important contribution to enhanced pertussis immunity. Show less
Belay, M.; Legesse, M.; Mihret, A.; Ottenhoff, T.H.M.; Franken, K.L.; Bjune, G.; Abebe, F. 2016
Markers for longevity, which reflect the health condition and predict healthspan are extremely scarce. Such markers are, however, valuable in aging research. Protein N-glycosylation is an important... Show moreMarkers for longevity, which reflect the health condition and predict healthspan are extremely scarce. Such markers are, however, valuable in aging research. Protein N-glycosylation is an important post-translational modification, which has been shown to be altered in several pathological conditions. N-glycosylation patters therefore reflect an individual__s health state, and have therefore the potential to become candidate biomarkers for healthspan. To investigate familial longevity, large sample cohorts are necessary, and since no high-throughput analysis methods were available to facilitate Large scale N-glycan analysis, several methods, comprising HPLC-FL, CGE-LIF and MALDI-MS have been developed. These methods were applied to samples from the Leiden Longevity Study. Several changes in N-glycosylation patterns associated with familial longevity were observed in this thesis. As the predictive value of these markers is rather low, it has to be concluded that multiple markers are needed for the prediction of complex phenotypes like familial longevity. It is anticipated that __ apart from allowing more detailed insight in the processes underlying aging and longevity- the developed methodology will as well be of considerable value to other fields of biomedical research. Show less
IgA nephropathy (IgAN) is the most common form of glomerulonephritis worldwide. The hallmark of the disease is depositions of polymeric IgA1 in the mesangium of the glomeuli. These depositions will... Show moreIgA nephropathy (IgAN) is the most common form of glomerulonephritis worldwide. The hallmark of the disease is depositions of polymeric IgA1 in the mesangium of the glomeuli. These depositions will lead to inflammation in the kidneys and eventually to deterioration of renal function. The pathogenesis of IgAN is not clear, but it is generally accepted that disturbances in the immune system of IgAN patients are responsible for this disease. In the current thesis we have investigated the immune response of IgAN patients in comparison with control persons. We have shown that IgAN patients have a hampered primary IgA immune response upon mucosal vaccination with a neoantigen, whereas a systemic vaccination with a neoantigen resulted in a similar immune response in both groups. We hypothesized that dendritic cells (DC), as professional antigen presenting cells could have an impaired function , or that less DC are present in the nasal mucosa. We were able to show that the number of DC present in the nasal mucosa of IgAN patients was not reduced as compared with controls. Using an in vitro model we studied the function of DC in the primary immune response and showed that DC of IgAN patients induced less IgA production in na_ve B cells than DC of control persons. Furthermore we studied the size distribution of the antigen specific IgA molecules in IgAN patients. In summary we showed that patients with IgAN have an impaired IgA production upon mucosal vaccination with a neoantigen and that at least part of this IgA hypo response is due to an impaired capacity of DC to induce IgA production, whereas the number of mucosal DC in IgAN patients is not reduced. Show less
Primary IgA nephropathy (IgAN) is the most common form of primary glomerulonephritis world wide and leads to end stage renal disease in 30-50% of patients. The hallmark of IgAN is the deposition... Show morePrimary IgA nephropathy (IgAN) is the most common form of primary glomerulonephritis world wide and leads to end stage renal disease in 30-50% of patients. The hallmark of IgAN is the deposition of IgA1 in the mesangial area of the kidney. Since the inflammatory response which leads to progressive renal disease, is triggered and sustained by the deposition of IgA in the renal mesangium, it is important to determine by which mechanisms binding to mesangial cells (MC) occurs. Most likely both the intrinsic renal features, as well as circulating factors, such as structural alterations in serum IgA molecules are thought to be involved in the pathogenesis of IgAN. In this thesis we concentrated on two aspects. First identify and further characterize the IgA binding receptors potentially playing a role in IgA nephropathy, including FcRI/CD89, Fc/µR. Furthermore, we characterized the different molecular forms of IgA with special attention to the specific glycosylation patterns. Finally, we found a clear deposition of SIgA in the glomeruli of IgA nephropathy patients. Altogether, the data presented in this thesis support a role for SIgA in the pathogenesis of a subpopulation of IgAN patients. Show less
