Rogier Q. Hintzen, MD, PhD

Hintzen-51x53px Professor and Head of MS Center ErasMS,
 Department of Immunology

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 Neuroimmunology Brain Research Group

Multiple sclerosis (MS) is a chronic inflammatory disorder of the central nervous system (CNS), which is characterized by ongoing autoimmunity and myelin breakdown, eventually leading to neuronal deficits. Although the cause of MS is currently unknown, the central dogma is that certain autoreactive CD4+ T helper and B-cell populations interact and are activated in the periphery, transmigrate into the CNS and recognize local antigens to mediate pathology. The prime focus of our workgroup is to uncover subsets, mechanisms and antigen specificity of human T and B cells contributing to MS disease activity.

Our research covers the following topics: 1) control of IFN-γ-producing Th17 subsets (Th17.1) by gender hormones, vitamin D and glucocorticoids, 2) IFN-γ-mediated induction, effector functions and specificity of T-bet+ B cells, 3) B-cell antigen processing and presentation to Th17 subsets, 4) the impact of Epstein-Barr virus (EBV) on B- and T-cell immunity, and 5) antigenic targets of T- and B-cell populations from the CNS. These topics are strongly interrelated and have one ultimate goal: more in-depth understanding of T- and B-cell interaction as main driving force of MS.

1) Control of MS-associated Th17.1 cells by sex hormones, vitamin D and glucocorticoids
Gender-/pregnancy-related steroids, vitamin D as well as glucocorticoids are members of the nuclear receptor ligand (NRL) family, which likely closely interact to reduce early MS disease activity, especially  in women. Ongoing work reveals that glucocorticoid-resistant Th1-like Th17 (Th17.1) cells contribute to early MS and are controlled by NRL interaction. Our aim is to compare the effects of NRL interaction on these highly pathogenic Th17.1 cells between men and women with MS, and how this can be exploited to reduce disease activity in distinct clinical subgroups. Synergistic effects of vitamin D, female steroids and glucocorticoids on Th17.1 activation will be assessed in vitro using advanced autologous co-cultures and transmigration assays. For these purposes, we will use PBMC from untreated, vitamin D-treated and pregnant MS patients.

2) Development, effector functions and specificity of T-bet+ B cells as key drivers of MS
Picture2-250x240-newPeripheral B-cell tolerance defects and ectopic germinal center formation are key pathogenic features of MS, which are associated with autoreactive T-bet+ B cells in mice. In our human B-cell studies, we found that blood CXCR3+ B cells preferentially recruit to CSF, meninges and brain tissue of MS patients. CXCR3+ B cells from MS patients also showed more T-bet upregulation during IL-21/CD40L-based cultures, which was required for IFN-γ- and TLR9-induced switching to IgG1. In this part of research, we are interested in a) the sensitivity of naive B cells for IFN--mediated differentiation into CXCR3+T-bet+ memory cells, and b) the reactivity of CXCR3+T-bet+ B cells to candidate MS autoantigens, including EBV and myelin-derived antigens. Experiments will be performed in the context of a newly discovered MS risk SNP in IFNGR2 (IFN-γ receptor β-chain).

3) Regulation and impact of BCR-mediated antigen processing and presentation in MS
In this part of our work, we are particularly interested in genes identified by genome-wide association studies that control the biology of HLA class II (HLA-II), the major genetic factor in MS. The success of B-cell depletion therapies in MS points to B cells as potent antigen-presenting cells. We will use RNAi and CRISPR-Cas9-based tools to explore gene function in relation to the HLA-II pathway using qPCR, flow cytometry, cell sorting and high-resolution microscopy.  Furthermore, we will perform antigen-specific B-and T-cell cultures to compare Th17 skewing between MS risk and non-risk carriers. Our aim here is to identify disease-related genes that are selectively regulated to trigger B-cell antigen presentation in MS.

4) Contributions of EBV to B-cell differentiation, survival and interaction with T cells in MS
Picture1-250x240-newEBV is a major MS risk factor that establishes lifelong latency in B cells. We argue that EBV is a key factor for breaking immune tolerance, causing activation of pathogenic B- and T-cell subsets in MS. Using high-throughput molecular arrays (expertise by groups of Dr. van Zelm and Dr. van der Burg), we will focus on the distribution of EBV DNA in B-cell subsets and how this influences their development and survival in MS. Additionally, we will add highly immunogenic EBNA1 peptides to IL-21/CD40L-based cultures with B cells from MS patients and controls to assess differences in EBV specificity using FACS and ELISA. Ex vivo and in vitro EBV-specific T-cell populations will be assessed using EBNA1-specific class I tetramers.


5) Phenotype and antigen specificity of B- and T-cell populations in the CNS of MS patients
First, we will study the effector phenotypes of B  and T cells derived from paired blood, CSF, meninges and brain tissues of MS patients and non-demented controls (Netherlands Brain Bank) using multicolor flow cytometry. The presence of distinct IgM+ and IgG+ memory B cells, as well as plasmablast/plasma cell subsets will be compared. To increase our knowledge on the local antigens recognized, we will use a novel BV-independent method (Prof. Spits,  Amsterdam) to immortalize intrathecal B cells from patients with MS, NMO and ADEM. Using this technology, the original function and specificity of B-cell clones are maintained. This gives us the unique opportunity to assess Ig reactivity against CNS autoantigens and viruses using ELISA and cell-based assays. Local CD4+ and CD8+ T cells will be separated and assessed for the production of IL-17, IFN-γ, GM-CSF and TNF-α. Ultimately, we want to sequence the TCR from MS lesion-specific T cells on a single-cell level.

 Group members

Rogier Hintzen, MD, PhD, Group leader
Marvin van Luijn, PhD, Supervisor
Liza Rijvers, PhD student
Jamie van Langelaar, PhD student
Steven Koetzier, PhD student
Malou Janssen, PhD student
Annet Wierenga-Wolf, Research Assistant
Marie-José Melief, Research Assistant 

 Selected Publications

(See for all publications Rogier Hintzen and Marvin van Luijn in Pubmed)

van Langelaar J, van der Vuurst de Vries RM, Janssen M, Wierenga-Wolf AF, Spilt IM, Siepman TA, Dankers W, Verjans GM, de Vries HE, Lubberts E, Hintzen RQ and van Luijn MM.
T helper 17.1 cells associate with multiple sclerosis disease activity: perspectives for early intervention.
Brain 2018 (in press).

van Nierop GP, van Luijn MM, Michels SS, Melief MJ, Janssen M, Langerak AW, Ouwendijk WJD, Hintzen RQ, Verjans GMGM.
Phenotypic and functional characterization of T cells in white matter lesions of multiple sclerosis patients.
Acta Neuropathol 2017; 134(3):383-401.

Kreft KL, Van Nierop GP, Scherbeijn SMJ, Janssen M, Verjans GMGM, Hintzen RQ.
Elevated EBNA-1 IgG in MS is associated with genetic MS risk variants.
Neurol Neuroimmunol Neuroinflamm 2017; 12;4(6):e406

van der Vuurst de Vries RM, Mescheriakova JY, Runia TF, Jafari N, Siepman TA, Hintzen RQ.
Soluble CD27 Levels in cerebrospinal fluid as a prognostic biomarker in clinically isolated syndrome.
JAMA Neurol 2017; 74(3):286-292.

van Luijn MM, van Meurs M, Stoop MP, Verbraak E, Wierenga-Wolf AF, Melief MJ, Kreft KL, Verdijk RM, 't Hart BA, Luider TM, Laman JD, Hintzen RQ.
Elevated expression of the cerebrospinal fluid disease markers chromogranin A and clusterin in astrocytes of multiple sclerosis white matter lesions.
J Neuropathol Exp Neurol 2016; 75(1):86-98.

Ghanbari M, Darweesh SK, de Looper HW, van Luijn MM, Hofman A, Ikram MA, Franco OH, Erkeland SJ, Dehghan A.
Genetic Variants in MicroRNAs and Their Binding Sites Are Associated with the Risk of Parkinson Disease.
Hum Mutat 2016; 37(3):292-300.

van Luijn MM, Kreft KL, Jongsma ML, Mes SW, Wierenga-Wolf AF, van Meurs M, Melief MJ, der Kant RV, Janssen L, Janssen H, Tan R, Priatel JJ, Neefjes J, Laman JD, Hintzen RQ.
Multiple sclerosis-associated CLEC16A controls HLA class II expression via late endosome biogenesis.
Brain 2015; 138(Pt 6):1531-47.

van Luijn MM, van den Ancker W, van Ham SM, van de Loosdrecht AA.
Class II-associated invariant chain peptide as predictive immune marker in minimal residual disease in acute myeloid leukemia.
Oncoimmunology 2015; 3(12):e941737.

Kreft KL, van Meurs M, Wierenga-Wolf AF, Melief MJ, van Strien ME, Hol EM, Oostra BA, Laman JD, Hintzen RQ.
Abundant kif21b is associated with accelerated progression in neurodegenerative diseases.
Acta Neuropathol Commun 2014; 3;2:144. 

Stern JN, Yaari G, Vander Heiden JA, Church G, Donahue WF, Hintzen RQ, Huttner AJ, Laman JD, Nagra RM, Nylander A, Pitt D, Ramanan S, Siddiqui BA, Vigneault F, Kleinstein SH, Hafler DA, O'Connor KC.
B cells populating the multiple sclerosis brain mature in the draining cervical lymph nodes.
Sci Transl Med 2014; 6;6(248):248ra107.

Patsopoulos NA, Barcellos LF, Hintzen RQ, Schaefer C, van Duijn CM, Noble JA, Raj T; IMSGC; ANZgene, Gourraud PA, Stranger BE, Oksenberg J, Olsson T, Taylor BV, Sawcer S, Hafler DA, Carrington M, De Jager PL, de Bakker PI.
Fine-mapping the genetic association of the major histocompatibility complex in multiple sclerosis: HLA and non-HLA effects.
PLoS Genet 2013; 9(11):e1003926.

International Multiple Sclerosis Genetics Consortium (IMSGC), Beecham AH, Patsopoulos NA, Xifara DK, Davis MF, Kemppinen A, Cotsapas C, Shah TS, Spencer C, Booth D, Goris A, Oturai A, Saarela J, Fontaine B, Hemmer B, Martin C, Zipp F, D'Alfonso S, Martinelli-Boneschi F, Taylor B, Harbo HF, Kockum I, Hillert J, Olsson T, Ban M, Oksenberg JR, Hintzen RQ et al.
Analysis of immune-related loci identifies 48 new susceptibility variants for multiple sclerosis.
Nat Genet 2013; 45(11):1353-60. 

van Luijn MM, van de Loosdrecht AA, Lampen MH, van Veelen PA, Zevenbergen A, Kester MG, de Ru AH, Ossenkoppele GJ, van Hall T, van Ham SM.
Promiscuous binding of invariant chain-derived CLIP peptide to distinct HLA-I molecules revealed in leukemic cells.
PLoS One 2012; 7(4):e34649.

Kreft KL, Verbraak E, Wierenga-Wolf AF, van Meurs M, Oostra BA, Laman JD, Hintzen RQ.
Decreased systemic IL-7 and soluble IL-7Rα in multiple sclerosis patients.
Genes Immun 2012; 13(7):587-92. 

Kreft KL, Verbraak E, Wierenga-Wolf AF, van Meurs M, Oostra BA, Laman JD, Hintzen RQ.
The IL-7Rα pathway is quantitatively and functionally altered in CD8 T cells in multiple sclerosis.
J Immunol 2012; 188(4):1874-83. 

van Luijn MM, van den Ancker W, Chamuleau ME, Zevenbergen A, Westers TM, Ossenkoppele GJ, van Ham SM, van de Loosdrecht AA.
Absence of class II-associated invariant chain peptide on leukemic blasts of patients promotes activation of autologous leukemia-reactive CD4+ T cells.
Cancer Res 2011; 71(7):2507-17.

van Luijn MM, Westers TM, Chamuleau ME, van Ham SM, Ossenkoppele GJ, van de Loosdrecht AA.
Class II-associated invariant chain peptide expression represents a novel parameter for flow cytometric detection of acute promyelocytic leukemia.
Am J Pathol 2011; 179(5):2157-61.