Monash University
Immunology and Pathology

Author Of 2 Presentations

Genetics and Epigenetics Poster Presentation

P0528 - T cell composition and polygenic multiple sclerosis risk: a population-based study in children (ID 943)

Speakers
Presentation Number
P0528
Presentation Topic
Genetics and Epigenetics

Abstract

Background

Multiple sclerosis (MS) is associated with extensive immunological alterations in adult patients. MS patients show changes in T cell composition, including increased CD4+/CD8+ ratios. However, it is unclear to which extent these changes in T cell composition are influenced by genetic risk for MS, and how this may precede a possible disease onset.

Objectives

In the current study we investigate the association between polygenic risk scores (PRSs) for MS and T cell subsets in a large population-based pediatric sample, to provide new understanding about the link between genetic risk for MS and disease pathophysiology.

Methods

We included participants from the population-based Generation R study who had genetic- and immunological data available. Children were sampled for immunological data around the age of 6 years (IQR: 5.9-6.2). Linear regression analyses were used to analyze the impact of MS-PRSs on absolute T cell numbers (n=1,261) and CD4+ and CD8+ T cell fractions (n=675) adjusted for important child- (age and sex) and environmental confounding factors (serum vitamin D levels and cytomegalovirus positivity).

Results

The MS-PRS showed a negative correlation with CD8+ T cell frequencies (β=-0.05, SE=0.015, ΔR2=0.020, p=2.88 × 10-4), which resulted in a positive association with CD4+/CD8+ ratios (β=0.07, SE=0.011, ΔR2=0.054, p=9.20 × 10-10). Interestingly, the latter was driven by 2 out of 196 genome-wide significant MS risk variants. Both from within the HLA class II region, risk variants rs3135388 and rs9271366 were positively associated with the CD4+/CD8+ ratio. No association was found with absolute total T cell numbers.

Conclusions

This study shows that higher genetic risk for MS is associated with T cell alterations at an early age. Our results show a possibility that MS genetics affect the T cell composition during childhood, which may contribute to increased risk of MS disease later in life.

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Pathogenesis – Immunology Poster Presentation

P0973 - Impact of Epstein-Barr virus infection on CXCR3+ B-cell development: lessons learned from immunotherapies in MS (ID 751)

Abstract

Background

Epstein-Barr virus (EBV) infection of B cells is strongly associated with the onset of several chronic inflammatory and autoimmune diseases such as multiple sclerosis (MS). In MS, a subset of memory B cells infiltrates the central nervous system (CNS) and further differentiates into antibody-secreting cells to mediate local pathology.

Objectives

Here, we aimed to decipher whether and how EBV impacts the development of such CNS-homing memory B cells in MS patients.

Methods

Chemokine receptor profiles were analyzed for ex vivo B cells in single-cell suspensions obtained from paired CNS compartments of 10 MS patients (NBB) using multicolor flow cytometry. The CNS infiltration capacity of memory B-cell subsets was confirmed using confluent brain endothelial monolayers. Similar analyses were performed for distinct memory subsets in the blood from 16 untreated, 32 natalizumab-treated and 9 bone-marrow transplant (BMT)-treated MS patients as well as matched healthy controls. An IL-21-/CD40L-based germinal center-like culture system was used to compare naive and memory B-cell differentiation. EBV copy numbers were determined in total and memory B cells using a multiplex BALF5-related qPCR assay.

Results

CXCR3-expressing B cells were selectively enriched in paired CSF, meningeal and brain tissues versus blood of MS patients. Treatment of patients with natalizumab resulted in an accumulation of CXCR3high IgG+ B cells in the blood, corresponding to their increased ability to cross CNS barriers in vitro. Naive B cells developed into plasmablasts under IFN-γ-mediated germinal center-like conditions and required additional TLR9 signaling for differentiation into CXCR3+ switched memory cells. In 3-7 months post- vs pre-BMT blood samples, EBV DNA load was elevated and positively correlated to the frequency of CXCR3+, and not CXCR4+ or CXCR5+, switched memory B cells. High EBV load in memory B cells from natalizumab-treated MS patients corresponded to an increased potential to develop into anti-EBNA1 IgG-producing CXCR3+ plasma cells (CD38++CD27++CD138+) during IFN-γ-mediated germinal center-like cultures.

Conclusions

This study implicates that persistent viral infections such as EBV potentiate brain-homing and antibody-producing CXCR3(T-bet)+ B cells in MS patients. These findings may mechanistically link EBV infection to anti-EBNA1 IgG production as being a predictor of disease onset and to the massive B-cell influx into the CNS after natalizumab discontinuation in MS.

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