OP0040 HIPPOCAMPAL IMMUNE CELL TRAFFICKING AND A MYELOID PREDOMINANT INFLAMMATORY RESPONSE WITH ENHANCED ANTIGEN PRESENTATION AND DECREASED LEVELS OF NEUROTRANSMITTERS UNDERLY THE NEUROPSYCHIATRIC PHENOTYPE OF THE NZW/NZB MURINE LUPUS MODEL
Authors: Nikolopoulos D. , Manolakou T. , Filia A. , Nakos-Bimpos M. , Polissidis A. , Boumpas D. 
Published: May 2021

Background:

Neuropsychiatric events are common in patients with systemic lupus erythematosus (SLE), yet the underlying pathogenesis remains ill-defined, as the access to brain tissue is limited. We have previously shown that NZW/NZB F1 murine lupus model recapitulates the neuropsychiatric lupus phenotype including depressive-like behavior, increased rates of anxiety, cognitive dysfunction and motor disturbances, both at pre-nephritic and nephritic stages of the disease.

Objectives:

To dissect specific regions in the brain, which account for this phenotype and elucidate inflammatory and non-inflammatory mechanisms involved.

Methods:

Four distinct brain regions (hippocampus, amygdala, striatum and pre-frontal cortex) were dissected from brains of female C57BL/6 (WT) and NZW/NZB F1 mice at the age of 3 months (pre-nephritic) and 6 months (nephritic stage) (n=5-8/condition/experiment). Since most of the behavioral phenotype corresponds to the hippocampus, we first examined in depth the hippocampal pathology by bulk RNA sequencing, measurements of neurotransmitters levels via high-performance liquid chromatography (HPLC) and by immunophenotyping via flow cytometry analyses. For comparisons, statistical significance was indicated as a two-sided P<0.05.

Results:

Transcriptomic analysis revealed aberrant immune mediated response in the hippocampus of 6 month-old lupus mice compared to WT. Specifically, inflammatory pathways including both innate and adaptive immune responses, increased cytokine production, increased antigen presentation and immune cell trafficking, along with increased apoptosis and decreased cell proliferation suggest that immune aberrancies may lead to neuronal damage. These aberrancies were present in mice at 3 month-old, yet were progressed with time being more prominent at 6 month of age in lupus hippocampus. The RNA sequencing date were validated by immunophenotyping on lupus hippocampus demonstrating increased reactive GFAP+ astrocytes both at 3 and 6-month old mice. Activated IBA1+ microglia and CD11b+CD45hi CNS myeloid cells were increased only at 6 months of age. Furthermore, increased immune cell infiltration from the periphery including lymphocytes (CD45+CD11b-) mainly T cells (CD4+/CD8+) and monocytes (CD45+CD11b+Ly6G-Ly6C+), was evident only in 6 month-old lupus hippocampus compared to WT. Importantly, microglia cells in lupus hippocampus at 6 but not at 3 month of age, exhibited increased expression of antigen presenting markers including CD80, CD86 and MHC-II indicating that microglia cells may carry out the antigen presentation process seen in transcriptomic data. Low levels of serotonin and noradrenaline were observed at both 3 and 6 months of age in lupus mice; these aberrancies were mainly attributed to decreased serotonin synthesis as evidenced by intact serotonin metabolism (no differences were observed at its metabolite: 5-hydroxyindoleacetic acid). Analysis of the remaining regions of the brain combined with studies of metabolic activities of various brain regions by PET-CT scanning is in progress.

Conclusion:

Immune cell trafficking from the periphery combined with marked inflammatory response in the hippocampus underlie the neuropsychiatric phenotype in NZW/B murine lupus. Our data indicate increased expression of activated myeloid cells -including microglia- in the hippocampus of lupus mice culminating in increased antigen presentation and decreased neurotransmitter levels.

References:

[1]Nikolopoulos, D., et al. “THU0223 THE NEUROPSYCHIATRIC PHENOTYPE OF NZB/W LUPUS-PRONE MOUSE MODEL AT PRE-NEPHRITIC AND NEPHRITIC STAGES OF THE DISEASE: MURINE MODEL RECAPITULATES HUMAN DISEASE.” (2020): 334-335.

Acknowledgements:

This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No 742390)

Disclosure of Interests:

None declared