Single-Cell Repertoire Analysis Demonstrates that Clonal Expansion Is a Prominent Feature of the B Cell Response in Multiple Sclerosis Cerebrospinal Fluid

Owens, Gregory P.; Ritchie, Alanna M.; Burgoon, Mark P.; Williamson, R. Anthony; Corboy, John R.; Gilden, Donald H.

doi:10.4049/jimmunol.171.5.2725

Cited by 201 publications

(169 citation statements)

References 29 publications

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“…CSF CD19 + CD138 + plasmablast heavy‐ (VH) and light‐chain (VL) variable region sequences were recovered by RT‐PCR and DNA sequencing as described previously 1, 7. Recombinant antibodies were produced in HEK293 cells (Invitrogen, Carlsbad, CA, R620‐07)7 and reactivity tested via immunofluorescence for binding to a permanent cell line expressing M23‐AQP4 8…”

Section: Methodsmentioning

confidence: 99%

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CNS Aquaporin‐4‐specific B cells connect with multiple B‐cell compartments in neuromyelitis optica spectrum disorder

Kowarik

Astling

Gasperi

et al. 2017

Ann Clin Transl Neurol

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ObjectivesNeuromyelitis optica spectrum disorder (NMOSD) is a severe inflammatory disorder of the central nervous system (CNS) targeted against aquaporin‐4 (AQP4). The origin and trafficking of AQP4‐specific B cells in NMOSD remains unknown.MethodsPeripheral (n = 7) and splenic B cells (n = 1) recovered from seven NMOSD patients were sorted into plasmablasts, naïve, memory, and CD27‐IgD‐ double negative (DN) B cells, and variable heavy chain (VH) transcriptome sequences were generated by deep sequencing. Peripheral blood (PB) VH repertoires were compared to the same patient's single‐cell cerebrospinal fluid (CSF) plasmablast (PB) VH transcriptome, CSF immunoglobulin (Ig) proteome, and serum Ig proteome. Recombinant antibodies were generated from paired CSF heavy‐ and light chains and tested for AQP4 reactivity.ResultsApproximately 9% of the CSF VH sequences aligned with PB memory B cells, DN B cells, and plasmablast VH sequences. AQP4‐specific VH sequences were observed in each peripheral B‐cell compartment. Lineage analysis of clonally related VH sequences indicates that CSF AQP4‐specific B cells are closely related to an expanded population of DN B cells that may undergo antigen‐specific B‐cell maturation within the CNS. CSF and serum Ig proteomes overlapped with the VH sequences from each B‐cell compartment; the majority of matches occurring between the PB VH sequences and serum Ig proteome.InterpretationDuring an acute NMOSD relapse, a dynamic exchange of B cells occurs between the periphery and CNS with AQP4‐specific CSF B cells emerging from postgerminal center memory B cells and plasmablasts. Expansion of the PB DN B‐cell compartment may be a potential biomarker of NMOSD activity.

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Section: Methodsmentioning

confidence: 99%

“…Recombinant antibodies were produced in HEK293 cells (Invitrogen, Carlsbad, CA, R620‐07)7 and reactivity tested via immunofluorescence for binding to a permanent cell line expressing M23‐AQP4 8…”

Section: Methodsmentioning

confidence: 99%

CNS Aquaporin‐4‐specific B cells connect with multiple B‐cell compartments in neuromyelitis optica spectrum disorder

Kowarik

Astling

Gasperi

et al. 2017

Ann Clin Transl Neurol

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“…The efficacy of the approach at the single cell level is comparable to other methods which focus on the amplification of less complex rearranged BCR genes. 28,29 T cells isolated from diseased tissues cannot only be characterized for their TCR-b rearrangement but also for the expression of additional genes.…”

Section: Discussionmentioning

confidence: 99%

High throughput analysis of TCR-β rearrangement and gene expression in single T cells

Zhou

Srivastava

Grummel

et al. 2006

Laboratory Investigation

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Analysis of T-cell receptor beta chain (TCR-b) rearrangement is essential to investigate T-cell responses in human autoimmune diseases, infection and cancer. Since the TCR-b locus contains 55 variable (V) region gene segments, multiple assays have been necessary to determine TCR-b rearrangements of individual T cells. We established a seminested rtPCR method for single T-cell analysis with two sets of degenerate primers covering 76 and 24% of the TCR-Vb genes, respectively. The specificity of the approach was validated by screening cDNAs obtained from T-cell clones (TCC) with defined TCR-b rearrangement. We applied the method successfully to profile TCR-b rearrangement of single T cells sorted from body fluids or dissected tissue. Concomitant analysis of other gene transcripts allowed determining phenotype and function of TCR-b-defined single T cells. Our fast, cost-efficient and high throughput approach will facilitate studies on T-cell responses in human diseases.

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“…More so, Owens et al found a significant over-representation of the H and L chains of Ig receptors on B-cells of MS patients. Effectively, this may increase the possibility of Ig recombination and antigen identification [100].…”

Section: Citationmentioning

confidence: 99%

Elucidating the Molecular Basis of Multiple Sclerosis and Understanding the Disease Pathophysiology

Eiman¹

2016

Immunome Res

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Multiple sclerosis (MS) is a complex, multifactorial autoimmune disorder of the central nervous system (CNS) that causes inflammation, demyelination and neurodegeneration. The increased prevalence of this disease in Arabian Gulf Countries (AGCs) has captivated the author. The following is a deliberative review of the disease with respect to its molecular basis. Briefly, it considers disease pathophysiology though the molecular composition of the cell; the genome, epigenome and mitochondrial genome; and relates these factors to environmental, etiological factors, including: vitamin D, UVR, EBV infection, smoking and obesity. All in all, this review aims to explain the reasons underlying the increasing prevalence of MS in AGCs.

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Single-Cell Repertoire Analysis Demonstrates that Clonal Expansion Is a Prominent Feature of the B Cell Response in Multiple Sclerosis Cerebrospinal Fluid

Cited by 201 publications

References 29 publications

CNS Aquaporin‐4‐specific B cells connect with multiple B‐cell compartments in neuromyelitis optica spectrum disorder

CNS Aquaporin‐4‐specific B cells connect with multiple B‐cell compartments in neuromyelitis optica spectrum disorder

High throughput analysis of TCR-β rearrangement and gene expression in single T cells

Elucidating the Molecular Basis of Multiple Sclerosis and Understanding the Disease Pathophysiology

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