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An optimized method for enumerating CNS derived memory B cells during viral-induced inflammation

Krista D DiSano1, Stephen A Stohlman2, Cornelia C Bergmann2

  • 1Department of Neurosciences NC30, Lerner Research Institute, Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH 44195, United States; School of Biomedical Sciences, Kent State University, Kent, OH 44242, United States.

Journal of Neuroscience Methods
|May 13, 2017
PubMed

Insights

Researchers developed a new assay to study memory B cells (Bmem) in the central nervous system (CNS). This method allows for better understanding of Bmem roles in neuroinflammation and CNS diseases.

Area of Science:

  • Neuroimmunology
  • Cellular immunology

Background:

  • Central nervous system (CNS) inflammation involves diverse B cell subsets.
  • Characterization of memory B cells (Bmem) in the inflamed CNS is limited due to challenges in their in vitro stimulation.
  • Reliable surface markers for murine Bmem are elusive compared to human Bmem.

Purpose of the Study:

  • To develop and optimize an in vitro stimulation assay for converting CNS-derived virus-specific Bmem into antibody-secreting cells (ASC).
  • To characterize the frequency and kinetics of Bmem and ASC in the CNS during viral encephalomyelitis.

Main Methods:

  • A modified limiting dilution in vitro stimulation assay was developed using a viral encephalomyelitis model.
  • Optimization involved varying stimulation duration, Toll-like receptor (TLR) activators, and culture supplements.
  • The optimized protocol used TLR7/8 agonist R848 with feeder cells for 2 days for optimal Bmem to ASC conversion.

Main Results:

  • Flow cytometry markers CD38 and CD73 showed diverse expression on CNS-derived B cell subsets compared to lymphoid tissue.
  • Increasing Bmem frequencies during chronic infection mirrored ASC kinetics.
  • Bmem prevailed in the brain, while ASC were more abundant in the spinal cord during chronic viral infection.

Conclusions:

  • The optimized Bmem assay allows simultaneous enumeration of antigen-specific Bmem and ASC in the CNS.
  • This method enables characterization of temporal changes in B cell populations during neuroinflammation.
  • The findings contribute to understanding B cell dynamics in CNS diseases.
Abstract

Keywords:
Antibody secreting cellsCentral nervous systemMemory B cellsVirus infection

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