May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Single–cell Analysis of the B–cell Repertoire in Optic Neuritis Patients Demonstrates Clonal Expansion
Author Affiliations & Notes
  • J.L. Bennett
    Neurology, U of Colorado Health Sci Ctr, Denver, CO
  • G.P. Owens
    Neurology, U of Colorado Health Sci Ctr, Denver, CO
  • D.H. Gilden
    Neurology, U of Colorado Health Sci Ctr, Denver, CO
  • K. Haubold
    Neurology, U of Colorado Health Sci Ctr, Denver, CO
  • Footnotes
    Commercial Relationships  J.L. Bennett, None; G.P. Owens, None; D.H. Gilden, None; K. Haubold, None.
  • Footnotes
    Support  none
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 1602. doi:
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      J.L. Bennett, G.P. Owens, D.H. Gilden, K. Haubold; Single–cell Analysis of the B–cell Repertoire in Optic Neuritis Patients Demonstrates Clonal Expansion . Invest. Ophthalmol. Vis. Sci. 2004;45(13):1602.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose: Increased amount of intrathecally produced IgG and oligoclonal bands (OCBs) are found at high frequency in optic neuritis (ON) cerebrospinal fluid (CSF). In infectious disorders of the CNS, OCBs are commonly directed against the infectious agent that causes disease. Recent molecular studies of immunoglobulins expressed in MS plaques, MS CSF, and measles–infected brain have revealed features of a targeted B cell response to antigenic stimulation. To determine whether the B cell repertoire in CSF from ON patients contains clonally expanded populations, we analyzed the heavy– and light–chain sequences of single B and plasma cells from ON CSF. Methods: Fluorescent activated cell sorting was used to isolate CD19+ and CD138+ B cells from the CSF of patients with ON or viral meningitis. Single B cells were lysed, and heavy (VH) and light (VL) chain variable regions amplified by RT–PCR. Sequence analysis was performed on amplified VH and VL sequences to determine the rearranged germline segment, J segment, and degree of homology among individual B cells. Results: Only two clonally related B cell populations were detected among the 53 IgG and 46 IgM VH sequences analyzed from viral meningitis CSF indicating a polyclonal response. In contrast, clonally expanded B cells and plasma cells were detected to varying degrees in the IgG repertoire of each of the ON donors: clonally expanded populations comprised up to 75% of the VH and 77% of the VL sequences in isolated CD19+ and CD138+ B cells. Analysis of the IgM B cell population yielded only a single clone in one patient. Conclusions: The features of the IgG repertoire obtained from the analysis of single ON B cells is consistent with a targeted response to antigen. Identification of overrepresented sequences will allow us to create recombinant antibodies that mirror those produced in vivo in ON patients. Such antibodies offer the potential to discover the relevant antigens in ON and MS.

Keywords: neuro–ophthalmology: optic nerve • inflammation • pathobiology 
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