PLoS ONE (Jan 2013)

Transient B-cell depletion with anti-CD20 in combination with proinsulin DNA vaccine or oral insulin: immunologic effects and efficacy in NOD mice.

  • Ghanashyam Sarikonda,
  • Sowbarnika Sachithanantham,
  • Yulia Manenkova,
  • Tinalyn Kupfer,
  • Amanda Posgai,
  • Clive Wasserfall,
  • Philip Bernstein,
  • Laura Straub,
  • Philippe P Pagni,
  • Darius Schneider,
  • Teresa Rodriguez Calvo,
  • Marilyne Coulombe,
  • Kevan Herold,
  • Ronald G Gill,
  • Mark Atkinson,
  • Gerald Nepom,
  • Mario Ehlers,
  • Teodora Staeva,
  • Hideki Garren,
  • Lawrence Steinman,
  • Andrew C Chan,
  • Matthias von Herrath

DOI
https://doi.org/10.1371/journal.pone.0054712
Journal volume & issue
Vol. 8, no. 2
p. e54712

Abstract

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A recent type 1 diabetes (T1D) clinical trial of rituximab (a B cell-depleting anti-CD20 antibody) achieved some therapeutic benefit in preserving C-peptide for a period of approximately nine months in patients with recently diagnosed diabetes. Our previous data in the NOD mouse demonstrated that co-administration of antigen (insulin) with anti-CD3 antibody (a T cell-directed immunomodulator) offers better protection than either entity alone, indicating that novel combination therapies that include a T1D-related autoantigen are possible. To accelerate the identification and development of novel combination therapies that can be advanced into the clinic, we have evaluated the combination of a mouse anti-CD20 antibody with either oral insulin or a proinsulin-expressing DNA vaccine. Anti-CD20 alone, given once or on 4 consecutive days, produced transient B cell depletion but did not prevent or reverse T1D in the NOD mouse. Oral insulin alone (twice weekly for 6 weeks) was also ineffective, while proinsulin DNA (weekly for up to 12 weeks) showed a trend toward modest efficacy. Combination of anti-CD20 with oral insulin was ineffective in reversing diabetes in NOD mice whose glycemia was controlled with SC insulin pellets; these experiments were performed in three independent labs. Combination of anti-CD20 with proinsulin DNA was also ineffective in diabetes reversal, but did show modest efficacy in diabetes prevention (p = 0.04). In the prevention studies, anti-CD20 plus proinsulin resulted in modest increases in Tregs in pancreatic lymph nodes and elevated levels of proinsulin-specific CD4+ T-cells that produced IL-4. Thus, combination therapy with anti-CD20 and either oral insulin or proinsulin does not protect hyperglycemic NOD mice, but the combination with proinsulin offers limited efficacy in T1D prevention, potentially by augmentation of proinsulin-specific IL-4 production.