CD33 recruitment inhibits IgE-mediated anaphylaxis and desensitizes mast cells to allergen
Shiteng Duan, Cynthia J. Koziol-White, William F. Jester Jr., Scott A. Smith, Corwin M. Nycholat, Matthew S. Macauley, Reynold A. Panettieri Jr., James C. Paulson
Shiteng Duan, Cynthia J. Koziol-White, William F. Jester Jr., Scott A. Smith, Corwin M. Nycholat, Matthew S. Macauley, Reynold A. Panettieri Jr., James C. Paulson
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Research Article Immunology

CD33 recruitment inhibits IgE-mediated anaphylaxis and desensitizes mast cells to allergen

Abstract

Allergen immunotherapy for patients with allergies begins with weekly escalating doses of allergen under medical supervision to monitor and treat IgE mast cell–mediated anaphylaxis. There is currently no treatment to safely desensitize mast cells to enable robust allergen immunotherapy with therapeutic levels of allergen. Here, we demonstrated that liposomal nanoparticles bearing an allergen and a high-affinity glycan ligand of the inhibitory receptor CD33 profoundly suppressed IgE-mediated activation of mast cells, prevented anaphylaxis in Tg mice with mast cells expressing human CD33, and desensitized mice to subsequent allergen challenge for several days. We showed that high levels of CD33 were consistently expressed on human skin mast cells and that the antigenic liposomes with CD33 ligand prevented IgE-mediated bronchoconstriction in slices of human lung. The results demonstrated the potential of exploiting CD33 to desensitize mast cells to provide a therapeutic window for administering allergen immunotherapy without triggering anaphylaxis.

Authors

Shiteng Duan, Cynthia J. Koziol-White, William F. Jester Jr., Scott A. Smith, Corwin M. Nycholat, Matthew S. Macauley, Reynold A. Panettieri Jr., James C. Paulson

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Figure 2

Tg mice with mast cells expressing functional human CD33.

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Tg mice with mast cells expressing functional human CD33. (A) Flow cytom...
(A) Flow cytometric analysis of GFP expression on representative peritoneal mast cells harvested from C57BL/6J, control-Tg (Mcpt5-Cre–Rosa26-Stopfl/fl-CD33+), and CD33-Tg (Mcpt5-Cre+/–Rosa26-Stopfl/fl-CD33+) mice. Mast cells were defined as PI–CD45+c-Kit+. Baseline GFP signal was determined by mast cells from C57BL/6J mice. (B) Quantification of the percentage of GFP+ peritoneal mast cells from mice of the 3 genotypes. Tg mice bearing 1 or 2 copies of CD33 were used. Both male and female mice 8 weeks or older were analyzed, with no difference observed. (C) Numbers of peritoneal mast cells from the same mice of the 3 genotypes as in B. (A–C) Results were compiled from 6 experiments. (D) Staining of peritoneal cells harvested from control-Tg or CD33-Tg mice with anti-CD33 (clone WM53) or isotype control, as analyzed by flow cytometry. (E) Binding of fluorescent liposome, with or without CD33L (20 μM), to peritoneal mast cells (c-Kit+FcεRI+CD45+). (F) Degranulation of CD33+ BMMCs induced by TNP-LP or TNP-LP-CD33L. (G) Cytokine induction of CD33+ BMMCs following treatment with TNP-LP (40 μM), TNP-LP-CD33L (40 μM), LP-CD33L (40 μM), or a mixture of TNP-LP and LP-CD33L (40 μM each). Supernatant from the unstimulated cells was subtracted as a background. (H) Degranulation of CD33+ BMMCs induced by TNP-LP or TNP-LP-CD33L (30 μM) in the presence of anti-CD33 (2 μg/ml). (I and J) Cytokine production of CD33+ BMMCs induced by TNP-LP or TNP-LP-CD33L (40 μM) in the presence of anti-CD33 (10 μg/ml). Supernatant from untreated cells was subtracted as a background. Results shown are representative of 3 (D–G) or 2 (H–J) independent experiments. (F–J) Values are plotted as the mean ± SD (n = 3 per condition). ***P < 0.001 and ****P < 0.0001, by 2-tailed Student’s t test (F) and 1-way ANOVA followed by Tukey’s test (C and G–J).