Depletion of major pathogenic cells in asthma by targeting CRTh2
Tao Huang, Meredith Hazen, Yonglei Shang, Meijuan Zhou, Xiumin Wu, Donghong Yan, Zhonghua Lin, Margaret Solon, Elizabeth Luis, Hai Ngu, Yongchang Shi, Arna Katewa, David F. Choy, Nandhini Ramamoorthi, Erick R. Castellanos, Mercedesz Balazs, Min Xu, Wyne P. Lee, Marissa L. Matsumoto, Jian Payandeh, Joseph R. Arron, Jo-Anne Hongo, Jianyong Wang, Isidro Hötzel, Cary D. Austin, Karin Reif
Tao Huang, Meredith Hazen, Yonglei Shang, Meijuan Zhou, Xiumin Wu, Donghong Yan, Zhonghua Lin, Margaret Solon, Elizabeth Luis, Hai Ngu, Yongchang Shi, Arna Katewa, David F. Choy, Nandhini Ramamoorthi, Erick R. Castellanos, Mercedesz Balazs, Min Xu, Wyne P. Lee, Marissa L. Matsumoto, Jian Payandeh, Joseph R. Arron, Jo-Anne Hongo, Jianyong Wang, Isidro Hötzel, Cary D. Austin, Karin Reif
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Research Article Immunology Therapeutics

Depletion of major pathogenic cells in asthma by targeting CRTh2

Abstract

Eosinophilic inflammation and Th2 cytokine production are central to the pathogenesis of asthma. Agents that target either eosinophils or single Th2 cytokines have shown benefits in subsets of biomarker-positive patients. More broadly effective treatment or disease-modifying effects may be achieved by eliminating more than one inflammatory stimulator. Here we present a strategy to concomitantly deplete Th2 T cells, eosinophils, basophils, and type-2 innate lymphoid cells (ILC2s) by generating monoclonal antibodies with enhanced effector function (19A2) that target CRTh2 present on all 4 cell types. Using human CRTh2 (hCRTh2) transgenic mice that mimic the expression pattern of hCRTh2 on innate immune cells but not Th2 cells, we demonstrate that anti-hCRTh2 antibodies specifically eliminate hCRTh2+ basophils, eosinophils, and ILC2s from lung and lymphoid organs in models of asthma and Nippostrongylus brasiliensis infection. Innate cell depletion was accompanied by a decrease of several Th2 cytokines and chemokines. hCRTh2-specific antibodies were also active on human Th2 cells in vivo in a human Th2-PBMC-SCID mouse model. We developed humanized hCRTh2-specific antibodies that potently induce antibody-dependent cell cytotoxicity (ADCC) of primary human eosinophils and basophils and replicated the in vivo depletion capacity of their murine parent. Therefore, depletion of hCRTh2+ basophils, eosinophils, ILC2, and Th2 cells with h19A2 hCRTh2–specific antibodies may be a novel and more efficacious treatment for asthma.

Authors

Tao Huang, Meredith Hazen, Yonglei Shang, Meijuan Zhou, Xiumin Wu, Donghong Yan, Zhonghua Lin, Margaret Solon, Elizabeth Luis, Hai Ngu, Yongchang Shi, Arna Katewa, David F. Choy, Nandhini Ramamoorthi, Erick R. Castellanos, Mercedesz Balazs, Min Xu, Wyne P. Lee, Marissa L. Matsumoto, Jian Payandeh, Joseph R. Arron, Jo-Anne Hongo, Jianyong Wang, Isidro Hötzel, Cary D. Austin, Karin Reif

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Figure 3

hCRTh2-reactive antibodies 19A2 and 8B1 specifically bind hCRTh2.

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hCRTh2-reactive antibodies 19A2 and 8B1 specifically bind hCRTh2. (A) Fl...
(A) Flow cytometry analyses of mouse anti-hCRTh2 Abs 19A2 and 8B1 binding to hCRTh2 expressed on 293 cells but not mock-transfected WT 293 cells. Anti-CRTh2 Ab concentrations were 20 μg/ml (solid line), 2 μg/ml (dashed line), and 0.2 μg/ml (dotted line); isotype control Ab was 20 μg/ml (tinted histogram). (B) Reactivity by flow cytometry of mouse anti-hCRTh2 antibodies 19A2 and 8B1 with primary human basophils or eosinophils from peripheral blood leukocytes. Anti-CRTh2 Ab concentrations were 10 μg/ml (solid line) or 1 μg/ml (dashed line); isotype control Ab was 10 μg/ml (tinted histogram). (C) Anti-CRTh2 Ab 8B1, but not 19A2, prevents PGD2-induced calcium mobilization. Calcium flux in response to PGD2 stimulation of in vitro polarized human Th2 cell cultures gated on CCR4+CCR6−CXCR3−CD4+ T cells was monitored by flow cytometry in the presence of anti-CRTh2 or isotype control antibodies at 100 nM. The CRTh2 receptor antagonist CAY10471 was included as a positive control. Data shown are representative of at least 3 independent experiments. PGD2, prostaglandin D2.