The role of Sp140 revealed in IgE and mast cell responses in Collaborative Cross mice
Kazufumi Matsushita, Xin Li, Yuki Nakamura, Danyue Dong, Kaori Mukai, Mindy Tsai, Stephen B. Montgomery, Stephen J. Galli
Kazufumi Matsushita, Xin Li, Yuki Nakamura, Danyue Dong, Kaori Mukai, Mindy Tsai, Stephen B. Montgomery, Stephen J. Galli
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Research Article Immunology Inflammation

The role of Sp140 revealed in IgE and mast cell responses in Collaborative Cross mice

Abstract

Mouse IgE and mast cell (MC) functions have been studied primarily using inbred strains. Here, we (a) identified effects of genetic background on mouse IgE and MC phenotypes, (b) defined the suitability of various strains for studying IgE and MC functions, and (c) began to study potentially novel genes involved in such functions. We screened 47 Collaborative Cross (CC) strains, as well as C57BL/6J and BALB/cJ mice, for strength of passive cutaneous anaphylaxis (PCA) and responses to the intestinal parasite Strongyloides venezuelensis (S.v.). CC mice exhibited a diversity in PCA strength and S.v. responses. Among strains tested, C57BL/6J and CC027 mice showed, respectively, moderate and uniquely potent MC activity. Quantitative trait locus analysis and RNA sequencing of BM-derived cultured MCs (BMCMCs) from CC027 mice suggested Sp140 as a candidate gene for MC activation. siRNA-mediated knock-down of Sp140 in BMCMCs decreased IgE-dependent histamine release and cytokine production. Our results demonstrated marked variations in IgE and MC activity in vivo, and in responses to S.v., across CC strains. C57BL/6J and CC027 represent useful models for studying MC functions. Additionally, we identified Sp140 as a gene that contributes to IgE-dependent MC activation.

Authors

Kazufumi Matsushita, Xin Li, Yuki Nakamura, Danyue Dong, Kaori Mukai, Mindy Tsai, Stephen B. Montgomery, Stephen J. Galli

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Figure 7

Sp140 contributes to IgE-dependent MC responses.

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Sp140 contributes to IgE-dependent MC responses. BM cells from C57BL/6J...
BM cells from C57BL/6J mice were cultured with IL-3 for 6–7 weeks to generate BMCMCs. Control siRNA or siRNA against Sp140 were transfected into the cells. (A) mRNA level for Sp140 24 hours after the siRNA transfection was determined by quantitative PCR. The Sp140 levels are indicated as relative to Actb. (B) siRNA-transfected cells were sensitized with mouse anti–DNP IgE mAb (1 μg/mL) overnight and then stimulated with DNP-HSA (10 ng/mL) for 1 hour. Histamine contents in cell lysates and the culture supernatants were determined by ELISA. Histamine release rates are indicated as percent release (supernatants) out of total histamine contents of the cells (cell lysate plus supernatant). (C–E) siRNA-transfected cells were stimulated as in B but for 6 hours. The levels of IL-4 (C), IL-6 (D), and IL-13 (E) in the culture supernatants were determined by ELISA. Each dot indicates different biological replicates. n = 7 pooled from 4 independent experiments. P values were determined by paired Student’s t test.