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Updated: May 28, 2026
An Efficient and High Yield Method for Isolation of Mouse Dendritic Cell Subsets
Published on: April 18, 2016
CD8α+ and CD8α- DC subsets from BCG-infected mice inhibit allergic Th2-cell responses by enhancing Th1-cell and
Xiaoling Gao1, Hong Bai, Jianjun Cheng
1Laboratory for Infection and Immunity, Departments of Medical Microbiology and Immunology, Faculty of Medicine, University of Manitoba, Winnipeg, Manitoba, Canada.
Insights
Mycobacterium bovis Bacille Calmette-Guérin (BCG) infection inhibits allergic diseases by modulating dendritic cell (DC) subsets. These distinct DC subsets promote immune deviation and regulation, offering new insights into allergy prevention.
Area of Science:
- Immunology
- Microbiology
- Allergy Research
Background:
- The hygiene hypothesis suggests infections can reduce allergic diseases, but mechanisms are unclear.
- Dendritic cells (DCs) are implicated in Mycobacterium bovis Bacille Calmette-Guérin (BCG)-mediated allergy inhibition via IL-12 and IL-10.
- BCG infection's impact on distinct DC subsets and their roles in allergy requires further investigation.
Purpose of the Study:
- To investigate how BCG infection modulates dendritic cell (DC) subset function.
- To determine if modulated DC subsets inhibit allergic responses through distinct mechanisms.
- To elucidate the roles of CD8α(+) and CD8α(-) DCs in modulating T-cell responses to ovalbumin (OVA).
Main Methods:
- Isolation and sorting of CD8α(+) and CD8α(-) DCs from BCG-infected mice.
- In vitro DC:T-cell co-culture assays to assess modulation of Th2-cell responses to OVA.
- In vivo adoptive transfer experiments to evaluate the impact of DC subsets on allergic responses.
Main Results:
- Both CD8α(+) and CD8α(-) DCs inhibited allergic Th2-cell responses.
- CD8α(+) DCs produced more IL-12, promoting Th1 cell responses.
- CD8α(-) DCs expressed more ICOS-L and IL-10, inducing regulatory T cells (Tregs).
Conclusions:
- BCG infection inhibits allergy through distinct mechanisms mediated by different DC subsets.
- CD8α(+) DCs drive immune deviation via Th1 induction.
- CD8α(-) DCs promote immune regulation through Treg induction.
Abstract:
The hygiene hypothesis has suggested an inhibitory effect of infections on allergic diseases, but the related mechanism remains unclear. We recently reported that DCs played a critical role in Mycobacterium bovis Bacille Calmette-Guérin (BCG)-mediated inhibition of allergy, which depended on IL-12 and IL-10-related mechanisms. Here, we tested the hypothesis that BCG infection could modulate the function of DC subsets, which might in turn inhibit allergic responses through different mechanisms. We sorted CD8α(+) and CD8α(-) DCs from BCG-infected mice and tested their ability to modulate Th2-cell responses to ovalbumin (OVA) using in vitro and in vivo approaches. We found that both DC subsets could inhibit the allergic Th2-cell response in both a DC:T-cell co-culture system and after adoptive transfer. These subsets exhibited different co-stimulatory marker expression and cytokine production patterns and were different in inducing Th1 and Treg cells. Specifically, we found that CD8α(+) DCs produced higher IL-12, inducing higher Th1 cell response, while CD8α(-) DCs expressed higher ICOS-L and produced higher IL-10, inducing CD4(+) CD25(+) FoxP3(+) Treg cells with IL-10 production and membrane-bound TGF-β expression. The finding suggests that one infection may inhibit allergy by both immune deviation and regulation mechanisms through modulation of DC subsets.
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