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Effector memory CD4+T cells in mesenteric lymph nodes mediate bone loss in food-allergic enteropathy model mice, creating IL-4 dominance

Abstract

Intestinal inflammation can be accompanied by osteoporosis, but their relationship, mediated by immune responses, remains unclear. Here, we investigated a non-IgE-mediated food-allergic enteropathy model of ovalbumin (OVA) 23-3 mice expressing OVA-specific T-cell-receptor transgenes. Mesenteric lymph nodes (MLNs) and their pathogenic CD4+T cells were important to enteropathy occurrence and exacerbation when the mice were fed an egg-white (EW) diet. EW-fed OVA23-3 mice also developed bone loss and increased CD44hiCD62LloCD4+T cells in the MLNs and bone marrow (BM); these changes were attenuated by MLN, but not spleen, resection. We fed an EW diet to F1 cross offspring from OVA23-3 mice and a mouse line expressing the photoconvertible protein KikGR to track MLN CD4+T cells. Photoconverted MLN CD44hiCD62LloCD4+T cells migrated predominantly to the BM; pit formation assay proved their ability to promote bone damage via osteoclasts. Significantly greater expression of IL-4 mRNA in MLN CD44hiCD62LloCD4+T cells and bone was observed in EW-fed OVA23-3 mice. Anti-IL-4 monoclonal antibody injection canceled bone loss in the primary inflammation phase in EW-fed mice, but less so in the chronic phase. This novel report shows the specific inflammatory relationship, via Th2-dominant-OVA-specific T cells and IL-4 production, between MLNs and bone, a distant organ, in food-allergic enteropathy.

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Fig. 1: Egg-white (EW)-fed OVA23-3 mice display bone loss.
Fig. 2: Egg-white (EW) feeding induces CD44hiCD62LloCD4+T cells in OVA23-3 mice throughout the experimental period.
Fig. 3: Mesenteric lymph nodes (MLNs), but not the spleen (SP), play an important role in the bone loss induced by egg-white (EW)-fed OVA23-3 mice.
Fig. 4: CD44hiCD62LloCD4+T cells promote bone damage independently of receptor activator of nuclear factor kappa-Β ligand (RANKL) expression.
Fig. 5: Egg-white (EW) feeding promotes weight loss, intestinal changes, bone loss, and migration of CD44hiCD62LloCD4+T cells from mesenteric lymph nodes (MLNs) to bone marrow (BM) in K23-3 mice.
Fig. 6: Analysis of cytokine mRNA expression patterns in egg-white (EW)-fed OVA23-3 mice shows predominant IL-4 and IL-1β production.
Fig. 7: Activated CD44hiCD62LloCD4+T cells in mesenteric lymph nodes (MLNs) show predominant IL-4 expression.
Fig. 8: Neutralization of IL-4 prevents weight loss, intestinal changes, and bone loss in R23-3/BALB mice.

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Acknowledgements

We thank Erika Hiraide, Mamiko Morinaga, Tomiko Asakura, Takashi Matsuwaki, Yoshikazu Saito, Takuya Miyakawa, and Jun Kunisawa (the University of Tokyo); staff of the Institute of Medical Science and the FACS Core Laboratory at the University of Tokyo; Naoyuki Takahashi (Matsumoto Dental University); Ken Kato, Atsushi Serizawa, and Takayuki Nara (Megmilk Snow Brand Co., Ltd.); Toshimitsu Yoshioka and Takashi Fujita (Bean Stalk Snow Co., Ltd.); and Akemi Ito (Ito Bone Histomorphometry Institute) for their technical advice and support.

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H.N.A. and A.O.O. provided substantial contributions to the conception of the work. M.T. provided KikGR mice. S.K. and Y.I. provided recombinant IL-1β and S.M. provided RE2 mAb and performed experiments. N.U., Y.N. and M.K. helped with the analysis of the bone. A.O.O., H.N.A., S.Y., S.U., M.T., K.S., S.N., Y.M., T.T., and T.I. performed the experiments and analyzed the data. H.N.A. and A.O.O. wrote the manuscripts. All authors discussed and interpreted the data.

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Correspondence to Haruyo Nakajima-Adachi.

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Competing interests

This work was supported by grants from the Kieikai Research Foundation (H.N.A., 2017S063, 2018T019), a Grant-in-Aid for Scientific Research (C) (H.N.A., 18K05502) from the Japan Society for the promotion of science, a grant from The Food Science Institute Foundation (Ryoushoku-kenkyukai; H.N.A., No. 2019A01), and grants from Megmilk Snow Brand Co., Ltd. (H.N.A., H.K., and S.H.). The authors declare no conflict of interest associated with this manuscript.

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Ono-Ohmachi, A., Yamada, S., Uno, S. et al. Effector memory CD4+T cells in mesenteric lymph nodes mediate bone loss in food-allergic enteropathy model mice, creating IL-4 dominance. Mucosal Immunol 14, 1335–1346 (2021). https://doi.org/10.1038/s41385-021-00434-2

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