Cryopreserved Interleukin-4-Treated Macrophages Attenuate Murine Colitis in an Integrin β7-Dependent Manner

Abstract

The adoptive transfer of alternatively activated macrophages (AAMs) has proven to attenuate inflammation in multiple mouse models of colitis; however, the effect of cryopreservation on AAMs, the ability of previously frozen AAMs to block dinitrobenzene sulfonic acid (DNBS) (Th1) and oxazolone (Th2) colitis and their migration postinjection remains unknown. Here we have found that while cryopreservation reduced mRNA expression of canonical markers of interleukin (IL)-4-treated macrophages [M(IL-4)], this step did not translate to reduced protein or activity, and the cells retained their capacity to drive the suppression of colitis. The anticolitic effect of M(IL-4) adoptive transfer required neither T or B cell nor peritoneal macrophages in the recipient. After injection into the peritoneal cavity, M(IL-4)s migrated to the spleen, mesenteric lymph nodes and colon of DNBS-treated mice. The chemokines CCL2, CCL4 and CX3CL1 were expressed in the colon during the course of DNBS-induced colitis. The expression of integrin β7 on transferred M(IL-4)s was required for their anticolitic effect, whereas the presence of the chemokine receptors CCR2 and CX3CR1 were dispensable in this model. Collectively, the data show that M(IL-4)s can be cryopreserved M(IL-4)s and subsequently used to suppress colitis in an integrin β7-dependent manner, and we suggest that these proof-of-concept studies may lead to new cellular therapies for human inflammatory bowel disease.

Macrophages treated with antigen from the tapeworm Hymenolepis diminuta condition CD25 + T cells to suppress colitis

“…Murine M (IL‐4) suppress colitis in RAG1 −/− mice induced by direct i.r. delivery of DNBS (31); however, intravenously administered M (HdAg) , although effectively blocking DNBS‐induced colitis in wild‐type mice, failed to do so in RAG1 −/− mice, indicative of a requirement for T or B cells or both in the recipient mice (Fig. 5).…”

“…The inability of intraperitoneally delivered M (HdAg) to inhibit colitis was unexpected. Macrophages shown to block colitis, including those elicited by AvCystatin, have increased arginase‐1 (20, 31, 54), but not the M (HdAg) ; arginase‐1 promotes wound healing and is implicated in macrophage‐driven inhibition of colitis (55). Yet, intravenous injection of M (HdAg) significantly reduced the severity of DNBS‐induced disease.…”

“…In other experiments, adoptive transfer of 1 × 10 5 CD4 + CD25 + cells or 9 × 10 5 CD4 + CD25 − cells retrieved from cocultures of splenic CD4 + cells from naive mice and HdAg‐treated macrophages was performed in the DNBS model of colitis. Finally, the impact of intravenously injected M (HdAg) in recombination‐activating gene (RAG) 1 −/− mice (breeding colony, University of Calgary) treated with DNBS [5 mg/mouse (C57/BL6 background)] was assessed (31).…”

Suppression of colitis by adoptive transfer of helminth antigen-treated dendritic cells requires interleukin-4 receptor-α signaling

“…IL-4 treated macrophages take on an alternatively activated phenotype (AAMs) characterized by expression of arginase1, Ym1, and Relm-α22, and the adoptive transfer of IL-4 AAMs can suppress DNBS-induced colitis26. Similarly, analysis of Ym1 and Relm-α expression reveals that IL-4 causes alternative activation in DCs (AADCs)21, raising the possibility that HD antigen was, through its induction of IL-4, inducing an AADC phenotype that could block colitis.…”

“…Finally, and directly linked to infection with helminth parasites and hence HD-DCs, IL-4 drives the development of alternatively activated macrophages (AAMs or M(IL-4)) that have been implicated in wound healing49 and the suppression of colitis-induced by DNBS, oxazolone and dextran sodium-sulfate (DSS)265051.…”