The amelioration of composite tissue allograft rejection by TIM-3-modified dendritic cell: Regulation of the balance of regulatory and effector T cells
Introduction
Understanding the immunology of tolerance and rejection has allowed composite tissue allotransplantation (CTA) to progress [1]. CTA has been introduced as a potential clinical treatment for complex reconstructive procedures [2]. At present, a particularly large number of people have been suffering from the loss of limbs, and CTA has tremendous potential for the reconstruction of such physiological defects [3]. However, CTA is considered to elicit a stronger response compared with solid organ transplants for the reason that CTA consists of heterogeneous tissues [2]. Hence, it is important to inhibit immunological rejection after CTA.
Previous research has shown that current immunosuppressive agents, including cyclosporine [4], rapamycin [5], and FK506 [6], prolong the survival of rat limb allografts [3]. Also, in order to reduce the degree of immunological rejection, recipients often have to take immunosuppressive agents after transplantation for a long time, which inevitably results in serious side effects, including drug toxicity, infection, malignancy, and even life-threatening side effects [3], [7]. Thus, long-term use of immunosuppressive agents for CTA recipients should be avoided, and a new therapeutic strategy should be established.
It has been confirmed that T cell-dependent immune responses play a central role in allograft rejection, and the balance between T cell activation and inhibition determines the ultimate fate of allografts [8], [9]. To be specific, complete T cell activation includes T cell proliferation, differentiation into effector or memory T cells, and cytokine release, which might finally result in allograft rejection [9]. Thus, exploring ways to disarm alloreactive T cells is crucial to survival of allograft, and manipulating activating or inhibiting signals to T cells represents a potential strategy to promote long-term allograft acceptance and survival [10], [11].
T cell Ig domain and mucin domain (TIM)-3, a transmembrane protein, is constitutively expressed on CD4+ Th1 and CD8+ Tc1 cells [12]. TIM-3 has previously been demonstrated as a central regulator of Th1 responses and immune tolerance. Research by the OB and FDA has reported that TIM-3 is a key regulatory molecule of alloimmunity through its ability to broadly modulate CD4+ T cell differentiation. Also, they found that blockade of TIM-3 increased allospecific effector T cells, enhanced Th1 and Th17 polarization, and resulted in a decrease in the overall number of allospecific Tregs [10]. Hence, TIM-3 may be an important molecule for decreasing immunological rejection during CTA. Also, it remains unknown whether TIM-3 overexpression can prolong allograft survival time by influencing leukomonocytes.
The family of TIMs has been described in mice [13]. In this study, we chose mice as the research model. Our results suggest that TIM-3 can prolong the survival time of allografts by inducing lymphocyte hyporesponsiveness. Furthermore, TIM-3 overexpression can induce CD4+ T cells to differentiate into Tregs. Hence, TIM-3 may be a potential therapeutic molecule for allograft rejection in CTA.
Section snippets
Materials
BALB/c and C57BL/6 mice were purchased from Jackson Laboratories (Bar Harbor, ME, USA). Atropine, RIPA lysis buffer, mitomycin C, hematoxylin, eosin, and lipopolysaccharide (LPS) were purchased from Sigma (St. Louis, MO, USA). DNase I and Collagenase were purchased from Hoffman-La Roche (Nutley, New Jersey, USA). Red blood cell (RBC) lysis buffer and One Step Staining Mouse Treg Flow™ Kit were purchased from Bio legend (San Diego, CA, USA). Rat anti-mouse CD40, CD80, CD86, and MHCII conjugated
Expression of TIM-3 in mDCs
Western blot was performed to confirm the transfection of TIM-3 into mDCs. Also, integrated OD of each target band compared with β-actin was analyzed by Gel-Pro analyzer 4.0 software. As shown in Fig. 1, TIM-3 levels were significantly higher in TIM-3-modified mDCs compared pTARGET–mDCs and normal mDCs. In addition, there was no difference in TIM-3 expression between pTARGET–mDCs and normal mDCs. This result validates the availability of TIM-3-modified mDCs.
Postoperative detection and observation
To explore the effects of TIM-3 on
Discussion
CTA represents an ideal method for the reconstruction or replacement of tissues following traumatic loss or tumor resection and for the repair of congenital abnormalities [19]. Actually, CTA can be used in any field of plastic surgery [31]. After CTA, long-term therapeutic immunosuppression is needed, which inevitably results in serious side effects [3], [7]. Thus, CTA is restricted by the risks presented by long-term therapeutic immunosuppression and a new therapeutic strategy should be
Conflict of interests
The authors declare that there are no conflicts of interest.
Acknowledgements
This study was supported by National Natural Science Funds of China (Grant No. 81401593).
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2018, Molecular ImmunologyCitation Excerpt :Tim-3 is a membrane protein initially characterized as a negative regulator of Th1 immunity (Anderson et al., 2016). Recently, Tim-3 has been also identified to play crucial roles in activated T helper, cytotoxic T cell, and macrophages (Wang et al., 2016b; Wang et al., 2015; Tripathi et al., 2015). Evidence suggests that Tim-3 contributes to some chronic diseases, such as atherosclerosis and chronic viral infection (Foks et al., 2013; Hou et al., 2012; Dong et al., 2017).
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These authors contributed equally to this work.