Elsevier

Transplantation Proceedings

Volume 42, Issue 9, November 2010, Pages 3784-3792
Transplantation Proceedings

Experimental study
Immunosuppression
Küpffer Cells Promote Acute Rejection via Induction of Th17 Differentiation in Rat Liver Allografts

https://doi.org/10.1016/j.transproceed.2010.08.030Get rights and content

Abstract

Background

Th17, a newly identified CD4+ T-cell subset, has been implicated in transplant rejection. Differentiation of Th17 cells is associated with transforming growth factor–β (TGF-β) and interleukin-6 (IL-6), which are the main products of Küpffer cells.

Objective

To determine whether Küpffer cells promote acute liver allograft rejection by inducing Th17 cell differentiation.

Methods

A rat model of allogeneic liver transplantation using Dark Agouti (DA) to Brown Norway (BN) rats was established with or without gadolinium chloride (GdCl3) pretreatment. Isogeneic liver transplantation (BN to BN) was performed as a control. Concentrations of cytokines secreted by Küpffer cells or Th17-related cytokines detected in the liver and peripheral blood were analyzed using immunohistochemistry assays, flow cytometry, and enzyme-linked immunosorbent assays. Survival differences were compared between treatment groups. In vitro, Küpffer cells from liver grafts were isolated and co-cultured with naïve CD4 T cells.

Results

Both Küpffer cells and Th17 cells infiltrated liver allografts, accompanied by an increase in concentrations of IL-6 and TGF-β. Pretreatment with GdCl3 attenuated intragraft infiltration of Küpffer cells and Th17 cells, and decreased IL-6 and TGF-β concentrations. Liver function improved after pretreatment, and mean (SD) survival time was prolonged, compared with the control group (16.33 [0.96] days vs 11.50 [0.99] days, respectively; P < .01). In vitro, Küpffer cells from livers with allografts secreted significantly higher concentrations of IL-6 and TGF-β and induced Th17 differentiation more effectively compared with livers with isografts (30.8% vs 8.1%, respectively).

Conclusion

Küpffer cells have the potential to induce Th17 cells by secreting IL-6 and TGF-β, and as a result, promote acute liver allograft rejection.

Section snippets

Animals

Inbred male Dark Agouti (DA) and Brown Norway (BN) rats aged 12 weeks and weighing 200 to 250 g were purchased from Beijing Vital River Company (Beijing, China). The animals were housed under standard animal care conditions, and received food and water ad libitum. The study was approved by the China Association of Laboratory Animal Care and the Institutional Animal Care Committee.

Rats were assigned to 3 treatment groups: an isograft group (BN to BN liver transplantation), an allograft group (DA

Increased Infiltration of Küpffer cells and Th17 cells Into Liver Allografts

The rejection of allogeneic liver grafts (DA to BN) was compared with that of isografts (BN to BN). No signs of rejection were observed in isograft recipients, whereas allograft hosts exhibited severe acute rejection accompanied by elevated serum concentrations of ALT and AST. Extensive intragraft mononuclear cell infiltration and severe liver damage were also observed in allograft recipients (data not shown). To analyze intragraft infiltration by Küpffer cells and IL-17–producing cells in

Discussion

The role of Küpffer cells in liver allograft rejection has been controversial. Some studies have demonstrated that Küpffer cells participate in the rejection reaction by mediating potent proinflammatory activities and acting as antigen-presenting cells.22, 23, 24 In contrast, other studies have demonstrated that Küpffer cells have a key role in development of immune tolerance by inducing T-cell apoptosis or producing IL-10 and TGF-β.25, 26 In the present study, an increased number of

Acknowledgements

This study was supported by grant 2009CB522403 from the National Basic Research Program (973) of China, grant 30730085 from the Key Program of the National Natural Science Foundation of China, and grant 30972795 from the National Natural Science Foundation of China.

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