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Updated: Oct 1, 2026
In Vitro and In Vivo Assessment of T, B and Myeloid Cells Suppressive Activity and Humoral Responses from Transplant Recipients
Published on: August 12, 2017
CD154 is expressed during treatment with calcineurin inhibitors after organ transplantation
Miranda M L van Rijen1, Mark L Kuijf, Herold J Metselaar
1Department of Gastroenterology and Hepatology, University Hospital Rotterdam, Rotterdam, The Netherlands.
Background:
CD154 (CD40 ligand) monoclonal antibody prevents allograft rejection in rodents and monkeys. Inasmuch as calcineurin inhibitors (CI) inhibit CD154 expression by pharmacologic agents in vitro, we investigated whether CD154 is also inhibited by CI in vivo and in vitro during allogeneic stimulation.
Methods:
CD154 expression was determined by immunohistochemistry and flow cytometry in human lymph nodes and spleen sections from rhesus monkeys with or without CI treatment. The effect of CI on induction of CD154 expression was studied by stimulating lymphocytes with phorbol 12-myristate 13-acetate (PMA) and ionomycin or with allogeneic monocyte-derived mature dendritic cells.
Results:
Lymph nodes from patients with or without CI cyclosporine (CsA) or FK506 (FK) treatment showed comparable CD154 expression, which was present on the cell surface of T cells. CD154-expressing cells were also present in spleens from monkeys treated with CsA in comparable numbers to those in the nontreated group. Moreover, in several liver transplant rejection biopsies taken during CI therapy CD154-expressing cells were observed. In vitro, CsA and FK strongly inhibited induction of CD154 expression on peripheral blood mononuclear cells by pharmacologic stimuli. Maximum inhibition was found at 50 ng/ml CsA and 20 ng/ml FK. CD154 expression induced by dendritic cells in peripheral blood mononuclear cells or spleen cells was also almost completely inhibited by CsA.
Conclusion:
Although CI strongly suppressed pharmacologic and allogeneic induction of CD154 expression on T cells in vitro at concentrations at approximately clinical trough levels, CD154 is prominently expressed during CI therapy in lymphoid tissue and (sporadically) in liver allografts. This suggests that the CD154-CD40 pathway remains functional during CI therapy, which may contribute to allograft rejections in the clinical setting.
Insights
Calcineurin inhibitors (CI) suppress CD154 (CD40 ligand) expression in vitro, but CD154 remains present in lymphoid tissues during CI therapy. This suggests the CD154-CD40 pathway may contribute to allograft rejection despite immunosuppression.
Area of Science:
- Immunology
- Transplantation immunology
Background:
- CD154 (CD40 ligand) monoclonal antibody is crucial for preventing allograft rejection.
- Calcineurin inhibitors (CI) are known to inhibit CD154 expression pharmacologically in vitro.
Purpose of the Study:
- To investigate whether CI inhibit CD154 expression in vivo and in vitro during allogeneic stimulation.
- To determine the functional status of the CD154-CD40 pathway during CI therapy.
Main Methods:
- CD154 expression analyzed by immunohistochemistry and flow cytometry in human and rhesus monkey tissues.
- In vitro studies involved stimulating lymphocytes with pharmacologic agents (PMA/ionomycin) or allogeneic dendritic cells, with or without CI (cyclosporine/FK506).
Main Results:
- CD154 expression was comparable in lymph nodes and spleens of CI-treated and untreated subjects.
- CD154-expressing cells were observed in liver transplant rejection biopsies during CI therapy.
- In vitro, CI significantly inhibited CD154 induction by both pharmacologic stimuli and dendritic cells.
Conclusions:
- Despite strong in vitro inhibition by CI, CD154 is prominently expressed in lymphoid tissues during CI therapy.
- The CD154-CD40 pathway appears functional during CI therapy, potentially contributing to allograft rejection.
- Findings suggest alternative or complementary pathways may be involved in CI-resistant allograft rejection.
