Role for CD40–CD40 ligand interactions in the immune response to solid tumours
Introduction
CD40 is a 40 kDa type I glycoprotein member of the nerve growth factor receptor/tumour necrosis factor family. It is found on numerous types of cells including B lymphocytes, dendritic and follicular dendritic cells, macrophages, haematopoietic progenitors, endothelial and epithelial cells, fibroblasts and carcinoma cells (Stout and Suttles, 1996, Grewal and Flavell, 1997, Van Kooten and Banchereau, 1997, Alexandroff et al., 1998). Interaction with its ligand (CD40L, a type II membrane protein of 33 kDa and a member of the tumour necrosis factor gene family) results in potent activation and survival stimuli. These include the production of various cytokines (e.g. GM-CSF, IL-1, IL-4, IL-6, IL-8, IL-10, IL-12, RANTES, TNFα) and up-regulation of adhesion and costimulatory molecules (ICAM-1, CD23, CD62E, CD80, CD86, CD106, etc.). Upon activation, CD40L is transiently expressed on the surface of CD4+ and CD8+ lymphocytes, mast cells, NK cells and γδ T lymphocytes. Later it is rapidly shed in the supernatant in trimeric form where it retains its full biological activity (Armitage, 1992). Furthermore, CD40L is expressed on resting T lymphocytes at a minimal but sufficient level to exert a biological effect (Armant et al., 1996). CD40 delivers potent co-stimulatory signals to effector cells through CD40L and has been suggested to be responsible for the initiation of immune responses (Grewal and Flavell, 1997). Co-immunisation with CD40L markedly enhances antibody and CTL-mediated immune responses (Mendoza et al., 1997). It is not surprising therefore, that CD40-CD40L mediated interactions play a pivotal role in immune regulation including normal development of B lymphocytes, response to infections, rejection of transplants and immunosurveillance (Borrow et al., 1996, Mackey et al., 1997).
CD40 is well expressed by a variety of human carcinomas including bladder, breast and ovarian cancers as well as on melanoma cells. In contrast to leucocytes where the role of CD40–CD40L pair has been systematically investigated, their function in the biology of solid tumours remains surprisingly obscure even despite the fact that CD40 was initially described on carcinomas (Paulie et al., 1985). However, recent data suggests that its role on tumour cells may differ from that ascribed in immune cells. For example, it has been suggested that ligation of cell surface CD40 on certain tumour cells may convey a pro-apoptotic or inhibitory signal (Eliopoulos et al., 1996, Hess and Engelmann, 1996, Lollini et al., 1998, Afford et al., 1999) while on other tumour cell populations, CD40 ligation results in increased cell survival and activation (Jakobson et al., 1998, Voorzanger-Rousselot et al., 1998, Sbih-Lammali et al., 1999). In the current study we investigated the functional role of CD40 on the surface of solid tumour cells in the induction of adhesion and costimulatory molecules and the expression of pro-inflammatory cytokines. In addition, we examined the ability of rhCD40L to induce growth inhibition and apoptosis of carcinoma and melanoma cells as well as the importance of CD40-CD40L interactions in T lymphocyte activation.
Section snippets
Reagents
The antibodies used were obtained from the following sources: non-conjugated or fluorescein isothiocyanate (FITC) conjugated murine anti-human CD40 from Serotec (Oxford, UK) and Ancell (Bayport, USA), respectively; non-conjugated and FITC bound anti-ICAM-1, non-conjugated anti-FAS and isotype control mouse IgG1 antibodies, R&D Systems (Abingdon, UK); FITC and phycoerythrin conjugated mouse isotype-control and FITC conjugated anti FAS from Dako (Carpinteria, CA, USA); biotinylated anti-FAS
CD40 expression on carcinoma cells
The expression of CD40 by the cell lines used in this study is summarised in Table 1. Examination of CD40-specific mRNA revealed the presence of transcripts in the majority of cell lines studied. With the exception of the bladder tumour cell line RT4, cognate expression of cell surface CD40 was observed. When cells were treated with IFNγ an increase in cell surface CD40 was observed only for cells which constitutively expressed the protein (Fig. 1 and Table 2). In contrast to CD40, CD40L
Discussion
In this study expression of CD40 (but not CD40L) was demonstrated on a panel of carcinoma and melanoma cell lines. The expression of CD40 was augmented but not induced by IFNγ. Ligation of CD40 with antibody or recombinant ligand resulted in enhanced levels of ICAM-1 and FAS on tumour cell surfaces and secretion of a number of cytokines. In addition apoptosis and cell growth arrest were induced. CD40 positive tumour cells stimulated the generation of specific alloreactive T cells, whilst CD40
Acknowledgements
We thank R. Carter for useful discussion, J. Black and I. Ansell for technical assistance and A. Wood for help in producing retrovirus. Trimeric CD40L and anti-CD40L antibodies (M91, M92) were generously provided by Immunex. This work was supported in part by the University of Edinburgh Faculty of Medicine Cancer Research Fund and the Scottish Hospital Endowment Research Trust.
References (40)
- et al.
Autocrine regulation of ICAM-1 expression on bladder cancer cell-lines: evidence for the role of IL-1 alpha
Immunol. Lett.
(1994) - et al.
Tumor immunology: false hopes-new horizons?
Immunol. Today
(1998) - et al.
Expression and function of CD40 on Hodgkin and Reed-Sternberg cells and the possible relevance to Hodgkin's disease
Blood
(1994) - et al.
Neuronal apoptosis induced by HIV-1 gp120 and the chemokine SDF-1 is mediated by the chemokine receptor CXCR4
Curr. Biol.
(1998) - et al.
Inhibition of human breast carcinoma growth by soluble recombinant human CD40 ligand
Blood
(1999) - et al.
Expression of adhesion molecules by bladder cancer cells: Modulation by interferon-gamma and tumour necrosis factor-alpha
J. Urol.
(1992) - et al.
The many roles of CD40 in cell-mediated inflammatory response
Immunol. Today
(1996) - et al.
Resistance to cytotoxic chemotherapy induced by CD40 ligand in lymphoma cells
Blood
(1998) - et al.
CD40 activation induces apoptosis in cultured human hepatocytes via induction of cell surface Fas ligand expression and amplifies Fas-mediated hepatocyte death during allograft rejection
J. Exp. Med.
(1999) - et al.
Functional CD40 ligand expression on T lymphocytes in the absence of T cell receptor engagement: involvement in interleukin-2-induced interleukin-12 and interferon-gamma production
Eur. J. Immunol.
(1996)
Identification of a source of biologically active CD40 ligand
Eur. J. Immunol.
CD40L-deficient mice show deficits in antiviral immunity and have an impaired memory CD8+ CTL response
J. Exp. Med.
What is the role of CD40 in cancer immunotherapy
Immunol. Today
CD40-induced growth inhibition in epithelial cells is mimicked by Epstein-Barr virus-encoded LPM1: involvement of TRAF3 as a common mediator
Oncogene
Induction of melanoma antigen-specific cytotoxic T lymphocytes in vitro by stimulation with B7-expressing human melanoma cell lines
J. Immunother.
CD40 ligation on human cord blood CD34+ hematopoietic progenitors induces their proliferation and differentiation into functional dendritic cells
J. Exp. Med.
CD40 antibody evokes a cytotoxic T-cell response that eradicates lymphoma and bypasses T-cell help
Nat. Med.
Fas ligation induces apoptosis of CD40-activated human B lymphocytes
J. Exp. Med.
A central role of CD40 ligand in the regulation of CD4+ T cell responses
Immunol. Today
A novel function of CD40: induction of cell death in transformed cells
J. Exp. Med.
Cited by (76)
Harnessing the potential of CD40 agonism in cancer therapy
2024, Cytokine and Growth Factor ReviewsTurning tumour cells into antigen presenting cells: The next step to improve cancer immunotherapy?
2016, European Journal of CancerEfficacy of Zhuling polyporus polysaccharide with BCG to inhibit bladder carcinoma
2015, Carbohydrate PolymersCitation Excerpt :CD40 is a surface receptor best known for its capacity to initiate multifaceted activation signals in normal B cells and DCs. CD40-mediated interactions play an important role in the response to infections and cancer by affecting the development, activation, proliferation, and differentiation of a variety of immune cells (Alexandroff et al., 2000). Abnormal CD40 signaling seems to be directly associated with the pathogenesis of chronic inflammatory diseases associated with cancer (Feng et al., 2012).
Immunotherapeutic prospects and progress in bladder cancer
2024, Journal of Cellular and Molecular MedicineNext Generation CD40 Agonistic Antibodies for Cancer Immunotherapy
2022, Frontiers in Immunology