Eradication of lymphoma by CD8 T cells following anti-CD40 monoclonal antibody therapy is critically dependent on CD27 costimulation

Abstract

Growing evidence points to the potential of agonistic anti-CD40 mAbs as adjuvants for vaccination against cancer. These appear to act by maturing dendritic cells (DCs) and allowing them to prime CD8 cytotoxic T lymphocytes (CTLs). Although it is well established that optimal T-cell priming requires costimulation via B7:CD28, recent studies emphasize the contribution of TNF receptors to this process. To understand how anti-CD40 mAbs trigger effective antitumor immunity, we investigated the role of TNFR superfamily members CD27 and 4-1BB in the generation of this immunity and showed that, although partially dependent on 4-1BB:4-1BBL engagement, it is completely reliant on CD27:CD70 interactions. Importantly, blocking CD70, and to some extent 4-1BBL, during anti-CD40 treatment prevented accumulation of tumor-reactive T cells and subsequent tumor protection. However, it did not influence changes in DC number, phenotype, nor the activity of CTLs once immunity was established. We conclude that CD27: IntroductionInteractions between members of the TNF receptor (TNFR) superfamily and their ligands play an important role in providing costimulation at several stages during the development of an effective antigen-specific CD8 T-cell response. [1][2][3] Early in the response, the ligation of CD40 on dendritic cells (DCs) by its ligand, CD154, induces the maturation of DCs and potentiates their ability to stimulate antigen-specific naive CD8 T cells. [4][5][6] Conversely, the absence of DC maturation, for example, during presentation of self-or tumor-associated antigens, leads to the induction of T-cell tolerance. 7 Thus, antigen presentation by immature DCs maintains peripheral tolerance to self-tissues as well as tumors. Agonistic anti-CD40 mAb, which is a potent mimic of the natural ligand, CD154, has been shown to promote T-cell-mediated immunity in a number of settings, including vaccination, and treatment of tumors. [8][9][10][11] The success achieved with agonistic anti-CD40 mAbs in preclinical models has recently led to clinical evaluation of anti-human CD40 mAbs as a potential treatment for cancer. 12,13 It is assumed that anti-CD40 mAbs trigger the maturation, or licensing of DCs which subsequently leads to the priming of tumor-specific CD8 T cells. Identifying the critical changes in DCs during their CD40-triggered maturation is therefore key to understanding the mechanism of action of anti-CD40 mAbs. CD40-induced maturation of DCs is characterized by an increase in their expression of adhesion and costimulatory molecules, including ICAM-1, B7.1, B7.2, CD70, and 4-1BB ligand (4-1BBL) as well as cytokines. [14][15][16][17][18][19] Although initial antigen-specific cytotoxic T lymphocyte (CTL) activation and proliferation depends on the CD28:B7 engagement, 20,21 subsequent expansion and survival of effector and memory T cells are controlled by additional costimulatory interactions and cytokines. Two receptors that appear central in maintaining CD8 T-cell responses are the TNFR superfamily members 4-1BB (CD137) 3,22...

Agonist Anti-Human CD27 Monoclonal Antibody Induces T Cell Activation and Tumor Immunity in Human CD27–Transgenic Mice

“…FreeStyle CHO-S cells (Invitrogen) were transiently transfected to express the 1F5 N297S mutant Ab, which was subsequently purified from culture supernatants with a MabSelect SuRe Pro A column (GE Healthcare). The rat anti-murine CD27 (mCD27) mAb AT124-1 was described previously (27). The endotoxin level for all Abs used in functional assays or in vivo studies was ,1.0 EU/mg, as measured by the Gel Clot LAL method (Lonza).…”

“…Surprisingly, a CD70 mAb that blocked the CD70/ CD27 costimulatory pathway completely abolished the therapeutic effect of anti-CD40 mAb treatment in a BCL1 lymphoma model, demonstrating that CD27 signaling was necessary for the efficacy of anti-CD40 in this tumor model. This led to the development and testing of a rat anti-mouse CD27 mAb, AT124-1, which indeed provided similar antilymphoma efficacy as did the anti-CD40 mAb (27). The antitumor efficacy of CD27 targeting was later reproduced in the EG7 thymoma model by Sakanishi and Yagita (28) using novel anti-mouse CD27 mAbs.…”

“…The therapeutic effect of an anti-CD27 agonistic mAb was first shown in studies (27) investigating the mechanisms leading to antitumor responses following treatment with an agonist anti-CD40 mAb. Surprisingly, a CD70 mAb that blocked the CD70/ CD27 costimulatory pathway completely abolished the therapeutic effect of anti-CD40 mAb treatment in a BCL1 lymphoma model, demonstrating that CD27 signaling was necessary for the efficacy of anti-CD40 in this tumor model.…”

CD27 costimulation contributes substantially to the expansion of functional memory CD8+T cells after peptide immunization

“…The blocking anti-CD70 mAb (TAN 1.6, rat IgG2a) that does not deplete CD70-expressing cells in vivo was described previously [4,11]. Purified mAb against A31 (Mc39-16, rat IgG2a) and BCL1 lymphoma (Mc106A5, rat IgG2a) idiotypes (used as controls), CD80 (1610A1, hamster 1gG), CD86 (GL-1, rat IgG2a), CD40 (3/23, rat IgG2a), Fcγ receptors II/III (2.4G2), CD3 (145.2C11), and FITC-conjugated anti-human Fc (SB2H2) were prepared in house and kindly provided by Prof. M. Glennie (University of Southampton).…”

“…Indeed, CD27 signals are essential for maximal T-cell priming by protein Ag, cellular vaccines, or infectious agents [3,[5][6][7][8][9], and for the establishment of T-cell memory after influenza infection [6]. Furthermore, DC-dependent expansion of CD8 + T cells and tumor protection triggered by Ag and anti-CD40 both require CD70, and anti-CD40 can be replaced with agonist anti-CD27 mAb to recapitulate the antitumor response [4,5,10,11]. Together these data suggest that the signal delivered from a CD40-licensed DC to a naive CD8 + T-cell requires T-cellexpressed CD27, a finding recently confirmed by others [12].…”