Serial immunomonitoring of cancer patients receiving combined antagonistic anti-CD40 and chemotherapy reveals consistent and cyclical modulation of T cell and dendritic cell parameters

Abstract

BackgroundCD40 signalling can synergise with chemotherapy in preclinical cancer models, and early clinical studies are promising. We set out to define the immunological changes associated with this therapeutic combination to identify biomarkers for a response to the therapy. Here, we present serial immunomonitoring examining dendritic cell and T cell subpopulations over sequential courses of chemoimmunotherapy.MethodsFifteen patients with mesothelioma received up to six 21-day cycles of pemetrexed plus cisplatin chemotherapy and anti-CD40 (CP-870,893). Peripheral blood was collected weekly, and analysed by flow cytometry. Longitudinal immunophenotyping data was analysed by linear mixed modelling, allowing for variation between patients. Exploratory analyses testing for any correlation between overall survival and immunophenotyping data were undertaken up to the third cycle of treatment.ResultsLarge statistically significant cyclical variations in the proportions of BDCA-1+, BDCA-2+ and BDCA-3+ dendritic cells were observed, although all subsets returned to baseline levels after each cycle and no significant changes were observed between start and end of treatment. Expression levels of CD40 and HLA-DR on dendritic cells were also cyclically modulated, again without significant change between start and end of treatment. CD8 and CD4 T cell populations, along with regulatory T cells, effector T cells, and markers of proliferation and activation, showed similar patterns of statistically significant cyclical modulation in response to therapy without changes between start and end of treatment. Exploratory analysis of endpoints revealed that patients with a higher than average proportion of BDCA-2+ dendritic cells (p = 0.010) or a higher than average proportion of activated (ICOS+) CD8 T cells (0.022) in pretreatment blood samples had better overall survival. A higher than average proportion of BDCA-3+ dendritic cells was associated with poorer overall survival at both the second (p = 0.008) and third (p = 0.014) dose of anti-CD40.ConclusionsSubstantial cyclical variations in DC and T cell populations during sequential cycles of chemoimmunotherapy highlight the critical importance of timing of immunological biomarker assessments in interpretation of results and the value of linear mixed modelling in interpretation of longitudinal change over a full treatment course.Trial registrationAustralia New Zealand Clinical Trials Registry number ACTRN12609000294257 (18th May 2009).

CD40 Agonist Antibodies in Cancer Immunotherapy

“…Comparing the residual B cells to those at baseline, investigators note increased expression of CD80, CD86, CD54, and MHC classes I and II, consistent with enhanced antigen presentation capability. Similar findings on activation of circulating DCs before and after CD40 mAbs have also been reported (69).…”

The Immune Revolution: A Case for Priming, Not Checkpoint

“…Immune pharmacodynamics studies have provided evidence of CD40-induced activation of B cells and DC (Johnson et al, 2015; McDonnell et al, 2017; Ruter et al, 2010; Vonderheide et al, 2007). These effects are dose dependent and transient.…”

Trial watch: chemotherapy-induced immunogenic cell death in immuno-oncology

“…In this context, cancer cells potentially undergoing ICD can be examined for the release or exposure of ICD-associated DAMPs (see below), 12,15,21,149,[151][152][153] and/or co-cultured with myeloid cells such as dendritic cells (DCs) 21,119,[154][155][156] which are ultimately assessed for: (1) phagocytic activity, [157][158][159][160][161][162][163][164][165] (2) surface activation markers (e.g. CD80, CD83, CD86, CD83, CD40 and/or MHC Class II molecules), 166 (3) secretory activity, with specific reference to interleukin 1 beta (IL1B), IL6, IL12 and tumor necrosis factor (TNF), [167][168][169][170][171][172] and (4) T-cell crosspriming. [173][174][175][176][177][178][179][180][181] Of note, performing these ex vivo experiments with human cells generally involves allogeneic settings, since DCs or T cells derived from healthy individuals are typically not HLA-matched to human cancer cell lines.…”