Abstract 1773P
Background
Among immune related adverse events associated with anti PD-1 antibody, acute interstitial pneumonitis (AIP) is frequent and can be lethal. We analyzed alveolar immune infiltrates from bronchoalveolar lavages (BAL) of patients with AIP related to ICB.
Methods
Fresh BAL samples from patients with anti-PD-1 induced AIP were collected. Cells were analyzed by flow cytometry (BD, LSRFortessa X20®), mass cytometry (CyTOF, Fluidigm, Helios®), and single-cell RNA and TCR sequencing (scRNAseq, 10x genomics, chromium®).
Results
Ten BAL from patients with anti-PD-1-induced AIP were explored for analysis of cellular alveolar infiltration, by flow cytometry (n=7), CyTOF (n=6) and scRNAseq (n=2). Immune infiltrates were characterized by predominance of CD3+ T lymphocytes (68%, +/- 24%SD), and mostly CD8+ T cells (64% +/- 26%SD). These were mainly CD8+ effector and effector memory (TEM) T cells, which expressed predominantly CD27+CD28loCD38+CD69+KLRG1-PD-1+TIGIT+. Single-cell RNAseq analyzes showed that those CD8+ T cells expressed high levels of GZMH, GZMA, CST7, CTSW, NKG7, CCL4, CCL5, CD27, PDCD1, TIGIT, CD69 and ITGAE genes. They also expressed transcription factors IKZF6, ZNF683, TOX but not TCF7, EOMES or TBX21. Analysis of the TCR repertoire of those CD8+ T cells revealed oligoclonal expansion. CD14+CD163+CD1c- macrophages, partially double positive for CD206 and CCR2, were the main population among CD11c+ HLA-DR+ cells. CD206+ macrophages represented one third of total immune cells. They were positive for CD64, CD40 and partially for CD86 and PD-L1. Single-cell RNAseq of these cells found high levels of expression for MRC1, CD163, LILRB2, FCN1, S100A8, S100A9, CXCL9 and CXCL10 genes.
Conclusions
Alveolar immune infiltrates in patients with anti-PD1 induced AIP were characterized by a predominance and oligoclonal expansion of cytotoxic resident memory CD8+ T cells. Macrophages exhibited characteristics of cells derived from circulating monocytes and not from resident alveolar macrophages. Those results should help the diagnosis and treatment of lung toxicities induced by anti-PD-1 immunotherapies.
Clinical trial identification
Editorial acknowledgement
Legal entity responsible for the study
Gustave Roussy.
Funding
Fondation Gustave Roussy, Fondation Malakoff Médéric.
Disclosure
J-C. Soria: Financial Interests, Personal, Stocks/Shares: AstraZeneca; Financial Interests, Personal, Stocks/Shares: Gritstone Oncology; Financial Interests, Personal, Stocks/Shares: Relay Therapeutics; Non-Financial Interests, Personal, Member of the Board of Directors: Hookipa Pharmaceuticals; Non-Financial Interests, Personal, Full or part-time Employment, Sept 2017 to Dec 2019: AstraZeneca. L. Zitvogel: Non-Financial Interests, Personal, Member of the Board of Directors: Transgene; Non-Financial Interests, Personal, Advisory Board: Transgene; Non-Financial Interests, Personal, Advisory Board: Lytix Biopharma; Non-Financial Interests, Personal, Advisory Board: EpiVax; Non-Financial Interests, Personal, Advisory Board: NeoVax; Non-Financial Interests, Personal, Advisory Board: Vedanta; Non-Financial Interests, Institutional, Other, Research contracts: BMS; Non-Financial Interests, Institutional, Other, Research contracts: Incyte; Non-Financial Interests, Institutional, Other, Research contracts: GSK; Non-Financial Interests, Institutional, Other, Research contracts: Transgene; Non-Financial Interests, Institutional, Other, Research contracts: Innovate Biopharma; Non-Financial Interests, Institutional, Other, Research contracts: Pilege; Non-Financial Interests, Institutional, Other, Research contracts: Kaleido; Financial Interests, Personal, Leadership Role: everImmune. A. Marabelle: Financial Interests, Personal and Institutional, Research Grant: Merus; Financial Interests, Personal and Institutional, Research Grant: BMS; Financial Interests, Personal and Institutional, Research Grant: Boehringer Ingelheim; Financial Interests, Personal and Institutional, Research Grant: Transgene; Financial Interests, Personal and Institutional, Research Grant: Fondation MSD Avenir; Financial Interests, Personal and Institutional, Research Grant: Sanofi; Financial Interests, Personal, Stocks/Shares: Pegascy SAS; Financial Interests, Personal, Stocks/Shares: Centessa Pharmaceuticals; Financial Interests, Personal, Stocks/Shares: HiFiBio; Financial Interests, Personal, Stocks/Shares: Shattuck Labs; Financial Interests, Personal, Invited Speaker: Roche; Financial Interests, Personal, Invited Speaker: BMS; Financial Interests, Personal, Invited Speaker: MSD; Financial Interests, Personal, Invited Speaker: Merk Serono; Financial Interests, Personal, Invited Speaker: AstraZeneca; Financial Interests, Personal, Invited Speaker: Amgen; Financial Interests, Personal, Invited Speaker: Sanofi; Financial Interests, Personal, Invited Speaker: Servier. All other authors have declared no conflicts of interest.