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429 Tumor-Infiltrating Normal B Lymphocytes Have Remarkable Prognostic Effects and Are Crucial for Antitumor Immune Responses in Diffuse Large B-Cell Lymphoma

Program: Oral and Poster Abstracts
Type: Oral
Session: 622. Lymphomas: Translational – Non-Genetic: Elucidating Biomarkers and Mechanisms of Therapy in Lymphoma
Hematology Disease Topics & Pathways:
Research, Translational Research
Sunday, December 10, 2023: 10:00 AM

Zijun Y. Xu-Monette, PhD1*, Yong Li, PhD2, Thomas Snyder3*, Tiantian Yu, PhD4*, Tingxun LU4*, Alexandar Tzankov5*, Carlo Visco, MD6*, Govind Bhagat, MD7, Wenbin Qian8*, Karen Dybkaer, PhD, MSC9*, April Chiu, MD10*, Wayne Tam, MD11, Youli Zu, MD, PhD12*, Eric D. Hsi, MD13, Frederick Hagemeister14*, Heounjeong Go15*, Maurilio Ponzoni, MD16, Andrés José María Ferreri, MD17, Michael Møller18*, Benjamin M. Parsons, DO19, Joannes H.J.M. Van Krieken, PhD, MD20*, Yingjun Wang, MD21*, Miguel Angel Piris, PhD, MD22*, Jane Winter23, Qingyan Au24*, Ilan Kirsch25*, Mingzhi Zhang, MD26*, John D Shaughnessy, Jr, PhD27*, Bing Xu28* and Ken H. Young, MD, PhD29

1Department of Pathology, Duke University Medical Center, Durham, NC
2Medicine, Baylor College of Medcine, Houston, TX
3Adaptive Biotechnologies, Seattle, WA
4Duke University, Durham, NC
5Medical University Basel, Basel, Switzerland
6University of Verona, Verona, ITA
7Department of Pathology and Cell Biology, Columbia University Irving Medical Center, New York
8The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China
9Aalborg University Hospital, Aalborg, Denmark
10Mayo Clinic, Rochester, MN
11Well Cornell Medical College, New York, NY
12Houston Methodist Hospital, Houston, TX
13Wake Forest Baptist Medical Center, Winston Salem, NC
14Department of Lymphoma/Myeloma, The University of Texas MD Anderson Cancer Cente, Houston, TX
15Asan Medical Center Children's Hospital, Seoul, KOR
16Pathology Unit, San Raffaele H. Scientific Institute, Milano, Italy
17Unit of Lymphoid Malignancies, Department of Onco-Haematology, IRCCS San Raffaele Scientific Institute, Milano, Italy
18Odense University Hospital, Odense, Denmark
19Gundersen Lutheran Health System, La Crosse, WI
20University Hospital Nijmegen, Nijmegen, NLD
21The first affiliated hospital of Zhengzhou University, Zhengzhou, CHN
22Hospital Universitario Marques Valdecilla, Santander, Cantabria, ESP
23Robert H. Lurie Comp. Cancer Center, Chicago, IL
24NeoGenomics Laboratories, Aliso Viejo, CA
25Adaptive Biotechnologies, Seattle
26the First Affiliated Hospital of Zhengzhou University, Zhengzhou, CHN
27University of Arkansas For Medical Sciences, Little Rock, AR
28The First Affiliated Hospital of Xiamen University and Institute of Hematology, Xiamen, China
29Duke Cancer Institute, Duke University Medical Center, Durham, NC

Introduction: In recent years, increasing evidence suggests an important role of tumor-infiltrating B lymphocytes (TIL-Bs) in cancer immunity. In various types of solid tumors, high density of TIL-Bs and presence of mature tertiary lymphoid structures consistently correlate with better prognosis and response to immune checkpoint blockade therapies. Diffuse large B-cell lymphoma (DLBCL) is a type of aggressive lymphoma arising from mature B cells. TIL-Bs have been identified in DLBCL based on single-cell gene-expression data in several previous studies, but their prognostic significance in DLBCL is unknown. In this study, we determined the clinical significance of TIL-B abundance and investigated the potential underlying mechanisms.

Patients and Methods: This abstract includes a discovery study in which we distinguished TIL-Bs from malignant B-cells by immunoglobulin gene (IG) clonotype analysis and a validation study using flow cytometry. In the discovery study, a molecularly well-characterized DLBCL cohort assembled by the International DLBCL R-CHOP Consortium Program was analyzed. DNA samples from DLBCL patients were sequenced by high-throughput ultra-deep sequencing assays using multiplexed forward and reverse primers. Based on the IG heavy chain (IGH) sequencing results, malignant B-cell and TIL-B VDJ clonotypes were identified for 138 patients, and the frequencies and numbers of TIL-B clonotypes in individual sequence repertoires were calculated. In 79 DLBCL cases, a DJ-only sequence was predominant and no productive VDJ sequences met the criteria for diagnostic sequences. Total frequencies of productive VDJ sequences in the sequence repertoires were then used to access high and low TIL-B frequencies. TIL-B frequencies were also evaluated by IG light chain clonotypes for these 217 patients and additional 52 patients. The TIL-B frequencies were correlated with patient survival, gene-expression profiling (GEP) data GSE31312, and frequencies of DLBCL-infiltrating immune cells quantified by fluorescent multiplex immunohistochemistry (mIHC). To validate the prognostic significance of TIL-B frequencies, we assembled a validation cohort of 94 DLBCL patients treated at Duke University Medical Center. TIL-Bs in tissues were quantified by flow cytometry immunophenotyping. Ratios of TIL-Bs to abnormal B cells were assessed for prognostic effects.

Results: First, we found that high frequencies of TIL-Bs and clonotypic diversity were associated with significantly favorable prognostic effects in 138 DLBCL patients with their malignant B-cell IGH-VDJ clonotypes identified (Figure A). The prognostic effects of TIL-Bs outweighed those of the germinal center B-cell-like/activated B-cell-like classification (Figure B). Second, patients with high frequencies of TIL-Bs compared with those without had significant upregulation of immune response genes by GEP (Figure C), higher CD4 T cell infiltration quantified by fluorescent mIHC (Figure D), and enrichment of EcoTyper-defined specific cell states, including significantly increased frequencies of B cell state S2 (pre-memory), CD4 T cell state S3 (naïve), follicular helper T cell state S3, and endothelial cell state S3, whereas lower frequencies of B cell state S5 (plasmablast) and follicular helper T cell state S1. Third, high TIL-B abundance in 79 'DJ-only' clonotype cases and additional 52 cases assessed by light chain clonotype analysis showed similar favorable prognostic effects and associations with upregulated immune gene expression and CD4 T cell infiltration. Fourth, the identified GEP signatures, prominent with genes involved in T-dependent B cell activation and T cell activation and differentiation, showed significant prognostic effects in two GEO datasets. Fifth, TIL-B abundance showed significant prognostic effects in DLBCL cases infiltrated with T-cells in both the discovery study (Figure E) and validation study (Figure F, flow cytometry cohort).

Conclusions: TIL-B frequencies and clonotypic diversity have remarkable prognostic and biological effects in DLBCL, which has significant implications for developing novel therapeutic strategies. Our study identified a new prognostic biomarker in DLBCL and an important player in anti-lymphoma immune responses, providing a new point of view to analyze the tumor biology in aggressive B-cell lymphoma and precisely identify prognostic determinants.

Disclosures: Snyder: Adaptive Biotechnologies: Ended employment in the past 24 months. Hsi: Novartis: Consultancy. Ponzoni: ABBVIE: Honoraria; Roche Ventana Diagnostics: Consultancy, Honoraria. Ferreri: Adienne: Speakers Bureau; ADC Therapeutics, Amgen, BeiGene, BMS, Genmab, Gilead, Hutchison Medipharma, Novartis, Pharmacyclics, PentixaPharm, Pfizer, Roche: Research Funding; Ospedale San Raffaele srl.: Patents & Royalties; Gilead, Incyte, Novartis, PentixaPharm, Roche: Consultancy. Au: NeoGenomics Laboratories: Current Employment. Kirsch: Adaptive Biotechnologies: Consultancy. Young: Flagship Biosciences: Consultancy; Arima Genomics: Consultancy.

*signifies non-member of ASH