Regulation of antigen-specific T cell infiltration and spatial architecture in multiple myeloma and premalignancy
M. Hope Robinson, … , Kavita M. Dhodapkar, Madhav V. Dhodapkar
M. Hope Robinson, … , Kavita M. Dhodapkar, Madhav V. Dhodapkar
Published August 1, 2023
Citation Information: J Clin Invest. 2023;133(15):e167629. https://doi.org/10.1172/JCI167629.
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Research Article Hematology Immunology

Regulation of antigen-specific T cell infiltration and spatial architecture in multiple myeloma and premalignancy

Abstract

Entry of antigen-specific T cells into human tumors is critical for immunotherapy, but the underlying mechanisms are poorly understood. Here, we combined high-dimensional spatial analyses with in vitro and in vivo modeling to study the mechanisms underlying immune infiltration in human multiple myeloma (MM) and its precursor monoclonal gammopathy of undetermined significance (MGUS). Clustered tumor growth was a feature of MM but not MGUS biopsies, and this growth pattern was reproduced in humanized mouse models. MM biopsies exhibited intralesional as well as spatial heterogeneity, with coexistence of T cell–rich and T cell–sparse regions and the presence of areas of T cell exclusion. In vitro studies demonstrated that T cell entry into MM clusters was regulated by agonistic signals and CD2-CD58 interactions. Upon adoptive transfer, antigen-specific T cells localized to the tumor site but required in situ DC–mediated antigen presentation for tumor entry. C-type lectin domain family 9 member A–positive (CLEC9A+) DCs appeared to mark portals of entry for gradients of T cell infiltration in MM biopsies, and their proximity to T cell factor 1–positive (TCF1+) T cells correlated with disease state and risk status. These data illustrate a role for tumor-associated DCs and in situ activation in promoting the infiltration of antigen-specific T cells in MM and provide insights into spatial alterations in tumor/immune cells with malignant evolution.

Authors

M. Hope Robinson, Nancy Y. Villa, David L. Jaye, Ajay K. Nooka, Alyssa Duffy, Samuel S. McCachren, Julia Manalo, Jeffrey M. Switchenko, Sierra Barnes, Sayalee Potdar, Maryam I. Azeem, Ava A. Horvat, Vaunita C. Parihar, Jingjing Gong, Yan Liang, Geoffrey H. Smith, Vikas A. Gupta, Lawrence H. Boise, Jonathan L. Kaufman, Craig C. Hofmeister, Nisha S. Joseph, Sagar Lonial, Kavita M. Dhodapkar, Madhav V. Dhodapkar

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Figure 3

DC-mediated antigen presentation and T cell entry.

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DC-mediated antigen presentation and T cell entry. (A–C) Effects of DCs ...
(A–C) Effects of DCs on in vitro T cell infiltration. U266-MP colonies were injected with MP-pulsed HLA-A2+ mo-DCs (or unpulsed DCs as controls) followed by injection of MP-specific HLA-A2-tetramer+ T cells (tetramer– T cells as a control) after 4 hours. T cell entry was quantified after overnight culture. For some experiments clusters were also labeled with PI to assess tumor cell lysis. (A) Effect of DC-mediated antigen (Ag) presentation on entry of antigen-specific T cells. n = 59–85. (B) PI staining showing killing of MM colonies. n = 54–89. (C) Effect of CD58 blockade on entry of antigen-specific T cells. n = 21–44. Data show the mean ± SEM. Each dot represents a distinct tumor cluster, and data were pooled from a minimum of 3 independent experiments. ****P < 0.0001, by Kruskal-Wallis test with Dunn’s multiple-comparison test (A–C). (D–F) Effects of DCs on in vivo T cell infiltration. U266-MP-rluc cells were first engrafted intrafemorally into MISTRG6 mice either as tumor cells alone, or with MP-pulsed, mo-DCs from an HLA-A2+ donor. Tumor growth was documented by IVIS, and mice were injected with T cells from the same donor, which had been previously expanded ex vivo using MP-pulsed DCs. mIF images are representative of 5 experiments with 3–7 mice per group. (D) Flow cytometric analysis showing selective enrichment of A2-MP-tetramer+ T cells at the tumor site. Note that most of the T cells in the spleen are tetramer–. (E) IHC images showing that T cells localizing to the tumor site did not efficiently enter the tumor clusters in tumors without human DCs (original magnification, ×24), but these T cells did so when the tumors contained DCs (original magnification, ×20). (F) Representative T cell clusters in bones of mice with tumor-associated DCs (original magnification, ×39.3). Tet, tetramer.