PO.TB10.06 · 肿瘤生物学
淋巴渗出液是肿瘤相关免疫细胞的新型来源
Lymphatic exudate is a novel source of tumor-associated immune cells
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
自体肿瘤浸润淋巴细胞（TIL）疗法已在多种实体瘤类型中显示出疗效，但该疗法需要手术及大量的制备工作。淋巴渗出液（“淋巴液”）通常在肿瘤切除术后从手术引流管中常规收集。虽然通常被丢弃，但我们已证明这种新型近端生物体液富含ctDNA，且对头颈部鳞状细胞癌（HNSCC）患者的复发具有预后价值。在此，我们研究淋巴液作为肿瘤相关淋巴细胞的一种新型、可获取的来源。
术后24小时从11例HPV-的HNSCC患者和14例膀胱癌（BLC）患者中，使用K₂EDTA采血管收集淋巴液。从淋巴液中分离细胞，并通过流式细胞术表征免疫细胞群。对17组具有匹配肿瘤的患者样本（11例HNSCC，6例BLC）进行了批量T细胞受体β（TCR-b）测序。2例BLC患者未通过测序质控。比较了肿瘤与淋巴液之间的TCR谱，并使用Simpson克隆性指数评估克隆多样性。
淋巴液细胞显示出高活力（>85%）。免疫亚群见表1。同时检测到CD4⁺和CD8⁺ T细胞，CD4/CD8比值可变（HNSCC为1.05-4.9，BLC为0.11-8.869）。批量TCR-b分析显示淋巴液与肿瘤之间存在大量克隆重叠（HNSCC平均13.1%[0-33.1%]，BLC平均9.1%[0.3-23.8%]），超过了外周血通常所报道的重叠程度。淋巴液TCR表现出克隆扩增（Simpson克隆性均值：HNSCC=0.58%，BLC=0.64%）。根据手术引流速率（中位数1-3.5 mL/小时）及每毫升CD3+细胞的中位数，估计每天可从引流管中回收约4.8×10⁶-1.68×10⁷个T细胞。
表1. 按适应证经FACS测定的平均细胞频率；免疫亚群以CD45的百分比表示 HNSCC BLC CD45+ 95.36% 85.85% T细胞 3.92% 15.08% NKT细胞 0.78% 2.63% B细胞 0.55% 3.60% 单核细胞 0.37% 7.90%
术后淋巴液含有具活力的免疫细胞群，且与肿瘤浸润淋巴细胞存在大量TCR重叠。淋巴液可在无需转移灶切除相关挑战的情况下实现肿瘤相关T细胞的分离。淋巴液T细胞可能代表一种用于过继细胞疗法的新型、丰富、低成本且易于获取的来源。
查看英文原文 English abstract
Autologous tumor-infiltrating lymphocyte (TIL) therapy has demonstrated efficacy in multiple solid tumor types, but therapy requires surgery and extensive manufacturing. Lymphatic exudate (“lymph”) is routinely collected from surgical drains following tumor resection. Although usually discarded, we have shown that this novel proximal biofluid is rich in ctDNA and prognostic of recurrence in head and neck squamous carcinoma (HNSCC) patients. Here, we investigate lymph as a novel, accessible source of tumor-associated lymphocytes.
Lymph was collected 24 hours post-surgery from 11 HPV- HNSCC and 14 bladder cancer (BLC) patients in K₂EDTA blood collection tubes. Cells were isolated from lymph and immune populations were characterized by flow cytometry. Bulk T-cell receptor b (TCR-b) sequencing was performed on 17 patient sets with matched tumor (11 HNSCC, 6 BLC). Two BLC patients failed sequencing QC. TCR repertoires were compared between tumor and lymph and Simpsons clonality was used to assess clonal diversity.
Lymph cells displayed high viability (>85%). Immune subpopulations are described in Table 1. Both CD4⁺ and CD8⁺ T cells were detected with variable CD4/CD8 ratios (1.05-4.9 [HNSCC] and 0.11-8.869 [BLC]). Bulk TCR-b analysis showed substantial clonal overlap between lymph and tumor (HNSCC mean 13.1% [0-33.1%] and BLC 9.1% [0.3-23.8%]), exceeding typical overlap described for peripheral blood. Lymph TCRs exhibited clonal expansion (Simpsons clonality means: HNSCC = 0.58% and BLC = 0.64%). Based on surgical drainage rates (median 1-3.5 mL/hour) and median number of CD3+ cells per mL, an estimated 4.8 x 10 6 - 1.68 x 10 7 T cells could be recovered daily from drains.
Table 1. Mean cell frequencies by indication measured by FACS; immune subsets expressed as % of CD45 HNSCC BLC CD45+ 95.36% 85.85% T cells 3.92% 15.08% NKT cells 0.78% 2.63% B cells 0.55% 3.60% Monocytes 0.37% 7.90%
Post-surgical lymph contains viable immune populations with substantial TCR overlap with tumor infiltrating lymphocytes. Lymph could enable the isolation of tumor-associated T cells without the challenges around metastasectomy. Lymph T cells may represent a novel, abundant, low cost, and easily accessible source for adoptive cell therapy.
利益披露 Disclosure
S. Lazare, None..
M. A. Harmon, None..
A. Tellis, None..
Z. Costliow, None..
Z. Gu, None..
A. Benson, None..
S. Espinoza, None..
A. Crittenden, None..
M. Rivera, None..
M. Abern, None..
S. Patel, None..
K. Pohar, None..
G. Agarwal, None..
R. Correa, None.