Charmsaz Soren, Gross Nicole, Jaffee Elizabeth, Ho Won Jin
JCI Insight. 2021 Apr 8;6(7):143283. doi: 10.1172/jci.insight.143283.
With the advent of cancer immunology, mass cytometry has been increasingly employed to characterize the responses to cancer therapies and the tumor microenvironment (TME). One of its most notable applications is efficient multiplexing of samples into batches by dedicating a number of metal isotope channels to barcodes, enabling robust data acquisition and analysis. Barcoding is most effective when markers are present in all cells of interest. While CD45 has been shown to be a reliable marker for barcoding all immune cells in a given sample, a strategy to reliably barcode mouse cancer cells has not been demonstrated. To this end, we identified CD29 and CD98 as markers widely expressed by commonly used mouse cancer cell lines. We conjugated anti-CD29 and anti-CD98 antibodies to cadmium or indium metals and validated their utility in 10-plex barcoding of live cells. Finally, we established a potentially novel barcoding system incorporating the combination of CD29, CD98, and CD45 to multiplex 10 tumors from s.c. MC38 and KPC tumor models, while successfully recapitulating the known contrast in the PD1-PDL1 axis between the 2 models. The ability to barcode tumor cells along with immune cells empowers the interrogation of the tumor-immune interactions in mouse TME studies.
随着癌症免疫学的出现，质谱流式细胞术越来越多地用于表征对癌症治疗的反应和肿瘤微环境（TME）。其最显著的应用之一是通过将一些金属同位素通道用于条形码，将样本高效地多路复用成批次，从而实现强大的数据采集和分析。当标记物存在于所有感兴趣的细胞中时，条形码技术最为有效。虽然CD45已被证明是在给定样本中对所有免疫细胞进行条形码标记的可靠标记物，但尚未证明一种可靠地对小鼠癌细胞进行条形码标记的策略。为此，我们将CD29和CD98鉴定为常用小鼠癌细胞系广泛表达的标记物。我们将抗CD29和抗CD98抗体与镉或铟金属偶联，并验证了它们在活细胞10重条形码标记中的效用。最后，我们建立了一个潜在的新型条形码系统，该系统结合了CD29、CD98和CD45，对来自皮下MC38和KPC肿瘤模型的10个肿瘤进行多路复用，同时成功再现了这两种模型之间已知的PD1-PDL1轴对比。对肿瘤细胞和免疫细胞进行条形码标记的能力有助于在小鼠TME研究中探究肿瘤-免疫相互作用。
