Using Mass Cytometry to Analyze the Tumor-Infiltrating Lymphocytes in Human Melanoma

Tantalo, Daniela; Nguyen, Thu Quynh; Yeang, Han Xian Aw; Zhu, Joe Jiang; Macdonald, Sean; Wang, Minyu; Silva, Harini de; D’Souza, Criselle; Pizzolla, Angela; Neeson, Paul J.

doi:10.1007/978-1-0716-1205-7_38

Cited by 2 publications

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“…From the obtained single-cell suspensions, 3×10 6 cells per sample were added to a 1.5 mL centrifuge tube for live and dead cell staining and mass spectrometry staining experiments. 19 …”

Section: Methodsmentioning

confidence: 99%

Hypoxia inhibits the iMo/cDC2/CD8+ TRMs immune axis in the tumor microenvironment of human esophageal cancer

Wu,

Yu,

Liang

et al. 2024

J Immunother Cancer

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BackgroundEsophageal cancer (ESCA) is a form of malignant tumor associated with chronic inflammation and immune dysregulation. However, the specific immune status and key mechanisms of immune regulation in this disease require further exploration.MethodsTo investigate the features of the human ESCA tumor immune microenvironment and its possible regulation, we performed mass cytometry by time of flight, single-cell RNA sequencing, multicolor fluorescence staining of tissue, and flow cytometry analyses on tumor and paracancerous tissue from treatment-naïve patients.ResultsWe depicted the immune landscape of the ESCA and revealed that CD8+(tissue-resident memory CD8+T cells (CD8+TRMs) were closely related to disease progression. We also revealed the heterogeneity of CD8+TRMs in the ESCA tumor microenvironment (TME), which was associated with their differentiation and function. Moreover, the subset of CD8+TRMs in tumor (called tTRMs) that expressed high levels of granzyme B and immune checkpoints was markedly decreased in the TME of advanced ESCA. We showed that tTRMs are tumor effector cells preactivated in the TME. We then demonstrated that conventional dendritic cells (cDC2s) derived from intermediate monocytes (iMos) are essential for maintaining the proliferation of CD8+TRMs in the TME. Our preliminary study showed that hypoxia can promote the apoptosis of iMos and impede the maturation of cDC2s, which in turn reduces the proliferative capacity of CD8+TRMs, thereby contributing to the progression of cancer.ConclusionsOur study revealed the essential antitumor roles of CD8+TRMs and preliminarily explored the regulation of the iMo/cDC2/CD8+TRM immune axis in the human ESCA TME.

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“…From the obtained single-cell suspensions, 3×10 6 cells per sample were added to a 1.5 mL centrifuge tube for live and dead cell staining and mass spectrometry staining experiments. 19 …”

Section: Methodsmentioning

confidence: 99%

Hypoxia inhibits the iMo/cDC2/CD8+ TRMs immune axis in the tumor microenvironment of human esophageal cancer

Wu,

Yu,

Liang

et al. 2024

J Immunother Cancer

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CyTOF® for the Masses

2022

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Mass cytometry has revolutionized immunophenotyping, particularly in exploratory settings where simultaneous breadth and depth of characterization of immune populations is needed with limited samples such as in preclinical and clinical tumor immunotherapy. Mass cytometry is also a powerful tool for single-cell immunological assays, especially for complex and simultaneous characterization of diverse intratumoral immune subsets or immunotherapeutic cell populations. Through the elimination of spectral overlap seen in optical flow cytometry by replacement of fluorescent labels with metal isotopes, mass cytometry allows, on average, robust analysis of 60 individual parameters simultaneously. This is, however, associated with significantly increased complexity in the design, execution, and interpretation of mass cytometry experiments. To address the key pitfalls associated with the fragmentation, complexity, and analysis of data in mass cytometry for immunologists who are novices to these techniques, we have developed a comprehensive resource guide. Included in this review are experiment and panel design, antibody conjugations, sample staining, sample acquisition, and data pre-processing and analysis. Where feasible multiple resources for the same process are compared, allowing researchers experienced in flow cytometry but with minimal mass cytometry expertise to develop a data-driven and streamlined project workflow. It is our hope that this manuscript will prove a useful resource for both beginning and advanced users of mass cytometry.

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Using Mass Cytometry to Analyze the Tumor-Infiltrating Lymphocytes in Human Melanoma

Cited by 2 publications

References 1 publication

Hypoxia inhibits the iMo/cDC2/CD8+ TRMs immune axis in the tumor microenvironment of human esophageal cancer

Hypoxia inhibits the iMo/cDC2/CD8+ TRMs immune axis in the tumor microenvironment of human esophageal cancer

CyTOF® for the Masses

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