Publications

Comprehensive immune monitoring of clinical trials to advance human immunotherapy

Hartmann et al. Cell Rep (2019)

Immune cell types and subsets detectable with the 33-marker panel backbone

Immune monitoring has been used to study how drugs work and to identify signals within the immune system that can predict whether a drug treatment is likely to succeed. Conventional flow cytometry has traditionally been used for immune monitoring of clinical trials, but a typical panel taps out at around 6-10 markers. Matthew Spitzer, UCSF Associate Professor and scientific cofounder of Teiko Bio, is a senior author on this paper where they team developed a 33-marker mass cytometry panel with 10 open channels for customization that can be used to characterize all major human immune cell lineages.

Panel Design

The researchers selected phenotypic markers to identify and classify major immune cell populations, including T cells and their subpopulations, B cells at various stages of maturation, and subsets of NK cells, among others. They also included functional state markers to determine activation states, proliferation, exhaustion states, and levels of immune checkpoint molecules.

Full list of the 33 immune profiling markers and their functions, organized by immune cell type

Panel validation

To validate the panel’s coverage of immune subsets and replicability, the researchers profiled peripheral blood mononuclear cells (PBMCs) from healthy donors, and successfully assigned 98% of the cells to their corresponding immune cell lineages and functional subsets. Next, the panel was tested on PBMC samples from five healthy donors that were sent to two different research centers, where they were run using different instruments, reagents, and operators. The results showed a strong correlation between the two runs, with an r-value of 0.96. Additionally, this mass cytometry panel demonstrated high agreement with four flow cytometry panels that used the same markers, with an r-value of 0.98.

Panel clinical utilization in GvHD

Researchers validated the effectiveness of this panel on bone marrow from 15 patients with leukemia who underwent bone marrow transplantation and found that a reduction in CD27- B cells and the frequency of naive CD4+ T cells were associated with the development of graft-versus-host disease. This demonstrates the clinical utility of the panel in predicting and monitoring responses to treatment.

Immune profiling of bone marrow transplant patients shows differences in frequencies of specific B cell and T cell susbets between those with GvHD and those without

The paper highlights the value of utilizing a standardized mass cytometry panel for comprehensive immune profiling of all major immune subsets in clinical trial samples. This approach not only supports clinical development but also enables the identification of important biomarkers that may inform personalized treatment strategies.

Reference Citation: Hartmann FJ, Babdor J, Gherardini PF, Amir ED, Jones K, Sahaf B, Marquez DM, Krutzik P, O’Donnell E, Sigal N, Maecker HT, Meyer E, Spitzer MH, Bendall SC. Comprehensive Immune Monitoring of Clinical Trials to Advance Human Immunotherapy. Cell Rep. 2019 Jul 16;28(3):819-831.e4. doi: 10.1016/j.celrep.2019.06.049.

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A ring of grey and black in the center split into 3 sections with the words Flow, Mass, Spectral going around it clockwise. A dashed line coming out from the ring separates the rest of the space into three sections. Surrounding the ring are six colored circles each with a white icon. The circles directly opposite each other are the same color, but different shades (red opposite pink, blue opposite light blue, green opposite light green). Red shows a pencil, pink shows a grid-like matrix, Blue shows four dials, light blue shows only two; green shows a heavy metal spectrum with distinct lines, light green shows fluorescence spectrum with overlap. The green circle is in the Mass segment, the red overlaps between Mass and Flow, the Blue overlaps between Mass and Spectral. The Mass segment is the only one without any light colored circles.

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