Publications

Distinct injury responsive regulatory T cells identified by multi-dimensional phenotyping

Guo et al. Front Immunol (2022)

Mass cytometry in combination with RNA and TCR sequencing identified a distinct CD44high subset of Tregs that activate and proliferate in response to injury

Imbalances in regulatory T cells (Tregs) can predispose people to secondary infections or inflammation-mediated complications following severe injury. Researchers in the Lederer Lab at Brigham and Women’s Hospital and Harvard Medical School previously identified a distinct subtype of Tregs that are activated in response to injury, identified by high expression of the marker CD44 (CD44high Tregs). Here, the team used mass cytometry in combination with RNA sequencing and TCR repertoire analytical methods to characterize injury responsive Treg subsets in mice to understand how this balance is changed by injury.

RNA sequencing identified unique gene signatures of CD44high Tregs, including genes associated with cell migration and trafficking to sites of inflammation. TCR repertoire analysis also showed that injury increased the diversity of CD44high Tregs more than other Treg subsets. The group then used a 39-marker mass cytometry panel to validate these gene signatures at the protein level. In support of their findings, unsupervised analysis of the mass cytometry data identified two clusters corresponding to the CD44high and CD44low Treg subsets, with only the CD44high Treg cluster increasing in abundance 7 days after injury.

Using a multi-platform approach, the group was able to answer their research hypothesis in an unbiased manner. Further, by identifying and validating the differential expression of protein markers on CD44high Tregs using mass cytometry, the researchers may have identified targets for fine-tuning the balance of these populations to guide patients safely through recovery from injury.

Reference citation: Guo F, Hancock B, Griffith A, Lin H, Howard K, Keegan J, Zhang F, Chicoine A, Cahill L, Ng J and Lederer J. 2022. Distinct Injury Responsive Regulatory T Cells Identified by Multi-Dimensional Phenotyping. Front. Immunol. 13:833100. doi: 10.3389/fimmu.2022.833100

The complementary capabilities of single-cell RNA seq and high-dimensional cytometry

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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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