New year's work! Study on mass spectrometry flow cytometry of immune cells in helminth infection

Highlight of the paper: This study uses mass spectrometry (cytof) to study the European population (EU), uninfected Indonesian population (ID urban) and infected Indonesian population (ID) Through the data integration analysis of cell phenotype and function, the heterogeneity of human immune cells and immune regulatory network caused by worm infection were comprehensively evaluated Mass spectrometry flow cytometry breaks through the limitations of channel number, detection accuracy and analysis dimension in traditional research On the basis of improving the number and efficiency of single experiment, it makes the experimental data more abundant and complete More than 60% of the world's population is at risk of worm infection It is estimated that soil borne worms alone cause more than 1.5 billion infections, 24% of the population However, most people have chronic infection for up to 20 years [1] Therefore, it is very important for the prevention and treatment of helminth disease to elaborate the regulatory mechanism of immune cells in worm infection more comprehensively and systematically A large number of studies have shown that worms can regulate the response of mouse type 2 immune cells (ilc2) However, due to the low expression of ilc2 cells in human peripheral blood, there are some challenges in the research of human type 2 immune regulation caused by worms In 2020, Professor Maria yazdanbakhsh's team from Leiden University Medical Center in the Netherlands published a paper entitled "helminth infections drive ergonomicity in human type 2" on Science Translational Medicine And regulatory cells In this paper, the immune subsets including ilc2 and their functions in the peripheral blood of European (EU), uninfected Indonesian (ID urban) and infected Indonesian (ID rural) before and after treatment were detected and analyzed in detail by using the multi-channel detection and multi-dimensional Shengxin analysis of mass flow cytometry The results of this study provide a new strategy for more precise disease intervention aimed at these cells First of all, the researchers conducted hsne clustering analysis on the flow cytometry data of peripheral blood mass spectrum detected by EU and ID rural population before and after infection treatment Comprehensive data showed that immune cells were divided into CD4 +, There are six subsets of CD8 +, TCR γ δ, B cells, ILCs and myeloid cells, and the phenotype and function of these six subsets are heterogeneous (Fig Later, the researchers found that worm infection can increase the percentage of Th2 CD4 + cells (Fig.2 a-b) By further clustering analysis of Th2 cells, 22 cell subpopulations (fig.2c) were obtained These cells can express CD7 and klrg1 in reverse or not It is speculated that worm infection can make the effector cells in a terminal state and make CD161 + cells in a transitional state In addition, ID post can produce more CD7 + Th2 cells with low expression of GATA3 and crht2, suggesting that worm infection will reduce the production of type 2 immune factor cells (fig.2d) Cluster analysis of ILCs subsets showed that the proportion of ilc2 (CD127 + c161 + CRTH2 +) cells in ID post immune cells was significantly higher than that in EU (fig.3a), and worm infection could also reduce the proportion of klrg1 + CCR6 + ilc2s (cluster 5) cells in ilc2 subsets (fig.3b) Treg, as a functional subgroup of suppressive T cells, plays an important role in worm specific immune response It was found that the percentage of CTLA-4 + Treg and CD161 + Treg cells in ID pre was higher than that in EU, and the ratio decreased after treatment (Fig.3 C-D) Worm infection can also stimulate the expression of CTLA-4 + HLA-DR + CD38 + ICOS + Treg cells (Fig.3) E) The results showed that the subpopulation was closely related to the immune response after worm infection Further study showed that the expression of IL-4 / IL-5 / IL-13 + cells was mainly in CD8 +, CD4 +, ilc2 cells and TCR γ δ cells (fig.4a) In addition, the proportion of immune 2 cytokines in ID pre group was higher, indicating that worm infection can stimulate more immune cells to produce cytokines After one year of treatment, the proportion decreased (Fig 4b), which was mainly caused by the decrease of CD4 +, ilc2 and CD8 + cells In addition, it was found that IL-4 / 5 / 13 + cell / CD8 + in CD8 + cells was linearly correlated with CD45RO + CCR7 − CD161 − CD56 − CD8 + T cells (r = 0.87) (fig.4c) It was found that worm infection could increase the percentage of CD11c + and IL-10 + ifn-y-il-2-tnf-a - / IL-10 + CD4 + cell subsets (Fig.5) Finally, the researchers analyzed the European population (EU), the Indonesian urban population and the Indonesian rural population by mass spectrometry and found that the European population and the Indonesian urban population have similar immune characteristics, indicating that the type 2 of worm infection and the change of regulatory immune cells are not due to ethnic differences To sum up, the heterogeneity and function of rare cell subsets (such as ilc2s, TC2 cells and bregs) and important cell subsets (Th2 cells and Treg cells) of immune regulation in worm infected patients were studied and analyzed by using the advantages of mass flow cytometry, which provided a new strategy for more precise disease intervention targeting these cells.