Extended Data Fig. 9: Flow cytometry gating for phenotyping analysis of B/innate immune cells. | Nature Medicine

Extended Data Fig. 9: Flow cytometry gating for phenotyping analysis of B/innate immune cells.

From: Safety, immunogenicity and efficacy of PfSPZ Vaccine against malaria in infants in western Kenya: a double-blind, randomized, placebo-controlled phase 2 trial

Extended Data Fig. 9

Cellular gating tree using the 25-color panel outlined in Supplementry Table 4. (A) Lineage gating includes gating on single cells, leukocytes, live cells, and then CD19 and CD3 are used to separate live B cells, T cells, and NK/myeloid cells. (B) T cell gating to identify γδ T cells (pan-γδ TCR) and mucosal-associated invariant T (MAIT) cells (CD161+TCRVα7.2+). (C) Gating for B cell populations. B cells were further divided into memory B cells (CD27+IgD+), immature B cells (CD10+CD21+), and atypical B cells (CD27-CD21+). Memory B cells were further divided into plasmablasts (CD38+CD20-), plasmodium (CSP)-specific B cells (CSP-probe+), and B cell isotypes using IgG and IgM antibodies. (D) Gating tree to identify various subsets of NK/myeloid cell populations. NK cell populations were first divided by CD56 expression. From the CD56- cell gate, monocyte subsets were characterized using CD14 and CD16. CD14-CD16- that were HLA-DR+ were used to identify dendritic cell (DC) populations including plasmacytoid DCs (pDCs) (CD123+CD11c-) myeloid-derived DCs (mDCs) (CD123-CD11c+). mDCs subsets were identified using CD1c and CD141.

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