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Mass cytometry immunophenotyping data of two-week-old mouse pups' spleens depleted of maternal cells

Citation

Castellan, Flore (2022), Mass cytometry immunophenotyping data of two-week-old mouse pups' spleens depleted of maternal cells , Dryad, Dataset, https://doi.org/10.5061/dryad.7sqv9s4tz

Abstract

The maternal cells transferred into the fetus during gestation persist long after birth in the progeny. These maternal cells have been hypothesized to promote the maturation of the fetal immune system in utero but there are still significant gaps in our knowledge of their potential roles after birth. To provide insights into these maternal cells' postnatal functional roles, we set up a transgenic mouse model to specifically eliminate maternal cells in the neonates by diphtheria toxin injection and confirmed significant depletion in the spleens. We then performed immunophenotyping of the spleens of two-week-old pups by mass cytometry to pinpoint the immune profile differences driven by the depletion of maternal cells in early postnatal life. We observed a heightened expression of markers related to activation and maturation in some natural killer and T cell populations. We hypothesize these results to indicate a potential postnatal regulation of lymphocytic responses by maternal cells. Together, our findings highlight an immunological influence of maternal microchimeric cells postnatally, possibly protecting against adverse hypersensitivity reactions of the neonate at a crucial time of new encounters with self and environmental antigens.

Methods

To compare immune profiles in pups with or without maternal cells, B6CF1 control pups from crossing B6 females with Balb/c males (n=6; Ref), and DTR(-/-) sample pups from B6 DTR(+/-) females and Balb/c males (n=6; MMcDepleted) were injected i.p. every other day from postnatal day 3 until sacrifice day 15 with diphtheria toxin (8ng/g body weight). The pups from each group were obtained from the first litter of the dams and were all female. Spleen cell suspensions were prepared by mechanical dissociation using 70 μm nylon mesh (Axel), treated with red blood cells lysis buffer (Ammonium-Chloride-Potassium) and washed in staining buffer (HBSS(-), 2mM EDTA, 1% BSA), frozen at -80°C in Cell Banker 1 Plus, thawed and stained on the same day across experimental groups. The cells were stained using Maxpar Mouse Sp/LN Phenotyping Panel Kit (Fluidigm) containing the following Abs: Ly6G/C (RB6-8C5), CD11c (N418), CD69 (H1.2F3), CD45 (30-F11), CD11b (M1/70), CD19 (6D5), CD25 (3C7), CD3e (145-2C11), Ter-119 (TER119), CD62L (MEL-14), CD8a (53-6.7), TCRβ (H57-597), NK1.1 (PK136), CD44 (IM7), CD4 (RM4-5), B220 (RA3-6B2); with Cisplatin 198Pt used as a dead cells’ marker. Cells were fixed in 1.6% PFA, transported at 4 °C to Fluidigm Tokyo and analyzed on the Helios mass cytometry system. Normalization with beads was performed during acquisition. 

Usage Notes

The data files can be opened in R using the package openCyto, among others.  

Funding

Naito Foundation, Award: 6279:37

Takeda Science Foundation

Japan Society for the Promotion of Science, Award: 17K19547

Japan Society for the Promotion of Science, Award: 21K19254