MAIT Cells pp 55-70 | Cite as

Isolation of Immune Cells from Placental Tissues and Phenotypic and Functional Analysis of MAIT Cells

  • Martin SoldersEmail author
  • Laia Gorchs
  • Helen Kaipe
Part of the Methods in Molecular Biology book series (MIMB, volume 2098)


The placenta is an immunological paradox since maternal immune cells infiltrating placental tissues need to be tolerant toward the fetus but still retain immunity against potential infections. This makes the placenta an interesting tissue for studying immunological processes. Mucosal-associated invariant T (MAIT) cells are a subset of T cells that respond to bacterially derived metabolites of riboflavin synthesis. Upon activation, MAIT cells respond by secretion of inflammatory cytokines and by directed killing of infected cells by the use of granzymes and perforin. In this protocol, we describe methods for the isolation of immune cells from the placental intervillous space and adjacent tissues such as the umbilical cord, decidua parietalis, and decidua basalis. We further describe how to stimulate MAIT cells in mixed cell suspensions of mononuclear cells with bacteria, and how to analyze the phenotypic and functional responses with flow cytometry.

Key words

MAIT cells Placenta Intervillous blood Decidua 



H.K. was supported by the Swedish Research Council, the Swedish Childhood Cancer Foundation, the Cancer Society in Stockholm, the Swedish Cancer Foundation, Stockholm County Council, and Karolinska Institutet. MS was supported by Karolinska Institutet and Stockholm County Council.


  1. 1.
    Kay HH, Nelson DM, Wang Y (2011) The placenta from development to disease. Chichester, West SussexGoogle Scholar
  2. 2.
    Bartmann C, Segerer SE, Rieger L, Kapp M, Sutterlin M, Kammerer U (2014) Quantification of the predominant immune cell populations in decidua throughout human pregnancy. Am J Reprod Immunol (New York, NY : 1989) 71(2):109–119. Scholar
  3. 3.
    Solders M, Gorchs L, Gidlof S, Tiblad E, Lundell AC, Kaipe H (2017) Maternal adaptive immune cells in decidua parietalis display a more activated and coinhibitory phenotype compared to decidua basalis. Stem Cells Int 2017:8010961. Scholar
  4. 4.
    Solders M, Gorchs L, Erkers T, Lundell AC, Nava S, Gidlof S, Tiblad E, Magalhaes I, Kaipe H (2017) MAIT cells accumulate in placental intervillous space and display a highly cytotoxic phenotype upon bacterial stimulation. Sci Rep 7(1):6123. Scholar
  5. 5.
    Lashley LE, van der Hoorn ML, van der Mast BJ, Tilburgs T, van der Lee N, van der Keur C, van Beelen E, Roelen DL, Claas FH, Scherjon SA (2011) Changes in cytokine production and composition of peripheral blood leukocytes during pregnancy are not associated with a difference in the proliferative immune response to the fetus. Hum Immunol 72(10):805–811. Scholar
  6. 6.
    Tilburgs T, Scherjon SA, van der Mast BJ, Haasnoot GW, Versteeg VDV-MM, Roelen DL, van Rood JJ, Claas FH (2009) Fetal-maternal HLA-C mismatch is associated with decidual T cell activation and induction of functional T regulatory cells. J Reprod Immunol 82(2):148–157. Scholar
  7. 7.
    Apps R, Murphy SP, Fernando R, Gardner L, Ahad T, Moffett A (2009) Human leucocyte antigen (HLA) expression of primary trophoblast cells and placental cell lines, determined using single antigen beads to characterize allotype specificities of anti-HLA antibodies. Immunology 127(1):26–39. Scholar
  8. 8.
    Persson G, Melsted WN, Nilsson LL, Hviid TVF (2017) HLA class Ib in pregnancy and pregnancy-related disorders. Immunogenetics 69(8–9):581–595. Scholar
  9. 9.
    Kjer-Nielsen L, Patel O, Corbett AJ, Le Nours J, Meehan B, Liu L, Bhati M, Chen Z, Kostenko L, Reantragoon R, Williamson NA, Purcell AW, Dudek NL, McConville MJ, O’Hair RA, Khairallah GN, Godfrey DI, Fairlie DP, Rossjohn J, McCluskey J (2012) MR1 presents microbial vitamin B metabolites to MAIT cells. Nature 491(7426):717–723. Scholar
  10. 10.
    Treiner E, Duban L, Bahram S, Radosavljevic M, Wanner V, Tilloy F, Affaticati P, Gilfillan S, Lantz O (2003) Selection of evolutionarily conserved mucosal-associated invariant T cells by MR1. Nature 422(6928):164–169. Scholar
  11. 11.
    Lepore M, Kalinichenko A, Colone A, Paleja B, Singhal A, Tschumi A, Lee B, Poidinger M, Zolezzi F, Quagliata L, Sander P, Newell E, Bertoletti A, Terracciano L, De Libero G, Mori L (2014) Parallel T-cell cloning and deep sequencing of human MAIT cells reveal stable oligoclonal TCRbeta repertoire. Nat Commun 5:3866. Scholar
  12. 12.
    Dias J, Leeansyah E, Sandberg JK (2017) Multiple layers of heterogeneity and subset diversity in human MAIT cell responses to distinct microorganisms and to innate cytokines. Proc Natl Acad Sci U S A. Scholar
  13. 13.
    Solders M, Gorchs L, Tiblad E, Gidlof S, Leeansyah E, Dias J, Sandberg JK, Magalhaes I, Lundell AC, Kaipe H (2019) Recruitment of MAIT cells to the intervillous space of the placenta by placenta-derived chemokines. Front Immunol 10:1300. Scholar
  14. 14.
    Gold MC, Eid T, Smyk-Pearson S, Eberling Y, Swarbrick GM, Langley SM, Streeter PR, Lewinsohn DA, Lewinsohn DM (2013) Human thymic MR1-restricted MAIT cells are innate pathogen-reactive effectors that adapt following thymic egress. Mucosal Immunol 6(1):35–44. Scholar
  15. 15.
    Ostblom A, Adlerberth I, Wold AE, Nowrouzian FL (2011) Pathogenicity island markers, virulence determinants malX and usp, and the capacity of Escherichia coli to persist in infants’ commensal microbiotas. Appl Environ Microbiol 77(7):2303–2308. Scholar
  16. 16.
    Kurioka A, Ussher JE, Cosgrove C, Clough C, Fergusson JR, Smith K, Kang YH, Walker LJ, Hansen TH, Willberg CB, Klenerman P (2015) MAIT cells are licensed through granzyme exchange to kill bacterially sensitized targets. Mucosal Immunol 8(2):429–440. Scholar
  17. 17.
    Le Bourhis L, Dusseaux M, Bohineust A, Bessoles S, Martin E, Premel V, Core M, Sleurs D, Serriari NE, Treiner E, Hivroz C, Sansonetti P, Gougeon ML, Soudais C, Lantz O (2013) MAIT cells detect and efficiently lyse bacterially-infected epithelial cells. PLoS Pathog 9(10):e1003681. Scholar
  18. 18.
    Gold MC, Cerri S, Smyk-Pearson S, Cansler ME, Vogt TM, Delepine J, Winata E, Swarbrick GM, Chua WJ, Yu YY, Lantz O, Cook MS, Null MD, Jacoby DB, Harriff MJ, Lewinsohn DA, Hansen TH, Lewinsohn DM (2010) Human mucosal associated invariant T cells detect bacterially infected cells. PLoS Biol 8(6):e1000407. Scholar
  19. 19.
    Le Bourhis L, Martin E, Peguillet I, Guihot A, Froux N, Core M, Levy E, Dusseaux M, Meyssonnier V, Premel V, Ngo C, Riteau B, Duban L, Robert D, Huang S, Rottman M, Soudais C, Lantz O (2010) Antimicrobial activity of mucosal-associated invariant T cells. Nat Immunol 11(8):701–708. Scholar
  20. 20.
    Chatila T, Silverman L, Miller R, Geha R (1989) Mechanisms of T cell activation by the calcium ionophore ionomycin. J Immunol 143(4):1283–1289PubMedGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Department of Laboratory MedicineKarolinska InstitutetStockholmSweden
  2. 2.Clinical Immunology and Transfusion MedicineKarolinska University HospitalStockholmSweden

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