| Title | Flow cytometric measurement of intracellular Th1 and Th2 cytokine production by human villous and extravillous cytotrophoblast. | | Author(s) | Sacks GP, Clover LM, Bainbridge DR, Redman CW, Sargent IL | | Institution | Nuffield Department of Obstetrics and Gynaecology, University of Oxford, John Radcliffe Hospital, Oxford, OX3 9DU, UK. | | Source | Placenta 2001 Jul; 22(6):550-9. | | MeSH | Cell Separation
Centrifugation, Density Gradient
Chorion
Chorionic Villi
Cytokines
Female
Flow Cytometry
Histocompatibility Antigens Class I
Humans
Interferon Type II
Interleukin-10
Interleukin-12
Interleukin-4
Keratin
Placenta
Pregnancy
Pregnancy Trimester, First
Pregnancy Trimester, Third
RNA, Messenger
Receptor, Epidermal Growth Factor
Receptor, erbB-2
Research Support, Non-U.S. Gov't
Trophoblasts
Tumor Necrosis Factor-alpha
| | Abstract | A wide variety of cytokines are present at the maternal-fetal interface, but the extreme cellular complexity of the placenta has made it difficult to determine which cytokines are produced by which cells. Hence novel flow cytometric methods have been applied to determine intracellular cytokine production by specific cell-types in placental cell suspensions. Cell suspensions were prepared from first and third trimester chorionic villi and third trimester amniochorion by enzymatic digestion and Percoll density gradient centrifugation. After overnight incubation in the presence of monensin, cells were fixed, permeabilized and labelled with antibodies for villous cytotrophoblast (cytokeratin+, MHC class I-), extravillous cytotrophoblast (cytokeratin+, MHC class 1+) and leucocytes (CD45+). These cell types were further characterized by their expression of EGFR (proliferative cytotrophoblast) and c-erbB2 (invasive cytotrophoblast). Production of IL-4, IL-10, TNF-alpha, IFN-gamma and IL-12 was determined by simultaneous labelling with the appropriate monoclonal antibodies. Only IL-4 was detected consistently in all samples of cytotrophoblast. IL-10 was not detected but IL-10 mRNA was demonstrated in third trimester chorionic villus digests by RT-PCR. Although IL-4 secretion has not been demonstrated, these data suggest that, in vivo there may be a "Th2 type cytokine bias" orchestrated by the trophoblast. It is proposed that other cytokines (including IL-10 and TNF-alpha) are produced by decidual leukocytes, and not cytotrophoblast, at the maternal-fetal interface. | | Language | eng | | Pub Type(s) | Journal Article
| | PubMed ID | 11440543 |
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