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siRNA Transfection and EMSA Analyses on Freshly Isolated Human Villous Cytotrophoblasts
Published on: September 20, 2016
Chemokine receptor expression by human syncytiotrophoblast
G C Douglas1, T L Thirkill, V Sideris
1Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Tupper Hall, Davis, CA 95616-8643, USA. gcdouglas@ucdavis.edu
Abstract:
Despite their potential importance in placental HIV infection and placental immune function, nothing is known about the expression of chemokine receptors by human syncytiotrophoblast cells. Immunocytochemical analysis revealed that primary cultures of term syncytiotrophoblast cells express CCR1, CCR3, CXCR4, and CCR6. Immunohistochemical examination of cryosections of term placental villous tissue confirmed the expression of CCR3, CXCR4, and CCR6 by trophoblast cells. The primary syncytiotrophoblast cultures showed no reactivity with antibodies against CCR5. In the villous tissue sections, CCR5 was detected in stromal cells and blood vessel walls but was not found in trophoblast cells. RT-PCR analysis of RNA extracted from cultured syncytiotrophoblast cells confirmed that the cells express message for CCR1, CCR3, CXCR4, CCR6 and CCR10. No transcripts corresponding to CCR2b, CCR5, or CCR8 were detected. Other experiments showed that exposure of syncytiotrophoblast cells to soluble SDF-1alpha elicited a calcium mobilization response, consistent with the expression of functional CXCR4. Thus, human syncytiotrophoblast cells express CXCR4, a known co-receptor for TCL-tropic HIV-1 isolates but do not express CCR5, the major co-receptor for M-tropic isolates. In addition to implications for the maternal-fetal transmission of HIV, the expression of chemokine receptors by syncytiotrophoblast cells could be important in other aspects of placental immune function.
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