COV434 granulosa cell line: take it or leave it?

Journal of Reproductive Immunology 101–102 (2014) 18–39 31 Methods:The trophoblastmodel cell line JEG-3was cultured on a chamber slide and either untreated or treated with LPS, poly(I:C) and both together for 24h, and stained HLA-C, -E, and -G molecules using two-colour sequential staining. Images were acquired using a Nikon confocal microscope and analysed by using a sparse colocalisation algorithm, as previously described (Asma Jabeen et al., 2013). Results: Treatment of LPS alone caused a significant increase in HLA-G/HLA-E colocalisations (38%) compared with an untreated control sample (20%). In the case of double infection, a rise in thenumber ofHLA-G/HLA-E colocalisations remained statistically insignificant, whereas poly(I:C) alone did not cause any detectable change. Colocalisations of HLA-E/HLA-C remained unaffected by LPS, poly(I:C) or double treatment on trophoblast cells. Similarly, HLA-G/HLA-C colocalisation was insignificantly increased by poly(I:C) alone and double treatment, but remained unaffected by LPS alone. Conclusion: Our data indicate a significant upregulation of HLA-G/HLA-E heterodimer formation in the face of LPS infection alone; there was no significant effect of LPS, poly(I:C) alone or dual infection on HLA-G/HLA-C and HLA-E/HLA-C colocalisation.