Abstract
IntroductionA substantial part of the human genome originates from transposable elements, remnants of ancient retroviral infections. Roughly 8% of the human genome consists of about 400,000 LTR elements including human endogenous retrovirus (HERV) sequences. Mainly, the interplay between epigenetic and post-transcriptional mechanisms is thought to silence HERV expression in most physiological contexts. Interestingly, aberrant reactivation of several HERV-H loci appears specific to colorectal carcinoma (CRC).ResultsThe expression of HERV-H Gag proteins (Gag-H) was assessed using novel monoclonal mouse anti Gag-H antibodies. In a flow cytometry screen four antibody clones were tested on a panel of primary CRC cell lines and the most well performing ones were subsequently validated in western blot analysis. Finally, Gag-H protein expression was analyzed by immune histology on cell line cytospins and on clinical samples. There, we found a heterogeneous staining pattern with no background staining of endothelial, stromal and infiltrating immune cells but diffuse staining of the cytoplasm for positive tumor and normal crypt cells of the colonic epithelium.ConclusionTaken together, the Gag-H antibody clone(s) present a valuable tool for staining of cells with colonic origin and thus form the basis for future more detailed investigations. The observed Gag-H protein staining in colonic epithelium crypt cells demands profound analyses of a potential role for Gag-H in the normal physiology of the human gut.
Highlights
A substantial part of the human genome originates from transposable elements, remnants of ancient retroviral infections
8% of the human genome consists of about 400,000 LTR elements including human endogenous retrovirus (HERV) sequences
In a flow cytometry screen four antibody clones were tested on a panel of primary CRC cell lines and the most well performing ones were subsequently validated in western blot analysis
Summary
A substantial part of the human genome originates from transposable elements, remnants of ancient retroviral infections. 8% of the human genome consists of about 400,000 LTR elements including human endogenous retrovirus (HERV) sequences. The interplay between epigenetic and post-transcriptional mechanisms is thought to silence HERV expression in most physiological contexts. Data Availability Statement: All relevant data are within the paper and its Supporting Information files. De, funded CSM with a PostDoc scholarship. CSM received a “Mildred Scheel PostDoc Stipendium” from the German Cancer Aid The funder provided support, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the
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