Generation, Characterization and Application of Antibodies Directed against HERV-H Gag Protein in Colorectal SamplesReport as inadecuate




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Introduction

A 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.

Results

The 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.

Conclusion

Taken together, the Gag-H antibody clones 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.



Author: Christina S. Mullins, Maja Hühns, Mathias Krohn, Sven Peters, Valérie Cheynet, Guy Oriol, Michèle Guillotte, Sandrine Ducrot,

Source: http://plos.srce.hr/



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