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

PLoS One. 2016 Apr 27;11(4):e0153349. doi: 10.1371/journal.pone.0153349. eCollection 2016.

Abstract

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 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / immunology*
  • Cell Line
  • Colon / immunology*
  • Colon / virology*
  • Colorectal Neoplasms / immunology
  • Colorectal Neoplasms / virology
  • Cytoplasm / immunology
  • Cytoplasm / virology
  • Endogenous Retroviruses / immunology*
  • Female
  • Gene Products, gag / immunology*
  • Genome, Human / genetics
  • HEK293 Cells
  • Humans
  • Intestinal Mucosa / immunology
  • Intestinal Mucosa / virology
  • Mice
  • Mice, Inbred BALB C
  • Recombinant Proteins / immunology

Substances

  • Antibodies, Monoclonal
  • Gene Products, gag
  • Recombinant Proteins

Grants and funding

Deutsche Krebshilfe, http://www.krebshilfe.de, funded CSM with a PostDoc scholarship. This work was supported by bioMérieux SA in the form of salaries for authors [VC, GO, MG, SD and FM]. CSM received a “Mildred Scheel PostDoc Stipendium” from the German Cancer Aid (DKH e.V. #110943). 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 manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.