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Charest-Morin X, Pépin R, Gagné-Henley A, et al. C-C chemokine receptor-7 mediated endocytosis of antibody cargoes into intact cells. Front Pharmacol. 2013;4:122doi: 10.3389/fphar.2013.00122.
Charest-Morin, X., Pépin, R., Gagné-Henley, A., Morissette, G., Lodge, R., & Marceau, F. (2013). C-C chemokine receptor-7 mediated endocytosis of antibody cargoes into intact cells. Frontiers in pharmacology, 4122. https://doi.org/10.3389/fphar.2013.00122
Charest-Morin, Xavier, et al. "C-C chemokine receptor-7 mediated endocytosis of antibody cargoes into intact cells." Frontiers in pharmacology vol. 4 (2013): 122. doi: https://doi.org/10.3389/fphar.2013.00122
Charest-Morin X, Pépin R, Gagné-Henley A, Morissette G, Lodge R, Marceau F. C-C chemokine receptor-7 mediated endocytosis of antibody cargoes into intact cells. Front Pharmacol. 2013 Sep 24;4:122. doi: 10.3389/fphar.2013.00122. eCollection 2013. PMID: 24068998; PMCID: PMC3781320.
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Front Pharmacol. 2013 Sep 24;4:122. doi: 10.3389/fphar.2013.00122. eCollection 2013.

C-C chemokine receptor-7 mediated endocytosis of antibody cargoes into intact cells.

Frontiers in pharmacology

Xavier Charest-Morin, Rémy Pépin, Angélique Gagné-Henley, Guillaume Morissette, Robert Lodge, François Marceau

Affiliations

  1. Centre de Recherche en Rhumatologie et Immunologie, Centre Hospitalier Universitaire de Québec Québec, QC, Canada.

PMID: 24068998 PMCID: PMC3781320 DOI: 10.3389/fphar.2013.00122

Abstract

The C-C chemokine receptor-7 (CCR7) is a G protein coupled receptor that has a role in leukocyte homing, but that is also expressed in aggressive tumor cells. Preclinical research supports that CCR7 is a valid target in oncology. In view of the increasing availability of therapeutic monoclonal antibodies that carry cytotoxic cargoes, we studied the feasibility of forcing intact cells to internalize known monoclonal antibodies by exploiting the cycle of endocytosis and recycling triggered by the CCR7 agonist CCL19. Firstly, an anti-CCR7 antibody (CD197; clone 150503) labeled surface recombinant CCR7 expressed in intact HEK 293a cells and the fluorescent antibody was internalized following CCL19 treatment. Secondly, a recombinant myc-tagged CCL19 construction was exploited along the anti-myc monoclonal antibody 4A6. The myc-tagged ligand was produced as a conditioned medium of transfected HEK 293a cells that contained the equivalent of 430 ng/ml of immunoreactive CCL19 (average value, ELISA determination). CCL19-myc, but not authentic CCL19, carried the fluorophore-labeled antibody 4A6 into other recipient cells that expressed recombinant CCR7 (microscopy, cytofluorometry). The immune complexes were apparent in endosomal structures, co-localized well with the small GTPase Rab5 and progressed toward Rab7-positive endosomes. A dominant negative form of Rab5 (GDP-locked) inhibited this endocytosis. Further, endosomes in CCL19-myc- or CCL19-stimulated cells were positive for β-arrestin2, but rarely for β-arrestin1. Following treatment with CCL19-myc and the 4A6 antibody, the melanoma cell line A375 that expresses endogenous CCR7 was specifically stained using a secondary peroxidase-conjugated antibody. Agonist-stimulated CCR7 can transport antibody-based cargoes, with possible therapeutic applications in oncology.

Keywords: CCL19; CCR7; Rab5; Rab7; melanoma; β-arrestin

References

  1. J Immunol. 2006 Aug 15;177(4):2314-23 - PubMed
  2. J Virol. 2013 Jul;87(14):7893-901 - PubMed
  3. Biochemistry. 2004 Mar 30;43(12):3670-8 - PubMed
  4. Cancer Res. 2002 May 15;62(10):2937-41 - PubMed
  5. Adv Otorhinolaryngol. 2005;62:121-33 - PubMed
  6. Oncogene. 2007 May 10;26(21):2997-3005 - PubMed
  7. Science. 2013 Jan 18;339(6117):328-32 - PubMed
  8. Proc Natl Acad Sci U S A. 2010 Sep 28;107(39):16934-9 - PubMed
  9. Nature. 2001 Mar 1;410(6824):50-6 - PubMed
  10. Blood. 2002 Feb 15;99(4):1109-16 - PubMed
  11. J Immunol. 2008 Oct 1;181(7):4723-32 - PubMed
  12. EMBO J. 1994 Mar 15;13(6):1287-96 - PubMed
  13. Peptides. 2012 Apr;34(2):433-46 - PubMed
  14. FASEB J. 2007 Oct;21(12):3240-9 - PubMed
  15. Cancer Cell. 2007 Jun;11(6):526-38 - PubMed
  16. Eur J Immunol. 2012 Aug;42(8):1949-55 - PubMed
  17. Eur J Pharmacol. 2011 Oct 1;668(1-2):107-14 - PubMed
  18. Biol Chem. 2013 Mar;394(3):379-83 - PubMed
  19. Pharmacol Res. 2013 May;71:44-52 - PubMed
  20. J Biol Chem. 2001 Sep 7;276(36):33964-8 - PubMed
  21. Curr Top Microbiol Immunol. 2010;341:37-58 - PubMed
  22. Cancer Res. 2010 Oct 15;70(20):8138-48 - PubMed
  23. N Engl J Med. 2012 Nov 8;367(19):1847-8 - PubMed

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