Display options
Cite
Fletcher AL, Malhotra D, Acton SE, et al. Reproducible isolation of lymph node stromal cells reveals site-dependent differences in fibroblastic reticular cells. Front Immunol. 2011;2:35doi: 10.3389/fimmu.2011.00035.
Fletcher, A. L., Malhotra, D., Acton, S. E., Lukacs-Kornek, V., Bellemare-Pelletier, A., Curry, M., Armant, M., & Turley, S. J. (2011). Reproducible isolation of lymph node stromal cells reveals site-dependent differences in fibroblastic reticular cells. Frontiers in immunology, 235. https://doi.org/10.3389/fimmu.2011.00035
Fletcher, Anne L, et al. "Reproducible isolation of lymph node stromal cells reveals site-dependent differences in fibroblastic reticular cells." Frontiers in immunology vol. 2 (2011): 35. doi: https://doi.org/10.3389/fimmu.2011.00035
Fletcher AL, Malhotra D, Acton SE, Lukacs-Kornek V, Bellemare-Pelletier A, Curry M, Armant M, Turley SJ. Reproducible isolation of lymph node stromal cells reveals site-dependent differences in fibroblastic reticular cells. Front Immunol. 2011 Sep 12;2:35. doi: 10.3389/fimmu.2011.00035. eCollection 2011. PMID: 22566825; PMCID: PMC3342056.
Copy Download .nbib Format: NLM
Share it on

Front Immunol. 2011 Sep 12;2:35. doi: 10.3389/fimmu.2011.00035. eCollection 2011.

Reproducible isolation of lymph node stromal cells reveals site-dependent differences in fibroblastic reticular cells.

Frontiers in immunology

Anne L Fletcher, Deepali Malhotra, Sophie E Acton, Veronika Lukacs-Kornek, Angelique Bellemare-Pelletier, Mark Curry, Myriam Armant, Shannon J Turley

Affiliations

  1. Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute Boston, MA, USA.

PMID: 22566825 PMCID: PMC3342056 DOI: 10.3389/fimmu.2011.00035

Abstract

Within lymph nodes, non-hematopoietic stromal cells organize and interact with leukocytes in an immunologically important manner. In addition to organizing T and B cell segregation and expressing lymphocyte survival factors, several recent studies have shown that lymph node stromal cells shape the naïve T cell repertoire, expressing self-antigens which delete self-reactive T cells in a unique and non-redundant fashion. A fundamental role in peripheral tolerance, in addition to an otherwise extensive functional portfolio, necessitates closer study of lymph node stromal cell subsets using modern immunological techniques; however this has not routinely been possible in the field, due to difficulties reproducibly isolating these rare subsets. Techniques were therefore developed for successful ex vivo and in vitro manipulation and characterization of lymph node stroma. Here we discuss and validate these techniques in mice and humans, and apply them to address several unanswered questions regarding lymph node composition. We explored the steady-state stromal composition of lymph nodes isolated from mice and humans, and found that marginal reticular cells and lymphatic endothelial cells required lymphocytes for their normal maturation in mice. We also report alterations in the proportion and number of fibroblastic reticular cells (FRCs) between skin-draining and mesenteric lymph nodes. Similarly, transcriptional profiling of FRCs revealed changes in cytokine production from these sites. Together, these methods permit highly reproducible stromal cell isolation, sorting, and culture.

Keywords: endothelial cells; fibroblastic reticular cells; human lymph nodes; lymph node stromal cells; mouse lymph nodes; stromal cells; tolerance

References

  1. Prog Mol Biol Transl Sci. 2010;92:177-205 - PubMed
  2. J Exp Med. 2010 Apr 12;207(4):681-8 - PubMed
  3. J Exp Med. 2004 Sep 20;200(6):783-95 - PubMed
  4. Immunol Rev. 2000 Aug;176:181-93 - PubMed
  5. Circ Res. 2009 Aug 14;105(4):393-401 - PubMed
  6. Immunity. 1997 Apr;6(4):491-500 - PubMed
  7. Nat Rev Immunol. 2009 Jul;9(7):491-502 - PubMed
  8. J Invest Dermatol. 2006 Mar;126(3):561-8 - PubMed
  9. Immunity. 2011 Feb 25;34(2):237-46 - PubMed
  10. J Exp Med. 2010 Apr 12;207(4):689-97 - PubMed
  11. Int Immunol. 2004 Aug;16(8):1133-42 - PubMed
  12. Essays Biochem. 2006;42:105-17 - PubMed
  13. Birth Defects Res A Clin Mol Teratol. 2006 Jan;76(1):39-45 - PubMed
  14. Neuropathol Appl Neurobiol. 2010 Aug;36(5):368-87 - PubMed
  15. Nat Immunol. 2004 Jun;5(6):597-605 - PubMed
  16. Biostatistics. 2003 Apr;4(2):249-64 - PubMed
  17. Am J Pathol. 2011 Apr;178(4):1662-75 - PubMed
  18. J Immunol. 2004 Sep 1;173(5):2968-75 - PubMed
  19. Blood. 1996 Mar 15;87(6):2095-147 - PubMed
  20. Immunity. 2006 Dec;25(6):989-1001 - PubMed
  21. Nat Immunol. 2007 Nov;8(11):1255-65 - PubMed
  22. Nat Immunol. 2007 Oct;8(10):1076-85 - PubMed
  23. FASEB J. 1998 Sep;12(12):1135-42 - PubMed
  24. Genome Biol. 2003;4(5):P3 - PubMed
  25. Immunity. 2005 Jan;22(1):19-29 - PubMed
  26. Anat Rec. 1978 Feb;190(2):201-15 - PubMed
  27. Nat Rev Immunol. 2003 Apr;3(4):292-303 - PubMed
  28. J Immunol. 2007 Jul 15;179(2):993-1003 - PubMed
  29. J Pathol. 2009 Jan;217(1):21-31 - PubMed
  30. Science. 2008 Aug 8;321(5890):843-7 - PubMed
  31. EMBO J. 2000 May 2;19(9):2015-23 - PubMed
  32. Arch Biochem Biophys. 1993 Mar;301(2):221-7 - PubMed
  33. Nature. 2000 Jul 20;406(6793):309-14 - PubMed
  34. Trends Immunol. 2011 Jan;32(1):12-8 - PubMed
  35. Nat Immunol. 2008 Jun;9(6):667-75 - PubMed
  36. J Immunol. 2008 Sep 15;181(6):3887-96 - PubMed
  37. Nat Rev Immunol. 2008 Oct;8(10):764-75 - PubMed
  38. Gastroenterology. 2008 Apr;134(4):1028-37 - PubMed
  39. Am J Physiol Heart Circ Physiol. 2005 Jun;288(6):H2946-54 - PubMed
  40. J Anat. 1999 Jan;194 ( Pt 1):143-6 - PubMed
  41. J Biol Chem. 2002 Oct 18;277(42):39561-5 - PubMed
  42. Toxicol Pathol. 2006;34(5):409-24 - PubMed
  43. Eur J Immunol. 2011 Mar;41(3):624-33 - PubMed
  44. Proc Natl Acad Sci U S A. 2000 Nov 7;97(23):12694-9 - PubMed
  45. Semin Immunol. 2009 Feb;21(1):8-13 - PubMed
  46. Nat Immunol. 2007 Feb;8(2):181-90 - PubMed
  47. Nat Immunol. 2009 Sep;10(9):1026-33 - PubMed
  48. J Exp Med. 2002 May 20;195(10):1257-66 - PubMed
  49. J Exp Med. 1998 Jan 19;187(2):205-16 - PubMed
  50. J Immunol. 2008 Aug 1;181(3):1898-907 - PubMed
  51. J Immunol. 2008 Nov 1;181(9):6189-200 - PubMed

Publication Types

Grant support