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Isolation and Purification of Colon Lamina Propria Dendritic Cells from Mice with Colitis

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Abstract

Dendritic cells are prime antigen presenting cells for stimulation of T cell immune responses. These cells are present in trace amounts in normal tissue. At sites of disease the increased frequency of these cells interacting with T cells may provide the basis for the release of pro-inflammatory mediators and contribute to localised cell and tissue damage. Studies on dendritic cells in the colon lamina propria of inflammatory bowel disease (IBD) mice have been limited due to the difficulties encountered in the isolation and purification of sufficient numbers of these cells. This is the first detailed, reproducible method provided in the literature for the isolation of colon lamina propria dendritic cells from mice with colitis, yielding optimum purity of cells and sufficient numbers to advance the study of dendritic cell function in the colons of mice. The most frequently used identification marker of murine DC is the CD11c surface antigen. We have adapted, combined, and improved procedures developed for the isolation of other cell types, to develop an efficient procedure for the isolation of dendritic cells from colon tissue. This protocol describes a step-by-step method for optimising the purity and recovery of lamina propria CD11c+ dendritic cells from mice colons.

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Abbreviations

DC:

dendritic cells

PBS:

Dulbecco’s phosphate buffered saline

FACS:

fluorescent activated cell sorting

IBD:

inflammatory bowel disease

LPMC:

lamina propria mononuclear cells

MACS:

magnetic activated cell sorting

References

  • J. Banchereau R.M. Steinman (1998) ArticleTitleDendritic cells and the control of immunity Nature 392 245–252 Occurrence Handle10.1038/32588 Occurrence Handle9521319 Occurrence Handle1:CAS:528:DyaK1cXitFWrsr8%3D

    Article  PubMed  CAS  Google Scholar 

  • J.J. Cebra P.J. Gearhart R. Kamat S.M. Robertson J. Tseng (1977) ArticleTitleThe secretory immunoglobulin A response in the gut Biochem. Soc. Trans. (proc) 5 1565–1569 Occurrence Handle1:CAS:528:DyaE1cXoslCgug%3D%3D

    CAS  Google Scholar 

  • M.D. Davies D.M. Parrott (1981) ArticleTitlePreparation and purification of lymphocytes from the epithelium and lamina propria of murine small intestine Gut 22 481–488 Occurrence Handle6973508 Occurrence Handle1:STN:280:Bi6B283gsFQ%3D

    PubMed  CAS  Google Scholar 

  • S.B. Dillon T.T. MacDonald (1984) ArticleTitleFunctional properties of lymphocytes isolated from murine small intestinal epithelium Immunology 52 501–509 Occurrence Handle6611297 Occurrence Handle1:STN:280:BiuB28zhsFY%3D

    PubMed  CAS  Google Scholar 

  • M.L. Drakes L. Lu V.M. Subbotin A.W. Thomson (1997) ArticleTitle In vivo administration of flt3 ligand markedly stimulates generation of dendritic cell progenitors from mouse liver J. Immunol. 159 4268–4278 Occurrence Handle9379022 Occurrence Handle1:CAS:528:DyaK2sXmvVyjur8%3D

    PubMed  CAS  Google Scholar 

  • F.D. Finkelman A. Lees R. Birnbaum W.C. Gause S.C. Morris (1996) ArticleTitleDendritic cells can present antigen in vivo in a tolerogenic or immunogenic fashion J. Immunol. 157 1406–1414 Occurrence Handle8759720 Occurrence Handle1:CAS:528:DyaK28XkvFWiu7Y%3D

    PubMed  CAS  Google Scholar 

  • D.N. Hart (1997) ArticleTitleDendritic cells: unique leukocyte populations which control the primary immune response Blood 90 3245–3287 Occurrence Handle9345009 Occurrence Handle1:CAS:528:DyaK2sXntVSjsbs%3D

    PubMed  CAS  Google Scholar 

  • T. Krajina F. Leithauser P. Moller Z. Trobonjaca J. Reimann (2003) ArticleTitleColonic lamina propria dendritic cells in mice with CD4+ T cell-induced colitis Eur. J. Immunol. 33 1073–1083 Occurrence Handle10.1002/eji.200323518 Occurrence Handle12672074 Occurrence Handle1:CAS:528:DC%2BD3sXjtVaksr4%3D

    Article  PubMed  CAS  Google Scholar 

  • F. Leithauser Z. Trobonjaca P. Moller J. Reimann (2001) ArticleTitleClustering of colonic lamina propria CD4(+) T cells to subepithelial dendritic cell aggregates precedes the development of colitis in a murine adoptive transfer model Lab. Invest. 81 1339–1349 Occurrence Handle11598147 Occurrence Handle1:STN:280:DC%2BD3MrltFOgug%3D%3D

    PubMed  CAS  Google Scholar 

  • Y.J. Liu (2001) ArticleTitleDendritic cell subsets and lineages, and their functions in innate and adaptive immunity Cell 106 259–262 Occurrence Handle11509173 Occurrence Handle1:CAS:528:DC%2BD3MXmtFKnsLw%3D

    PubMed  CAS  Google Scholar 

  • P.J. Morrissey K. Charrier S. Braddy D. Liggitt J.D. Watson (1993) ArticleTitleCD4+ T cells that express high levels of CD45RB induce wasting disease when transferred into congenic severe combined immunodeficient mice. Disease development is prevented by cotransfer of purified CD4+ T cells J. Exp. Med. 178 237–244 Occurrence Handle10.1084/jem.178.1.237 Occurrence Handle8100269 Occurrence Handle1:STN:280:ByyA3c7mtVQ%3D

    Article  PubMed  CAS  Google Scholar 

  • F. Powrie M.W. Leach S. Mauze L.B. Caddle R.L. Coffman (1993) ArticleTitlePhenotypically distinct subsets of CD4+ T cells induce or protect from chronic intestinal inflammation in C. B-17 scid mice Int. Immunol. 5 1461–1471 Occurrence Handle7903159 Occurrence Handle1:STN:280:ByuD1czjslA%3D

    PubMed  CAS  Google Scholar 

  • V.G. Pillarisetty A.B. Shah G. Miller J.I. Bleier R.P. DeMatteo (2004) ArticleTitleLiver dendritic cells are less immunogenic than spleen dendritic cells because of differences in subtype composition J. Immunol. 172 1009–1017 Occurrence Handle14707074 Occurrence Handle1:CAS:528:DC%2BD2cXnsl2j

    PubMed  CAS  Google Scholar 

  • M. Rescigno G. Rotta B. Valzasina P. Ricciardi-Castagnoli (2001) ArticleTitleDendritic cells shuttle microbes across gut epithelial monolayers Immunobiology 204 572–581 Occurrence Handle10.1078/0171-2985-00094 Occurrence Handle11846220 Occurrence Handle1:STN:280:DC%2BD387hsVShuw%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • D.M. Spencer G.M. Veldman S. Banerjee J. Willis A.D. Levine (2002) ArticleTitleDistinct inflammatory mechanisms mediate early versus late colitis in mice Gastroenterology 122 94–105 Occurrence Handle10.1053/gast.2002.30308 Occurrence Handle11781285

    Article  PubMed  Google Scholar 

  • A.J. Stagg A.L. Hart S.C. Knight M.A. Kamm (2003) ArticleTitleThe dendritic cell: its role in intestinal inflammation and relationship with gut bacteria Gut 52 1522–1529 Occurrence Handle10.1136/gut.52.10.1522 Occurrence Handle12970149 Occurrence Handle1:CAS:528:DC%2BD3sXotl2ks74%3D

    Article  PubMed  CAS  Google Scholar 

  • S. Tseng (1982) ArticleTitleExpression of immunoglobulin isotypes by lymphoid cells of mouse intestinal lamina propria Cell. Immunol. 73 324–336 Occurrence Handle10.1016/0008-8749(82)90459-2 Occurrence Handle6760992 Occurrence Handle1:STN:280:BiyC38bkvFw%3D

    Article  PubMed  CAS  Google Scholar 

  • P.J. Heijden ParticleVan der W. Stok (1987) ArticleTitleImproved procedure for the isolation of functionally active lymphoid cells from the murine intestine J. Immunol. Methods 103 161–167 Occurrence Handle2889781

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Pediatrics, Case Western Reserve University School of Medicine, Cleveland, OH

    Maureen L. Drakes, Steven J. Czinn & Thomas G. Blanchard

  2. Pediatrics, Rm. 737, Rainbow Babies & Children’s Hospital, 11100 Euclid Avenue, Cleveland, OH, 44106

    Maureen L. Drakes

Authors
  1. Maureen L. Drakes
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  2. Steven J. Czinn
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  3. Thomas G. Blanchard
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Correspondence to Maureen L. Drakes.

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Drakes, M.L., Czinn, S.J. & Blanchard, T.G. Isolation and Purification of Colon Lamina Propria Dendritic Cells from Mice with Colitis. Cytotechnology 46, 151–161 (2004). https://doi.org/10.1007/s10616-005-2552-6

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  • DOI: https://doi.org/10.1007/s10616-005-2552-6

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