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A simple and enzyme-free method for processing infiltrating lymphocytes from small mouse tumors for ELISpot analysis.

Publication ,  Journal Article
Swartz, AM; Reap, E; Norberg, P; Schmittling, R; Janetzki, S; Sanchez-Perez, L; Sampson, JH
Published in: J Immunol Methods
August 2018

The ELISpot assay prevails as one of the most sensitive and meaningful assays for the detection of antigen-specific, effector immune responses. Acquisition of cellular analyte for ELISpot analysis is typically not problematic when derived from tissues enriched in lymphocytes (e.g., lymphoid organs and blood); however, cell processing becomes more difficult when lymphocytes represent only a very minor population relative to the source tissue, especially when the source tissue is in limited supply (e.g., small mouse tumors). Traditional enzymatic-based methods for dissociating tumors often result in poor yields, inconsistent lymphocyte enrichment, and can have deleterious effects on lymphocyte phenotype and function. To address these limitations, we have developed an enzyme-free protocol for processing tumor infiltrating lymphocytes (TILs) from small mouse tumors, which enables the enumeration of antigen-specific effector lymphocytes using ELISpot analysis. This procedure is predicated on the dissociation of tumor tissue using gentle agitation with a paddle blender followed by a brief in vitro culture period to remove adherent cells, as well as to revive lymphocytes from a non-responsive state. Although this method is demonstrated with mouse intracerebral tumors, we have found that this protocol is applicable to peripheral tumors and may likely extend to alternative tissue sources wherein lymphocytes exist in low numbers.

Duke Scholars

Published In

J Immunol Methods

DOI

EISSN

1872-7905

Publication Date

August 2018

Volume

459

Start / End Page

90 / 93

Location

Netherlands

Related Subject Headings

  • T-Lymphocytes
  • Neoplasms
  • Mice, Inbred C57BL
  • Mice
  • Lymphocytes, Tumor-Infiltrating
  • Immunology
  • Enzymes
  • Enzyme-Linked Immunospot Assay
  • Cell Separation
  • Animals
 

Citation

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Swartz, A. M., Reap, E., Norberg, P., Schmittling, R., Janetzki, S., Sanchez-Perez, L., & Sampson, J. H. (2018). A simple and enzyme-free method for processing infiltrating lymphocytes from small mouse tumors for ELISpot analysis. J Immunol Methods, 459, 90–93. https://doi.org/10.1016/j.jim.2018.05.015
Swartz, Adam M., Elizabeth Reap, Pamela Norberg, Robert Schmittling, Sylvia Janetzki, Luis Sanchez-Perez, and John H. Sampson. “A simple and enzyme-free method for processing infiltrating lymphocytes from small mouse tumors for ELISpot analysis.J Immunol Methods 459 (August 2018): 90–93. https://doi.org/10.1016/j.jim.2018.05.015.
Swartz AM, Reap E, Norberg P, Schmittling R, Janetzki S, Sanchez-Perez L, et al. A simple and enzyme-free method for processing infiltrating lymphocytes from small mouse tumors for ELISpot analysis. J Immunol Methods. 2018 Aug;459:90–3.
Swartz, Adam M., et al. “A simple and enzyme-free method for processing infiltrating lymphocytes from small mouse tumors for ELISpot analysis.J Immunol Methods, vol. 459, Aug. 2018, pp. 90–93. Pubmed, doi:10.1016/j.jim.2018.05.015.
Swartz AM, Reap E, Norberg P, Schmittling R, Janetzki S, Sanchez-Perez L, Sampson JH. A simple and enzyme-free method for processing infiltrating lymphocytes from small mouse tumors for ELISpot analysis. J Immunol Methods. 2018 Aug;459:90–93.
Journal cover image

Published In

J Immunol Methods

DOI

EISSN

1872-7905

Publication Date

August 2018

Volume

459

Start / End Page

90 / 93

Location

Netherlands

Related Subject Headings

  • T-Lymphocytes
  • Neoplasms
  • Mice, Inbred C57BL
  • Mice
  • Lymphocytes, Tumor-Infiltrating
  • Immunology
  • Enzymes
  • Enzyme-Linked Immunospot Assay
  • Cell Separation
  • Animals