High-throughput gene expression profiling of memory differentiation in primary human T cells
Background: The differentiation of naive T and B cells into memory lymphocytes is essential for immunity to pathogens. Therapeutic manipulation of this cellular differentiation program could improve vaccine efficacy and the in vitro expansion of memory cells. However, chemical screens to identify co...
| Main Authors: | , , , , , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
| Published: |
BioMed Central Ltd
2010
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| Online Access: | http://hdl.handle.net/1721.1/59279 |
| _version_ | 1811088678472122368 |
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| author | Angelosanto, Jill Brosnahan, Kathleen Ross, Kenneth Hahn, Cynthia Russell, Kate Drury, Linda Norton, Stephanie Nadler, Lee Stegmaier, Kimberly Haining, W. Nicholas |
| author2 | Broad Institute of MIT and Harvard |
| author_facet | Broad Institute of MIT and Harvard Angelosanto, Jill Brosnahan, Kathleen Ross, Kenneth Hahn, Cynthia Russell, Kate Drury, Linda Norton, Stephanie Nadler, Lee Stegmaier, Kimberly Haining, W. Nicholas |
| author_sort | Angelosanto, Jill |
| collection | MIT |
| description | Background: The differentiation of naive T and B cells into memory lymphocytes is essential for immunity to pathogens. Therapeutic manipulation of this cellular differentiation program could improve vaccine efficacy and the in vitro expansion of memory cells. However, chemical screens to identify compounds that induce memory differentiation have been limited by 1) the lack of reporter-gene or functional assays that can distinguish naive and memory-phenotype T cells at high throughput and 2) a suitable cell-line representative of naive T cells. Results: Here, we describe a method for gene-expression based screening that allows primary naive and memory-phenotype lymphocytes to be discriminated based on complex genes signatures corresponding to these differentiation states. We used ligation-mediated amplification and a fluorescent, bead-based detection system to quantify simultaneously 55 transcripts representing naive and memory-phenotype signatures in purified populations of human T cells. The use of a multi-gene panel allowed better resolution than any constituent single gene. The method was precise, correlated well with Affymetrix microarray data, and could be easily scaled up for high-throughput. Conclusion: This method provides a generic solution for high-throughput differentiation screens in primary human T cells where no single-gene or functional assay is available. This screening platform will allow the identification of small molecules, genes or soluble factors that direct memory differentiation in naive human lymphocytes. |
| first_indexed | 2024-09-23T14:05:48Z |
| format | Article |
| id | mit-1721.1/59279 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T14:05:48Z |
| publishDate | 2010 |
| publisher | BioMed Central Ltd |
| record_format | dspace |
| spelling | mit-1721.1/592792022-09-28T18:24:29Z High-throughput gene expression profiling of memory differentiation in primary human T cells Angelosanto, Jill Brosnahan, Kathleen Ross, Kenneth Hahn, Cynthia Russell, Kate Drury, Linda Norton, Stephanie Nadler, Lee Stegmaier, Kimberly Haining, W. Nicholas Broad Institute of MIT and Harvard Ross, Kenneth Norton, Stephanie Stegmaier, Kimberly Background: The differentiation of naive T and B cells into memory lymphocytes is essential for immunity to pathogens. Therapeutic manipulation of this cellular differentiation program could improve vaccine efficacy and the in vitro expansion of memory cells. However, chemical screens to identify compounds that induce memory differentiation have been limited by 1) the lack of reporter-gene or functional assays that can distinguish naive and memory-phenotype T cells at high throughput and 2) a suitable cell-line representative of naive T cells. Results: Here, we describe a method for gene-expression based screening that allows primary naive and memory-phenotype lymphocytes to be discriminated based on complex genes signatures corresponding to these differentiation states. We used ligation-mediated amplification and a fluorescent, bead-based detection system to quantify simultaneously 55 transcripts representing naive and memory-phenotype signatures in purified populations of human T cells. The use of a multi-gene panel allowed better resolution than any constituent single gene. The method was precise, correlated well with Affymetrix microarray data, and could be easily scaled up for high-throughput. Conclusion: This method provides a generic solution for high-throughput differentiation screens in primary human T cells where no single-gene or functional assay is available. This screening platform will allow the identification of small molecules, genes or soluble factors that direct memory differentiation in naive human lymphocytes. 2010-10-13T15:17:50Z 2010-10-13T15:17:50Z 2008-08 2008-05 2010-09-03T16:13:43Z Article http://purl.org/eprint/type/JournalArticle 1471-2172 http://hdl.handle.net/1721.1/59279 BMC Immunology. 2008 Aug 01;9(1):44 en http://dx.doi.org/10.1186/1471-2172-9-44 BMC Immunology Creative Commons Attribution http://creativecommons.org/licenses/by/2.0 Haining et al.; licensee BioMed Central Ltd. application/pdf BioMed Central Ltd BioMed Central Ltd |
| spellingShingle | Angelosanto, Jill Brosnahan, Kathleen Ross, Kenneth Hahn, Cynthia Russell, Kate Drury, Linda Norton, Stephanie Nadler, Lee Stegmaier, Kimberly Haining, W. Nicholas High-throughput gene expression profiling of memory differentiation in primary human T cells |
| title | High-throughput gene expression profiling of memory differentiation in primary human T cells |
| title_full | High-throughput gene expression profiling of memory differentiation in primary human T cells |
| title_fullStr | High-throughput gene expression profiling of memory differentiation in primary human T cells |
| title_full_unstemmed | High-throughput gene expression profiling of memory differentiation in primary human T cells |
| title_short | High-throughput gene expression profiling of memory differentiation in primary human T cells |
| title_sort | high throughput gene expression profiling of memory differentiation in primary human t cells |
| url | http://hdl.handle.net/1721.1/59279 |
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