Evaluation of gene expression profiles of immature dendritic cells prepared from peripheral blood mononuclear cells

Jeong Won Shin, Ping Jin, Yong Fan, Stefanie Slezak, Virginia David-Ocampo, Hanh M. Khuu, Elizabeth J. Read, Ena Wang, Francesco M. Marincola, David F. Stroncek

Research output: Contribution to journalArticle

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Abstract

BACKGROUND: Dendritic cells (DCs) generated ex vivo from peripheral blood monocytes or mobilized CD34+ cells and intended for clinical immunotherapy are typically characterized by morphologic, phenotypic, and functional assays. Assay results are highly dependent on conditions used to prepare the cells, so there is no standard assay battery for clinical DC products. This study evaluated gene expression profiling for characterization of immature DCs prepared from monocytes that had been elutriated from normal donor peripheral blood mononuclear cells (PBMNCs) immediately after collection or after storage at 4°C for 48 hours. STUDY DESIGN AND METHODS: RNA was isolated from fresh and 48-hour-stored PBMNCs, elutriated monocytes, elutriated lymphocytes, and immature DCs from five healthy subjects and was analyzed with a cDNA gene expression microarray with 17,500 genes. RESULTS: Unsupervised hierarchical clustering separated the 40 products into four groups: one with all 10 immature DCs, one with all 10 elutriated lymphocytes, one with 7 PBMNCs, and one with 10 elutriated monocytes and 3 PBMNCs. Within each of the four groups, however, fresh and stored products, or products derived from fresh or stored products, clustered together. Comparison of genes differentially expressed by fresh versus stored products (paired t tests, p < 0.005) found 273 genes that differed between fresh and stored PBMCs, 429 between lymphocytes elutriated from fresh versus stored PBMNCs, 711 between monocytes elutriated from fresh versus stored PBMNCs, and 3 between immature DCs prepared from monocytes elutriated from fresh versus stored PBMCs. CONCLUSIONS: This study demonstrates the potential utility of gene expression profiling for characterization of cell therapy products.

Original languageEnglish
Pages (from-to)647-657
Number of pages11
JournalTransfusion
Volume48
Issue number4
DOIs
Publication statusPublished - Apr 2008
Externally publishedYes

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Transcriptome
Dendritic Cells
Monocytes
Blood Cells
Gene Expression Profiling
Lymphocytes
Genes
Cell- and Tissue-Based Therapy
Immunotherapy
Cluster Analysis
Healthy Volunteers
Complementary DNA
Tissue Donors
RNA
Gene Expression

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Hematology

Cite this

Shin, J. W., Jin, P., Fan, Y., Slezak, S., David-Ocampo, V., Khuu, H. M., ... Stroncek, D. F. (2008). Evaluation of gene expression profiles of immature dendritic cells prepared from peripheral blood mononuclear cells. Transfusion, 48(4), 647-657. https://doi.org/10.1111/j.1537-2995.2007.01615.x

Evaluation of gene expression profiles of immature dendritic cells prepared from peripheral blood mononuclear cells. / Shin, Jeong Won; Jin, Ping; Fan, Yong; Slezak, Stefanie; David-Ocampo, Virginia; Khuu, Hanh M.; Read, Elizabeth J.; Wang, Ena; Marincola, Francesco M.; Stroncek, David F.

In: Transfusion, Vol. 48, No. 4, 04.2008, p. 647-657.

Research output: Contribution to journalArticle

Shin, JW, Jin, P, Fan, Y, Slezak, S, David-Ocampo, V, Khuu, HM, Read, EJ, Wang, E, Marincola, FM & Stroncek, DF 2008, 'Evaluation of gene expression profiles of immature dendritic cells prepared from peripheral blood mononuclear cells', Transfusion, vol. 48, no. 4, pp. 647-657. https://doi.org/10.1111/j.1537-2995.2007.01615.x
Shin, Jeong Won ; Jin, Ping ; Fan, Yong ; Slezak, Stefanie ; David-Ocampo, Virginia ; Khuu, Hanh M. ; Read, Elizabeth J. ; Wang, Ena ; Marincola, Francesco M. ; Stroncek, David F. / Evaluation of gene expression profiles of immature dendritic cells prepared from peripheral blood mononuclear cells. In: Transfusion. 2008 ; Vol. 48, No. 4. pp. 647-657.
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AU - Slezak, Stefanie

AU - David-Ocampo, Virginia

AU - Khuu, Hanh M.

AU - Read, Elizabeth J.

AU - Wang, Ena

AU - Marincola, Francesco M.

AU - Stroncek, David F.

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