Calcium Ionophore-Treated Peripheral Blood Monocytes and Dendritic Cells Rapidly Display Characteristics of Activated Dendritic Cells

Brian J. Czerniecki, Charles Carter, Licia Rivoltini, Gary K. Koski, Hyun I. Kim, David E. Weng, James G. Roros, Yasmine M. Hijazi, Shuwen Xu, Steven A. Rosenberg, Peter A. Cohen

Research output: Contribution to journalArticle

Abstract

Human peripheral blood contains a small subpopulation of immature dendritic cells (iDC) distinguished from circulating monocytes by their low expression of CD14. We utilized leukapheresis and countercurrent centrifugal elutriation to obtain myeloid origin mononuclear cell (MOMC) fractions of monocytes and iDC for study. These subpopulations were ultrastructurally and immunophenotypically similar before culture. After a 20- to 96-h culture either alone, with recombinant human granulocyte-monocyte CSF, or with endotoxin, greater up-regulation of costimulatory molecule expression was observed among iDC than among monocytes, and only iDC expressed the activation molecule CD83. Treatment with rhIL-4 caused many MOMC to develop morphologic properties of dendritic cells within 96 h, but costimulatory molecule up-regulation and CD14 down-regulation were heterogeneous, and CD83 expression was infrequent. In contrast, calcium ionophore (Cl) treatment induced rapid and consistent effects in MOMC from both healthy volunteers and cancer patients, including down-regulated CD14 expression, acquisition of dendritic cell morphologic properties, up-regulated MHC and costimulatory molecule expression, and de novo CD83 expression. Many such effects occurred within 20 h of treatment. Cl treatment activated purified CD14 + monocytes and also enhanced the spontaneous activation of purified CD14 -/dim iDC in culture. Unfractionated MOMC, purified monocytes, and purified iDC displayed equivalently enhanced T cell-sensitizing efficiency following Cl treatment. CD4 + T cell sensitization to keyhole limpet hemocyanin and CD8 + T cell sensitization to MART-1 melanoma-associated peptide were achieved in a single culture stimulation. Therefore, circulating monocytes and iDC can be induced by Cl to manifest properties of activated DC, providing large numbers of efficient, nontransformed autologous APC for T cell sensitization strategies.

Original languageEnglish
Pages (from-to)3823-3837
Number of pages15
JournalJournal of Immunology
Volume159
Issue number8
Publication statusPublished - Oct 15 1997

ASJC Scopus subject areas

  • Immunology

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    Czerniecki, B. J., Carter, C., Rivoltini, L., Koski, G. K., Kim, H. I., Weng, D. E., Roros, J. G., Hijazi, Y. M., Xu, S., Rosenberg, S. A., & Cohen, P. A. (1997). Calcium Ionophore-Treated Peripheral Blood Monocytes and Dendritic Cells Rapidly Display Characteristics of Activated Dendritic Cells. Journal of Immunology, 159(8), 3823-3837.