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

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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 (US)
Pages (from-to)3823-3837
Number of pages15
JournalJournal of Immunology
Volume159
Issue number8
StatePublished - Oct 15 1997
Externally publishedYes

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Calcium Ionophores
Dendritic Cells
Monocytes
Blood Cells
T-Lymphocytes
Up-Regulation
Leukapheresis
Therapeutics
Granulocytes
Endotoxins
Melanoma
Healthy Volunteers
Down-Regulation
Cell Culture Techniques
Peptides

ASJC Scopus subject areas

  • Immunology

Cite this

Czerniecki, B. J., Carter, C., Rivoltini, L., Koski, G. K., Kim, H. I., Weng, D. E., ... 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.

Calcium Ionophore-Treated Peripheral Blood Monocytes and Dendritic Cells Rapidly Display Characteristics of Activated Dendritic Cells. / Czerniecki, Brian J.; Carter, Charles; Rivoltini, Licia; Koski, Gary K.; Kim, Hyun I.; Weng, David E.; Roros, James G.; Hijazi, Yasmine M.; Xu, Shuwen; Rosenberg, Steven A.; Cohen, Peter A.

In: Journal of Immunology, Vol. 159, No. 8, 15.10.1997, p. 3823-3837.

Research output: Contribution to journalArticle

Czerniecki, BJ, Carter, C, Rivoltini, L, Koski, GK, Kim, HI, Weng, DE, Roros, JG, Hijazi, YM, Xu, S, Rosenberg, SA & Cohen, PA 1997, 'Calcium Ionophore-Treated Peripheral Blood Monocytes and Dendritic Cells Rapidly Display Characteristics of Activated Dendritic Cells', Journal of Immunology, vol. 159, no. 8, pp. 3823-3837.
Czerniecki BJ, Carter C, Rivoltini L, Koski GK, Kim HI, Weng DE et al. Calcium Ionophore-Treated Peripheral Blood Monocytes and Dendritic Cells Rapidly Display Characteristics of Activated Dendritic Cells. Journal of Immunology. 1997 Oct 15;159(8):3823-3837.
Czerniecki, Brian J. ; Carter, Charles ; Rivoltini, Licia ; Koski, Gary K. ; Kim, Hyun I. ; Weng, David E. ; Roros, James G. ; Hijazi, Yasmine M. ; Xu, Shuwen ; Rosenberg, Steven A. ; Cohen, Peter A. / Calcium Ionophore-Treated Peripheral Blood Monocytes and Dendritic Cells Rapidly Display Characteristics of Activated Dendritic Cells. In: Journal of Immunology. 1997 ; Vol. 159, No. 8. pp. 3823-3837.
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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.",
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AU - Roros, James G.

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AU - Rosenberg, Steven A.

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