PMID- 10687141
OWN - NLM
STAT- MEDLINE
DCOM- 20000511
LR  - 20191103
IS  - 1524-9557 (Print)
IS  - 1524-9557 (Linking)
VI  - 23
IP  - 1
DP  - 2000 Jan
TI  - Immunomodulatory dendritic cells generated from nonfractionated bulk peripheral 
      blood mononuclear cell cultures induce growth of cytotoxic T cells against renal 
      cell carcinoma.
PG  - 83-93
AB  - Dendritic cells (DCs) loaded with tumor antigens have the potential to become a 
      powerful tool for clinical cancer treatment. Recently, the authors showed that a 
      tumor-specific immune response can be elicited in culture via stimulation with 
      autologous renal tumor lysate (Tuly)-loaded DCs that were generated from 
      cytokine-cultured adherent peripheral blood mononuclear cells (PBMCs). Here, the 
      authors show that immunomodulatory DCs can be generated directly from 
      nonfractionated bulk PBMC cultures. Kinetic studies of DC differentiation and 
      maturation in PBMC cultures were performed by monitoring the acquisition of 
      DC-associated molecules using fluorescence-activated cell sorting analysis to 
      determine the percentage of positive immunostained cells and the mean relative 
      linear fluorescence intensity (MRLFI). Compared with conventional adherent CD14+ 
      cultures, which have mostly natural killer, T, and B cells removed before 
      cytokine culture, bulk PBMC cultures exhibited an early loss of CD14+ cells (day 
      0 = 78.8%, day 2 = 29.6% versus day 0 = 74%, day 2 = 75%) with an increase in 
      yield of mature DCs (DC19- CD83+) (day 5 = 17%, day 6 = 21%, day 7 = 22% versus 
      day 5 = 11%, day 6 = 15%, day 7 = 23%). Although a comparable percentage of DCs 
      expressing CD86+ (B7-2), CD40+, and HLA-DR+ were detected in both cultures, 
      higher expression levels were detected in DCs derived from bulk culture (CD86 = 
      MRLFI 3665.1 versus 2662.1 on day 6; CD40 = MRLFI 1786 versus 681.2 on day 6; 
      HLA-DR = MRLFI 6018.2 versus 3444.9 on day 2). Cytokines involved in DC 
      maturation were determined by polymerase chain reaction demonstrating 
      interleukin-6 (IL-6), IL-12, interferon-gamma, granulocyte-macrophage 
      colony-stimulating factor, and tumor necrosis factor-alpha mRNA expression by 
      bulk culture cells during the entire 9-day culture period. This same cytokine 
      mRNA profile was not found in the conventional adherent DC culture. Autologous 
      renal Tuly (30 micrograms protein/10(7) PBMCs) enhanced human leukocyte antigen 
      expression by DCs (class I = 7367.6 versus 4085.4 MRFLI; class II = 8277.2 versus 
      6175.7 MRFLI) and upregulated cytokine mRNAs levels. Concurrently, CD3+ CD56-, 
      CD3+ CD25+, and CD3+ TCR+ cell populations increased and cytotoxicity against 
      autologous renal cell carcinoma tumor target was induced. Specific cytotoxicity 
      was augmented when cultures were boosted continuously with IL-2 (20 U/mL 
      biological response modifier program) plus Tuly stimulation. These results 
      suggest that nonadherent PBMCs may participate in enhancing DC maturation. 
      Besides the simplicity of this culture technique, bulk DC cultures potentially 
      may be used with the same efficiency as conventional purified DCs. Furthermore, 
      bulk culture-derived DCs may be used directly in vivo as a tumor vaccine, or for 
      further ex vivo expansion of co-cultured cytotoxic T cells to be used for 
      adoptive immunotherapy.
FAU - Hinkel, A
AU  - Hinkel A
AD  - Department of Urology, University of California, Los Angeles, USA.
FAU - Tso, C L
AU  - Tso CL
FAU - Gitlitz, B J
AU  - Gitlitz BJ
FAU - Neagos, N
AU  - Neagos N
FAU - Schmid, I
AU  - Schmid I
FAU - Paik, S H
AU  - Paik SH
FAU - deKernion, J
AU  - deKernion J
FAU - Figlin, R
AU  - Figlin R
FAU - Belldegrun, A
AU  - Belldegrun A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Immunother
JT  - Journal of immunotherapy (Hagerstown, Md. : 1997)
JID - 9706083
RN  - 0 (Antigens, CD)
RN  - 0 (Biomarkers)
RN  - 0 (Cytokines)
SB  - IM
MH  - Antigen-Presenting Cells/immunology
MH  - Antigens, CD/biosynthesis
MH  - Biomarkers
MH  - Carcinoma, Renal Cell/blood/*immunology
MH  - Cell Culture Techniques/methods
MH  - Cell Differentiation
MH  - Cell Separation
MH  - Chemical Fractionation
MH  - Cytokines/metabolism
MH  - Dendritic Cells/cytology/*immunology
MH  - Humans
MH  - Kidney Neoplasms/blood/*immunology
MH  - Leukocytes, Mononuclear/cytology/immunology
MH  - T-Lymphocytes, Cytotoxic/*immunology
MH  - Tumor Cells, Cultured
EDAT- 2000/02/25 09:00
MHDA- 2000/05/16 09:00
CRDT- 2000/02/25 09:00
PHST- 2000/02/25 09:00 [pubmed]
PHST- 2000/05/16 09:00 [medline]
PHST- 2000/02/25 09:00 [entrez]
AID - 10.1097/00002371-200001000-00011 [doi]
PST - ppublish
SO  - J Immunother. 2000 Jan;23(1):83-93. doi: 10.1097/00002371-200001000-00011.