Skip Navigation

JNCI Journal of the National Cancer Institute 2004 96(17):1331-1342; doi:10.1093/jnci/djh252
© 2004 by Oxford University Press
This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (17)
Right arrow Request Permissions
Google Scholar
Right arrow Articles by Lesinski, G. B.
Right arrow Articles by Carson, W. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lesinski, G. B.
Right arrow Articles by Carson, W. E., III
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© 2004 Oxford University Press

ARTICLE

Multiparametric Flow Cytometric Analysis of Inter-Patient Variation in STAT1 Phosphorylation Following Interferon Alfa Immunotherapy

Gregory B. Lesinski, Sri Vidya Kondadasula, Tim Crespin, Lei Shen, Kari Kendra, Michael Walker, William E. Carson, III

Affiliations of authors: Departments of Human Cancer Genetics (GBL, SVK, WEC), Epidemiology and Biometrics (LS), Medical Oncology (KK), and Surgery (MW, WEC), Arthur G. James Cancer Hospital and Richard J. Solove Research Institute, The Ohio State University, Columbus, OH; Primetrics, Hilliard, OH (TC)

Correspondence to: William E. Carson III, MD, Division of Surgical Oncology, The Ohio State University, N924 Doan Hall, 410 W. 10th Ave., Columbus, OH 43210 (e-mail: carson-1{at}medctr.osu.edu)

Background: Regulation of gene expression by signal transducer and activator of transcription 1 (STAT1) within host tissues mediates the antitumor effects of interferon alfa (IFN {alpha}). We used a novel flow cytometric assay to examine phosphorylation-mediated activation of STAT1 within immune effector cell subsets following in vitro or in vivo IFN {alpha} treatments. Methods: Peripheral blood mononuclear cells (PBMCs) isolated from healthy donors (n = 17) or melanoma patients (n = 19) were treated in vitro with interferon alfa-2b (IFN {alpha}-2b) or phosphate-buffered saline (PBS) and subjected to multiparametric flow cytometry to measure the levels of phosphorylated STAT1 (P-STAT1) within immune cell subsets. We similarly analyzed PBMCs isolated from melanoma patients before and 1 hour after immunotherapy with IFN {alpha}-2b. All statistical tests were two-sided. Results: P-STAT1 levels in all major immune cell subsets increased within 15 minutes of in vitro IFN {alpha}-2b treatment of PBMCs; the increase was most pronounced in T lymphocytes and monocytes. Relatively low doses of IFN {alpha}-2b (i.e., 102–103 IU/mL) induced maximal STAT1 activation in vitro. Compared with melanoma patients, healthy donors had higher basal levels of P-STAT1 (specific fluorescence [Fsp]; i.e., FspPBS, the level of P-STAT1 in PBS-treated cells) in total PBMCs, natural killer (NK) cells, and T cells (mean FspPBS in total PBMCs: 5.5 in healthy donors versus 1.6 in patients, difference = 3.9, 95% confidence interval [CI] = 1.4 to 6.5, P = .004; mean FspPBS in NK cells: 4.6 in healthy donors versus 0.9 in patients, difference = 3.7, 95% CI = 1.7 to 5.7, P = .001; mean FspPBS in T cells: 6.8 in healthy donors versus 0.9 in patients, difference = 5.9, 95% CI = 2.5 to 9.3, P = .002). P-STAT1 was detected in the NK and T cells of two patients who received IFN {alpha}-2b immunotherapy (20 MU/m2 [MU = million units], administered by intravenous injection). P-STAT1 levels in the PBMCs of a patient treated sequentially with 5 MU/m2 and 10 MU/m2 IFN {alpha}-2b (administered by subcutaneous injection) also increased in response to treatments with IFN {alpha}-2b but did not increase further with the increased dosage of IFN {alpha}-2b. Conclusion: This flow cytometry method can be used to monitor STAT1 activation within subsets of immune cells from patients undergoing IFN {alpha} immunotherapy.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 BloodHome page
A. Gowda, J. Roda, S.-R. A. Hussain, A. Ramanunni, T. Joshi, S. Schmidt, X. Zhang, A. Lehman, D. Jarjoura, W. E. Carson, et al.
IL-21 mediates apoptosis through up-regulation of the BH3 family member BIM and enhances both direct and antibody-dependent cellular cytotoxicity in primary chronic lymphocytic leukemia cells in vitro
Blood, May 1, 2008; 111(9): 4723 - 4730.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
J. M. Zimmerer, A. M. Lehman, A. S. Ruppert, C. W. Noble, T. Olencki, M. J. Walker, K. Kendra, and W. E. Carson III
IFN-{alpha}-2b-Induced Signal Transduction and Gene Regulation in Patient Peripheral Blood Mononuclear Cells Is Not Enhanced by a Dose Increase from 5 to 10 Megaunits/m2
Clin. Cancer Res., March 1, 2008; 14(5): 1438 - 1445.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
G. B. Lesinski, J. Trefry, M. Brasdovich, S. V. Kondadasula, K. Sackey, J. M. Zimmerer, A. R. Chaudhury, L. Yu, X. Zhang, T. R. Crespin, et al.
Melanoma Cells Exhibit Variable Signal Transducer and Activator of Transcription 1 Phosphorylation and a Reduced Response to IFN-{alpha} Compared with Immune Effector Cells
Clin. Cancer Res., September 1, 2007; 13(17): 5010 - 5019.
[Abstract] [Full Text] [PDF]

Home page
 J. Exp. Med.Home page
K. M. Dhodapkar, D. Banerjee, J. Connolly, A. Kukreja, E. Matayeva, M. C. Veri, J. V. Ravetch, R. M. Steinman, and M. V. Dhodapkar
Selective blockade of the inhibitory Fc{gamma} receptor (Fc{gamma}RIIB) in human dendritic cells and monocytes induces a type I interferon response program
J. Exp. Med., June 11, 2007; 204(6): 1359 - 1369.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
J. M. Zimmerer, G. B. Lesinski, S. V. Kondadasula, V. I. Karpa, A. Lehman, A. RayChaudhury, B. Becknell, and W. E. Carson III
IFN-{alpha}-Induced Signal Transduction, Gene Expression, and Antitumor Activity of Immune Effector Cells Are Negatively Regulated by Suppressor of Cytokine Signaling Proteins
J. Immunol., April 15, 2007; 178(8): 4832 - 4845.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
K. A. Varker, S. V. Kondadasula, M. R. Go, G. B. Lesinski, R. Ghosh-Berkebile, A. Lehman, J. P. Monk, T. Olencki, K. Kendra, and W. E. Carson III
Multiparametric Flow Cytometric Analysis of Signal Transducer and Activator of Transcription 5 Phosphorylation in Immune Cell Subsets In vitro and following Interleukin-2 Immunotherapy.
Clin. Cancer Res., October 1, 2006; 12(19): 5850 - 5858.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
J. M. Roda, R. Parihar, A. Lehman, A. Mani, S. Tridandapani, and W. E. Carson III
Interleukin-21 Enhances NK Cell Activation in Response to Antibody-Coated Targets
J. Immunol., July 1, 2006; 177(1): 120 - 129.
[Abstract] [Full Text] [PDF]

Home page
 JCOHome page
C. F. Eisenbeis, G. B. Lesinski, M. Anghelina, R. Parihar, D. Valentino, J. Liu, P. Nadella, P. Sundaram, D. C. Young, M. Sznol, et al.
Phase I Study of the Sequential Combination of Interleukin-12 and Interferon Alfa-2b in Advanced Cancer: Evidence for Modulation of Interferon Signaling Pathways by Interleukin-12
J. Clin. Oncol., December 1, 2005; 23(34): 8835 - 8844.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.