Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2016 Nov 17;128(20):2465-2468.
doi: 10.1182/blood-2016-06-724856. Epub 2016 Oct 3.

High incidence of activating STAT5B mutations in CD4-positive T-cell large granular lymphocyte leukemia

Emma I Andersson  1 , Takahiro Tanahashi  2 , Nodoka Sekiguchi  3   4 , Vanessa Rebecca Gasparini  1 , Sabrina Bortoluzzi  1 , Toru Kawakami  3 , Kazuyuki Matsuda  5 , Takeki Mitsui  6 , Samuli Eldfors  7 , Stefania Bortoluzzi  8 , Alessandro Coppe  8 , Andrea Binatti  8 , Sonja Lagström  7 , Pekka Ellonen  7 , Noriyasu Fukushima  9 , Sayaka Nishina  3 , Noriko Senoo  3 , Hitoshi Sakai  4 , Hideyuki Nakazawa  3 , Yok-Lam Kwong  10 , Thomas P Loughran  11 , Jaroslaw P Maciejewski  12 , Satu Mustjoki  1   13 , Fumihiro Ishida  2   3   14
Affiliations

High incidence of activating STAT5B mutations in CD4-positive T-cell large granular lymphocyte leukemia

Emma I Andersson et al. Blood. .
No abstract available

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
STAT5B mutation characterization. (A) Linear representation of the STAT5B protein structure. Previously known LGL leukemia mutations in STAT5B are marked in the SH2 domain, whereas the novel Q706L and S715F mutations in the transactivation domains are marked with green boxes. Multiple tyrosine and serine phosphorylation sites are marked in red. (B) STAT5B reporter assay results. Mutated STAT5B constructs (pCMV6-XL6 STAT5B) were generated through site-directed mutagenesis followed by transfection and expression of WT and mutated STAT5B (Q706L, S715F, N642H) in HeLa cells together with a STAT5B reporter. Dual-reporter luciferase assay was used to determine activation and phosphorylation of mutated STAT5B. The experiment was repeated 3 times. Columns represent mean of the fold-change activity. Error bars indicate the standard error of the mean (SEM), and the statistical significance was calculated with a 1-way analysis of variance (ANOVA; *P < .05, **P < .001). (C) To investigate the phosphorylation status of the variants, HeLa cells transfected with the abovementioned variants were analyzed by western blot with a phosphoSTAT5 (Tyr694) specific antibody. Protein lysates of the different variants were separated on an sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel and transferred to a nitrocellulose membrane. STAT5 protein levels of the different variants were used to normalize for the transfection efficacy. β-Actin was used as a loading control. (D) Transfected HeLa cells were stimulated with 100 ng/mL interferon-α for 6 hours. A dual-reporter luciferase assay was used to determine activation and phosphorylation of mutated STAT5B. The experiment was repeated 2 times. Columns represent mean of the fold-change activity. Error bars indicate the SEM, and the statistical significance was calculated with a 1-way ANOVA (*P < .05, **P < .001). (E) Typical morphology of a representative LGL cell in a STAT5B mutated T-LGL patient. Scale bar, 15 μm. (F) Morphology of lymphocyte expressing CD4, CD56, and TCRαβ in a healthy individual. CD4+CD56+ and TCRαβ-type lymphocytes were sorted by the FACS method and stained with Wright-Giemsa stain. A representative cell is shown. Scale bar, 15 μm.
See this image and copyright information in PMC

References

    1. Loughran TP., Jr Clonal diseases of large granular lymphocytes. Blood. 1993;82(1):1-14. - PubMed
    1. Zhang D, Loughran TP Jr. Large granular lymphocytic leukemia: molecular pathogenesis, clinical manifestations, and treatment. Hematol Am Soc Hematol Educ Program. 2012;2012:652-659. - PubMed
    1. Sabattini E, Bacci F, Sagramoso C, Pileri SA. WHO classification of tumours of haematopoietic and lymphoid tissues in 2008: an overview. Pathologica. 2010;102(3):83-87. - PubMed
    1. Koskela HL, Eldfors S, Ellonen P, et al. . Somatic STAT3 mutations in large granular lymphocytic leukemia. N Engl J Med. 2012;366(20):1905-1913. - PMC - PubMed
    1. Jerez A, Clemente MJ, Makishima H, et al. . STAT3 mutations unify the pathogenesis of chronic lymphoproliferative disorders of NK cells and T-cell large granular lymphocyte leukemia. Blood. 2012;120(15):3048-3057. - PMC - PubMed

Publication types