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A Novel Modification of a Flow Cytometric Assay of Phosphorylated STAT1 in Whole Blood Monocytes for Immunomonitoring of Patients on IFNα Regimen
Author(s) -
Vakkila J.,
Nieminen U.,
Siitonen S.,
Turunen U.,
Halme L.,
Nuutinen H.,
Mustonen H.,
Puolakkainen P.,
Färkkilä M.,
Repo H.
Publication year - 2008
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.2007.02028.x
Subject(s) - regimen , flow cytometry , stat1 , immunology , whole blood , medicine , phosphorylation , monocyte , chemistry , interferon , biochemistry
We explored whether episodes stimulating leucocytes in vivo could be tracked from whole blood samples by monitoring activation of STAT1 by flow cytometry. The method was tested in hepatitis C patients ( n = 9) that were on interferon (IFN)α regimen. CD14 + monocytes responded strongly to IFNα/γ being sensitive indicators for recent immune activation. At 45 min after s.c. IFNα 91% of monocytes were phosphorylated STAT1 + . The frequency of responding cells decreased to a base level within 6 h. Monocytes, however, had a long‐term deficient phosphorylated STAT1 response to IFNα in vitro that in patients on standard IFNα regimen lasted for 48 h. In patients on pegylated IFNα the phosphorylated STAT1 response was completely absent. We conclude that whole blood analysis of STAT1 activation by flow cytometry is applicable to monitor immune cells in patient material.