Fluorescently activated cell sorting followed by microarray profiling of helper T cell subtypes from human peripheral blood.
BACKGROUND
Peripheral blood samples have been subjected to comprehensive gene expression profiling to identify biomarkers for a wide range of diseases. However, blood samples include red blood cells, white blood cells, and platelets. White blood cells comprise polymorphonuclear leukocytes, monocytes, and various types of lymphocytes. Blood is not distinguishable, irrespective of whether the expression profiles reflect alterations in (a) gene expression patterns in each cell type or (b) the proportion of cell types in blood. CD4+ Th cells are classified into two functionally distinct subclasses, namely Th1 and Th2 cells, on the basis of the unique characteristics of their secreted cytokines and their roles in the immune system. Th1 and Th2 cells play an important role not only in the pathogenesis of human inflammatory, allergic, and autoimmune diseases, but also in diseases that are not considered to be immune or inflammatory disorders. However, analyses of minor cellular components such as CD4+ cell subpopulations have not been performed, partly because of the limited number of these cells in collected samples.


RESULTS
We describe fluorescently activated cell sorting followed by microarray (FACS-array) technology as a useful experimental strategy for characterizing the expression profiles of specific immune cells in the circulation. We performed reproducible gene expression profiling of Th1 and Th2, respectively. Our data suggest that this procedure provides reliable information on the gene expression profiles of certain small immune cell populations. Moreover, our data suggest that GZMK, GZMH, EOMES, IGFBP3, and STOM may be novel markers for distinguishing Th1 cells from Th2 cells, whereas IL17RB and CNTNAP1 can be Th2-specific markers.


CONCLUSIONS
Our approach may help in identifying aberrations and novel therapeutic or diagnostic targets for diseases that affect Th1 or Th2 responses and elucidating the involvement of a subpopulation of immune cells in some diseases.
Department of Disaster Psychiatry, Internal Research Institute of Disaster Science, Tohoku University, Sendai, Japan; Department of Biological Psychiatry, Tohoku University Graduate School of Medicine, Sendai, Japan.
Chiaki
C
Ono
Department of Disaster Psychiatry, Internal Research Institute of Disaster Science, Tohoku University, Sendai, Japan; Department of Biological Psychiatry, Tohoku University Graduate School of Medicine, Sendai, Japan.
Zhiqian
Z
Yu
Department of Disaster Psychiatry, Internal Research Institute of Disaster Science, Tohoku University, Sendai, Japan; Department of Biological Psychiatry, Tohoku University Graduate School of Medicine, Sendai, Japan.
Yoshiyuki
Y
Kasahara
Department of Disaster Psychiatry, Internal Research Institute of Disaster Science, Tohoku University, Sendai, Japan; Department of Biological Psychiatry, Tohoku University Graduate School of Medicine, Sendai, Japan.
Yoshie
Y
Kikuchi
Department of Microbiology and Immunology, Tohoku University Graduate School of Medicine, Sendai, Japan; Tohoku Medical Megabank Organization, Tohoku University, Sendai, Japan.
Naoto
N
Ishii
Department of Disaster Psychiatry, Internal Research Institute of Disaster Science, Tohoku University, Sendai, Japan; Department of Biological Psychiatry, Tohoku University Graduate School of Medicine, Sendai, Japan; Tohoku Medical Megabank Organization, Tohoku University, Sendai, Japan.
Hiroaki
H
Tomita
D000328:Adult; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D020869:Gene Expression Profiling; D006801:Humans; D008297:Male; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D018417:Th1 Cells / Q000166:cytology* / Q000378:metabolism*; D018418:Th2 Cells / Q000166:cytology* / Q000378:metabolism*
D012313:RNA