title: Ex vivo TCR-induced leukocyte gene expression of inflammatory mediators is increased in type 1 diabetic patients but not in overweight children.
abstract: BACKGROUND
  Abnormal systemic concentrations of proinflammatory cytokines/chemokines have been implicated in the development of long-term cardiovascular complications in type 1 diabetes (T1DM) and obesity. Whether leukocyte white blood cell (WBC) gene expression of these proinflammatory mediators contributes to their increased systemic levels, however, remains unclear, especially in the pediatric patient populations. This study examines mRNA changes of 9 cytokines and chemokines in WBCs following ex vivo immunostimulation from 9 T1DM (13.4 +/- 0.5 year, 4F/5 M), 23 overweight (OW, 12.3 +/- 0.5 year, 10F/13M, BMI% 97.1 +/- 0.5 and > 90.0), and 21 healthy (CL, 13.8 +/- 0.7 year, 9F/12 M, BMI% 59.6 +/- 4.6 and < 85.0) children.
  
  
  METHODS
  All subjects had been maintained in euglycemic conditions for at least 90 min before blood draws. Whole blood was then sampled and incubated with anti-T-cell receptor (TCR) antibody or heat-aggregated IgG (HAG) to stimulate T-cell and Fc receptors (FcR), respectively. After lysis of leukocytes, mRNA levels of six tumor necrosis factor superfamily cytokines (TNFSF2, 5, 6, 7, 9, 14) and three chemokines (CCL8, 20, and CXCL10) were measured using RT-PCR.
  
  
  RESULTS
  Following TCR stimulation, T1DM displayed significantly greater mRNA responses than CL for TNFSF5, 7, 9, and CCL8, and CXCL10; TNFSF9, CCL8, and CXCL10 were also significantly higher in T1DM than OW; no difference was observed between OW and CL. FcR stimulation induced similar responses across groups.
  
  
  CONCLUSIONS
  Leukocytes of T1DM children displayed exaggerated gene expression in response to ex vivo TCR induction of five key proinflammatory cytokines/chemokines. This elevated leukocyte gene expression may be one of the pathophysiological contributors to the development of vascular complications in T1DM.
authors: Department of Pharmacology, School of Medicine, University of California-Irvine, Irvine, CA 92612, USA. jsrosa@uci.edu
authors: Jaime S
authors: JS
authors: Rosa
authors: Masato
authors: M
authors: Mitsuhashi
authors: Stacy R
authors: SR
authors: Oliver
authors: Mieko
authors: M
authors: Ogura
authors: Rebecca L
authors: RL
authors: Flores
authors: Andria M
authors: AM
authors: Pontello
authors: Pietro R
authors: PR
authors: Galassetti
mesh_terms: D000293:Adolescent; D000906:Antibodies / Q000494:pharmacology; D018925:Chemokines / Q000235:genetics*; D002648:Child; D016207:Cytokines / Q000235:genetics*; D003922:Diabetes Mellitus, Type 1 / Q000097:blood / Q000235:genetics*; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D007074:Immunoglobulin G / Q000494:pharmacology; D007962:Leukocytes / Q000187:drug effects / Q000502:physiology*; D008297:Male; D050177:Overweight / Q000097:blood / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D011948:Receptors, Antigen, T-Cell / Q000276:immunology*; D011961:Receptors, Fc / Q000187:drug effects / Q000276:immunology; D012016:Reference Values; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology
chemical_list: D000906:Antibodies; D018925:Chemokines; D016207:Cytokines; D007074:Immunoglobulin G; D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell; D011961:Receptors, Fc