Prevalence of ZAP-70, LAT, SLP-76, and DNA methyltransferase 1 expression in CD4+ T cells of patients with systemic lupus erythematosus.
T cells from systemic lupus erythematosus (SLE) patients exhibit defective function of CD4(+) T cells that can be responsible for improper activation of B cells and antibody biosynthesis against host antigens. We compared the level of ZAP-70, LAT, and SLP-76, transcripts and proteins in CD4(+) T cells from SLE patients (n = 22) and healthy individuals (n = 15). We also determined DNA methyltransferase 1 (DNMT1) protein content in CD4(+) T cells of SLE patients. The CD4(+) T cells were isolated by positive biomagnetic separation technique. The quantitative analysis of messenger RNA (mRNA) was performed by reverse transcription and real-time quantitative polymerase chain reaction (RQ-PCR) SYBR Green I system. The protein level in the CD4(+) T cells was determined by Western blotting analysis. We found that the LAT protein level was significantly higher in SLE CD4(+) T cells than in controls (P = 0.006). Western blot analysis revealed that ZAP-70 protein content in SLE CD4(+) T cells may be reciprocally correlated with disease activity expressed in SLEDAI scale (R = -0.623, P = 0.002) or number of affected organ systems (R = -0.549, P = 0.008). We also observed reciprocal correlation between DNMT1 protein content in CD4(+) T cells and disease activity scored with SLEDAI scale (R = -0.779, P = 0.001) or number of affected organ systems (R = -0.617, P = 0.019), respectively. Our findings might indicate that LAT, ZAP-70, and DNMT1 protein levels in CD4(+) T cells can be associated with SLE disease.
Januchowski
Radosław
R
Department of Biochemistry and Molecular Biology, University of Medical Sciences, 6 Swiecickiego St., 60-781, Poznań, Poland.
Wudarski
Mariusz
M
Chwalińska-Sadowska
Hanna
H
Jagodzinski
Paweł P
PP
D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism; D000328:Adult; D000368:Aged; D015496:CD4-Positive T-Lymphocytes / Q000201:enzymology* / Q000276:immunology; D005260:Female; D015870:Gene Expression; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000097:blood* / Q000276:immunology / Q000503:physiopathology; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008875:Middle Aged; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism*; D051746:ZAP-70 Protein-Tyrosine Kinase / Q000235:genetics / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; C411744:DMAP1 protein, human; C110810:LAT protein, human; D008565:Membrane Proteins; D010750:Phosphoproteins; D012333:RNA, Messenger; D012097:Repressor Proteins; C092947:SLP-76 signal Transducing adaptor proteins; D011494:Protein Kinases; D051746:ZAP-70 Protein-Tyrosine Kinase; C497261:ZAP70 protein, human