Sp3 expression in immune cells: a quantitative study.
We previously reported the lack of expression of the bifunctional transcription factor Sp3 in peripheral blood mononuclear cells from most patients with multiple sclerosis (MS) (Grekova et al, 1996). An RT-PCR technique was developed to evaluate Sp3 mRNA levels in peripheral blood mononuclear cell subsets. Semi-quantitative and quantitative competitive RT-PCR assays were used to compare the level of Sp3 expression among subjects and among immune cell subsets. The competitor DNA fragment contained a deletion from the normal Sp3 cDNA sequence. The wild-type Sp3 cDNA and the competitor DNA fragment amplified with equal efficiency, and the two PCR products were distinguished by size. These studies demonstrated that normal CD4(+) and CD8(+) T cells, B cells, and macrophages expressed comparable amounts of Sp3 mRNA. No Sp3 expression could be detected in normal natural killer cells nor in any of these cell types from Sp3-negative MS patients. We propose that transcription of the Sp3 gene is blocked in immune cells from most patients with MS and that this contributes to the development of central nervous system inflammation in the disease.
Grekova
Maria C
MC
Department of Microbiology and Immunology, Georgetown University Medical Center, Washington DC 20007, USA.
Salerno
Kilian
K
Mikkilineni
Radha
R
Richert
John R
JR
D000199:Actins / Q000235:genetics; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000235:genetics*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008969:Molecular Sequence Data; D009103:Multiple Sclerosis / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051705:Sp3 Transcription Factor; D014157:Transcription Factors / Q000235:genetics*
D000199:Actins; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; C497121:SP3 protein, human; D014157:Transcription Factors; D051705:Sp3 Transcription Factor