[Gene expression of the interferon and cell-immunity systems in human blood samples].
The interferon (IF) and cell-apoptosis (CA) systems are interrelated and regulate the protective reactions in body by means of a complex of biologically active proteins. The transcription levels of mRNA were determined by the RT-PCR semi-quantitative method in order to compare the constitutive expression levels of IF (alpha, beta, gamma) genes, IF-dependent enzymes of 2'5'-oligoadenylatesynthetyase (OAS), RNAase L, dsRNA-protein kinase (dsPK) and CA effectors (Fas-Ag, bcl-2 and gamma-actin) in human blood microsamples. cDNA dilutions, different numbers of amplification cycles (PCR with specific pairs of primers) as well as PCR-products' dilutions for dot-hybridization with specific probes were made use of to detect and evaluate levels of 9 mRNAs. The constitutive levels of gene expression of the IF and CA systems were found to differ essentially from others (1000-fold). The studied mRNA types were shared between 5 groups according to their transcription levels: very high--alpha-IF, high--RNAase L, medium--gamma-actin and bcl-2, low--beta-IF and very low (detectable after induction only)--gamma-IF, OAS, dsPK and Fas-Ag. The used detection method has a sufficiently high sensitivity and can be recommended for studies of IF inductors with unknown action mechanisms.
Sokolova
T M
TM
Bibikova
O V
OV
Bystrov
N S
NS
Uryvaev
L V
LV
D015088:2',5'-Oligoadenylate Synthetase / Q000096:biosynthesis / Q000235:genetics; D000199:Actins / Q000096:biosynthesis / Q000235:genetics; D000328:Adult; D017209:Apoptosis / Q000235:genetics*; D001773:Blood Cells / Q000502:physiology*; D001782:Blood Donors; D017931:DNA Primers; D004722:Endoribonucleases / Q000096:biosynthesis / Q000235:genetics; D005260:Female; D015870:Gene Expression; D006801:Humans; D007372:Interferons / Q000096:biosynthesis / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019892:eIF-2 Kinase / Q000096:biosynthesis / Q000235:genetics; D019014:fas Receptor / Q000096:biosynthesis / Q000235:genetics
D000199:Actins; D017931:DNA Primers; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D019014:fas Receptor; D007372:Interferons; D019892:eIF-2 Kinase; D015088:2',5'-Oligoadenylate Synthetase; D004722:Endoribonucleases; C031956:2-5A-dependent ribonuclease