Intravenous infusion of E coli increases mRNA for E-selectin but not P-selectin in baboon tissues. A lethal dose of E coli was infused intravenously into a baboon. A control baboon received no E coli but was otherwise treated identically. After 4 hours, the animals were killed, and the organs were immediately collected for RNA isolation. The levels of mRNA for P-selectin, E-selectin, and β-actin were measured by a competitive RT-PCR method in which serial dilutions of a DNA mimic containing flanking sequences matching the target DNA were added to first-strand cDNA prepared from a fixed concentration of tissue RNA. The amount of mimic that allows amplification of equal quantitities of mimic and target DNA provides a measure of the level of target DNA, and thus mRNA, in the sample. Shown is representative agarose gel electrophoresis of the PCR products from baboon heart. In this example, mimic cDNA at a concentration of 10−3 attomoles/μL allowed equal amplification of mimic and P-selectin cDNA in both the control baboon and the baboon that received E coli. Mimic cDNA at a concentration of 10−1 attomoles/μL allowed equal amplification of mimic and E-selectin cDNA in the E coli baboon, whereas no E-selectin cDNA was amplified in the control baboon even at the lowest concentration of mimic (10−5 attomoles/μL). Mimic cDNA at a concentration of 10−1 attomoles/μL allowed equal amplification of mimic and β-actin cDNA in both the control baboon and the E coli baboon. Identical results were obtained with another pair of baboons, one of which received E coli.
