Spontaneous inflammatory cytokine gene expression in normal human peripheral blood mononuclear cells.
Cytokines regulate cellular immune activity and are produced by a variety of cells, especially lymphocytes, monocytes, and macrophages. Measurement of cytokine levels has yielded useful information on the pathological process of different diseases such as AIDS, endotoxic shock, sepsis, asthma, and cancer. It may also be of use in the monitoring of disease progression and/or inflammation. To determine spontaneous cytokine gene expression in whole blood and PBMCs, whole blood was obtained from healthy volunteers and total mRNA was isolated from PBMCs. The kinetics of response were determined by sequential testing of cytokine gene expression by RT-PCR analysis. Our results demonstrated that isolated and incubated PBMCs expressed TNF-alpha and high levels of IL-1beta, IL-6, IL-8, and IL-10. In contrast, WB only expressed the mRNA cytokines of TNF-alpha and IL-8 (p < 0.05). These results suggest that spontaneous myriad mRNA cytokine expression can be avoided with the use of WB incubation and the rapid collection of PBMCs. Furthermore, this method should be employed in all cases where the levels of cytokine gene expression can be evaluated.
Molina
Moisés Armides Franco
MA
Laboratorio de Inmunología y Virología, Departamento de Microbiología e Inmunología. Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León, Nuevo León, México. moyfranco@hotmail.com
Gamboa
Edgar Mendoza
EM
Tello
Paloma Castillo
PC
Benavides
Pablo Zapata
PZ
León
Leonardo Castillo
LC
Guerra
Reyes Tamez
RT
Padilla
Cristina Rodríguez
CR
D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics*; D005786:Gene Expression Regulation; D006801:Humans; D007249:Inflammation / Q000235:genetics; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics
D016207:Cytokines; D012333:RNA, Messenger