Newly designed primer sets available for evaluating various cytokines and iNOS mRNA expression in guinea pig lung tissues by RT-PCR.
Guinea pigs are often used as an animal model of human tuberculosis (TB). However, there are few methods available for pursuing the immunological processes involved in guinea pig TB. In this study, we developed for the first time systematic reverse transcription (RT)-PCR for evaluation of guinea pig mRNA expression. RT-PCR primer sets were newly designed for detection of cytokines and inducible nitric oxide synthase (iNOS) mRNA in guinea pig TB. Interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta, interleukin (IL)-1beta, IL-2, IL-10, IL-12p40, granulocyte-macrophage-colony stimulating factor (GM-CSF) and iNOS mRNA expression were detected significantly and reproducibly when these primer sets were used. The data by real-time PCR were comparable with those of RT-PCR. We showed that these RT-PCR primer sets could be used to examine mRNA expression semi-quantitatively in guinea pig tissues, and conclude that these newly designed primer sets for conventional RT-PCR will be useful for studying the immunological processes in guinea pig tuberculosis experiments to investigate and evaluate efficacy of new vaccines or anti-mycobacterial drugs.
Yamada
Hiroyuki
H
Pathology Division, Mycobacterium Reference Center, The Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association, Tokyo.
Udagawa
Tadashi
T
Mizuno
Satoru
S
Hiramatsu
Kumiko
K
Sugawara
Isamu
I
D000818:Animals; D016207:Cytokines / Q000032:analysis* / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry; D004195:Disease Models, Animal; D005260:Female; D006168:Guinea Pigs; D006801:Humans; D008168:Lung / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000032:analysis* / Q000235:genetics; D052247:Nitric Oxide Synthase Type II; D012333:RNA, Messenger / Q000032:analysis*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014376:Tuberculosis / Q000276:immunology
D016207:Cytokines; D017931:DNA Primers; D015342:DNA Probes; D012333:RNA, Messenger; C496319:NOS2 protein, human; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II