[Use of the real time RT-PCR for immune related gene expression quantitation in experimentally infected Neospora caninum bovine calves].
Neospora caninum is one of the main causes of abortion and natimortality in cattle. Host immune defense is capable to inhibit tachyzoite activity during acute infection, but there is no action against bradyzoites in tissue cysts. Activation and modulation of this response is controlled by cell mediators. The real-time RT-PCR technique was employed to detect some of those mediators during N. caninum infection. Holstein and Nelore calves intramuscularly infected with tachyzoites and uninfected controls were slaughtered at the sixth day post-infection and popliteal lymph node, liver and brain cortex samples were analyzed. Real-time RT-PCR detected gene expression in all tissues. No significant variation of GAPDH gene expression was detected among groups, its amplification efficiency was similar to the other genes tested and it was used as the endogenous control for the analysis. Comparisons between infected and uninfected groups allowed the relative gene expression quantification. IFN-gamma and TNF-alpha genes showed increased expression in some samples. iNOS and TGF-beta1 genes had some non-significant variations and IL-4 and IL-10 stayed practically unaltered.
Nishi
Sandra Mayumi
SM
Departamento de Medicina Veterinária Preventiva e Saúde Animal, Faculdade de Medicina Veterinária e Zootecnia--FMVZ, Universidade de São Paulo--USP, Brazil. smnishi@usp.br.
Viero
Luciana Mori
LM
Soares
Rodrigo Martins
RM
Maiorka
Paulo Cesar
PC
Gennari
Solange Maria
SM
D000818:Animals; D002417:Cattle; D003048:Coccidiosis / Q000235:genetics* / Q000276:immunology*; D015870:Gene Expression / Q000276:immunology*; D018688:Neospora; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors