Project Number: 1265-32000-087-07
Project Type: Reimbursable

Start Date: Oct 06, 2004
End Date: Aug 31, 2008

Objective:
Assess gene expression patterns of immune markers in pigs infected or vaccinated with porcine reproductive and respiratory syndrome virus (PPRSV) for various NC229 (Swine PRRSV project) collaborators. 1) With UNE test 60 RNA and cDNA preparations from lung tissue collected from pigs infected with PRRSV and determine whether specific immune markers are associated with differences in resistant versus susceptible pig anti-PRRSv responses. 2) With SD State immune responses from boars infected with PRRSV will be compared. With UIL gene expression patterns of RNA prepared from blood cells from pigs vaccinated for PRRSV will be assessed and effect of in vitro culture time compared.

Approach:
Samples of lung tissue from 60 pigs, 30 infected with PRRSV and 30 control littermates, will be shipped from UNE to Lunney's laboratory. The samples will include 3-4 pigs of each population from each of the high and low tails of the principal component distribution, to represent extreme differences in PRRSV response, and their control littermates. RNA will be isolated from each sample. Individual samples will have cDNA prepared and then be subjected to RT-PCR for immune gene expression patterns. Data will be summarized and evaluated before it is uncoded by UNE for identification of different groups of pigs. Statistical comparisons will be performed to determine differences between populations and between phenotypic classes in expression of 12 specific immune markers will be characterized. The markers, for which arrays exist in Lunney's laboratory, are control [RPL32], IFNG, IFNA, IL15, TNFA, STAT1, IL6, IL1B, CSF2 [GM-CSF], IL8, IL10, and IL12B (p40). For SD State and UIL samples of PBMC or tissues will be sent frozen, or stored in RNLater, to the Lunney lab. RNA will be isolated from each sample, cDNAs prepared, and real-time PCR assays will be used to determine expression of specific immune markers. The markers, for which arrays already exist in Lunney's laboratory, include control [RPL32], IFNG, IFNA, IL15, TNFA, STAT1, IL6, IL1B, CSF2 [GM-CSF], IL8, IL10, and IL12B (p40), as well as some of the >250 other immune markers available through CRIS 64 collaborative research efforts. BSL-2 1/27/06.