Cytokines and acute phase proteins associated with acute swine influenza infection in pigs

Abstract

This study set out to investigate the cytokines and acute phase proteins (APPs) associated with the acute stages of experimentally-induced swine influenza virus (SIV) infection in 3-week-old, colostrum-deprived, caesarean-derived piglets. The piglets were inoculated intratracheally with 10(7.5) 50% egg infective dose [EID(50)] Swine/Belgium/1/98 (H1N1) SIV and were euthanased at time-points between 0 and 120h post-inoculation (PI). Broncho-alveolar lavage fluid (BALF), lung homogenates and sera were examined for inflammatory mediators by bioassay or ELISA. Interferon (IFN)-α, interleukin (IL)-6, IL-1 and tumour necrosis factor (TNF)-α peaked in BALF 24-30h PI, when virus titres and the severity of clinical signs were maximal. Whereas IFN-γ and IL-12, but not IL-18, increased in tandem in BALF, serum cytokine concentrations were either undetectable or were up to 100-fold lower. The APP C-reactive protein (CRP) and haptoglobin peaked 24h later than the cytokines and reached higher levels in serum than in BALF. In contrast, lipopolysaccharide (LPS)-binding protein (LBP) only increased in BALF. Lung virus titres tightly correlated with BALF IFN-α, IL-6, IL-1, TNF-α, IFN-γ and IL-12, as well as with serum IL-6, IFN-α and IFN-γ. Signs of disease correlated with the same cytokines in BALF and serum, as well as with BALF LBP and serum CRP. The findings suggest that IFN-γ and IL-12 play a role in the pathogenesis of SIV and that APPs are induced by cytokines. This influenza infection model may have value in assessing the therapeutic potential of cytokine antagonists.

Fig. 1

Temporal profiles of IFN-α (■), IL-6 (♦), TNF-α (▴) and IL-1 (●) in BALF (A), lung tissue (B) and serum (C) post-inoculation (PI) with swine influenza virus. Values represent means of 2 or 3 pigs (+standard error of mean [SEM]), and are expressed as biological units (U)/mL (for BALF and serum) or U/g (lung tissue).

Fig. 2

Temporal profiles of IL-12 (■), IFN-γ (♦) and IL-18 (▴) in BALF (A), lung tissue (B) and serum (C) post-inoculation (PI) with swine influenza virus. Values represent means of 2 or 3 pigs (+standard error of mean [SEM]), and are expressed as pg/mL (for BALF and serum) or pg/g (lung tissue).

Fig. 3

Temporal profiles of HG (■), LBP (♦) and CRP (▴) in BALF (A) and serum (B) post-inoculation (PI) with swine influenza virus. Values represent means of 2 or 3 pigs (+standard error of mean [SEM]), and are expressed as μg/mL (HG) or ng/mL (LBP and CRP).