Recombinant Human Serum Amyloid A [His]

Circulating serum amyloid A (SAA) is increased in various inflammatory conditions. The human SAA protein family comprises the acute phase SAA1/SAA2, known to activate a large set of innate and adaptive immune cells, and the constitutive SAA4. The liver synthesis of SAA1/SAA2 is well-established but there is still an open debate on extrahepatic SAA expression especially in macrophages. We aimed to investigate the ability of human primary monocytes and monocyte-derived macrophages to express SAA1, SAA2 and SAA4 at both the transcriptional and protein levels, as previous studies almost exclusively dealt with monocytic cell lines. Monocytes and derived macrophages from healthy donors were stimulated under various conditions. In parallel with SAA, pro-inflammatory IL1A, IL1B and IL6 cytokine expression was assessed. While LPS alone was non-effective, a combined LPS/dexamethasone treatment induced SAA1 and to a lesser extent SAA2 transcription in human monocytes and macrophages. In contrast, as expected, pro-inflammatory cytokine expression was strongly induced following stimulation with LPS, an effect which was dampened in the presence of dexamethasone. Furthermore, in monocytes polarized towards a pro-inflammatory M1 phenotype, SAA expression in response to LPS/dexamethasone was potentiated; a result mainly seen for SAA1. However, a major discrepancy was observed between SAA mRNA and intracellular protein levels under the experimental conditions used. Our results demonstrate that human monocytes and macrophages can express SAA genes, mainly SAA1 in response to an inflammatory environment. While SAA is considered as a member of a large cytokine network, its expression in the monocytes-macrophages in response to LPS-dexamethasone is strikingly different from that observed for classic pro-inflammatory cytokines. As monocytes-macrophages are major players in chronic inflammatory diseases, it may be hypothesized that SAA production from macrophages may contribute to the local inflammatory microenvironment, especially when macrophages are compactly organized in granulomas as in sarcoidosis.

Several isoforms of serum amyloid A (SAA) have been identified so far and because the plasma concentration of it increases dramatically, it is used as an indicator of inflammation in animals. In many terrestrial mammals, the circulating isoforms are SAA1 and SAA2, which are synthesized in the liver. Extra-hepatically synthesized SAA3, however, is a predominantly local SAA isoform with a characteristic N-terminal TFLK motif and a highly alkaline isoelectric point (pI). The aim of this study was to characterize the circulating SAA isoforms in bottlenose dolphins (dSAA) by determining the deduced amino acid sequence isolated from liver and the pI of plasma from healthy dolphins and those with inflammation. The deduced amino acid sequences of dSAA showed characteristics of SAA3 with an N-terminal TFLK motif, a predicted alkaline pI and were phylogenetically clustered with the SAA3 group rather than the SAA1 and SAA2 groups. Various tissues contained dSAA mRNA with the highest levels being detected in the liver. Isoelectric focusing and western blot analysis showed that one highly alkaline SAA was markedly detected in plasma obtained from dolphins affected by inflammation. These results suggest that, unlike other mammals, the circulating SAA in dolphins exhibits SAA3 properties, as is the case in pigs. (C) 2012 Elsevier B.V. All rights reserved.