Pentraxin 3 (PTX3) is a soluble pattern recognition receptor (PRR), which is produced by severalkinds of cells, such as neutrophils, dendritic cells, macrophages, and epithelial cells.PTX3 is known to play an important protective effect against Aspergillus. Genetic linkage ingene-targeted mice and human PTX3 plays a non-redundant role in the immune protectionagainst specific pathogens, especially Aspergillus. Recent studies have shown that the polymorphismof PTX3 is associated with increased susceptibility to invasive aspergillosis (IA). Inthis review, we provide an overview of these studies that underline the potential of PTX3 indiagnosis and therapy of IA.

Pentraxin 3 (PTX3) is a soluble pattern recognition receptor (PRR), which is produced by severalkinds of cells, such as neutrophils, dendritic cells, macrophages, and epithelial cells.PTX3 is known to play an important protective effect against Aspergillus. Genetic linkage ingene-targeted mice and human PTX3 plays a non-redundant role in the immune protectionagainst specific pathogens, especially Aspergillus. Recent studies have shown that the polymorphismof PTX3 is associated with increased susceptibility to invasive aspergillosis (IA). Inthis review, we provide an overview of these studies that underline the potential of PTX3 indiagnosis and therapy of IA.

OBJECTIVETo develop an improved substrate for indirect immunofluorescence test (IIF) for detecting anti-Ro60/Sjogren's syndrome A (Ro/SSA) autoantibodies.

METHODS60-kDa Ro/SSA autoantigens (Ro60) cDNAs were obtained from human placental cDNA library using polymerase chain reaction (PCR) and were cloned into the mammalian expression vector-pEGFP-C1. Then, the recombinant plasmids were transfected into HEp-2 cells. We confirmed the overexpression, localization and antigenicity of fusion proteins in transfected cells by means of immunoblotting, confocal fluorescence microscopy and IIF. HEp-2 and HEp-Ro60 were analyzed by IIF using a panel of 10 precipitin-positive anti-Ro human sera simultaneously.

RESULTSStable expression of Ro60-green fluorescent protein (Ro60-GFP) fusion proteins were maintained ten more generations. Ro60-GFP kept the antigenicity of Ro while demonstrating its own characteristic immunofluorescent pattern in HEp-Ro60 cells. The transfectants dramatically increased the sensitivity of IIF testing (a mean increase of 6.7-fold in endpoint titer). Eight over ten (8/10) positive anti-Ro sera showed characteristic immunofluorescent patterns for HEp-Ro60, including two sera that were anti-nuclear antibodies (ANA) negative for untransfected HEp-2. IIF-ANA in all healthy sera was negative for HEp-Ro60.

CONCLUSIONSAs a new substrate for IIF, the Ro60 transfectants can be used to detect anti-Ro antibodies. In addition, transfected HEp-2 cells keep the immunofluorescent properties of HEp-2 cells in IIF-ANA tests and can be employed as a substrate for routine IIF-ANA detection.

Antibodies, Antinuclear ; blood ; Autoantigens ; Cell Line ; Fluorescent Antibody Technique, Indirect ; Humans ; Molecular Weight ; RNA, Small Cytoplasmic ; Recombinant Fusion Proteins ; immunology ; Ribonucleoproteins ; immunology ; Transfection