The possibility that cofactor proteins are only involved in the assay but have no pathogenic role sera may be due to the immunostimulatory effect and the polyclonal activation often observed in the course of EBV infection

The possibility that cofactor proteins are only involved in the assay but have no pathogenic role sera may be due to the immunostimulatory effect and the polyclonal activation often observed in the course of EBV infection. The majority of autoimmune-type aPl are dependent on the presence of 2-GPI [42,43] and, possibly, other plasma proteins, such as prothrombin and annexin V [21,22]. syndrome involving the phospholipid-binding plasma proteins. infection. Exclusion criteria were: age under 18 years, pregnancy or lactation, alcoholism, severe concomitant diseases, penicillin allergy, disorders of clotting, oesophageal reflux disease, gastric ulcer or resection of belly for gastric neoplasm, antibiotics or bismuth salts therapy during the month prior to inclusion, as well as proton pump inhibitor therapy during the last week. In all cases the identification of contamination was based on the histological examination of endoscopic biopsy specimens and confirmed by CLO test (Delta West Pty Ltd, W. Australia); 30 healthy blood donors. Detection of aCl by immunostaining on TLC plates Lathosterol Immunostaining was performed as previously explained [25]. Briefly, this assay was performed using 2 g of five different phospholipid antigens: CL, phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine and phosphatidylinositol (PI), purchased by Sigma Chemical Co. (St Louis, MO). Phospholipids were separated by thin-layer Lathosterol chromatography, using aluminium-backed silica gel 60 (20 20) high performance thin layer chromatography (HPTLC) plates (Merck, Darmstadt, Germany). Chromatography was performed in chloroform:methanol:CH3COOH:water (100:75:7:4) (v/v/v/v). The dried chromatograms were soaked for 90 s in a 05% (w/v) answer of poly(isobutyl methacrylate) beads (Polysciences, Warrington, PA) dissolved in hexane. After air flow drying, the chromatograms were incubated for 1 h at 25C in 05% (w/v) gelatin/PBS. The blocking answer was removed and replaced by a washing buffer (PBS). The chromatograms were then incubated for 1 h at 25C with sera, diluted 1:100 in 05% (w/v) gelatin/PBS. Sera were removed and chromatograms were washed three times for 10 min with PBS. Horseradish peroxidase-conjugated goat anti-human IgG (Sigma), diluted 1:500 in 05% (w/v) gelatin/PBS, was added and incubated at 25C for 1 h. The colour reaction was obtained Lathosterol by adding 200 mg of sodium nitroprusside (Sigma) and 80 mg of contamination504 (8%)0000Healthy blood donors3000000 Open in a separate windows aCl, Anti-cardiolipin antibodies; aPC, anti-phosphatidylcholine antibodies; aPE, anti-phosphatidylethanolamine antibodies; aPI, anti-phosphatidylinositol antibodies; IM, infectious mononucleosis; SLE, systemic lupus erythematosus; PAPS, main anti-phospholipid antibody syndrome. aCl were detected in 12 out of 18 (666%) SLE(APS?) sera; two of these sera reacted with phosphatidylserine and PE and none with PC or PI. In the PAPS group the prevalence of aCl antibodies was 523% Lathosterol (11 out of 21 sera); six of these sera reacted with phosphatidylserine, five with PE and none with PC or PI (Fig. 1, Table 1). Among the patients with contamination only four sera reacted with Cl and none with the other phospholipids under test. None of the sera from 30 healthy blood donors showed any presence of aPl. Detection of anti-cofactor protein antibodies None of the patients with IM experienced detectable levels of anti-2-GPI antibodies. However, anti-2-GPI antibodies were detected in seven sera from your PAPS group (333%) and in SLE(APS?) patients (166%) (Fig. 2). None of the sera from patients with contamination or from your healthy blood donors showed the presence of anti-2-GPI antibodies. Open in a separate window Fig. 2 Anti-cofactor protein antibody pattern in infectious and autoimmune sera. The occurrence of anti-cofactor protein antibodies in infectious mononucleosis (IM), systemic lupus erythematosus (SLE), primary anti-phospholipid antibody syndrome (PAPS), infection patients and healthy blood donors was detected by ELISA. The prevalence of anti-annexin V antibodies in patients with IM was 260%, as against 619% found in PAPS patients and 222% in SLE(APS?) patients. Anti-annexin V antibodies were Lathosterol also found in one patient with infection (2%) and in one healthy Rabbit Polyclonal to SCFD1 blood donor (33%). Anti-protein S antibodies were detected in 10.