It suggests that LPS processed by macrophages results in the presentation of IC of LPS, which we showed previously to be a target for MBL [3]

It suggests that LPS processed by macrophages results in the presentation of IC of LPS, which we showed previously to be a target for MBL [3]. We also confirmed the presence ofYersiniaLPS-reacting antibodies in synovial fluids from patients diagnosed with JIA (Physique 3). importance of LPS-associated ECA for the antigenicity of endotoxin. Furthermore, we confirmedin vitrothatYersiniaLPS processing leads to the exposure of its core region and enhanced potency of match lectin pathway activation. == 1. Introduction == Yersinia enterocoliticaO:3 (YeO3) is usually a causative agent of gastrointestinal infections but may also cause sepsis, with a mortality rate above 50% [1]. It is characterized by certain unique features like the ability to multiply at an extensive range of temperatures (from <4C to >40C) and by temperature-regulated expression of some virulence factors. Furthermore, in contrast to the majority ofEnterobacteriaceae, its lipopolysaccharide (LPS, endotoxin) is composed of lipid A-inner core (IC) oligosaccharide backbone, substituted either with a long O-specific polysaccharide (OPS) chain (lipid A-IC-OPS) or an outer core oligosaccharide (OC) (lipid A-IC-OC) [25]. IC may be substituted with enterobacterial common antigen (ECA) polysaccharide (lipid A-IC-ECA-OPS) in OPS-carrying molecules, called ECALPS. In most Gram-negative bacteria endotoxins, OPS is usually attached to the outer core (lipid A-IC-OC-OPS); therefore, the molecule contains both OC and OPS. The composition of YeO3 LPS Ac-LEHD-AFC is usually influenced by the heat of growth. The lower favour the synthesis of molecules containing OPS while the higher culture heat of bacteria those decorated by shorter OC. No experimental data concerning YeO3 LPS biosynthesisin vivoare Mouse monoclonal to ESR1 available; however, both OPS and core region are essential for bacterial virulence. They modulate the activity of the LPS harmful theory (lipid A) and influence bacterial serum resistance. For example, shortening of OPS at 37C is usually associated with the increased ability ofYersiniaAil factor (outer membrane protein) to bind inhibitors of classical, lectin, and option match pathways (C4b-binding protein and H-factor) [6]. Additionally, substituting the inner core with OC or OPS/ECA prevents the conversation of mannose-binding lectin (MBL) conversation with IC heptose residues and associated match lectin pathway activation, YeO3 endotoxin IC, OC, or Ac-LEHD-AFC OPS are receptors for bacteriophages, considered potential diagnostic and therapeutic agents [7]. In some cases of yersinioses, post-infection manifestations like erythema nodosum or myocarditis are observed. Moreover, as mentioned,Y. enterocoliticamay cause sepsis, as a rare complication after blood transfusion (as it is able to survive in stored blood preparations) [8]. Infections caused byY. enterocoliticastrains of serotypes Ac-LEHD-AFC O:3 and O:9 are often complicated with the development of reactive arthritis (ReA) or juvenile idiopathic arthritis (JIA), mainly in HLA-B27 (human leukocyte antigen B27)-positive patients [9]. Although ReA was considered a sterile disease with no microbes found in the joints, immune complexes ofYersiniaantigens,Yersinia-specific antibodies of IgM, IgG, and IgA, classes as well LPS were found in sera and synovial fluids from patients diagnosed withYersinia-triggered ReA, even several years after infection [10]. We report here presence ofYersiniaLPS-reactive antibodies, recognizing lipid A-Kdo, core oligosaccharide, or OPS in synovial fluids from patients suffering from JIA. == 2. Materials and Methods == == 2.1. Clinical Material == Synovial fluid samples from 39 paediatric patients aged 3-18 years (mean 11.2 0.1) were collected at the Department of Rheumatology, St Louis Voivodeship Specialist Children’s Hospital (Cracow, Poland). Female patients accounted for 62%. Patients were subdivided into subgroups based on positive (JIA Ye+) or negative (JIA Ye-) results of the recomWell test (Mikrogen Diagnostics, Neuried, Germany), allowing for detection of IgG, IgM, and IgA antibodies and recognizingYersiniaouter membrane proteins (Yops) (Table 1). The results for 39 sera Ac-LEHD-AFC were available and reported earlier by Kasperkiewicz et al. [3]. Twelve of them (30.7%) were found to be positive. The study was approved by the Bioethics Committee of the Regional Medical Chamber in Cracow, and informed parental consent was obtained. This work confirms the provisions of the Declaration of Helsinki. == Table 1. == The comparison of clinical data for JIA patients positive and negative forYersiniaYops antibodies. : MannWithneyUtest; : Fisher’s exact test. == 2.2. Bacterial Strains, Growth Conditions, and LPS Isolation == The bacteria used in this work are listed inTable 2. They have grown aerobically at 37C in LB medium, in the presence of kanamycin or chloramphenicol when required. The LPS from smoothYersinia enterocoliticaO:3 (6471/76-c) andSalmonella entericaserovar Montevideo SH94 strains were isolated by Ac-LEHD-AFC the hot water method/water method [11]. In contrast, the LPS of the rough strains (YeO3-c-R1, YeO3-c-Rfb-R7, YeO3-c-R1-M205, and YeO3-c-OCR-ECA) were separated by the hot phenol/water extraction followed by the phenol/chloroform/petroleum.