Palusiak Agata, Sidorczyk Zygmunt
Department of General Microbiology, Institute of Microbiology, Biotechnology and Immunology, University of Łódź, Banacha 12/16, 90-237 Łódź, Poland.
Arch Immunol Ther Exp (Warsz). 2009 Jul-Aug;57(4):303-10. doi: 10.1007/s00005-009-0034-9. Epub 2009 Jul 4.
Both smooth and rough Proteus sp. strains can be found. The latter are characterized by their lack of an O-polysaccharide chain in the lipopolysaccharide (LPS) molecule, which makes them suitable for obtaining anti-core sera. Using this kind of material enables identifying fragments of the Proteus LPS core region that might be involved in cross-reactions. To date only a few similar epitopes have been established for the genus Proteus.
Polyclonal rabbit antisera directed against three rough strains of Proteus sp. were tested by enzyme-linked immunosorbent assay (ELISA) with a set of LPSs. The reactivity of the selected cross-reactive and homologous systems was checked by the Western blot technique and by a passive immunohemolysis assay preceded by the absorption of each antiserum with appropriate cross-reactive and homologous alkalized LPSs.
On the basis of the ELISA results, 19 cross-reactive antigens were selected among which both smooth and rough LPS forms were found. All the observed reactions involved the core region of the LPS. Using the antisera absorbed with the appropriate LPSs allowed identification of four groups of antigens with serologically identical core regions.
Comparing the results of the serological studies with the known chemical structures of the core regions of the LPSs used enabled the identification of a few core oligosaccharide fragments probably involved in the observed cross-reactions. All were located in the most distal part of LPS core region, which made them more easily recognized by specific antibodies.
变形杆菌属菌株既有光滑型也有粗糙型。后者的特征是其脂多糖(LPS)分子中缺乏O - 多糖链,这使得它们适合用于制备抗核心血清。使用这种材料能够鉴定出可能参与交叉反应的变形杆菌LPS核心区域片段。迄今为止,变形杆菌属仅确定了少数几个类似的表位。
用一组LPS通过酶联免疫吸附测定(ELISA)检测针对三种变形杆菌粗糙菌株的兔多克隆抗血清。通过蛋白质印迹技术以及在每种抗血清用适当的交叉反应性和同源性碱化LPS吸收后进行的被动免疫溶血测定,检查所选交叉反应性和同源性系统的反应性。
根据ELISA结果,选择了19种交叉反应性抗原,其中发现了光滑型和粗糙型LPS形式。所有观察到的反应都涉及LPS的核心区域。使用用适当LPS吸收的抗血清能够鉴定出四组具有血清学相同核心区域的抗原。
将血清学研究结果与所用LPS核心区域的已知化学结构进行比较,能够鉴定出一些可能参与观察到的交叉反应的核心寡糖片段。所有这些片段都位于LPS核心区域的最远端,这使得它们更容易被特异性抗体识别。