Lairini K, Stenbaek E, Lacouture S, Gottschalk M
Groupe de Recherche sur les Maladies Infectieuses du Porc (GREMIP), Faculté de Médecine Vétérinaire, Université de Montréal, Qué., Canada.
Vet Microbiol. 1995 Oct;46(4):369-81. doi: 10.1016/0378-1135(94)00139-n.
Three monoclonal antibodies (mAbs) against Actinobacillus pleuropneumoniae serotype 1, designated 4.2 A11 B5, 5.1 G8 F10 and 1.5 C5 F4 (IgG3, IgG2b and IgM respectively), were produced and characterized. mAbs 4.2 A11 B5 and 5.1 G8 F10 were directed against different epitopes located in the O chain of the LPS. Both clones also recognized reference strains of A. pleuropneumoniae serotypes 9 and 11. The mAb 1.5 C5 F4 reacted with the reference strain of A. pleuropneumoniae serotype 1, with the encapsulated strain 4045 (but not with its non-capsulated mutant) and with A. pleuropneumoniae serotype 1 purified capsular polysaccharides (CPS). The epitope was sensitive to periodate oxidation, heat-labile, and located in the capsular material of A. pleuropneumoniae serotype 1, as demonstrated by immunoblotting. Treatment of the CPS with 5% ammonium hydroxide eliminated the reaction, which may indicate that the epitope recognized by 1.5 C5 F4 mAb is a O-acetyl containing determinant. When different A. pleuropneumoniae field strains were tested, the percentage of strains recognized by the mAbs varied with the mAb and the test used. Cross-reactions associated with the LPS of some A. pleuropneumoniae serotype 5 field strains could be observed with the 4.2 A11 B5 mAb. Of the three mAbs characterized, 1.5 C5 F4 seemed to be the most suitable for A. pleuropneumoniae serotype 1 detection since it reacted with 99% of serotype 1 field strains and it did not recognize any of the strains belonging to other serotypes.
制备并鉴定了三种抗胸膜肺炎放线杆菌1型的单克隆抗体（mAb），分别命名为4.2 A11 B5、5.1 G8 F10和1.5 C5 F4（分别为IgG3、IgG2b和IgM）。单克隆抗体4.2 A11 B5和5.1 G8 F10针对位于脂多糖O链上的不同表位。这两个克隆还识别胸膜肺炎放线杆菌9型和11型的参考菌株。单克隆抗体1.5 C5 F4与胸膜肺炎放线杆菌1型参考菌株、包膜菌株4045（但不与其无包膜突变体反应）以及胸膜肺炎放线杆菌1型纯化的荚膜多糖（CPS）反应。通过免疫印迹证明，该表位对高碘酸盐氧化敏感、热不稳定，且位于胸膜肺炎放线杆菌1型的荚膜物质中。用5%氢氧化铵处理CPS消除了反应，这可能表明1.5 C5 F4单克隆抗体识别的表位是一个含O-乙酰基的决定簇。当检测不同的胸膜肺炎放线杆菌田间菌株时，单克隆抗体识别的菌株百分比因单克隆抗体和所用检测方法而异。用4.2 A11 B5单克隆抗体可观察到一些胸膜肺炎放线杆菌5型田间菌株的脂多糖相关交叉反应。在所鉴定的三种单克隆抗体中，1.5 C5 F4似乎最适合用于胸膜肺炎放线杆菌1型检测，因为它与99%的1型田间菌株反应，且不识别任何其他血清型的菌株。
