Kariyawasam S, Wilkie B N, Hunter D B, Gyles C L
Department of Pathobiology, Ontario Veterinary College, University of Guelph, Canada.
Avian Dis. 2002 Jul-Sep;46(3):668-78. doi: 10.1637/0005-2086(2002)046[0668:SAMART]2.0.CO;2.
The immune response to four cell surface antigens of avian pathogenic Escherichia coli (APEC) was investigated as the first step in identifying vaccine candidates. F1 pilus adhesin, P pilus adhesin, aerobactin receptor protein, and lipopolysaccharide (LPS) from an O78 E. coli (strain EC99) were used as antigens. The proteins were purified as 6xhistidine-tagged recombinant proteins and LPS was purified from a phenol/water extract. Groups of 12 broiler chickens were vaccinated intranasally with the EC99 strain and challenged with the same strain 10 days later via the intra-air sac route. The chickens that survived were euthanatized 10 days postchallenge. Scores were assigned to infected chickens on the basis of lesions and recovery of the challenge E. coli. The immunoglobulin (Ig) IgG, IgA, and IgM antibodies to the four antigens were measured in serum and air sac washings in an enzyme-linked immunosorbent assay. Among the chickens that were not vaccinated prior to challenge, two died and three of the survivors were ill, whereas, of the chickens that were vaccinated prior to challenge, one died and one of the survivors became ill. After the intranasal vaccination, high antibody activity against all four antigens was associated with each Ig isotype in serum and air sac washings. IgG was the predominant isotype of Ig in air sac washings as detected by radial immunodiffusion. Chickens that were not ill after challenge had greater IgG, IgA, and IgM antibody activity against all four antigens in serum and air sac washings than did sick chickens. Thus, all of the antigens tested appear to be suitable candidates for a vaccine to protect chickens from respiratory tract infections caused by APEC.
作为确定候选疫苗的第一步,对禽致病性大肠杆菌(APEC)的四种细胞表面抗原的免疫反应进行了研究。来自O78大肠杆菌(菌株EC99)的F1菌毛黏附素、P菌毛黏附素、气杆菌素受体蛋白和脂多糖(LPS)被用作抗原。这些蛋白质被纯化为带有6x组氨酸标签的重组蛋白,LPS则从酚/水提取物中纯化得到。将12只肉鸡分为一组,通过鼻内接种EC99菌株,并在10天后通过气囊内途径用同一菌株进行攻毒。存活的鸡在攻毒后10天进行安乐死。根据病变情况和攻毒大肠杆菌的恢复情况对感染鸡进行评分。在酶联免疫吸附测定中,检测血清和气囊肿洗液中针对这四种抗原的免疫球蛋白(Ig)IgG、IgA和IgM抗体。在攻毒前未接种疫苗的鸡中,有2只死亡,3只存活鸡患病;而在攻毒前接种疫苗的鸡中,有1只死亡,1只存活鸡患病。鼻内接种后,血清和气囊肿洗液中针对所有四种抗原的高抗体活性与每种Ig同种型相关。通过放射免疫扩散检测,IgG是气囊肿洗液中主要的Ig同种型。攻毒后未患病的鸡血清和气囊肿洗液中针对所有四种抗原的IgG、IgA和IgM抗体活性高于患病鸡。因此,所有测试的抗原似乎都是保护鸡免受APEC引起的呼吸道感染的疫苗的合适候选物。