Zigterman G J, Verheul A F, Ernste E B, Rombouts R F, De Reuver M J, Jansze M, Snippe H, Willers J M
Laboratory of Microbiology, Department of Experimental Immunology, Utrecht, The Netherlands.
J Immunol Methods. 1988 Jan 21;106(1):101-7. doi: 10.1016/0022-1759(88)90277-3.
A sensitive ELISA has been developed to study immune responses in mice against Streptococcus pneumoniae type 3 capsular polysaccharide (S3PS) and hexasaccharide (HS)-protein conjugates derived therefrom. An advantage of the described system is that the same microtiter plates can be used for both ELISA and ELISPOT tests with a standardized washing procedure and diluent composition. S3PS induced predominantly IgM antibodies and minute amounts of IgG as measured by ELISA in serum. This was accompanied by large numbers (greater than 14000) of IgM spot-forming cells in the spleen. A shift towards IgG production was achieved by addition of lipid A. HS-protein conjugates induced predominantly IgG antibodies after booster immunization(s). Furthermore these conjugates induced large numbers (greater than 40000) of IgG spot-forming cells (SFC) in the spleen. ELISA and ELISPOT assays on microtiter plates are both reliable and highly reproducible assays for the evaluation of immune responses to S. pneumoniae antigens.
已开发出一种灵敏的酶联免疫吸附测定法（ELISA），用于研究小鼠针对3型肺炎链球菌荚膜多糖（S3PS）及其衍生的六糖（HS）-蛋白缀合物的免疫反应。所述系统的一个优点是，相同的微量滴定板可用于ELISA和酶联免疫斑点试验（ELISPOT），且具有标准化的洗涤程序和稀释剂组成。通过ELISA测定，S3PS在血清中主要诱导IgM抗体和微量IgG。与此同时，脾脏中出现大量（超过14000个）IgM斑点形成细胞。通过添加脂多糖实现了向IgG产生的转变。在加强免疫后，HS-蛋白缀合物主要诱导IgG抗体。此外，这些缀合物在脾脏中诱导大量（超过40000个）IgG斑点形成细胞（SFC）。微量滴定板上的ELISA和ELISPOT测定法都是评估对肺炎链球菌抗原免疫反应的可靠且高度可重复的测定法。
