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Volumn 288, Issue 5474, 2000, Pages 2222-2226

A primitive T cell-independent mechanism of intestinal mucosal IgA responses to commensal bacteria

Author keywords

[No Author keywords available]

Indexed keywords

BACTERIAL ANTIGEN; IMMUNOGLOBULIN A;

EID: 0034705397     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.288.5474.2222     Document Type: Article
Times cited : (921)

References (32)
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    • note
    • 2). Cells were washed off, and plates were incubated successively with 1:1000 affinity-purified goat anti-mouse IgA (Sigma) and 1:1000 alkaline phosphatase-conjugated donkey anti-goat IgG (Jackson ImmunoResearch) before development of alkaline phosphatase color reactions.
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    • note
    • a (PharMingen) were diluted in 100 μl of PBS containing 3% BSA 1:3 over seven to nine steps starting from 1 in 30, 1 in 3, or 3 μg/ml, respectively. Lower dilutions of serum were used in some experiments to compare overall binding between serum and intestinal washings, although the technique of intestinal washing does inherently dilute the secretory IgA sample. After washing, the secondary antibody was goat affinity-purified, peroxidase-conjugated anti-mouse IgA (Sigma; 1:1000 in 100 μl of PBS containing 3% BSA), and the enzyme reaction was developed. Bacterial protein or LPS binding was measured similarly after coating with 1 μg/ml of pure OmpF protein (24), 5 μg/ml of pure LPS isolated from cecal wild-type E. cloacae [O. Westphal and K. Jann, Adv. Carbohydr. Chem. 25, 83 (1966)] or from E. coli O26B6 (Sigma), 1 μg/ml of purified chimeric Xa-CAT or 1 μg/ml of purified FimH (26).
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    • lo) and injected in basal salts solution. Chimeras were analyzed after 5 to 6 months.
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    • note
    • / mice, to P. Sebbel and R. Glockshuber for the purified FimH protein, to I. Bloomfield for E. coli strain AAEC356, to G. Dougan for advice about in vivo expression, to B. Odermatt for immunohistology and to B. Ludewig, K. McCoy, K. Maloy, and A. Lamarre for helpful discussion.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.