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Volumn 272, Issue 5258, 1996, Pages 104-107

Protective effect of rotavirus VP6-specific IgA monoclonal antibodies that lack neutralizing activity

Author keywords

[No Author keywords available]

Indexed keywords

IMMUNOGLOBULIN A ANTIBODY; MONOCLONAL ANTIBODY; VIRUS ANTIBODY; VIRUS CAPSID ANTIGEN;

EID: 0029962378     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.272.5258.104     Document Type: Article
Times cited : (468)

References (33)
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    • Mazanec, M.B.1    Coudret, C.L.2    Fletcher, D.R.3
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    • note
    • Four to six days after oral immunization with infectious wild-type munne rotavirus strains EC, EHP, or EW (9), Peyer's patch lymphocytes were obtained from 4-month-old mice and fused with FOX-NY myeloma cells (21), as previously described (22) Rotavirus-positive clones were identified with an immunohistochemical staining procedure and were isolated and characterized as previously described (23) The protein specificity of the clones was determined by ELISA with the use of recombinant baculovirus antigen or solid-phase immunoisolation techniques (or both) using radiolabeled rotavirus-infected cell lysates (22, 24) The isotype of the individual mAbs was determined as previously described (22), and the molecular weight was determined on non-denaturing PAGE gels as previously described (25), In vitro neutralization assays of individual mAbs were carried out as previously described (23).
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    • unpublished data
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    • note
    • 4 shedding dose 50s of wild-type EC munne rotavirus) and 1 hour after challenge (100 μl per animal). Subsequently, ascites were fed every 8 hours for 8 days Stool samples were collected and antigen shedding was measured by ELISA.
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    • 6 shedding dose 50s) plus 25 μl of 7D9 ascites (1:4 dilution); (ii) 25 μl of activated EC plus 25 μl of a cocktail of four high-titer neutralizing IgG antibodies directed at VP4 and VP7 (27, 28) (mAbs 4F8, 159, 2G4, and 57-4, each 1:4 diluted); (iii) 25 μl of activated EC plus 25 μl of phosphate-buffered saline (PBS), and (iv) 25 μl of mouse myeloma IgA (undiluted, 1 mg/ml) (Sigma) plus 25 μl of activated EC After injection, the loops were returned to the abdominal cavity and the abdominal wall was closed The loops were removed 12 hours later and immunostained for EC with a rabbit hyperimmune serum against rotaviruses (Fig 4).
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    • note
    • Twenty-five microliters of activated EC plus 25 μl of PBS were injected into the intestinal loops of a mouse that had received the 7D9 hybridoma transplant 2 weeks before. The loop was removed and immunostained for EC 12 hours after inoculation as described [(18) and Fig. 4 legend].
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    • note
    • Supported by NIH grants R37AI21362 and DK38707, by a VA Merit Review grant, and by funds from the World Health Organization. H.B.G. is a VA Medical Investigator


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.