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Volumn 288, Issue 5473, 2000, Pages 2051-2054

Role of CD47 as a marker of self on red blood cells

Author keywords

[No Author keywords available]

Indexed keywords

CD47 ANTIGEN; REGULATOR PROTEIN;

EID: 0034674421     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.288.5473.2051     Document Type: Article
Times cited : (1452)

References (37)
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    • note
    • -/- mice (33) on a mixed 129/C56BL/6 background were a gift from H. Molina (Washington University, St. Louis, MO). Animals were kept in accordance with NIH and local guidelines and maintained in a specific pathogen-free barrier facility.
  • 17
    • 0343767681 scopus 로고    scopus 로고
    • note
    • -/- recipients (detection by staining for CD47 and flow cytometry) gave similar results.
  • 20
    • 0342896656 scopus 로고    scopus 로고
    • note
    • Dichloromethylene diphosphonate (clodronate; a gift from Roche Diagnostics/Boehringer Mannheim GmbH) liposomes was prepared as described (T8). Clodronate (200 μl) or NaCl liposomes were given to recipient mice by tail-vein injection at 48 and 24 hours before RBC transfer. Immunohistochemical analyses revealed a complete macrophage depletion in recipient spleens 48 hours after a single injection of clodronate liposomes.
  • 21
    • 0343767679 scopus 로고    scopus 로고
    • note
    • + cells were visualized with Tyramide Signal Amplification (NEN Life Science Products, Boston, MA).
  • 23
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    • S. Adams et al. J. Immunol. 161, 1853 (1998).
    • (1998) J. Immunol. , vol.161 , pp. 1853
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    • 0343331790 scopus 로고    scopus 로고
    • note
    • -/- RBCs were added to each well for another 60 min. Nonadherent cells were washed off with warm phosphate-buffered saline (PBS)-0.1% HSA, and noningested RBCs were lysed with ACK lysis buffer. Phagocytosis index (the number of RBCs phagocytosed per 100 adherent macrophages) was determined by fluorescence microscopy (Zeiss Axioscope).
  • 27
    • 0343767676 scopus 로고    scopus 로고
    • note
    • -/- RBCs were added, then allowed to settle for 3 min at 4 °C, after which sodium pervanadate (Sigma) was added to 2 mM and cells were transferred to a water bath at 37 °C At the times indicated, the medium was quickly aspirated, and cells were lysed in cold lysis buffer [50 mM tris-HCl (pH 7.5), 150 mM NaCl, 1% NP-40 (Pierce, Rockford, IL), 1 mM diisopropyl, aprotinin (10 μg/ml), 2 mM sodium pervanadate]. Lysates were cleared by centrifugation and incubated for 2 hours at 4 °C with anti-SIRP monoclonal antibody (mAb) P84 bound to 20 μl of goat anti-rat Sepharose (ICN, Costa Mesa, CA) for 2 hours at 4 °C. After two washes in lysis buffer, immunoprecipitated proteins were eluted with reducing SDS-polyacrylamide gel electrophoresis (PACE) sample buffer, separated on 8% SDS-PACE, and transferred to a polyvinylidene difluoride membrane (Millipore, Bedford, MA). Tyrosine phosphorylation was detected by Western blot with anti-phosptiotyrosine mAb 4G10, followed by HRP-conjugated secondary antibody and enhanced chemoluminescence as described (36).
  • 29
    • 0342462584 scopus 로고    scopus 로고
    • unpublished data
    • F. P. Lindberg et al., unpublished data.
    • Lindberg, F.P.1
  • 31
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    • M. Seiffert et al. Blood 94, 3633 (1999).
    • (1999) Blood , vol.94 , pp. 3633
    • Seiffert, M.1
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    • P. Mombaerts et al., Cell 68, 869 (1992).
    • (1992) Cell , vol.68 , pp. 869
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  • 37
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    • note
    • We thank T. Steinberg, W. Yokoyama, J. Atkinson, and E. Brown for critically reading the manuscript; D. Chaplin, H. Molina, and P. Leenen for reagents; and E. Ford for technical assistance. Supported by grants from the NIH (GM57S73-01), the American Diabetes Association, the Washington University-Monsanto Research Agreement, a pilot grant from the Howard Hughes Medical Institute, and the Medical Research Service of the Depatment of Veterans Affairs. P.-A.O is supported by a postdoctoral fellowship from the Umeå University-Washington University exchange program and by the Swedish Medical Research Council.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.