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Volumn 285, Issue 5431, 1999, Pages 1261-1265

Rapid infection of oral mucosal-associated lymphoid tissue with simian immunodeficiency virus

Author keywords

[No Author keywords available]

Indexed keywords

VIRUS ANTIGEN;

EID: 0033588005     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.285.5431.1261     Document Type: Review
Times cited : (211)

References (38)
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    • note
    • 50 of the same virus stock and monitoring them with virologic assays for several weeks and by histology in one animal on day 4. At necropsy lymphoid and nonlymphoid organs were sampled from various body locations for virus isolation, histology, immunohistochemistry, and in situ hybridization.
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    • note
    • + T cell percentage by fluorescence-activated cell sorting (FACS).
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    • 6 dpm of probe per milliliter) overnight at 45°C in a moist chamber. After several washings in standard saline citrate (SSC), the sections were digested with ribonuclease at 37°C for 40 min, washed again in 2 × SSC dehydrated, and dipped in Kodak NTB-2 emulsion. Exposure at 4°C was for 2 to 3 days for frozen sections and 7 days for paraffin sections. After development in Kodak D-19, sections were counter-stained with hemalaun, mounted, and examined with an Axiophot Zeiss microscope equipped with epiluminescent illumination. Viral RNA - positive cells were counted with a 20× objective, a 3CD color camera, and a PC-based image analysis system (KS400; Kontrol, Esching, Germany). Positive cells had >20 silver grains, corresponding to a sixfold excess over background. Four entire sections were counted for each recorded value to obtain a mean number of infected cells per section and per unit area. The percentage of infected cells was then recorded for the epithelial, germinal center, and extrafollicular lymphoid tissue.
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    • note
    • Immunolabeling was performed on paraffin-embedded and cryostat sections according to the alkaline phosphatase anti-alkaline phosphatase method. Antibodies included CD68 (Dako, macrophages), CK1 (Dako, cytokeratin), p55 (provided by E. Langhoff, mature DCs), CD1a (Immunotech, immature DCs), CD4 (Leu3a, Becton Dickinson; OKT4, Ortho Diagnostics; and NCL, CD-4 1F6 Novocastra; all together). CO8 (Leu2a, Becton Dickinson, and C8/144B, Dako; together), and polyclonal CD3 (Dako, visualized with the peroxidase anti-peroxidase method). Immunolabeling was performed before in situ hybridization.
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    • note
    • 35S-labeled, single-stranded, antisense RNA probe of SIV-mac239 (Lofstrand Labs, Gaithersburg, MD) was obtained in collaboration with S. Gartner (Yale University, New Haven, CT) through the NIH Research Program. Grant support was provided by the NIH (AI 42129 and 40874 to R.M.S., and AI 40877 to M.P.), the German Ministry of Education and Research (BMBF, grant number 01KI-9767/9), the Korber Foundation (Hamburg), and the Fogarty Foundation (RO3TW00792).


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.