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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Virus and Prion Research » Research » Publications at this Location » Publication #85329
Title: IN VITRO ACTIVATION OF VIRUS-SPECIFIC T-LYMPHOCYTES FROM CATTLE INFECTED WITH BVDV

Author
item QUADE, M - IOWA STATE UNIVERSITY
item ROTH, J - IOWA STATE UNIVERSITY
item Bolin, Steven - Steve

Submitted to: Research Workers in Animal Diseases Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 11/11/1997
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: The purpose of this experiment was to detect BVDV antigen specific activation of T lymphocytes in vitro. Peripheral blood mononuclear cells (PBMC) were isolated from two BVDV-infected cattle and from two noninfected control cattle. Isolated PBMC were cultured in medium with or without homologous-live BVDV Con A. Cultured PBMC were subjected to lymphocyte blastogenesis testing and 2-color flow cytometry. For blastogenesis, cells were pulsed with 3**H-thymidine on day 5 of culture and harvested on day 6. PBMC from BVDV-infected animals had an average stimulation index of 7.3 when cultured with BVDV; the stimulation index of PBMC from uninfected animals was 0.36 when cultured with BVDV. For flow cytometry, cells were removed from culture wells after 3 days, washed, and incubated with monoclonal antibody against bovine CD3 and either interleukin 2 receptor (IL2r) or MHC II. Activated lymphocytes were identified by increased expression of IL2r and/or MHC II. Cells were stained with propidium iodide to discriminate between live and dead cells, and the two color fluorescence of live cells was analyzed. The percentage of CD3+ cells from infected cattle expressing IL2r increased from 51% to 71% when cultured with BVDV, and the percentage of CD3+ cells expressing MHC II increased from 61% to 92.5% when cultured with BVDV. The average fluorescence intensity for both IL2r and MHC II also increased.