Selected isolates of bovine viral diarrhea (BVD) virus propagated on bovine turbinate cells: Virus titer and soluble antigen production as factors in immunogenicity of killed BVD virus

1978 ◽  
Vol 58 (2) ◽  
pp. 119-125 ◽  
Author(s):  
A. W. McClurkin ◽  
M. F. Coria
Keyword(s):  
Virus Titer ◽  
Bovine Viral Diarrhea ◽  
Soluble Antigen ◽  
Viral Diarrhea ◽  
Antigen Production ◽  
Bovine Turbinate

Immunogold-Silver Staining (IGSS) of (NCP and CP) BVDV Infected Subcellular Fraction Bands

1997 ◽  
Vol 3 (S2) ◽  
pp. 155-156
Author(s):  
C.E. Hearne ◽  
D.L. Johnson ◽  
H. Van Campen
Keyword(s):  
Subcellular Fraction ◽  
Membrane Vesicles ◽  
Bovine Viral Diarrhea ◽  
Fixed Angle ◽  
Cell Monolayers ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Transmission Electron ◽  
Viral Diarrhea Virus ◽  
Bovine Turbinate

Pre-embedding immunogold-silver (IGSS) techniques are useful to localize antigens in cell monolayers and agarose embedded cell suspensions for transmission electron microscopy. Procedural centrifugations, however, present a challenge when attempting to localize antigens in subcellular fractions. Using a Beckman Airfuge Ultracentrifuge to concentrate the subcellar fraction bands and resuspending the organelles in agarose simplifies IGSS processing and resin embedding procedures.Control bovine turbinate (BT), and BT cells infected with cytopathic (cp NADL) and non-cytopathic (ncp NY-1) strains of bovine viral diarrhea virus (BVDV) were fractioned according to Bienz et al (1992). Bands containing membrane vesicles (Fig 2) were collected and each fraction band was pelleted at 169,000g for 20 min using an A-95 fixed angle rotor in a Beckman Airfuge Ultracentrifuge. Each fraction pellet was resuspended in 50μl of 30% agarose, solidified, and trimmed to < lmm. IGSS procedures were carried out according to Nanoprobes, Inc., Stoney Brook, NY, and Hearne & Van Campen (1996).

Soluble antigen of bovine viral diarrhea virus

1965 ◽  
Vol 15 (2) ◽  
pp. 159-168 ◽  
Author(s):  
Donald E. Gutekunst ◽  
Winston A. Malmquist
Keyword(s):  
Bovine Viral Diarrhea Virus ◽  
Bovine Viral Diarrhea ◽  
Soluble Antigen ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Diarrhea Virus

Immunogold-Silver Staining (IGSS) of Agarose Embedded (NCP) Bvdv-Infected Cell Suspensions

1996 ◽  
Vol 54 ◽  
pp. 902-903
Author(s):  
C.E. Hearne ◽  
H. Van Campen
Keyword(s):  
Silver Staining ◽  
Phosphate Buffer ◽  
Cell Suspensions ◽  
Bovine Viral Diarrhea ◽  
Require Time ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Cell Fractions ◽  
Viral Diarrhea Virus ◽  
Bovine Turbinate

The use of pre-embedding immunogold-silver staining (IGSS) in electron microscopy of cell culture monolayers has been reported by various authors and its usefulness to localize antigens has been documented. Embedding cell suspensions or cell fractions for IGSS can require time-consuming centrifugation steps. Embedding cell suspensions in agarose simplifies IGSS processing and embedding procedures.Bovine turbinate cells (BTs) were grown to confluence in 25cm3 culture flasks. BTs were infected with a non-cytopathic strain (ncp) of bovine viral diarrhea virus (BVDV) while control cells were sham inoculated. At 18 h pi, cells were detached with trypsin. All following procedures were carried out at RT. The cells were fixed for 30 min in 2.0% paraformaldehyde + 0.2% glutaraldehyde in 0.1 M. phosphate buffer, pH 7.4, and rinsed 3 times in 0.1 M phosphate buffer, pH 7.4 (each centrifuged for 10 min at 300g). The cells were resuspended in 1 ml of buffer, transferred to an eppendorf tube and centrifuged for 10 min at 300g. The excess buffer was removed, 1-2 drops of warm, molten 1.0% agarose added, and the mixture centrifuged for 1 min at 300g to pellet the cells to the tip of the tube. The solidified cell-agarose mixture was removed, cut into <0.5mm3pieces, and placed in phosphate buffer.

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