The University of Georgia College of Veterinary Medicine

eorgia Veterinary Scholars Program

GVSP Summer 2008 Scholars

Georgia Veterinary Scholar	Faculty Mentor

Cheryl Bettis University of Georgia Class of 2011	Drs. Tomas Krunkosky & Amelia Woolums

Establishment of Bovine Primary Airway Cultures and Their Responses to BRSV Infection

Cheryl Bettis*. Jarrett, Carla; Woolums, Amelia; Ard, Mary; Berghaus, Londa; Krunkosky, Thomas; Dept. of Anatomy and Radiology, Large Animal Medicine, Pathology, College of Veterinary Medicine, University of
Georgia, Athens GA 30602

Bovine respiratory disease complex (BRDC) is a polymicrobial infection of cattle caused by viral and bacterial coinfection and exacerbated by a variety of stressors. Bovine respiratory syncytial virus (BRSV) is a major contributor to this complex. Study of bovine airway epithelium is difficult in vivo; therefore, establishing an invitro culture system will allow examination of airway epithelial cell responses. Goals of this project included 1) establishing a fully differentiated primary cell culture of bovine bronchial epithelium (BBE), 2) infection of a
“control” airway epithelium with BRSV, 3) infection of the bronchial epithelium with BRSV, and 4) chemokine/adhesion molecule profiling of cell responses. Bovine nasal turbinate (BT) cells were used as a positive control for virus infectivity. Immunohistochemistry and confocal microscopy were performed on cells labeled with anti-BRSV F-protein antibody (to confirm virus infectivity), B-tubulin (to identify cilia), DAPI (to identify nuclei), and phalloidin (to identify F-actin). BRSV-infected BT cells exhibited syncytial formation, a classic pathologic feature of this virus. BBE cells were isolated from calves using bronchial rushings,
proliferated in tissue culture flasks, and allowed to differentiate in an air-liquid interface culture system. Confocal, SEM, and TEM microscopy were utilized to validate differentiation and infection. The BBE cells were exposed to BRSV in both differentiated and undifferentiated states and examined for infection and cell
response. Chemokine and adhesion molecule expression was assessed by real time PCR. This model will be a useful tool to determine how respiratory epithelial cell responses contribute to the pathogenesis of BRDC.

Research Support: UGA Dept. of Anatomy and Radiology and Vet. Med. experiment Station Student Support: Merck-Merial and UGA Veterinary Medical Experiment Station