Effects of Pasteurella haemolytica cytotoxin on ovine peripheral blood leucocytes and lymphocytes obtained from gastric lymph

Abstract

Ovine peripheral blood leucocytes were separated on discontinuous Percoll density gradients into fractions rich in lymphocytes (PBLy) and polymorphonuclear leucocytes. Supernatant fluid from a dialysis sac culture of Pasteurella haemolytica biotype A serotype 1 (A1) was cytotoxic to these leucocytes in vitro. PBLy retained viability after storage in liquid nitrogen and could be employed in cytotoxicity assays. However, sheep cannulated via the common gastric lymph duct were an excellent source of large numbers of homogeneous lymphoid cells (GLy) which also stored well in liquid nitrogen. As both freshly collected and stored GLy were killed by culture supernatant fluid GLy offer advantages as target cells for further characterisation of the extracellular cytotoxin produced by P. haemolytica. From the results obtained, it is considered that all ovine leucocytes are susceptible to P. haemolytica cytotoxin.