Assessment of antibody-independent cellular cytotoxicity (AICC) of porcineneutrophilic granulocytes by quantitative flow cytometry. Lack of modulation by larval products of Oesophagostomum dentatum
Hj. Schuberth et al., Assessment of antibody-independent cellular cytotoxicity (AICC) of porcineneutrophilic granulocytes by quantitative flow cytometry. Lack of modulation by larval products of Oesophagostomum dentatum, J VET MED B, 47(8), 2000, pp. 607-617
Citations number
56
Categorie Soggetti
Veterinary Medicine/Animal Health
Journal title
JOURNAL OF VETERINARY MEDICINE SERIES B-INFECTIOUS DISEASES AND VETERINARYPUBLIC HEALTH
After infection of pigs by the larvae of Oesophagostomum dentatum, granulom
as are formed around the third-stage larvae in the submucosa of the gut whi
ch contain a considerable number of neutrophils. This has no obvious impact
on the larvae, which develop to fourth-stage larvae within these granuloma
s. We therefore asked, whether the products of O. dentatum larvae modulate
the functional capacity of porcine neutrophils. The antibody-independent ce
llular cytotoxicity (AICC) was chosen as a model system. This assay was dev
eloped for the pig and quantified using flow cytometry. Bovine lymphoblasto
id cells (cell line Anna TA1) served as targets. The measurement of cytotox
icity was based on the determination of absolute numbers of vital target ce
lls. This procedure proved to be reliable and required no additional labell
ing of target and/or effector cells. Porcine neutrophils, when stimulated w
ith phorbol 12-myristate 13-acetate (PMA; greater than or equal to 10 nmol/
l), killed target cells at effector:target ratios between 1 : 1 and 9 : 1.
AICC was not demonstrable after 4 h but could be observed between 16 h and
20 h after in vitro co-culture. Killing of targets required close physical
contact between effector and targets, since supernatants of PMA-stimulated
polymorphonuclear cells were not able to lyse the target cells. Homogenates
of third- and fourth-stage larvae of O. dentatum did not affect the vitali
ty of porcine granulocytes or target cells in vitro, nor did they modulate
the AICC capacity of porcine granulocytes.