Jp. Scharsack et al., Flow cytometric analysis of mitogen-induced activation of rainbow trout (Oncorhynchus mykiss) peripheral blood leucocytes, J VET MED B, 48(5), 2001, pp. 331-339
Citations number
21
Categorie Soggetti
Veterinary Medicine/Animal Health
Journal title
JOURNAL OF VETERINARY MEDICINE SERIES B-INFECTIOUS DISEASES AND VETERINARYPUBLIC HEALTH
Proliferation of rainbow trout peripheral blood leucocytes in vitro is usua
lly assessed by measuring incorporated tritiated thymidine. In this report
we monitored the in vitro proliferative response to the mitogen Concanavali
n A (Con A) by means of flow cytometry (FCM) and H-3-thymidine incorporatio
n. When analysed by FCM, blood leucocytes displayed two main cell populatio
ns with distinct forward and side scatter (FSC/SSC) characteristics: lympho
cytes with low FSC/SSC values and non-lymphoid leucocytes (NLL) with increa
sed FSC/SSC values. The nature of these cell types were confirmed by micros
copy. Interestingly, the FSC/SSC pattern of lymphocytes remained unchanged
after in vitro stimulation with Con A, whereas cells from the NLL populatio
n showed a marked shift towards increased FSC values. In stimulated culture
s, the increase of FSC values of the NLL population significantly correlate
d with contemporarily measured H-3-thymidine incorporation (r = 0.7, P < 0.
001). The mitogenic response of blood leucocytes originating from different
individual fish varied over wide ranges. It was found to be related to the
numbers of NLL present in the leucocyte sample. The present results show t
hat qualitative and quantitative FCM analysis of morphological parameters (
FSC/SSC) of blood leucocytes makes it possible to discriminate between leuc
ocyte populations of the rainbow trout and to monitor cell proliferation ex
periments.