J. Harenberg et al., PREFERENTIAL BINDING OF HEPARIN TO GRANULOCYTES OF VARIOUS SPECIES, American journal of veterinary research, 57(7), 1996, pp. 1016-1020
Objectives-To address the problem of heparin binding to leukocytes of
various animal species. Design-Leukocytes of the various species were
incubated with fluorescent-labeled, low molecular mass heparin (LMMH).
Fluorescence intensity on granulocytes, lymphocytes, and monocytes wa
s analyzed by flow cytometry analysis. Sample Population-leukocytes we
re prepared from EDTA-anticoagulated blood of human subjects, rats, ra
bbits, dogs, pigs, and sheep 3 times. Procedure-The leukocyte populati
ons were identified by their light scatter properties. In addition, ph
ycoerythrin-labeled CD4, CD13, and CD14 antibodies were used to identi
fy human lymphocytes, granulocytes, and monocytes; CD4, GR-1, and CD11
b antibodies were used for mouse, and CD45, RP 3, and ED 9 antibodies
were used for identification of rat leukocyte subpopulations. Results-
Granulocytes, monocytes, and lymphocytes of ail species bound LMMH in
dose-dependent manner. Binding of LMMH-tyramine (tyr)-fluorescein-5-is
othiocyanate (FITC) to granulocytes was higher in human subjects, rats
, rabbits, dogs, and pigs, compared with binding to monocytes and lymp
hocytes, Mouse and sheep granulocytes did not bind more heparin than m
onocytes or lymphocytes. Binding of LMMH-tyr-FITC was reversible in th
e presence of unlabeled heparin or LMMH. More than 99% of human, rat,
rabbit, dog, and sheep granulocyte populations were distinguished from
monocytes and lymphocytes by means of their fluorescence intensity ow
ing to LMMH-tyr-FITC, This separation was not obtained for mouse and p
ig granulocytes, Conclusion-Evidence of specific heparin binding to gr
anulocytes of many species indicates the relevance of fluorescent-labe
led LMMH for biological investigations. Clinical Relevance-Binding of
heparin and LMMH to granulocytes, lymphocytes, and monocytes may have
a substantial role in atherosclerosis, inflammation, malignancy, and i
mmunologic diseases.