Effects of three Hb-based oxygen-carrying solutions on neutrophil activation in vitro: quantitative measurement of the expression of adherence receptors.
BACKGROUND
Hb-based oxygen carriers (HbOCs) are currently under investigation as RBC substitutes. These solutions have vasoactive properties and may modify the behavior of the RBC in vitro. The effects of these agents on other blood cells have not been extensively investigated. The aim of this study was to evaluate the potential effect(s) of three differently modified HbOCs on the activation of PMNs.


METHODS
Whole blood from humans was incubated with Hb dextran-benzene-tetracarboxylate, alphaalpha-Hb, or o-raffinose-poly-Hb for 15, 30, and 60 minutes. The expression of adherence receptors CD62L, CD18, and CD11b, which reflects the activation state of the neutrophils, was assessed in vitro by a direct immunofluorescence method using calibration beads measured by flow cytometry.


RESULTS
The observed expression of each of the three adherence receptors for each solution at each time point was similar to that noted with the RPMI control. Decreases in CD62L and increases in CD18 expression were noted at 15 minutes of incubation but, when those values were compared to the values obtained with TNFalpha as the positive control of PMN activation, the profile of expression of adherence receptors obtained with HbOC solutions did not conform to the expected activation profile of PMNs.


CONCLUSIONS
Hb solutions modified by a variety of methods, as obtained and employed in this study, did not appear to activate neutrophils in vitro.
Toussaint
M
M
Department of Hematology and Physiology, Faculty of Pharmacy, Université de Nancy, France. marie.toussaint@pharma.u-nancy.fr
Latger-Cannard
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Caron
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Lecompte
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Stoltz
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Vigneron
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D001241:Aspirin / Q000031:analogs & derivatives* / Q000494:pharmacology*; D001802:Blood Substitutes / Q000494:pharmacology*; D002352:Carrier Proteins / Q000494:pharmacology*; D003911:Dextrans / Q000494:pharmacology*; D006454:Hemoglobins / Q000494:pharmacology*; D006801:Humans; D016023:Integrins / Q000096:biosynthesis; D007958:Leukocyte Count; D008875:Middle Aged; D018375:Neutrophil Activation / Q000187:drug effects*; D011887:Raffinose / Q000031:analogs & derivatives* / Q000494:pharmacology*; D012996:Solutions / Q000494:pharmacology*; D013997:Time Factors
D001802:Blood Substitutes; D002352:Carrier Proteins; D003911:Dextrans; D006454:Hemoglobins; D016023:Integrins; C097124:O-raffinose cross-linked human hemoglobin; D012996:Solutions; C095659:hemoglobin-dextran 10-benzene-tetracarboxylate; C083478:diaspirin-cross-linked hemoglobin; D011887:Raffinose; D001241:Aspirin