The Use of the Vitality and Chemotaxis of Human Polymorphonuclear Leukocytes for the In Vitro Estimation of the Acute Toxicity of the First Ten Chemicals from the MEIC List
Restricted accessResearch articleFirst published online January, 1993
The Use of the Vitality and Chemotaxis of Human Polymorphonuclear Leukocytes for the In Vitro Estimation of the Acute Toxicity of the First Ten Chemicals from the MEIC List
The acute cytotoxicity of the first ten MEIC chemicals has been estimated by others in various cell lines. In the present investigation, isolated human polymorphonuclear leukocytes (PMN) from ten healthy non-smoking laboratory personnel were used to assess in vitro toxicity of the same chemicals.
The cells were treated with different concentrations of the respective chemicals for three hours and their vitality and chemotaxis were tested. Vitality was measured by fluorescence microscopy after the addition of fluorescein diacetate and ethidium bromide. Living cells which took up and hydrolysed fluorescein diacetate, and dead cells, stained by ethidium bromide, were counted and the percentage of live cells was calculated.
Locomotion stimulated by the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (F-MLP), was measured in blind-well Boyden chambers and a chemotactic index was calculated.
The results were mathematically transformed to produce a linear curve, and then fitted by the linear least squares procedure, from which LC50 and IC50 values were obtained by interpolation. All the chemicals decreased the vitality and inhibited the chemotaxis of the PMN. Obviously the chemotactic test was more sensitive than the vitality one. A correlation (r = 0.933) was found between vitality and chemotaxis inhibition. Spearman rank correlation analysis revealed significant correlations between our results and those from in vitro experiments conducted in other laboratories, as well as with data concerning mouse, rat and human lethal doses.
FerranteA., and ThongY.H. (1980). Optimal conditions for simultaneous purification of mononuclear and polymorphonuclear leukocytes from human peripheral blood by the Hypaque-Ficoll method.Journal of Immunological Methods36, 109–117.
2.
AeschbacherM., ReinhardtC.A., and ZbindenG. (1986). A rapid cell membrane permeability test using fluorescent dyes and flow cytometry.Cell Biology and Toxicology2, 246–255.
3.
ValentinoM., GovernaM., MarchiseppeI., & VisonàI. (1991). Effects of lead polymorphonuclear leukocyte (PMN) functions in occupation-ally exposed workers.Archives of Toxicology65, 685–688.
4.
GovernaM., ValentinoM., VisonàI., and RoccoM. (1986). Impairment of human polymorphonuclear leukocyte chemotaxis by 2,5-hexanedione.Cell Biology and Toxicology2, 33–39.
5.
GovernaM., ValentinoM., & VisonàI. (1987). In vitro impairment of human granulocyte functions by lead.Archives of Toxicology59, 421–425.
6.
HillH.R., HoganN.A., and MitchellT.G. (1975). Evaluation of a cytocentrifuge method for measuring neutrophil granulocyte chemotaxis.Journal of Laboratory and Clinical Medicine86, 703–710.
7.
OleskeJ.M., De La CruzA., AhdichH., SorvinoD., La BraicoJ., CooperR., SinghR., LinR., and MinneforA. (1983). Effects of antibiotics on polymorphonuclear leukocyte chemiluminescence and chemotaxis.Journal of Antimicrobial Chemotherapy12, Suppl. C, 35–38.
8.
Van der AuweraP., and HussonM. (1989). Influence of rifampicin and ansamycin on motility and adherence of human neutrophils studied in vitro.Journal of Antimicrobial Chemotherapy24, 347–353.
9.
FumaruloR., and ArestaM. (1987). Cisplatin toxicity: effects on phagocyte functions.Inorganic Chimica Acta137, 99–103.
10.
MarcoliM., RicevutiG., MazzoneA., BekkeringM., LecchiniS., and FrigoG.M. (1988). Opioid-induced modification of granulocyte function.International Journal of Immunopharmacology10, 425–433.
11.
IpM., LomasD.A., ShawJ., BurnettD., and StockleyR.A. (1990). Effect of non-steroidal antiinflammatory drugs on neutrophil chemotaxis. An in vitro and in vivo study.British Journal of Rheumatology29, 363–367.
12.
ElferinkJ.G.R., and DeierkaufM. (1984). The effect of verapamil and other calcium antagonists on chemotaxis of polymorphonucelar leukocytes.Biochemical Pharmacology33, 35–39.
13.
HyersT.M., TricomiS.M., and LiaoJ.J. (1992). Inhibition of some human neutrophil functions by the cyclooxygenase inhibitor ketorolac tromethamine.Journal of Leukocyte Biology51, 490–495.
14.
RoquesC., FrayretM.N., LucJ., MichelG., PerruchetA.M., CaquilJ., and LevyD. (1991). Effect of an in vivo immunostimulant treatment on PMN functions: interaction with antibiotics in vitro.International Journal of Immunopharmacology13, 1,051–1,057.
15.
MiyagiM., AoyamaH., MorishitaM., and IwamotoY. (1992). Effects of sex hormones on chemotaxis of human peripheral polymorphonuclear leukocytes and monocytes.Journal of Periodontology63, 28–32.
16.
LevesqueL., GaudreaultR.C., and MarceauF. (1992). Comparison of two classes of non-peptide drugs as antagonists of neutrophil receptors for f-Met-Leu-Phe. Pyrazolons and iodinated radiographic contrast agents.Biochemical Pharmacology43, 553–560.
17.
TorreD., BrogginiM., BottaV., SampietroC., BusarelloR., and GarberiC. (1991). In vitro and ex vivo effects of recent and new macrolide antibiotics on chemotaxis of human polymorphonuclear leukocytes.Journal of Chemotherapy3, 236–239.
18.
SasagawaS. (1991). Inhibitory effects of local anaesthetics on migration, extracellular release of lysosomal enzyme, and superoxide anion production in human polymorphonuclear leukocytes.Immunopharmacology and Immunotoxicology13, 607–622.
19.
CapodicasaE., ScaringiL., RosatiE., De BellisF., SbaragliaG., MarconiP., & Del FaveroA. (1991). In vitro effects of teicoplanin, teicoplanin derivative MDL 62211 and vancomycin on human polymorphonuclear cell function.Antimicrobial Chemotherapy27, 619–626.
20.
DavisJ.M., AlbertJ.D., TracyK.L., CalvanoS.E., LowryS.F., ShiresG.T., and YurtR.W. (1991). Increased neutrophil mobilisation and decreased chemotaxis during Cortisol and epinephrine infusions.Journal of Trauma31, 725–731.
21.
SkaanildM.T., and ClausenJ. (1991). The use of human lymphocytes for the in vitro estimation of acute toxicity of ten chemicals from MEIC List.ATLA19, 187–190.
22.
PonsodaX., JoverR., CastellJ.V., & Gómez-LechónM.J. (1989). In vitro toxicity to two cellular systems of the first ten chemicals on the MEIC list.ATLA17, 218–223.
23.
EkwallB. (1990). Preliminary studies on the validity of in vitro measurement of drug toxicity using HeLa cells. II. Drug toxicity in the MIT-24 system compared with mouse and human lethal dosage of 52 drugs.Toxicology Letters5, 309–317.
24.
ZanettiC., De AngelisI., StammatiA.L., and ZuccoF. (1992). Evaluation of toxicity testing of 20 MEIC chemicals on Hep 2 cells using two viability endpoints.ATLA20, 120–125.
25.
EkwallB., BondessonI., CastellJ.V., Gómez-LechónM.J., HellbergS., HögbergJ., JoverR., PonsodaX., RomertL., StenbergK., and WalumE. (1989). Cytotoxicity evaluation of the first ten MEIC chemicals: acute lethal toxicity in man predicted by cytotoxicity in five cellular assays and by oral LD50 tests in rodents.ATLA17, 83–100.
26.
GovernaM., ValentinoM., RoccoM., BertoliE., and ZoleseG. (1987). Inhibition of chemotactic activity of PMN by arsenic: a membrane mediated effect. In Proceedings International Conference: Developments in analytical methods in pharmaceutical biomedical and forensic sciences (ed. PiemonteG., TagliaroF., FrigerioA., MargioM.), pp. 297–302. New York: Plenum Publishing Co.
27.
HuntJ.A., RemesA., and WilliamsD.F. (1992). Stimulation of neutrophil movement by metal ions.Journal of Biomedical Materials Research26, 819–828.
28.
ManganD., and SnyderI.S. (1978). The effect of acrylamide on human polymorphonuclear neutrophils in vitro.British Journal of Industrial Medicine35, 305–311.
29.
ArakawaY., and WadaO. (1984). Inhibition of neutrophil chemotaxis by organotin compounds.Biochemical and Biophysical Research Communications123, 543–548.
30.
WoodinA.M., and HarrisA. (1973). The inhibition of locomotion of the polymorphonuclear leukocyte by organophosphorus compounds.Experimental Cell Research77, 41–46.
31.
BergeretA., PougetE., TedoneR., MeygretT., CadotR., and DescotesJ. (1990). Neutrophil functions in lead-exposed workers.Human Exposure Toxicology9, 231–233.
32.
HermanowiczA., and KossmanS. (1984). Neutrophil function and infectious disease in workers occupationally exposed to phosphoorganic pesticides: role of mononuclear-derived chemotactic factor for neutrophils.Clinical Immunology and Immunopathology33, 13–22.
33.
BondessonI., EkwallB., HellbergS., RomertL., StenbergK., and WalumE. (1989). MEIC — a new international multicentre project to evaluate the relevance to human toxicity of in vitro cytotoxicity tests.Cell Biology and Toxicology5, 331–347.
