title: Systematic single cell analysis of migration and morphological changes of human neutrophils over stimulus concentration gradients.
abstract: To compare the responses of individual neutrophils to chemoattractants, migration pathway data were obtained using TAXIScan, an optically accessible/horizontal apparatus in which a concentration gradient is established reproducibly for a given stimulus. The observed linear-mode trajectory pattern of neutrophils toward N-formyl-methionyl-leucyl-phenylalanine (fMLP) or Interleukin (IL)-8/CXCL8 was distinguished from random migration patterns toward leukotriene (LT) B4 or platelet activating factor (PAF). The median values of velocity and directionality calculated for individual cells toward fMLP and IL-8 were both relatively similar and high, whereas the values toward LTB4 and PAF were widely dispersed over a lower range of directionality and from low to high ranges of velocity. The different patterns between the groups may be explained by unique morphology with single polarity toward fMLP and IL-8, and unstable morphology with multiple polarities toward LTB4 and PAF. Unique morphologies toward fMLP and IL-8 were not affected by coexisting LTB4 or PAF. On the other hand, the addition of suboptimum concentrations of fMLP or IL-8 to LTB4 or PAF induced a nearly maximum chemotactic response in most cells. These data suggest that exogenous formyl peptides and endogenous chemokines augment neutrophil accumulation at inflammation sites, whereas lipid mediators may play a role in supporting activation of the inflammatory cells for recruitment.
authors.affiliation: Department of Biochemistry, Kawasaki Medical School, 577 Matsushima, Kurashiki 701-0192, Japan. Electronic address: akiray@med.kawasaki-m.ac.jp.
authors.forename: Akira
authors.initials: A
authors.lastname: Yamauchi
authors.affiliation: Phoenix Medical Clinic, 3-41-6 Sendagaya, Tokyo 151-0051, Japan.
authors.forename: Mikako
authors.initials: M
authors.lastname: Degawa-Yamauchi
authors.affiliation: Department of Biochemistry, Kawasaki Medical School, 577 Matsushima, Kurashiki 701-0192, Japan.
authors.forename: Futoshi
authors.initials: F
authors.lastname: Kuribayashi
authors.affiliation: YU-ECI Research Center for Medical Science, Yeungnam University, Gyeongsan-City 712-749, Republic of Korea.
authors.forename: Shiro
authors.initials: S
authors.lastname: Kanegasaki
authors.affiliation: YU-ECI Research Center for Medical Science, Yeungnam University, Gyeongsan-City 712-749, Republic of Korea. Electronic address: ttsuchiy-tky@umin.ac.jp.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Tsuchiya
mesh_terms: D000328:Adult; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D005260:Female; D006801:Humans; D016209:Interleukin-8 / Q000494:pharmacology*; D007975:Leukotriene B4 / Q000494:pharmacology*; D008297:Male; D008875:Middle Aged; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D010972:Platelet Activating Factor / Q000494:pharmacology*; D061251:Primary Cell Culture; D059010:Single-Cell Analysis / Q000379:methods*
chemical_list: D016209:Interleukin-8; D010972:Platelet Activating Factor; D007975:Leukotriene B4; D009240:N-Formylmethionine Leucyl-Phenylalanine
keywords: Chemoattractants; Chemotaxis; Imaging; Lamellipodia; Polarity; TAXIScan