Cell-associated IL-8 in human blood monocytes: Analysis by flow cytometry

Seitaro Fujishima, Hidetoshi Nakamura, Yasuhiro Waki, Kenzo Soejima, Yuji Takeuchi, Motoyuki Ogawa, Masahide Shiozawa, Yoshiki Hiraoka, Takeo Kawashiro, Sadakazu Aiso, Minoru Kanazawa

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Several cell-associated cytokines, such as interleukin-1 (IL-1) and tumor necrosis factor, exist on the cell surface and are biologically active. Although extracellular IL-8, a potent chemotactic factor for primarily neutrophils, has been studied extensively, cell-associated IL-8 has barely been studied. In this study, we analyzed the intracellular and cell-surface IL-8 in human blood monocytes in vitro by using flow cytometry and predicted the biological activity of the cell-associated IL-8 in vivo. After fixation with paraformaldehyde, mononuclear cells were divided into two subgroups. One subgroup was left untreated to study cell-associated antigens, and the other subgroup was permeabilized with saponin to detect intracellular antigens. In lipopolysaccharide (LPS)-stimulated monocytes, IL-8 was detected solely intracellularly, whereas both the intracellular and cell-surface IL-1β was detectable. In a time-course study, the intracellular IL-8 increased in response to LPS stimulation, but the cell-surface IL-8 was undetectable throughout he course. In an LPS-stimulated monocytic cell line, both ELISA and flow cytometry detected the quantitative change of the intracellular IL-8. The dissmilar localization between IL-8 and IL-1β within cells was confirmed by the immunohistochemical analysis. In summary, LPS stimulation induced a time-dependent increase in intracellular but not cell-surface IL-8 in monocytes. Thus, it is unlikely that the cell-associated IL-8 is functioning physiologically. The semiquantitative flow cytometric procedure may be useful for simultaneous examination for cell-surface and intracellular cytokines.

Original languageEnglish
Pages (from-to)382-389
Number of pages8
JournalCytometry
Volume24
Issue number4
DOIs
Publication statusPublished - 1996 Aug 1
Externally publishedYes

Keywords

  • Cell-associated cytokine
  • Flow cytometry
  • Immunohistochemistry
  • Interleukin-1β
  • Interleukin-8
  • Saponin

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Biophysics
  • Hematology
  • Endocrinology
  • Cell Biology

Cite this

Fujishima, S., Nakamura, H., Waki, Y., Soejima, K., Takeuchi, Y., Ogawa, M., ... Kanazawa, M. (1996). Cell-associated IL-8 in human blood monocytes: Analysis by flow cytometry. Cytometry, 24(4), 382-389. https://doi.org/10.1002/(SICI)1097-0320(19960801)24:4<382::AID-CYTO10>3.0.CO;2-L

Cell-associated IL-8 in human blood monocytes : Analysis by flow cytometry. / Fujishima, Seitaro; Nakamura, Hidetoshi; Waki, Yasuhiro; Soejima, Kenzo; Takeuchi, Yuji; Ogawa, Motoyuki; Shiozawa, Masahide; Hiraoka, Yoshiki; Kawashiro, Takeo; Aiso, Sadakazu; Kanazawa, Minoru.

In: Cytometry, Vol. 24, No. 4, 01.08.1996, p. 382-389.

Research output: Contribution to journalArticle

Fujishima, S, Nakamura, H, Waki, Y, Soejima, K, Takeuchi, Y, Ogawa, M, Shiozawa, M, Hiraoka, Y, Kawashiro, T, Aiso, S & Kanazawa, M 1996, 'Cell-associated IL-8 in human blood monocytes: Analysis by flow cytometry', Cytometry, vol. 24, no. 4, pp. 382-389. https://doi.org/10.1002/(SICI)1097-0320(19960801)24:4<382::AID-CYTO10>3.0.CO;2-L
Fujishima, Seitaro ; Nakamura, Hidetoshi ; Waki, Yasuhiro ; Soejima, Kenzo ; Takeuchi, Yuji ; Ogawa, Motoyuki ; Shiozawa, Masahide ; Hiraoka, Yoshiki ; Kawashiro, Takeo ; Aiso, Sadakazu ; Kanazawa, Minoru. / Cell-associated IL-8 in human blood monocytes : Analysis by flow cytometry. In: Cytometry. 1996 ; Vol. 24, No. 4. pp. 382-389.
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