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In vitro induction of CCL2‐ and IL‐10‐producing PMN (PMN‐II) using band PMN isolated from exercised volunteers
Author(s) -
Kobayashi Makiko,
Shigematsu Kenji,
Herndon David N,
Suzuki Fujio
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.672.34
Subject(s) - ficoll , interleukin 8 , chemistry , in vitro , immunology , medicine , pharmacology , peripheral blood mononuclear cell , cytokine , biochemistry
M2Mφ are known to be inhibitory on the Mφ conversion from resident Mφ to M1Mφ (a major effector cell on host antibacterial innate immunities). As a M2Mφ inducer cell, PMN‐II have been demonstrated in severely burned mice with decreased antibacterial resistances. To analyze the immunosuppressive property of PMN‐II, in this study, we tried to induce PMN‐II in cultures of healthy volunteer PMN. By Ficoll‐Hypaque and dextran sedimentations, PMN were prepared from peripheral blood of healthy volunteers with or without 1 hr‐exercise. Band PMN and segmented PMN were isolated from the PMN preparation using biotin‐conjugated anti‐CD10 mAb and magnetic‐beads coated with streptavidin. Cells obtained (1 x 10 6 cells/ml) were cultured with SAC, GM‐CSF or VEGF. Culture fluids harvested 18 hrs after cultivation were assayed for CCL2 and IL‐10 (biomarkers for PMN‐II). In the results, segmented and band PMN preparations from volunteers without any exercise did not produce CCL2 and IL‐10 after the cultivation with various stimulants. However, in response to the stimulation with SAC, GM‐CSF or VEGF, band PMN (but not segmented PMN) isolated from volunteers 3 to 24 hrs after 1 hr‐exercise produced CCL2 and IL‐10 into their culture fluids. The highest production of CCL2 was shown when PMN prepared from volunteers 3 hrs after 1 hr‐exercise were stimulated with SAC and GM‐CSF. Also, the numbers of band PMN increased greatly in the peripheral blood of volunteers 3 to 24 hrs after having exercised. These results indicate that, by the stimulation with SAC, GM‐CSF or VEGF, PMN‐II can be inducible from band PMN of exercised volunteers. PMN‐II induced in vitro will be useful in the analytical studies of PMN‐II‐associated immunosuppression. This study was supported by SHC NA #8840.