April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Human Monocytes Heterogeneity And Its Implication In Uveitis Therapy
Author Affiliations & Notes
  • Baoying Liu
    Lab Immunology, National Eye Institute/NIH, Bethesda, Maryland
  • Diamond Ling
    Lab Immunology, National Eye Institute/NIH, Bethesda, Maryland
  • H. Nida Sen
    Lab Immunology, National Eye Institute/NIH, Bethesda, Maryland
  • Silu Lucy Tan
    Lab Immunology, National Eye Institute/NIH, Bethesda, Maryland
  • Zhiyu Li
    Lab Immunology, National Eye Institute/NIH, Bethesda, Maryland
  • Lai Wei
    Lab Immunology, National Eye Institute/NIH, Bethesda, Maryland
  • Robert B. Nussenblatt
    Lab Immunology, National Eye Institute/NIH, Bethesda, Maryland
  • Footnotes
    Commercial Relationships  Baoying Liu, None; Diamond Ling, None; H. Nida Sen, None; Silu Lucy Tan, None; Zhiyu Li, None; Lai Wei, None; Robert B. Nussenblatt, None
  • Footnotes
    Support  NIH intramural research program
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 4298. doi:
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    • Get Citation

      Baoying Liu, Diamond Ling, H. Nida Sen, Silu Lucy Tan, Zhiyu Li, Lai Wei, Robert B. Nussenblatt; Human Monocytes Heterogeneity And Its Implication In Uveitis Therapy. Invest. Ophthalmol. Vis. Sci. 2011;52(14):4298.

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Abstract

Purpose: : The expression of CD14 and CD16 defines three monocyte subsets:CD14 highCD16-, CD14highCD16+, and CD14lowCD16+ cells. The goals of this study are to characterize these 3 populations and evaluate the association between monocyte subsets and uveitis therapy.

Methods: : Human peripheral blood mononuclear cells (PBMCs) were isolated from the blood of healthy donors using a Ficoll gradient centrifugation protocol. Monocyte phenotying was performed using 4 color flow cytometry staining protocol. LPS and IFNγ were used in vitro to polarize classically-activated monocytes and IL-4 was used to polarize alternatively-activated monocytes. Real-time polymerase chain reactions (RT-PCR) were used to detect mRNA expression of cytokines and chemokines.

Results: : Classically-activated monocytes present higher percentage expression of CD14highCD16- than do alternative and unpolarized monocytes. RT-PCR showed that CD14highCD16- monocytes expressed higher IL-6, IL-8, IL-10, IL-1β, CXCL1, CXCL3, CXCL5 and CXCL6 levels than did CD14highCD16+ monocytes. The expression of CD163, a scavenger receptor on monocytes which is believed to be a regulatory monocyte marker, is lower in CD14lowCD16+ and CD14highCD16- cells than in CD14highCD16+ monocytes. Importantly, we observed a higher percentage of CD14highCD16+, a lower percentage of CD14lowCD16+ and CD14highCD16- populations in quite uveitis patients on immuno-suppressive therapy than inactive patients not on immunosuppressives and normal donors [p<0.05].

Conclusions: : CD14lowCD16+ and CD14highCD16- monocyte populations may represent subsets of proinflammatory monocytes. Our data suggest a potential role for monocyte phenotyping in understanding the effects of immuno-therapy in uveitis patients.

Keywords: autoimmune disease • uveitis-clinical/animal model 
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