Purification of natural antikeratin autoantibodies from normal human serum and their effect on human keratinocytes cultured in vitro

Cx, Li; Yh, Wan; Sm, Chi; Wang, G; Lc, Sun; Yg, Zhang; Xd, Zhao; Tw, Gao; Liu, YF

doi:10.1046/j.1365-2133.2001.04469.x

Cited by 9 publications

(2 citation statements)

References 33 publications

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“…Although AK autoantibodies are reported to have many physiological functions, 8,9 we believe that anti‐K16 autoantibodies may constitute a source of chronic inflammation in psoriatic lesions. Whether or not antikeratin monoclonal antibodies cause expression of inflammatory genes or cytokines remains to be elucidated.…”

Section: Reportmentioning

confidence: 90%

Effects of antikeratin 16 antibodies on the expression of Toll-like receptors 2 and 4 in keratinocytes

Wu¹,

Li²,

Zheng³

2009

Clinical and Experimental Dermatology

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Innate immunity plays an important role in psoriasis. The possible role of Toll-like receptors (TLRs) in the activation of innate immunity in psoriasis remains unknown. Serum antikeratin 16 (anti-K16) autoantibody levels are raised in patients with psoriasis. The aim of this study was to investigate the effects of anti-K16 antibodies on TLR2 and TLR4 expression in keratinocytes. Keratinocytes were incubated with mouse anti-K16 monoclonal antibodies. Levels of TLR2 and TLR4 were measured by quantitative real-time PCR and flow cytometry. TLR2 mRNA level was increased by 1.73-fold, 1.60-fold and 2.52-fold at 6, 24 and 36 h after incubation, respectively. TLR4 mRNA level was increased by 3.62-fold and 2.21-fold at 12 and 36 h, respectively. TLR2 protein expression was increased by 1.73-fold on the cell membrane and 2.22-fold in cell membrane and intracytoplasmic locations. There was no change in TLR4. The results of this study indicate that anti-K16 antibodies may be a modulator of TLRs.

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Section: Reportmentioning

confidence: 90%

Effects of antikeratin 16 antibodies on the expression of Toll-like receptors 2 and 4 in keratinocytes

Wu¹,

Li²,

Zheng³

2009

Clinical and Experimental Dermatology

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“…This disparity indicates the poor species-specificity of AK autoAbs. We have recently obtained AK autoAbs instead of purified AK16 autoAbs (34). Because they may recognize similar epitope, effects of mouse monoclonal antibodies may imitate the effects of human AK16 autoAbs to a certain degree.…”

Section: Discussionmentioning

confidence: 99%

Innate immune modulation of keratinocytes by antikeratin 16 antibodies

Wei

et al. 2008

Experimental Dermatology

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Cloning and characterization of antikeratin human antibodies using a semisynthetic phage antibody library

Wang

Liu

et al. 2004

Arch Dermatol Res

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Antikeratin autoantibodies (AK auto Abs) are very important elements of the human immune system. To improve the outcome of studies on AK auto Abs, it is necessary to generate antikeratin human monoclonal antibodies. The purpose of present study was to isolate antikeratin human monoclonal antibodies by panning a phage antibody library. A semisynthetic phage antibody library with capacity of 4.0x10(8) members was previously constructed. Panning of the library was performed against human epidermal keratin extracted from psoriatic scales. At the last round of the panning, individual colonies were grown in culture for expression of phage antibodies. Their binding activities and specificities to keratin were determined by ELISA, and positive clones were analyzed by DNA fingerprinting. The selected clones were induced with IPTG to express soluble Fab fragments, which were further characterized by ELISA, immunohistochemistry and Western blotting. Finally, DNA sequencing of the variable regions was performed. A human antibody clone which was able to express soluble Fab fragments and recognize Mr 46,000 keratin (K17) was isolated. DNA sequencing demonstrated that the VH and VL of the antibody came from the human VH1 and Vkappa2 families, respectively. We conclude that phage antibody library technology is a powerful way to generate human monoclonal antibodies. The antikeratin antibody we isolated in the present study would be useful in the research on AK auto Abs.

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Purification of natural antikeratin autoantibodies from normal human serum and their effect on human keratinocytes cultured in vitro

Abstract: The present research lays a solid foundation for further investigation into the biological significance of natural AK autoantibodies.

Cited by 9 publications

References 33 publications

Effects of antikeratin 16 antibodies on the expression of Toll-like receptors 2 and 4 in keratinocytes

Effects of antikeratin 16 antibodies on the expression of Toll-like receptors 2 and 4 in keratinocytes

Innate immune modulation of keratinocytes by antikeratin 16 antibodies

Cloning and characterization of antikeratin human antibodies using a semisynthetic phage antibody library

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