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Updated: Jul 4, 2026

Generation and Culturing of Primary Human Keratinocytes from Adult Skin
Published on: December 22, 2017
Innate immune modulation of keratinocytes by antikeratin 16 antibodies
Caixia Wu1, Chengxin Li, Liming Wei
1Department of Dermatology, Huashan Hospital, Fudan University, Shanghai, China.
Background:
Chronic inflammation of psoriatic lesions may be due to an exaggerated innate immune response. Toll-like receptors (TLRs) are expressed by keratinocytes in psoriasis. Antikeratin 16 autoantibodies (AK16 autoAbs) are increased in serum from patients with psoriasis. Whether the elevated AK16 autoAbs play a role in psoriasis by exaggerating innate immune response is still unknown.
Objective:
To prove that AK16 autoAbs are involved in psoriasis, by exaggerating the innate immune response of keratinocytes.
Methods:
Keratinocytes were incubated with mouse antikeratin 16 monoclonal antibodies (AK16 mAbs) for a given length of time. Levels of TLR2, TLR4, involucrin and nascent polypeptide-associated complex (NACA) mRNA were measured by quantitative RT-PCR. Levels of TLR2, TLR4, involucrin, NF-kappaB and actin-related protein 2 (ARP2) protein were measured by flow cytometry or Western blot. Effects of the mAbs on keratinocytes were studied using DNA synthesis and cell cycle analysis.
Results:
TLR2 mRNA increased 1.73-, 1.60- and 2.52-fold at 6, 24 and 36 h after incubation, respectively. TLR4 mRNA increased 3.62- and 2.21-fold after 12 and 36 h. Involucrin mRNA increased 2.33- and 2.0-fold after 12 and 36 h. NACA mRNA increased 5.93-, 3.35- and 3.54-fold after 12, 24 and 36 h. TLR2 protein increased 1.73-fold on the cell membrane and 2.22-fold on membrane plus intracytoplasm. NF-kappaB increased 2.64-fold after 6 h. Involucrin protein increased 4.5-fold, whereas Arp2 protein decreased 1.82-fold, after 36 h. The mAbs had an inhibitory effect on cultured keratinocytes.
Conclusion:
AK16 autoAbs may be involved in the chronic inflammation of psoriasis lesions by promoting TLR expression.
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