Abstract
Fibroblast growth factor 2 (FGF-2) is not only an angiogenic factor, but also a mitogen for epidermal keratinocytes. FGF-2 has been shown to be positively immunoreactive in the basal layer of psoriatic lesions. In previous work, we used the Escherichia coli (E. coli) expression system to biosynthesize a biologically active anti-FGF-2 nanobody (Nb) screened by phage display technology, but the low yield limited its clinical application. In this study, we aimed to increase the yield of anti-FGF-2 Nb, and evaluate its therapeutic potential for psoriasis by inhibiting FGF-2-mediated mitogenic signaling in psoriatic epidermal keratinocytes. We demonstrated a 16-fold improvement in the yield of anti-FGF-2 Nb produced in the Pichia pastoris (P. pastoris) compared to the E. coli expression system. In vitro, the FGF-2-induced HaCaT cell model (FHCM) was established to mimic the key feature of keratinocyte overproliferation in psoriasis. Anti-FGF-2 Nb was able to effectively inhibit the proliferation and migration of FHCM. In vivo, anti-FGF-2 Nb attenuated the severity of imiquimod (IMQ)-induced psoriatic lesions in mice, and also improved the inflammatory microenvironment by inhibiting the secretion of inflammatory cytokines (IL-1β, IL-6, IL-23, and TNF-α), chemokines (CXCL1 and CCL20), and neutrophil infiltration in skin lesions. These were mainly related to the suppression of FGF-2-mediated mitogenic signaling in psoriatic keratinocytes. In conclusion, we have improved the production of anti-FGF-2 Nb and demonstrated the modality of attenuating the abnormal proliferative behavior of psoriatic keratinocytes by inhibiting FGF-2-mediated mitogenic signaling, which offers the possibility of treating psoriasis.
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Data Availability
The datasets used and analyzed in the present study are available from the corresponding author on reasonable request.
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Acknowledgements
This work was supported by the Special Project for Research and Development in Key areas of Guangdong Province, “Green Biomanufacturing” Key Special Project (Grant No. 2022B1111070007).
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SX, HZ, and BXL conceived and designed experiments. ZLZ, JT, KTL, and SLW performed the experiments. ZLZ, BXL, and JT analyzed the data. ZLZ, BXL, and MJT wrote and revised the paper. SX was responsible for study supervision. All authors approved the final manuscript.
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Zhou, Z., Liao, B., Wang, S. et al. Improved Production of Anti-FGF-2 Nanobody Using Pichia pastoris and Its Effect on Antiproliferation of Keratinocytes and Alleviation of Psoriasis. Arch. Immunol. Ther. Exp. 71, 20 (2023). https://doi.org/10.1007/s00005-023-00685-w
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DOI: https://doi.org/10.1007/s00005-023-00685-w