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Podocytes Produce and Secrete Functional Complement C3 and Complement Factor H.

Frontiers in immunology (2020-09-15)
Anne K Mühlig, Lindsay S Keir, Jana C Abt, Hannah S Heidelbach, Rachel Horton, Gavin I Welsh, Catherine Meyer-Schwesinger, Christoph Licht, Richard J Coward, Lars Fester, Moin A Saleem, Jun Oh
ZUSAMMENFASSUNG

Podocytes are an important part of the glomerular filtration barrier and the key player in the development of proteinuria, which is an early feature of complement mediated renal diseases. Complement factors are mainly liver-born and present in circulation. Nevertheless, there is a growing body of evidence for additional sites of complement protein synthesis, including various cell types in the kidney. We hypothesized that podocytes are able to produce complement components and contribute to the local balance of complement activation and regulation. To investigate the relevant balance between inhibiting and activating sides, our studies focused on complement factor H (CFH), an important complement regulator, and on C3, the early key component for complement activation. We characterized human cultured podocytes for the expression and secretion of activating and regulating complement factors, and analyzed the secretion pathway and functional activity. We studied glomerular CFH and C3 expression in puromycin aminonucleoside (PAN) -treated rats, a model for proteinuria, and the physiological mRNA-expression of both factors in murine kidneys. We found, that C3 and CFH were expressed in cultured podocytes and expression levels differed from those in cultivated glomerular endothelial cells. The process of secretion in podocytes was stimulated with interferon gamma and located in the Golgi apparatus. Cultured podocytes could initiate the complement cascade by the splitting of C3, which can be shown by the generation of C3a, a functional C3 split product. C3 contributed to external complement activation. Podocyte-secreted CFH, in conjunction with factor I, was able to split C3b. Podocytes derived from a patient with a CFH mutation displayed impaired cell surface complement regulation. CFH and C3 were synthesized in podocytes of healthy C57Bl/6-mice and were upregulated in podocytes of PAN treated rats. These data show that podocytes produce functionally active complement components, and could therefore influence the local glomerular complement activation and regulation. This modulating effect should therefore be considered in all diseases where glomerular complement activation occurs. Furthermore, our data indicate a potential novel role of podocytes in the innate immune system.

MATERIALIEN
Produktnummer
Marke
Produktbeschreibung

Sigma-Aldrich
Anti-Laminin in Kaninchen hergestellte Antikörper, 0.5 mg/mL, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Puromycin-Aminonukleosid
Sigma-Aldrich
Anti-Tubulin antibody produced in rabbit, whole antiserum
Sigma-Aldrich
Anti-Faktor H Ziege pAb, liquid, Calbiochem®