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201904151249411

CD55-RT-PCR real time Product information​​

Quantification assay of complementopathies                 cod. BM-003

 

Principle of the test: Quantitative analysis of CD55 mRNA expression

Technology: Relative Quantitative Real Time PCR

Gene Target: CD55

Specimen: cDNA

Results: ΔΔCt method

Reporting Units: Arbitrary Units (AU)

Number of tests: 25 tests BM-003

Kit storage: -20°C

Necessary equipment: Thermocycler, 7500 Real Time PCR System

Status: Ready to use

CD55-RT-PCR real time cod. BM-003 

 

Quantification assay of CD55

 

·      CD55-RT-PCR real time Quantification complete kit 25 tests BM-003

 

The complement system is an essential part of the innate immune system that requires careful regulation to ensure responses are appropriately directed against harmful pathogens, while preventing collateral damage to normal host cells and tissues. While deficiency in some components of the complement pathway is associated with increased susceptibility to certain infections, it has also become clear that inappropriate activation of complement is an important contributor to human disease6. CD46 is a surface-expressed regulator acting as cofactor for serum FI, which cleaves C3b to iC3b, thereby irreversibly preventing the reassembly of AP amplifying activity. CD55 is a membrane-bound regulator factor that accelerates the decay of cell surface–assembled C3 and C5 convertases favouring the disassociation of Bb from C3bBb and C2b from C4b and inhibiting their re-aggregation, hence preventing formation of the final MAC7. The intracellular complement activation is important for regulation of the adaptive immune response. After interaction, T cells and antigen presenting cells (APCs) produce and release AP components C3, FB and FD. This is regulated by a transient cell surface expression of CD55. Locally produced C3a and C5a bind to their receptors and further stimulate T cells and APCs. CD46 is expressed on T cells and promotes the switch from Th1 to regulatory T cells (Tregs). In spite of the common belief that complement activation and its downstream effects increase the nephrotoxicity of otherwise innocent—lanthanic—mesangial IgA deposits, the data in favour of a simple relationship between C3 deposition and progression of IgAN have not been as convincing as expected.

We have established a novel quantitative analysis of CD55 mRNA expression. This method is based on real-time PCR. Relative quantification of mRNA expression of CD55 genes was achieved by normalization to the reference gene GAPDH. Relative quantification of target gene expression in patients was performed with the ΔΔCt method and the relative CD55 fold changes were determined. Results are expressed in corresponding arbitrary units (AU) User friendly and complete, the CD55-RT-PCR real time Quantification kit is suitable for any laboratory.

 

Reference

1.      Baines AC, Brodsky RA. Complementopathies. Blood Rev. 2017 Jul;31(4):213-223. doi: 10.1016/j.blre.2017.02.003.

2.      Coppo R, Peruzzi L, Loiacono E, Bergallo M, Krutova A, Russo ML, Cocchi E, Amore A, Lundberg S, Maixnerova D, Tesar V, Perkowska-Ptasinska A, Durlik M, Goumenos D, Gerolymos M, Galesic K, Toric L, Papagianni A, Stangou M, Mizerska-Wasia Membek M, Gesualdo L, Montemurno E, Benozzi L, Cusinato S, Hryszko T, Klinger M, Kaminska D, Krajewska M; VALIGA Study Group of the ERA-EDTA Immunonephrology Working Group. Defective gene expression of the membrane complement inhibitor CD46 in patients with progressive immunoglobulin A nephropathy. Nephrol Dial Transplant. 2018 Apr 9. doi: 10.1093/ndt/gfy064.