Elsevier

Transplant Immunology

Volume 6, Issue 3, September 1998, Pages 193-197
Transplant Immunology

Genotyping for polymorphisms in interferon-γ, interleukin-10, transforming growth factor-β1 and tumour necrosis factor-α genes: a technical report

https://doi.org/10.1016/S0966-3274(98)80045-2Get rights and content

Abstract

Polymorphic variants of cytokine genes are associated with acute and chronic transplant rejection. In this technical report, the methods currently used in our centre to genotype individuals for interferon-γ, interleukin-10, transforming growth facgor-β1 and tumour necrosis factor-α are described in detail. The DNA sequences of primers and probes, and conditions for polymerase chain reactions are given, and the allele and genotype frequencies in our control populations are summarized.

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    A total of 8 polymorphisms were studied in the TNF-α, TGF-β, IL-10, IL-6, and IFN-γ genes of both patients and healthy individuals. We focused on determining the following single nucleotide polymorphisms, which have been reported in the literature at our disposal (Perrey et al., 1998) to influence the degree of gene expression: TNF- α − 308 A/G; TGF-β1 codon 10 T/C and codon 25 G/C; IL-10-1082 A/G, − 819 C/T, − 592 C/A; IL-6 -174 G/C; IFN- γ + 874 A/T. The DNA of the individuals studied was isolated from peripheral blood mononuclear cells by an automated system based on magnetic beads (iPrep™, Invitrogene).

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