Figure 2.

 (A) Southern blot analysis of transcripts for interferon γ (IFN-γ) (upper blot) and β-actin (lower blot) in duodenal mucosa from five untreated coeliac disease (CD) patients cultured in vitro with medium alone (M) and with peptic-tryptic (PT)-gliadin in the absence (PT) or presence of human recombinant interleukin 10 (rhIL-10 50 ng/ml) (PT+IL-10) for 24 hours. Total RNA (700 ng) was used for cDNA preparation. An equivalent amount of cDNA per sample was amplified using specific primers for IFN-γ (26 cycles) and β-actin (18 cycles). Polymerase chain reaction products were then separated on a 1% agarose gel, blotted, and hybridised with oligonucleotide probes specific for IFN-γ and β-actin. In the presence of rhIL-10, gliadin induced IFN-γ mRNA production was reduced in 3/5 patients. (B) Southern blot analysis of transcripts for IFN-γ (upper blot) and β-actin (lower blot) in duodenal mucosa from six untreated CD patients cultured in vitro with medium alone (M) or in the presence of rhIL-10 (50 ng/ml) for 24 hours. Southern blot was performed as in (A). IFN-γ levels were not downregulated in biopsies cultured in the presence of rhIL-10 if compared with biopsies cultured in medium alone; paradoxically, in 4/6 patients there was upregulation of IFN-γ transcripts in the presence of rhIL-10.