Fig. 4

TNFα-driven intestinal inflammation during rLeptin treatment is reversed by anti-TNFα therapy. a–e Sustained inflammation under rLeptin substitution and previously acquired structural damage (stenosis and fistula-induced abscess) made proctocolectomy and ileal resection necessary 16 days after initiation of rLeptin therapy. The specimen (a) was analyzed histologically by H&E staining (scale bar depicts 100 µm) (b), showing severe inflammation, and by immunohistochemistry (IHC; c–e). c Microscopic pictures of IHC staining for TNFα and CD206 in gut tissue from the AGLCD patient and a Crohn’s disease (CD) patient (The scale bar depicts 20 µm, images were recorded using an AxioImager Z1 from Zeiss). d Additional pictures of IHC staining for different immune cell markers (recorded with the Vectra3 system from PerkinElmer, the scale bar displays 20 µm) and (e) quantification of cells staining positive for respective markers per 10 high power fields (For CD45, CD163 and ADRP stainings a total of 11 tissue samples from n = 7 biologically independent CD control patients was compared to the AGLCD patient (n = 1); for TNFα and CD206 stainings a total of 9 samples of n = 6 independent CD patients was obtained and compared to the AGLCD patient (n = 1); for CD11b, CD86 and iNOS stainings a total of 6 samples from n = 3 biologically independent CD patients was compared to the AGLCD patient (n = 1); in case that several samples were analyzed from the same patients, tissues were derived from different anatomical locations, blinded analysis). The box extends from the 25th to 75th percentiles. Whisker plots show the min (smallest) and max (largest) values. The line in the box denotes the median. f–i Following surgery, rLeptin treatment was continued and the patient presented with new inflammation beginning at the terminal ileostomy. The clinical decision was made to initiate anti-TNFα therapy while continuing rLeptin substitution, resulting in clinical and endoscopic remission of Crohn’s disease. f Comparison of pictures taken during endoscopy before starting anti-TNFα therapy and six months later. g–i Flow cytometric analysis of the effects of anti-TNFα therapy on (g) TNFα and (h) IFNγ production after ex vivo stimulation, as well as (I) the expression of the transcription factors FOXP3 and RORγt (“before anti-TNFα” combines data from three different time points). Error bars on column charts display the standard deviation (SD). The source data are provided as a Source Data file.