Journal: Journal for Immunotherapy of Cancer

Article Title: The IL-1/IL-1 receptor axis and tumor cell released inflammasome adaptor ASC are key regulators of TSLP secretion by cancer associated fibroblasts in pancreatic cancer

doi: 10.1186/s40425-019-0521-4

Figure Lengend Snippet: Tumor cell-derived IL-1 has a key role in TSLP secretion by CAFs. a TSLP secretion by CAFs ( n = 18) either untreated (NT) or treated with the indicated cytokines (20 ng/ml). Each symbol refers to a single experiment and represents the mean of duplicate determinations. Significance was determined using Mann Whitney test. b TSLP secretion by CAFs in the absence (NT) or in the presence of increasing doses of recombinant IL-1α, IL-1β and TNF-α. Significance was determined using Mann Whitney test. c TSLP secretion by CAFs ( n = 14) either untreated (NT) or treated with supernatants of necrotic IL-1 negative (IL-1-) ( n = 16) or positive (IL-1+) ( n = 26) tumor cells. Each symbol refers to a single experiment and represents the mean of duplicate determinations. Significance was determined using Mann Whitney test. d TSLP secretion by CAFs induced by necrotic tumor cell-derived supernatants in the absence (NT) or in the presence of the indicated neutralizing Abs or isotype control Abs or the IL-1RA (Anakinra). Representative experiments performed with the indicated tumor cell lines. Error bars represent standard deviations of triplicate values. Significance was determined using Student’s t -test. e Cumulative results: each dot refers to a single experiment and represents the percentage of inhibition calculated on the value of TSLP secretion in the presence of the isotype control antibody. f TSLP secretion by CAFs either untreated or stimulated with the supernatant of necrotic Hs766T cells after treatment with IL-1α/IL-1β siRNA or negative control siRNA. Treatment of CAFs with recombinant IL-1α + IL-1β (20 ng/ml each) in the absence or in the presence of anakinra were used as controls. Error bars represent standard deviations of triplicate values. Significance was determined using Student’s t -test. Values significantly different were indicated as: * p < 0.05, ** p < 0.01 and *** p < 0.001

Article Snippet: 2 × 10 6 Hs766T (IL-1 positive) cells or 2 × 10 6 CAFs or 2 × 10 6 Hs766T cells plus 2 × 10 6 CAFs were injected in the pancreas of immunodeficient NSG mice (The Jackson Laboratory).

Techniques: Derivative Assay, MANN-WHITNEY, Recombinant, Control, Inhibition, Negative Control

Journal: Journal for Immunotherapy of Cancer

Article Title: The IL-1/IL-1 receptor axis and tumor cell released inflammasome adaptor ASC are key regulators of TSLP secretion by cancer associated fibroblasts in pancreatic cancer

doi: 10.1186/s40425-019-0521-4

Figure Lengend Snippet: Treatment of immunodeficient NSG mice orthotopically transplanted with IL-1 positive Hs766T cells plus CAFs with anakinra reduces TSLP expression in vivo. a Treatment schedule. Mice were orthotopically injected on day 0 with Hs766T cells plus CAFs. Hs766T cells alone and CAFs alone were injected as controls. The next day mice transplanted with Hs766T cells plus CAFs started intraperitoneal injection with the two indicated doses of anakinra and mice were sacrificed on day 6 after daily administration. b Hematoxylin and eosin staining of the pancreas. Left, a tumor mass (T) localized within the normal pancreas (N) is evident. Right, at higher magnification, histological features of pancreatic cancer with cancer cells, fibroblasts and extracellular matrix can be recognized. c Human (left) and mouse (right) TSLP mRNA expression in pancreata of NSG mice implanted with CAFs alone ( n = 3), Hs766T cells alone ( n = 3) or CAFs plus Hs766T cells ( n = 3). Significance was determined using Student’s t -test. d TSLP expression in pancreata of NSG mice untreated (NT) ( n = 4) or treated with the two indicated doses of Anakinra ( n = 4 for each dose). Significance was determined using Student’s t -test. Values significantly different were indicated as: * p < 0.05

Article Snippet: 2 × 10 6 Hs766T (IL-1 positive) cells or 2 × 10 6 CAFs or 2 × 10 6 Hs766T cells plus 2 × 10 6 CAFs were injected in the pancreas of immunodeficient NSG mice (The Jackson Laboratory).

Techniques: Expressing, In Vivo, Injection, Staining