P088 TLR3 ameliorates colitis-associated colon tumourigenesis in mice

Abstract Background Toll-like receptor 3 (TLR3) signalling regulates innate and adaptive immune systems by the recognition of dsRNA. Activation of TLR3 signalling by poly(I:C) attenuates dextran sodium sulphate (DSS)-induced murine colitis. However, little information is available on the role of TLR3 signalling in the development of colitis-associated colon tumourigenesis. Methods Wild-type (WT) and TLR3-deficient (TLR3−/−) mice were intraperitoneally injected azoxymethane (AOM) 12.5 mg/kg on day 0 followed by three cycles of 2% DSS for 5 days and 2 weeks of free water consumption. Clinical indices such as weight change, colon length, the number of tumours, and the histologic severity of colitis were evaluated in each experiment. Immunohistochemical or immunofluorescence analyses for phospho-IκB kinase (IKK) and β-catenin were performed in colon tissues. To elucidate the antitumourigenic mechanism by colon inflammation, poly(I:C) or PBS was intraperitoneally injected in the AOM/DSS-induced tumourigenesis model in WT mice. To evaluate direct antitumor effect on tumourigenesis, as first experimental model, both WT and TLR3−/− mice were intraperitoneally injected AOM weekly for 12 weeks without DSS treatment. As the second experimental model, WT and TLR3−/− mice were received 2% DSS mixed with drinking water three times for 5 days every 2 weeks after one intraperitoneal AOM injection. Results TLR3−/− mice exhibited a higher tumour burden compared with wild-type mice. Body weight loss was greater in TLR3−/− mice than in WT mice. However, here was no significant difference in colon length and the severity of colitis between the two groups. Immunoreactivity for β-catenin was markedly increased in TLR3−/− mice. However, there was no difference in IKK expression. Activation of TLR3 by poly(I:C) was not associated with the reduced tumour development in WT mice. However, repeated AOM injections without DSS resulted in greater body weight loss in TLR3−/− mice than in WT mice, which was associated with the increased tumour development in TLR3−/−mice. Conclusion TLR3 signalling attenuated colitis-associated colon cancer development. Based on our experiments, TLR3 signalling inhibits colon tumourigenesis by direct antitumor activity rather than anti-inflammatory effect of colitis.

Download Full-text

Feeding of heifers for survival on whole and cracked sorghum grain under simulated drought conditions

Australian Journal of Experimental Agriculture ◽

10.1071/ea9680668 ◽

1968 ◽

Vol 8 (35) ◽

pp. 668 ◽

Cited By ~ 1

Author(s):

JG Morris

Keyword(s):

Weight Loss ◽

Body Weight ◽

Dry Matter ◽

Body Weight Loss ◽

Initial Body Weight ◽

Fresh Weight Basis ◽

Significant Difference ◽

Simulated Drought ◽

Sorghum Grain ◽

Body Energy

Groups of Hereford heifers of a mean (� SE.) body weight 187 � 3 kg were fed in yards a sole ration of sorghum grain + one per cent limestone for 26 weeks. Four groups were fed the equivalent of 1.36 kg of sorghum grain per head per day as either whole or cracked grain at either daily or twice weekly intervals. A fifth group was fed 1.64 kg of whole sorghum grain per head per day which provided a digestible dry matter intake comparable to that of 1.36 kg of cracked grain. When heifers were fed the same quantity of dry matter as whole and cracked grain, the rate of body weight loss of those fed whole grain was significantly greater than that of those fed cracked grain; and the rate of body weight loss of those fed daily was significantly greater than that of those fed twice weekly. With similar intakes of digestible dry matter from whole and cracked grain, there was no significant difference in the rate of body weight loss. Heifers from sub-groups of high initial body weight lost significantly more body weight than heifers from sub-groups of low initial body weight. The whole bodies of three heifers that died from undernutrition contained less than 0.3 per cent ether extract on a fresh weight basis, indicating complete exhaustion of body energy reserves.

Download Full-text

TFL Deletion Induces Incredible CXCL13 Secretion and Cachexia in Vavp-Bcl2Transgenic Mice

Blood ◽

10.1182/blood-2018-99-114351 ◽

2018 ◽

Vol 132 (Supplement 1) ◽

pp. 3908-3908

Author(s):

Kentaro Minagawa ◽

Kanako Wakahashi ◽

Fukui Chie ◽

Shinichiro Nishikawa ◽

Noboru Asada ◽

...

Keyword(s):

Weight Loss ◽

Body Weight ◽

T Cell ◽

B Cell ◽

Mrna Expression ◽

Cell Lines ◽

Cell Lineage ◽

Body Weight Loss ◽

Wild Type ◽

B Cell Lymphomas

Abstract Diffuse large B-cell lymphomas (DLBCL) are heterogeneous diseases caused by several genetic aberrations. The novel post-transcriptional regulator gene called transformed follicular lymphoma (TFL) was first identified from t(2;6)(p12;q23), which appeared during the transformation of FL to DLBCL (Minagawa et al. Br J Haematol 2007). Normal human lymphocytes generally express TFL, but it is defective in some leukemia/lymphoma cell lines. TFL overexpression in such cell lines inhibited cell growth, suggesting that TFL functions as a tumor suppressor (Minagawa et al. Mol Cancer Res 2009). TFL locates in mRNA processing body in the cytoplasm and has the unique CCCH-type zinc finger motif functioning as RNase. TFL regulates several cytokines, including IL-2, IL-6, IL-10, TNF-α, and IL-17a, via mRNA degradation. In an experimental autoimmune encephalitis model, TFL null mice (TFL-/-) demonstrated persistent paralysis, resulting from more infiltration of Th17 cells into CNS with markedly increased IL-17a mRNA levels. Therefore, a TFL-driven feedback mechanism for excessive inflammation is indispensable to suppress T-cell-mediated autoimmune diseases (Minagawa et al. J Immunol 2014). TFL deletion examined by FISH using a 110kbp DNA probe containing an entireTFL locus was found in 12.8% of mature B-cell lymphomas (n=86, FL=30, DLBCL=40). However, the pathological significance of TFL deletion has not yet been clarified. To investigate how TFL loss affects lymphoma biology, we developed VavP-bcl2 transgenic (Bcl2-Tg)/TFL-/-mice. Although the survival of TFL-/- was comparable to the wild-type, Bcl2-Tg/TFL-/- died about 19 weeks earlier than Bcl2-Tg (Fig. 1). Both strains developed lymphadenopathy and splenomegaly similarly. No different microscopic finding was noted in lymph nodes, spleen, or bone marrow (BM). No additional malignancy was found in Bcl2-Tg/TFL-/- on autopsy. However, significant body weight loss appeared by 30 weeks in Bcl2-Tg/TFL-/- but not in Bcl2-Tg (Fig. 3). To identify what causes earlier death in Bcl2-Tg/TFL-/-, we carefully examined the phenotypic change of BM lymphocytes. We found a unique B220-IgM+ population in Bcl2-Tg BM, which was not found in wild-type. We speculated that TFL deficiency in this population might drive the deterioration in Bcl2-Tg/TFL-/-. To identify which mRNA was dysregulated by TFL deficiency, we comprehensively analyzed mRNA expression profiles in B220-IgM+ cells in both strains using cDNA microarray chip. Among several genes upregulated at least threefold in Bcl2-Tg/TFL-/- than Bcl2-Tg, we paid attention to CXCL13, the mRNA expression of which in Bcl2-Tg/TFL-/- was 4.19-fold higher than that in Bcl2-Tg (p=0.03). In fact, CXCL13 concentration in BM extracellular fluid as well as plasma in Bcl2-Tg/TFL-/- showed incredible increase in a logarithmic scale (Fig. 2). As a noteworthy event, body weight loss in Bcl2-Tg/TFL-/- followed the increase of CXCL13 in plasma by 30 weeks (Fig. 3). To confirm that TFL post-transcriptionally regulates CXCL13 mRNA through the degradation of its 3′UTR, we performed a reporter assay with a plasmid vector containing 3′UTR of CXCL13 mRNA. Co-transfection with a TFL expression vector showed decreased luciferase activity compared to the control. This suggests that TFL directly regulates CXCL13 mRNA via its 3′UTR degradation. This regulation occurs more prominently in B-cell lineage rather than myeloid or T-cell lineage, whereas IL-2 mRNA regulation occurs promiscuously. CXCL13 secretion was significantly increased in the culture supernatant of BM cells but not spleen cells derived from Bcl2-Tg/TFL-/-. We further sorted several cell populations, including B220-IgM+ in BM, and cultured them for 96 h. CXCL13 secretion from B220-IgM+ population was increased significantly compared to other populations. Thus, we concluded that B220-IgM+ cells in BM are the main producer of CXCL13 in Bcl2-Tg/TFL-/-. Loss of TFL-driven attenuation for excessive inflammation in lymphoma-bearing mice could contribute to the short survival. It is of interest whether high plasma CXCL13 directly affects cachexia and early death in Bcl2-Tg/TFL-/-. TFL deletion in human lymphoma might contribute not only to malignant transformation but also to a major B symptom, i.e., weight loss. Our findings may open a new window for the predictive factor on the prognosis of B-cell lymphoma and/or new therapeutic intervention by targeting CXCL13. Disclosures No relevant conflicts of interest to declare.

Download Full-text

573 A novel human anti-PD1/IL15 bi-functional protein with robust anti-tumor activity and low systemic toxicity

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-sitc2020.0573 ◽

2020 ◽

Vol 8 (Suppl 3) ◽

pp. A607-A607

Author(s):

Dan Lu ◽

Zhanna Polonskaya ◽

Tzu-Pei Chang ◽

Stella Martomo ◽

Xenia Luna ◽

...

Keyword(s):

Cell Proliferation ◽

Weight Loss ◽

Body Weight ◽

Fusion Protein ◽

Body Weight Loss ◽

Systemic Toxicity ◽

Wild Type ◽

Functional Protein ◽

Bifunctional Protein ◽

Anti Tumor Activity

BackgroundIL-15 is a key cytokine promoting CD8+ T, NK, and NKT cell proliferation and has demonstrated clinical activity in cancer patients without evidence of any Treg stimulation.1 2 However, its short half-life and systemic toxicity limit its clinical utility. Kadmon has established an IL-15 fusion protein platform to extend the IL-15 serum half-life and direct its action to the tumor microenvironment.3 A major asset of this platform is anti-PD1/IL15 bifunctional protein. To test the bifunctionality hypothesis of this fusion protein in murine models, four different formats of the surrogate bi-functional proteins were engineered by fusing mouse IL-15 to a mouse-human chimeric anti-mouse PD1 antibody (m3A7). We presented earlier that the single IL-15 N-terminal fusion to anti-PD1 antibody (1N-IL-15/m3A7) showed significantly stronger anti-tumor activity in vivo mainly due to the cis-presentation to the PD1 and IL2Rβγ co-expressed on TILs. The cis-presentation potentially maximizes the bi-functionality of PD1 blockade and IL-15 stimulation.4 Here, the therapeutic single IL-15 N-terminal fusion antibodies containing a novel human PD1 antagonist antibody (38B2) and either wild-type IL15 (1N-IL-15/38B2) or mutated 65S-IL15 (65S-1N-IL-15/38B2) have been constructed; their anti-PD1 functions, IL15 stimulation and anti-tumor activities were evaluated.MethodsPurified 1N-IL-15/38B2 and 65S-1N-IL-15/38B2 were generated and characterized in vitro.4 The anti-tumor activities were examined in the human-PD-1/PD-L1 transgenic BALB/c mice subcutaneously transplanted with the human-PD-L1 positive CT26 colon carcinoma. The treatment was started when tumors reached 100 mm3 (IP, QW).ResultsAll 1N-IL-15/anti-PD1 fusions showed similar potencies in binding to the soluble and cell expressed human PD1 and blocking the hPDL-1 binding to hPD1. Comparing to wild-type 1N-IL-15/38B2, mutated 65S-1N-IL-15/38B2 showed lower stimulation (>150 folds) in the M07e, CTLL2, mouse spleen cells and hPBMC (mainly CD8T+ T cell) proliferation. When we treated hPDL1-CT26 tumor transplanted hPDL1-hPD1 transgenic mice with 65S-1N-IL-15/38B2 at 6 mg/kg, 80% of tumor growth inhibition (TGI) was achieved with no body-weight loss. Although wild-type 1N-IL-15/38B2 at 3 mg/kg demonstrated similar efficacy, a significant mouse body-weight loss was observed and 1/3 mice died after second injection. No anti-tumor activity was observed for 65S-1N-IL-15 non-target fusion in 6 mg/kg.ConclusionsThe previous observation of robust anti-tumor activity of surrogate 1N-IL-15/m3A7 in PD1 resistant LL2 model was replicated with the therapeutic bifunctional protein in this study. We also found that lower stimulation 65S-1N-IL-15/38B2 showed strong anti-tumor activity with significant low systemic toxicity; suggesting that the 65S mutation increased the therapeutic window of this bi-functional proteinReferencesGoldrath, AW etc. Cytokine requirements for acute and basal homeostatic proliferation of naive and memory CD8+ T cells. J Exp Med 2002; 195:1515–1522.Waldmann TA. Cytokines in Cancer Immunotherapy. Cold Spring Harb Perspect Biol 2018;103. Martomo, S. etc. ESMO 2019 1221P; SITC 2019 P#4854. Polonskaya Z. etc. AACR 2020 #2263

Download Full-text

Water Requirements of Gerbils and Kangaroo Rats in the Laboratory

Psychological Reports ◽

10.2466/pr0.1968.23.3f.1063 ◽

1968 ◽

Vol 23 (3_suppl) ◽

pp. 1063-1069 ◽

Cited By ~ 5

Author(s):

Robert Boice ◽

Janet G. Arledge

Keyword(s):

Weight Loss ◽

Drinking Water ◽

Body Weight ◽

Free Water ◽

Body Weight Loss ◽

Sunflower Seeds ◽

Water Requirements ◽

Kangaroo Rats

Both gerbils ( Meriones unquiculates) and kangaroo rats ( Dipodymus ordii) consumed free water (0.088 and 0.107 cc./gm. body weight/day; respectively) at reliable daily rates irrespective of two food types. When drinking water was withheld, sunflower seeds retarded body weight loss more than Lab Blox but Ss were dead or near death by the 19th day when weights were down approximately 30%.

Download Full-text

Effect of heat exposure on the thermoregulatory responses of selected naked neck chickens

Arquivo Brasileiro de Medicina Veterinária e Zootecnia ◽

10.1590/s0102-09352002000100006 ◽

2002 ◽

Vol 54 (1) ◽

pp. 35-41 ◽

Cited By ~ 9

Author(s):

C.M. Mazzi ◽

M.I.T. Ferro ◽

A.A.D. Coelho ◽

V.J.M. Savino ◽

M. Macari ◽

...

Keyword(s):

Weight Loss ◽

Body Weight ◽

Heat Stress ◽

Heat Exposure ◽

Body Weight Loss ◽

The Other ◽

Naked Neck ◽

Significant Difference ◽

Before And After ◽

Slight Advantage

The variation in cloacal temperature, body weight loss and expression of the 70 kDa heat shock protein (Hsp70) in three naked neck broiler genotypes during heat stress were studied. Twelve birds of each genotype (Na/Na, Na/na and na/na) were reared to market weight (approximately 2.1kg) at thermoneutral temperature. Six birds from each group served as controls and the remaining six underwent gradual heat stress (from 28ºC to 36ºC). Cloacal temperature and body weight were measured before and after exposure to heat. Liver samples were collected and Hsp70 levels were quantified using western blotting with monoclonal anti-chicken Hsp70 antibody. Heterozygous (Na/na) birds had a significantly lower cloacal temperature variation and less body weight loss during heat stress than the other genotypes. There was no significant difference in the Hsp70 levels among the genotypes. Heterozygous birds (Na/na) appeared to have a slight advantage over the other genotypes during gradual heat stress, perhaps because of a heterotic effect.

Download Full-text

Influence of housing conditions, number of farrowing and number of pigs in litter on weight loss in sows during lactation

Veterinarski glasnik ◽

10.2298/vetgl1306337s ◽

2013 ◽

Vol 67 (5-6) ◽

pp. 337-348

Author(s):

Z. Sladojevic ◽

D. Kasagic ◽

B. Kukolj ◽

Danijela Kirovski

Keyword(s):

Weight Loss ◽

Body Weight ◽

Body Weight Loss ◽

Housing Conditions ◽

Intensive Farming ◽

Group I ◽

Lower Body Weight ◽

Significant Difference ◽

The Difference ◽

Extensive Farming

The objective of this work was to investigate the influence of housing conditions, number of farrowing and number of pigs in litter on weight loss in sows during lactation. The experiment included 60 sows, half breed developed by cross breeding of Big Yorkshire and Swedish Landrace in lactation. Body weight of the sows (kg) was determined 2nd, 7th, 14th, 21st and 28th day after farrowing. On the basis of the obtained values, there was calculated the difference in body weight between two consecutive investigation periods. The sows were divided into two groups and held in different housing conditions: 30 of them were held in extensive, and other 30 sows in intensive way of farming. In regard to number of farrowing, the sows were divided into three groups: sows with one or two farrowing (group P1, n=20), with three or four farrowing (group P2, n=20), and sows that farrowed five or more times (P3,n=20). In regard to number of pigs in litter, the sows were divided into two groups: the first (Group I, n=30) with sows that had up to eight pigs, and the second (Group II, n=30) with sows that had nine or more pigs in litter. The obtained results showed that in all perids of the investigation during lactation, body weight in sows held in extensive farming conditions was statistically significantly higher compared to those from intensive farming conditions. On the other hand, the loss of body weight during lactation was significantly higher in sows from extensive in regard to intensive farming conditions in the first two weeks of lactation. The sows with bigger number of farrowing had greater body weight, compared to those with smaller number of farrowing. There was no statistically significant difference in body weight loss during lactation, between sows with different number of farrowing, but considering that the sows with less farrowings had significantly lower body weight, they consequently lost more weight in percentage. The sows with greater number of pigs had significantly smaller body weight in the end of lactation, that is on 21st and 28th day of lactation. The results of this investigation point out to the fact that mainly housing conditions, that is nutrition influence body weight loss, and after that comes the number of pigs in litter.

Download Full-text

Withdrawal of salt supplementation from adrenalectomized Wistar rats distinguishes between those animals with, and those without, adrenocortical insufficiency

Clinical Science ◽

10.1042/cs0710675 ◽

1986 ◽

Vol 71 (6) ◽

pp. 675-683 ◽

Cited By ~ 5

Author(s):

S. M. Gardiner ◽

T. Bennett ◽

I. A. MacDonald

Keyword(s):

Weight Loss ◽

Body Weight ◽

Free Water ◽

Body Weight Loss ◽

Plasma Aldosterone ◽

The Body ◽

Separate Experiment ◽

Adrenocortical Insufficiency ◽

Group 2 ◽

Group 1

1. The effects of replacing a 1% NaCl drinking solution with Na+-free water for 2 days on body weight and fluid and electrolyte balances were studied in adrenalectomized and sham-operated rats. 2. Eight weeks after operation, after the animals had been drinking Na+-free water for 2 days, some adrenalectomized animals (about 75%; designated group 1) experienced body weight losses which were outside the 99% confidence limits for the sham-operated rats (– 9.2 to + 5.3 g) whereas the remainder (designated group 2) were indistinguishable from the controls. 3. The body weight loss in group 1 was associated with negative fluid, Na+ and K+ balances. In group 2 rats, fluid balance was maintained as well as in the sham-operated rats, but their handling of Na+ and K+ was different. 4. In a separate experiment, plasma aldosterone, corticosterone, catecholamine and solute concentrations were measured in adrenalectomized rats from groups 1 and 2 (selected on the basis of body weight loss whilst drinking Na+-free water) and sham-operated rats, after drinking Na+-free water for 2 days. 5. In group 1 animals, plasma aldosterone levels were unmeasurable and corticosterone was extremely low (less than 5% of controls). In group 2, aldosterone was measurable but low, and corticosterone was higher than in group 1 but lower than in sham-operated rats. In line with these findings, animals in group 1, but not those in group 2, were hyponatraemic and hyperkalemia These results are consistent with the activation of latent adrenocortical tissue in the group 2 animals. 6. We suggest that monitoring body weight changes resulting from drinking Na+-free water for 2 days provides a useful, non-invasive test for reliably distinguishing between those adrenalectomized animals with and those without adrenocortical insufficiency.

Download Full-text