ABSTRACT
BACKGROUND AND AIMS: Tumour necrosis factor-α (TNF) antagonists have improved the management of inflammatory bowel disease (IBD), however, their usage and administration persist to be suboptimal. Here, we examined the relationship between tissue-specific TNF mRNA expression in mucosal biopsies from IBD patients and anti-TNF treatment response. METHODS: Archived tissue samples from patients with luminal IBD that had all been or were in treatment with anti-TNF were included (18 adults and 24 paediatric patients). Patients were stratified into three groups according to anti-TNF response: responders, primary non-responders (PNR) and secondary loss of response (SLOR). TNF mRNA was detected using RNAscope in situ hybridisation (ISH) and the expression was quantified using image analysis. RESULTS: The ISH analysis showed varying occurrence of TNF mRNA positive cells located in lamina propria and often with increased density in lymphoid follicles (LF). Consequently, expression estimates were obtained in whole tissue areas with and without LF. Significantly higher TNF mRNA expression levels were measured in adults compared to paediatric patients in both the analyses with and without LF (p = .015 and p = .016, respectively). Considering the relation to response, the adult and paediatric patients were evaluated separately. In adults, the TNF expression estimates were higher in PNRs compared to responders with and without LF (p = .017 and p = .024, respectively). CONCLUSION: Our data indicate that adult PNR have significantly higher TNF mRNA levels than responders. This suggests that higher anti-TNF dose may be considered for IBD patients with high TNF mRNA expression estimates from the start of treatment.
Subject(s)
Inflammatory Bowel Diseases , Tumor Necrosis Factor-alpha , Adult , Humans , Child , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolism , Tumor Necrosis Factor Inhibitors/therapeutic use , RNA, Messenger/genetics , Intestinal Mucosa/pathology , Inflammatory Bowel Diseases/pathologyABSTRACT
Objective To study the effects and mechanisms of growth hormone(GH) pretreatment on the inflammatory response of liver in rats after cardiopulmonary bypass(CPB).Methods The CPB model in rat was established.The rats were randomly divided into GH pretreatment group(GH group),CPB group,and sham group.The serum levels of endotoxin,TNF-?,and liver function were determined at different intervals respectively.Simultaneously,the liver tissues were collected for the detection of the CD14 mRNA and TNF-? mRNA by RT-PCR.Results Compared with pre-CPB,the serum levels of endotoxin,TNF-? were significantly increased,and the lesion of liver function was apparent in GH group and CPB group.Meanwhile,the expression of CD14 mRNA and TNF-? mRNA in liver tissues were also increased obviously in both groups.However,the serum levels of endotoxin and TNF-? and the expression of CD14 mRNA and TNF-? mRNA in liver tissues at the same time point in GH group were significantly lower than those in CPB group(P
ABSTRACT
It is well known that HL-60 cell, a human promyelocytic line, is differentiated into eosinophil-like cells in the presence of butyric acid, and thus the differentiated HL-60 cells have been used as a model system to study irnmunological properties of peripheral eosinophils which are thought to be terminally differentiated. To study whether HL-60 cells alter their capability of expressing cytokines during differentiation to eosinophil-like cells, we examined TNF mRNA levels in HL-60 cells treated with butyric acid by Ribonuclease Protection Assay (RPA). HL-60 cells were incubated for 3 days in the presence of butyric acid (0.5 mM), and stimulated with PMA and lipopolysaccharide (LPS). The levels of TNF mRNA decreased by 50 % and 95 % upon one and two days of post-treatment of butyric acid, respectively. The decreased pattern in TNF mRNA levels was also observed in HL-60 cells that have been treated with retinoic acid known as an inducer for differentiation of them. In accordance with these results, prominent azurophilic granules typical in eosinophils appeared in the cytoplasm of the differentiated HL-60 cells. The decreased expression of TNF mRNA was not attributable to the presence of serum, since increasing concentrations of serum had no effect. Furthermore, interleukin-5 (IL-5), which is known to be involved in activation and trafficking of eosinophils in vivo and in vitro, failed to affect TNF mRNA production when it was used in place of butyric acid. These data suggest that the differentiated HL-60 cells may have immunological resemblance to eosinophils in that they weakly produce the cytokine mRNA.
Subject(s)
Humans , Butyric Acid , Cytokines , Cytoplasm , Eosinophils , HL-60 Cells , Interleukin-5 , Ribonucleases , RNA, Messenger , TretinoinABSTRACT
Objective To study the time-course relationship of eotaxin mRNA and TNF-? mRNA expression in the lung tissue of sensitized mice after antigen challenge and the effects of Cryptoporus polysaccharide (CP) on these expression. Methods Eotaxin mRNA and TNF-? mRNA expression was determined by semi-quantitative RT-PCR. The function implications of eotaxin mRNA and TNF-? mRNA expression were examed by detecting the number of total leucocyte and eosinophils in bronchoalveolar lavage fluid (BALF). Results Peak level of TNF-? mRNA expression appeared at 8 h and eotaxin mRNA expression at 24 h in the lung tissue of sensitized mice after antigen challenge. Compared with the control group, total numbers of leukocyte and eosinophil in BALF increased in sensitized mice. CP 3, 10, and 30 mg/kg inhibited eotaxin mRNA and TNF-? mRNA expression, and inflammatory cells accumulation in airway of the sensitized mice in a dose-dependent manner. Conclusion The mechanism of inhibitory effects of CP on eotaxin mRNA and TNF-? mRNA expression may be related to the anti-allergic inflammation.
