ABSTRACT
Antibody-mediated rejection (AMR) is one of the major causes of graft loss after transplantation. Recently, the regulation of B cell differentiation and the prevention of donor-specific antibody (DSA) production have gained increased attention in transplant research. Herein, we established a secondary allogeneic in vivo skin transplant model to study the effects of romidepsin (FK228) on DSA. The survival of grafted skins was monitored daily. The serum levels of DSA and the number of relevant immunocytes in the recipient spleens were evaluated by flow cytometry. Then, we isolated and purified B cells from B6 mouse spleens in vitro by magnetic bead sorting. The B cells were cultured with interleukin-4 (IL-4) and anti-clusters of differentiation 40 (CD40) antibody with or without FK228 treatment. The immunoglobulin G1 (IgG1) and IgM levels in the supernatant were evaluated by enzyme-linked immunosorbent assay (ELISA). Quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and western blotting were conducted to determine the corresponding levels of messenger RNA (mRNA) and protein expression in cultured cells and the recipient spleens. The results showed that FK228 significantly improved the survival of allogeneic skin grafts. Moreover, FK228 inhibited DSA production in the serum along with the suppression of histone deacetylase 1 (HADC1) and HDAC2 and the upregulation of the acetylation of histones H2A and H3. It also inhibited the differentiation of B cells to plasma cells, decreased the transcription of positive regulatory domain-containing 1 (Prdm1) and X-box-binding protein 1 (Xbp1), and decreased the expression of phosphorylated inositol-requiring enzyme 1 α (p-IRE1α), XBP1, and B lymphocyte-induced maturation protein-1 (Blimp-1). In conclusion, FK228 could decrease the production of antibodies by B cells via inhibition of the IRE1α-XBP1 signaling pathway. Thus, FK228 is considered as a promising therapeutic agent for the clinical treatment of AMR.
Subject(s)
Animals , Mice , Depsipeptides , Endoribonucleases , Hematopoietic Stem Cell Transplantation , Histone Deacetylase Inhibitors/pharmacology , Protein Serine-Threonine Kinases , Skin TransplantationABSTRACT
Objective To summarize the effect of professional organ procurement team at Tongji Hospital in Chinese donation after citizen's death.Methods A retrospective analysis was done on the clinical data of 335 cases of organ acquisition by the professional organ procurement team of Tongji Hospital,including 27 infant donors and the efficiency of the team was assessed.Results Retrospective analysis on the clinical data of 335 cases of organ acquisition by the professional organ procurement team of Tongji Hospital,including cases of 27 infant donors and assessment on the efficiency of the team.Conclusion The establishment of a professional organ procurement team is to standardize the procurement procedure and shorten the time in pre-surgical preparation and operation,decreasing the errors and deviations caused by human factors in the procurement process.
ABSTRACT
Objective The detection results of two methods of quantitative methods and blood culture method were compared to explore the value of quantitative detection of serum procalcitonin.Methods From February 2014 to January 2015,the clinical data of 192 patients who were tested for two quantitative detection and blood culture were collected at the same time,and the results were eligible analyzed.One of the two quantitative detection methods was the electrochemical luminescence,and the other was the up-converting phosphor method.Results Compared with the result of blood culture,the positive rate was significantly higher in electrochemical luminescence and the up-con-verting phosphor method (χ2 =70.531,43.671,all P<0.05).The positive rates of up-converting phosphor and electrochemical luminescence were 66.1% and 75.5% respectively, and the difference between two quantitative methods was also statistically significant (χ2 =5.297,P<0.05).Method of electrochemical luminescence tested on higher sensitivity.When the level of PCT was less than threshold,for method of the electrochemical luminescence,the sensitivity on the blood culture was 93.7%,the specificity was 33.3%,the positive predictive value was 40.7%,the negative predictive value was 91.5%,the area under ROC curve was 0.628.For method of up-converting phosphor, the sensitivity on the blood culture was 90.5%,the specificity was 46.5%,the positive predictive value was 45.7%, the negative predictive value was 90.7%,the area under ROC curve was 0.554.Conclusion The electrochemical luminescence detection method of serum procalcitonin was better than up -converting phosphor method and blood culture.The electrochemical luminescence method which makes rapidly qualitative and accurately quantitative detec-tion,can give early diagnosis,medication guide in a short time,and predict the prognosis of disease.