ABSTRACT
【Objective】 To investigate the viability of rapamycin-treated rapamycin-treated dendritic cells (DCs) in intervening transfusion-related acute lung injury (TRALI) after infection. 【Methods】 1)The TRALI mouse model was induced by lipopolysaccharide (LPS) combined with anti-H2Kd antibody. The mice anal temperature and the wet/dry ratio of lung, kidney, spleen and brain tissues were measured. 2) Mouse bone marrow-derived DC cells were induced in vitro and treated with rapamycin (10nM) for 24h. 3) Mice were injected with or without rapamycin or rapamycin-treated DC, then injected with LPS intraperitoneally one hour later, finally injected with anti-H2Kd antibody 24 hours later to induce the onset of TRALI. The death situation of the mice was observed and recorded. The condition of mice after the onset of TRALI was analyzed by mouse body temperature, lung wet-dry ratio, and pleural effusion weight and lung histopathological sections. 【Results】 By comparing the induction effects of anti-H2Kd antibody solutions with different concentrations and volumes, the mouse model induced by 0.1mg/kg LPS combined with 4.5 mg/kg anti-H2Kd antibody (infusion volume of 100μL) was selected as the TRALI mouse model for this study. After the onset of TRALI, the wet/dry ratio of the lungs could be significantly increased and the body temperature could be significantly reduced in the model mice. After the intervention of TRALI mice with DCs treated with rapamycin, the mortality rate was significantly reduced, and the lung tissue lesions of the mice were significantly improved, whose protection effect was better than that of the rapamycin-treated group. Compared with the TRALI incidence group, the weight of pleural effusion in the intervention group was significantly reduced (P<0.05), but there was no significant difference in lung wet/dry ratio and body temperature. 【Conclusion】 The combination of LPS and antibodies can effectively induce a stable and typical TRALI mouse model, suggesting that the presence of infectious inflammation and blood transfusion-related inflammatory substances are the decisive factor for the pathogenesis of TRALI. Meanwhile, DCs treated with rapamycin have a protective effect on post-infection transfusion-related acute lung injury, which is expected to be a potential cell therapy strategy to intervene in the exacerbation of TRALI.
ABSTRACT
<p><b>OBJECTIVE</b>To explore the biological characteristics and the immuno-suppression function of tolerogenic dendritic cells (tDC) induced by tacrolimus.</p><p><b>METHODS</b>Human monocytes derived from peripheral blood were cultured in the cGMP-compliant CellGro DC medium supplemented with GM-CSF and IL-4 to obtain dendritic cells (DCs), and 0.1 μmol/L immunosuppressive drug tacrolimus was added to the culture medium at the third and fifth day to obtain tDCs. The molecular markers of them and the livability were assayed by flow cytometry. Then the tolerance functionality of tDCs induced by many agents and these tDCs modulated allogeneic CD4 T cells was determined via CFSE proliferation assay. And the research also analyzed the biological characters and immunosuppression function of tDCs induced by tacrolimus after storing.</p><p><b>RESULTS</b>tDCs induced by tacrolimus exhibit a typical tolerogenic phenotype, whose level of costimulatory molecules CD80, CD83, CD86 and HLA-DR is (2.95 ± 1.32)%, (2.33 ± 1.60)%, (90.02 ± 7.42)% and (91.80 ± 6.18)%, respectively. It's survival rate was (85.2 ± 4.72)%. And immunosuppressive drugs didn't influence the differentiation of tDCs from monocytes. tDCs induced by immunosuppressive drugs dexamethasone, cyclosporin A and tacrolimus had lower immunogenic than control DCs as CD4+ T proliferation rate of tDCs induced by tacrolimus is 0.42% and could not primed allogeneic CD4+ T cells proliferation. Functional analyses showed that tDCs induced by tacrolimus can more effectively suppressed mature DC-induced T cell proliferation than other tDCs, whose inhibition rate can reach (67.01 ± 19.73)%. Importantly, tDCs induced by tacrolimus had phenotypical and functional stability after storing.</p><p><b>CONCLUSION</b>tDCs induced by tacrolimus with tolerance functionality are a promising cellular therapeutic for immunomodulation.</p>
Subject(s)
Humans , Cell Proliferation , Cells, Cultured , Dendritic Cells , Allergy and Immunology , Immune Tolerance , Lymphocyte Culture Test, Mixed , Tacrolimus , PharmacologyABSTRACT
The aim of this study was to observe the difference in respect to the leukocyte reduction efficiency and quality of fresh-frozen plasma (FFP) from filtered whole blood between two types of in-line filters wherein only filter materials were surface modified by the two methods respectively. Whole blood was kept in refrigerator and filtered within 6 h of collection at ambient temperature. Samples were taken pre- and post filtration for analysis of WBC numbers, coagulation factors and complement activation (n = 8 for each type of filter). All filtered units contained < 2. 5 x 10(6) residual leucocytes. RBCs recovery was over 93%. No significant difference between group A and B was seen. But group B appeared to take longer time for filtration than did group A (9'29" vs. 8'01"). Neither group A nor group B showed statistically significant losses of total protein, album, IgG, IgM, fibrin, factors VIII, IX, vWF and C3 (P > 0.05). Factor V, XI and AT-III decreased significantly in two group filters. Group B showed more significant losses of IgA content and factor V activity than did group A, which appeared to be related to the difference in surface character between group A and group B filters. These two types of filters could remove leukocytes effectively, and no significant changes were observed in the quality of FFP from the filtered whole blood. It is presumed that the filter material with better bio-compatibility will give a high recovery of plasma protein and coagulation factors after filtration.