OBJECTIVETo investigate the effect of kirenol on bovine type II collagen (CII)-specific lymphocytes in vivo and in vitro, and explore the mechanism of kirenol-induced immunosuppression in antigen-specific lymphocytes.

METHODSTwenty-four Wistar rats were randomized into control group, collagen-induced arthritis (CIA) model group, kirenol group (2 mg/kg), and prednisolone group (2 mg/kg). After CII injection, the rats in the latter two groups received intragastric administration of kirenol and prednisolone for 30 days, and the spleens and draining lymph nodes of the rats were harvested to prepare single cell suspensions for measurement of the cytokine levels using ELISA. In the in vitro experiment, the lymphocytes from the control rats, with or without 20 µg/ml CII treatment in the presence of 0-80 µg/ml kirenol, were evaluated for cell proliferation and apoptosis using [(3)H]-thymidine incorporation and flow cytometry, respectively.

RESULTSCompared with those in CIA group, IFN-γ and TNF-α production was significantly reduced in splenocyte culture supernatant of kirenol group (P<0.05 and P<0.01, respectively), and the level of IL-10 and IL-4 was up-regulated (P<0.05 and P<0.01, respectively); IFN-γ and TNF-α secretion by the cultured lymph node cells (LNCs) significantly decreased (P<0.05 and P<0.001, respectively) and IL-10 and IL-4 production increased (P<0.05, P<0.001) in kirenol group. In the in vitro experiment, kirenol treatment caused obvious suppression of CII-induced LNC proliferation and dose-dependently induced antigen-specific apoptosis of the splenocytes and LNCs.

CONCLUSIONKirenol treatment reduces pro-inflammatory cytokine secretion, increases anti-inflammatory cytokine production, inhibits cell proliferation and induces apoptosis of CII-specific lymphocytes in vitro, suggesting the potential of kirenol as an immunosuppressant.

Animals ; Anti-Inflammatory Agents ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Arthritis, Rheumatoid ; chemically induced ; drug therapy ; immunology ; Cattle ; Cell Proliferation ; Cells, Cultured ; Collagen Type II ; immunology ; Cytokines ; immunology ; Diterpenes ; pharmacology ; therapeutic use ; Female ; Immunosuppressive Agents ; pharmacology ; therapeutic use ; Lymphocytes ; cytology ; drug effects ; immunology ; Rats ; Rats, Wistar

Objective To explore the effect of calycosin(CAL)on neuronal autophagy and apoptosis in rats with spinal cord injury(SCI)by regulating PI3K/Akt signaling pathway and its mechanism.Methods A total of 32 male or female adult SD rats were randomly divided into the sham group,the SCI group,the CAL low(20 mg/kg)dose group and the CAL high(40 mg/kg)dose group with 8 rats in each group.The rat model of moderate SCI was established by modified Allen's method.After successful modeling,rats were injected intraperitoneally immediately with different dosage of CAL or equal amount of saline once a day for 7 consecutive days.Basso,Beattie and Bresnahan(BBB)scores were used to evaluate the recovery of motor function of rats at 1,3 and 7 d after surgery.At 7 d after surgery,Nissl staining was used to detect the surviving number of motor neurons in anterior horn of spinal cord.Western blot assay was used to assess expression levels of p62,Beclin-1,microtubule-associated protein 1 light chain 3(LC3B),phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)pathway,Cleaved-Caspase-3,B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X protein(Bax)proteins.Immunofluorescence staining was used to measure the expression of LC3B in anterior neurons of spinal cord.Results Compared with the sham group,BBB scores,the surviving number of motor neurons and levels of Bcl-2,Bcl-2/Bax,p-PI3K,p-PI3K/PI3K,p-Akt and p-Akt/Akt were significantly decreased(P＜0.05),and levels of p62,Beclin-1,LC3B Ⅱ,LC3BⅡ/Ⅰ,Cleaved-Caspase-3 and Bax were significantly increased in the SCI group(P＜0.05).Compared with the SCI group,BBB scores,the survival of anterior horn motor neurons and levels of LC3B Ⅱ,LC3B Ⅱ/Ⅰ,Bcl-2,Bcl-2/Bax,p-PI3K,p-PI3K/PI3K,p-Akt and p-Akt/Akt were increased in the CAL low dose group and CAL high dose group,and levels of p62,Cleaved-Caspase-3 and Baxcould were significantly decreased(P＜0.05).Conclusion CAL could promote autophagy and inhibit apoptosis of neurons through activating PI3K/Akt signaling pathway,thereby conferring a protective role following SCI in rats.

[摘 要] 基因修饰T细胞疗法在肿瘤治疗领域取得突破性进展，主要包括嵌合抗原受体基因修饰T（chimeric antigen receptor engineered T，CAR-T）细胞和T细胞受体基因修饰T（T-cell receptor modified T，TCR-T）细胞。虽然CAR-T细胞疗法在血液系统肿瘤治疗领域呈现良好的临床治疗效果，但CAR-T细胞仅能识别肿瘤细胞膜抗原（占细胞全部抗原的比例约10%），而TCR-T细胞可以识别人白细胞抗原（human leukocyte antigen，HLA）提呈的细胞内抗原，因此TCR-T细胞可以识别更多种类的肿瘤抗原，进而实现对CAR-T细胞的合理补充。由于TCR-T细胞需要同时识别细胞内抗原和对应的HLA，而不同患者的HLA分型和表达的肿瘤抗原都可能存在巨大差异，因此有必要为每个/每类肿瘤患者定制个体化的TCR-T细胞，其中的关键为筛选特异识别肿瘤抗原的TCR。当前主要有筛选靶向“已知”肿瘤抗原TCR和筛选靶向“未知”肿瘤抗原TCR的两种策略，但其各有适用性，应针对每个患者制定适合的筛选方法，以制备多种肿瘤特异性TCR-T细胞，从而实现个体化TCR-T细胞的肿瘤治疗。

Background Organic ultraviolet (UV) filters are widely used in personal care products. So far, relevant studies on organic UV filters in indoor dust have been reported. Objective This study aims to establish a thermal desorption combined with gas chromatography-mass spectrometry (TD-GCMS) method to identify organic UV filters in indoor air collected from different indoor environments, so as to reveal the pollution levels and characteristics of organic UV filters in indoor environment. Methods Based on the standard indoor air sampling protocol, a total of 60 samples were collected from eight different kinds of indoor environments (male and female dormitory rooms, offices, labs, barber shops, printing shops, hotels, and private cars) on and nearby Minhang Campus of Shanghai Jiao Tong University from August to November, 2020. The concentrations of six common organic UV filters, including homosalate (HMS), 2-ethylhexyl salicylate (EHS), 3-(4-methylbenzylidene)-camphor (4-MBC), isoamyl 4-methoxycinnamate (IMC), octocrylene (OC), and octyl 4-methoxycinnamate (EHMC), in the air of different indoor environments were detected by TD-GCMS. Furthermore, the correlations of individual organic UV filters in different indoor environments were analysed. Results Under optimized detection conditions, the correlation coefficients of the quantitative standard curves of selected six organic UV filters were all at or above 0.997. The relative standard deviations of 1 mg·L⁻³ samples ranged from 1.74% to 7.11%, and the recoveries ranged from 67.17% to 106.5%. The relative standard deviations of 10 mg·L⁻³ samples ranged from 3.59% to 8.76%, and the recoveries ranged from 78.80% to 126.60%. The detection rates of the other five organic UV filters except IMC were all at or more than 92% in eight different kinds of indoor air. The median concentration of total organic UV filters was 75.17 ng·m⁻³, and EHS presented the highest median concentration of 28.55 ng·m⁻³. Regarding different indoor environments, the highest concentration of total organic UV filters was found in the female dormitory samples, 154.98 ng·m⁻³. The respective pair-analysis among HMS, EHMC, OC, and EHS of all indoor air samples reached a significant level of correlation (r=0.40-0.61, P<0.01). Conclusion The TD-GCMS method is satisfactory for the determination of organic UV filters in indoor air. EHS, EHMC, HMS, OC, and 4-MBC are identified in selected eight indoor environments, and they may have similar sources of pollution.