菠萝蜜低聚肽的免疫调节作用研究

目的:探究菠萝蜜低聚肽(jackfruit oligopeptides,JOPs)免疫调节功能。方法:采用SPF级雌性BALB/c小鼠为实验对象,进行8项免疫实验测定,系统评价不同干预剂量(0.20、0.40、0.80 g/kg·bw)JOPs对小鼠免疫器官相对重量、细胞免疫功能、体液免疫功能、单核-巨噬细胞功能、自然杀伤细胞(nature killer cell,NK细胞)活性以及T淋巴细胞亚群的影响。结果:与空白对照组相比,JOPs 0.40、0.80 g/kg·bw剂量组脾脏指数显著提高(P<0.05),且JOPs 0.40 g/kg·bw剂量组脾脏指数显著高于乳清蛋白组(P<0.05);与空白对照组及乳清蛋白组相比,JOPs 0.40、0.80 g/kg·bw剂量组ConA诱导的小鼠脾淋巴细胞增殖能力、足跖增厚度、溶血空斑数、碳廓清吞噬指数a、巨噬细胞吞噬率与吞噬指数及NK细胞活性显著提高(P<0.05),JOPs 0.80 g/kg·bw剂量组血清溶血素含量、CD3⁺T细胞数量、CD4⁺T细胞百分比及CD4⁺CD25⁺/CD4⁺、CD4⁺/CD8⁺比值显著提高(P<0.05);各JOPs剂量组对小鼠体重增长、胸腺指数均无显著性影响(P>0.05)。结论:JOPs可提高机体适应性免疫及固有免疫功能,起到免疫增强作用。     

Capability of Human Dendritic Cells Pulsed with Autologous Induced Pluripotent Stem Cell Lysate to Induce Cytotoxic T Lymphocytes against HLA-A33-Matched Cancer Cells

Irradiated murine induced-pluripotent stem cells (iPSCs) elicit the antitumor response in vivo. However, it is unclear whether human iPSCs would elicit antitumor effects. In the present study, we investigated the capability of human iPSC lysate (iPSL)-pulsed dendritic cells (DCs) (iPSL/DCs) to induce cancer-responsive cytotoxic T lymphocytes (CTLs) in vitro. iPSCs and DCs were induced from peripheral blood mononuclear cells isolated from a human leukocyte antigen (HLA)-A33 homozygous donor. The iPSL was pulsed with immature DCs, which were then stimulated to allow full maturation. The activated DCs were co-cultured with autologous CTLs and their responses to SW48 colorectal carcinoma cells (HLA-A32/A33), T47D breast cancer cells (HLA-A33/A33), and T98G glioblastoma cells (HLA-A02/A02) were tested with enzyme-linked immunospot (ELISPOT) assays. Comprehensive gene expression analysis revealed that the established iPSCs shared numerous tumor-associated antigens with the SW48 and T47D cells. Immunofluorescent analysis demonstrated that the fluorescent-labeled iPSL was captured by the immature DCs within 2 h. iPSL/DCs induced sufficient CTL numbers in 3 weeks for ELISPOT assays, which revealed that the induced CTLs responded to SW48 and T47D cells. Human iPSL/DCs induced cancer-responsive CTLs on HLA-A33-matched cancer cells in vitro and could be a promising universal cancer vaccine for treating and preventing cancer.     

Molecular Features and Diagnostic Challenges in Alpha/Beta T-Cell Large Granular Lymphocyte Leukemia

Large granular lymphocyte leukemia is a rare chronic lymphoproliferative disease of cytotoxic lymphocytes. The diagnosis, according to the WHO, is based on a persistent (>6 months) increase in the number of LGL cells in the peripheral blood without an identifiable cause. A further distinction is made between T-LGL and NK-LGL leukemia. The molecular sign of LGL leukemia is the mutation of STAT3 and other genes associated with the JAK/STAT pathway. The most common clinical features are neutropenia, anemia, and thrombocytopenia, and it is often associated with various autoimmune conditions. It usually has an indolent course. Due to the rarity of the disease, no specific treatment has yet been identified. Immunosuppressive therapy is used and may allow for disease control and long-term survival, but not eradication of the leukemic clone. Here, we discuss the clinical presentation, diagnostic challenges, pathophysiology, and different treatment options available for alpha/beta T-LGL leukemia, which is the most common disease (85%), in order to better understand and manage this often misunderstood disease.     

X射线低剂量多次辐射对小鼠外周血淋巴细胞的影响

为了评估低剂量多次辐射对健康机体产生的生物学风险,检测了X射线多次照射对BALB/c小鼠免疫系统的影响.采用X射线全身多次照射,第1天照射0.07 Gy,之后每天照射0.08 Gy共12 d,剂量率0.2 Gy/min,累积剂量为0、0.07、0.23、0.39、0.55、0.71、0.87和1.03 Gy时取样.照射24 h后取血,用流式细胞仪检测外周血中免疫细胞周期和死亡的变化.结果表明,外周血淋巴细胞的周期在0.39和1.03 Gy时,被阻滞在G2/M期,在0.07、0.23、0.71和0.87Gy时,被阻滞在G0/G1期;细胞的死亡随着累积剂量的增加而增加,在0.71Gy时达到峰值.经过X射线多次全身照射小鼠后,可引起外周血淋巴细胞周期和死亡比例发生变化,符合线性平方模型,造成其免疫系统一定的损伤.     

Immunoreactive properties of α-casein and κ-casein: Ex vivo and in vivo studies

The aim of this study was to evaluate the ex vivo and in vivo studies immune potential of α- and κ-casein. Ex vivo, naïve mouse splenocytes were stimulated with α- or κ-casein. After 120 h of culture, the proliferation index (PI), determined by 3-(4,5 dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and carboxyfluorescein diacetate N-succinimidyl ester (CFSE) staining, did not vary for either antigen, suggesting similar ex vivo immunogenic potential of both casein fractions. In vivo, BALB/ccmdb mice were sensitized with α- or κ-casein and then gavaged with primary antigen. Mice immunized with α-casein had higher levels of IgG (2^16.33) and IgA (2^10.22) in serum at the end of the experiment compared with mice immunized with κ-casein (2¹⁵ and 2^9.3 for IgG and IgA, respectively). The use of α-casein for mouse immunization and ex vivo lymphocyte stimulation resulted in higher concentrations of secreted cytokines (IL-4, IL-10) compared with κ-casein stimulation. This is consistent with increasing regulatory T cell (Treg) lymphocyte populations, independent of the antigen used for stimulation. In summary, the immunogenic potential of α- and κ-casein was similar. Humoral and cellular immune responses confirmed their strong, independent potential to induce B and T cells. We propose that the lymphocyte proliferation index be used as an initial screening for protein immunogenicity.     

不同剂量牛初乳粉对小鼠免疫调节作用的研究

本研究用牛初乳粉对小鼠进行喂养实验,20d后测定NK细胞杀伤活性、T淋巴细胞增殖活性、巨噬细胞的吞噬功能,研究了不同剂量的牛初乳粉对小鼠免疫调节作用效果。结果表明:剂量在36.46～1250mg/kg·d之间的牛初乳粉都能极显著地增强巨噬细胞的吞噬功能,NK细胞的杀伤活性和T淋巴细胞的增殖活性,其中78.13～156.25mg/kg·d之间对小鼠细胞免疫调节作用和巨噬细胞吞噬功能增强作用效果优于其他剂量。     

瑞士乳杆菌发酵乳中多肽的分离对小鼠体外淋巴细胞增殖的影响

目的:分离瑞士乳杆菌发酵乳中多肽片段,并对分离片段的的免疫调节作用进行研究。方法:利用瑞士乳杆菌LH0906发酵牛乳获得多肽样品,采用离心、超滤的方法对所得多肽进行粗提,利用凝胶层析色谱和反相层析色谱进一步分离发酵乳中的多肽,使用Folin-Lowry法测定发酵后样品中多肽含量,采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)比色法检测发酵乳中的多肽对小鼠脾脏淋巴细胞体外增殖的影响。结果:牛乳发酵液其对小鼠脾脏淋巴细胞增殖效果显著高于未发酵牛乳,并经过凝胶层析色谱和反相层析色谱分离发酵乳得到的峰S12-A对小鼠脾脏淋巴细胞体外增殖效果十分显著(P<0.01),峰S12-C对小鼠脾脏淋巴细胞体外增殖效果显著(P<0.05)。结论:牛乳经瑞士乳杆菌LH0906发酵后,分离得到的多肽片段对小鼠脾脏淋巴细胞体外增殖具有显著的增强作用。     

In vivo modulatory effects of chitooligosaccharide nanoparticles on mouse serum cytokines and splenocytes

Water-soluble chitosan derivative-based nanoparticles (carboxymethyl chitosan-chitooligosaccharide nanoparticles (CMC-COS NP) and sulphated chitosan-chitooligosaccharide nanoparticles (SC-COS NP)) were prepared by the formation of polyelectrolyte complexes. SC-COS NP and CMC-COS NP both induced the proliferation of mouse fibroblasts, whereas they elicited dose-dependent inhibitory effects on the proliferation of both HeLa and mouse B16 melanoma cells. Injection of SC-COS NP and CMC-COS NP modulated serum Th cytokines and stimulated the proliferation of splenic lymphocytes (CD4⁺ and CD8⁺ T cells, CD19⁺ B cells and NK cells) in mice, indicating the ability of these particles to regulate both humoral and cell-mediated immune responses. These properties demonstrated their promising potential for application as biomedical materials.     

Immunophenotyping of lymphocyte subsets in the third eyelid tissue in dogs (Canis familiaris): morphological, microvascular, and secretory aspects of this ocular adnexa

The third eyelid is an important adnexa of the eye. The objective of this study was to evaluate (i) morphological aspects (ii) vascularization, and (iii) the immunophenotype of lymphocyte subsets in the third eyelid of dogs. Flow cytometric analysis revealed the presence of three patterns concerning the immunophenotype of the third eyelid tissue. Dogs without ocular insult or deficient tear production might belong to one of the following immunophenotype patterns: I--the number of T-cells that expressed CD3(+) CD8(+) was higher than the number of cells that expressed CD3(+)CD4(+). II--the number of cells CD3(+)C4(+) was higher than the number of cells CD3(+)CD8(+) and in this case a higher number of cells that expressed CD19 were identified. III--Proximity of values of the cells that expressed CD3(+)CD4(+) and CD3(+)CD8(+). These data might suggest that the number of lymphocyte T cells alone should not be considered a direct indicator of the presence of an immune-based inflammation. Besides, a particular population of T-cells does not indicate a particular inflammatory state. The morphological study of the third eyelid revealed a rather uncommon angioarchitecture. The artery that irrigates the eyelid crosses almost the entire length of this structure to achieve its free border, and only then, ramificates deeply towards an inner segmental level. This spatial microvascular arrangement probably results from an adaptation to the fact that the third eyelid, in the medial cantus of the eye, is inwardly compressed into a tiny space. Efficient vascularization is achieved by allowing the first ramifications of the third eyelid artery to run straight to the top. Accini secretor cells of the third eyelid show a mucin content while tubuloacinar cells are mainly serous.