Sequential delivery of immunomodulatory cytokines to facilitate the M1-to-M2 transition of macrophages and enhance vascularization of bone scaffolds

In normal tissue repair, macrophages exhibit a pro-inflammatory phenotype (M1) at early stages and a pro-healing phenotype (M2) at later stages. We have previously shown that M1 macrophages initiate angiogenesis while M2 macrophages promote vessel maturation. Therefore, we reasoned that scaffolds that promote sequential M1 and M2 polarization of infiltrating macrophages should result in enhanced angiogenesis and healing. To this end, we first analyzed the in vitro kinetics of macrophage phenotype switch using flow cytometry, gene expression, and cytokine secretion analysis. Then, we designed scaffolds for bone regeneration based on modifications of decellularized bone for a short release of interferon-gamma (IFNg) to promote the M1 phenotype, followed by a more sustained release of interleukin-4 (IL4) to promote the M2 phenotype. To achieve this sequential release profile, IFNg was physically adsorbed onto the scaffolds, while IL4 was attached via biotin-streptavidin binding. Interestingly, despite the strong interactions between biotin and streptavidin, release studies showed that biotinylated IL4 was released over 6 days. These scaffolds promoted sequential M1 and M2 polarization of primary human macrophages as measured by gene expression of ten M1 and M2 markers and secretion of four cytokines, although the overlapping phases of IFNg and IL4 release tempered polarization to some extent. Murine subcutaneous implantation model showed increased vascularization in scaffolds releasing IFNg compared to controls. This study demonstrates that scaffolds for tissue engineering can be designed to harness the angiogenic behavior of host macrophages towards scaffold vascularization.     

中药“425”对血吸虫卵肉芽肿病变免疫调节作用的机理研究

本文试验结果发现，应用“４２５”制剂可明显增强宿主脾细胞在体外自发性的及在ＬＰＳ刺激下的增生反应，明显抑制其在ＰＨＡ刺激下的增生反应，而用药后宿主脾细胞在特异性抗原刺激下的增生反应未见明显改变；虽然用药组感染鼠的脾脏明显小于对照组，但血清及虫卵肉芽肿培养上清中特异性抗体ＩｇＧ水平未见明显改变。研究结果表明，“４２５”制剂具有双重非特异性免疫调节作用。该制剂抑制宿主肝组织内虫卵肉芽肿病变的机理除了在于其能非特异地增强宿主Ｂ细胞介导的体液免疫应答外，该制剂对宿主体内Ｔ细胞介导的细胞免疫应答的非特异性抑制作用可能起了更重要的作用。     

调节性B淋巴细胞在视神经脊髓炎谱系疾病患者外周血中的表达和意义

背景 调节性B淋巴细胞(Breg)是一类具有负向免疫调节作用的细胞,在多种自身免疫性疾病中表达异常,视神经脊髓炎谱系疾病(NMOSD)系自身免疫性疾病,目前关于Breg在NMOSD患者外周血中的表达和意义的研究较少。目的 研究Breg在NMOSD患者外周血中的表达和意义。方法 选取2019年6月—2021年12月新疆医科大学第一附属医院神经内科收治的NMOSD患者55例作为NMOSD组,选取同期本院体检正常的健康志愿者50例作为对照组。运用流式细胞仪检测两组研究对象外周血中CD19⁺CD24^hiCD38^hi Breg、CD19⁺CD24^hiCD27⁺Breg的表达。采用酶联免疫吸附测定(ELISA)法检测和比较两组研究对象血清中白介素10(IL-10)、白介素35(IL-35)和转化生长因子β1(TGF-β1)的水平。结果 NMOSD组患者CD19⁺CD24^hiCD38^hi Breg、CD1...     

Clearance of Theiler's virus infection depends on the ability to generate a CD8+ T cell response against a single immunodominant viral peptide

Theiler's murine encephalomyelitis virus (TMEV) induces a chronic demyelinating disease in the central nervous system of susceptible mice. Resistance to persistent TMEV infection maps to he D locus of the major histocompatibility complex suggesting a prominent role of antiviral CTL in the protective immune response. Introduction of the D(b) gene into the FVB strain confers resistance to this otherwise susceptible mouse line. Infection of the FVB/D(b) mouse with TMEV provides a model where antiviral resistance is determined by a response elicited by a single class I molecule. Resistant mice of the H-2(b) haplotype mount a vigorous H-2D(b)-restricted immunodominant response to the VP2 capsid protein. To investigate the extent of the contribution of the immunodominant T cell population in resistance to TMEV, FVB/D(b) mice were depleted of VP2-specific CD8(+) T cells by peptide treatment prior to virus infection. Peptide-treated mice were not able to clear the virus and developed extensive demyelination. These findings demonstrate that the D(b)-restricted CD8(+) T cells specific for a single viral peptide can confer resistance to TMEV infection. Our ability to manipulate this cellular response provides a model for investigating the mechanisms mediating protection against virus infection by CD8(+) T cells.     

骨髓间充质干细胞异体移植免疫反应的研究进展

骨髓间充质干细胞（BMSC）具有独特的生物学特性，其免疫抑制作用引起了人们广泛的兴趣。大量资料表明，MSC能在同种异基因，甚至异种基因的环境中长期存活，并且保持它的多向分化潜能。这一特性使MSC异体移植预防和治疗免疫性疾病成为可能。因此有必要从问充质干细胞的低免疫原性、灯免疫细胞及补体系统的调节和诱导抑制性免疫微环境等方面探讨BMSC避免同种异体移植排斥反应的可能机制。     

Inhibition of the development of immediate hypersensitivity by staphylococcal enterotoxin B

We investigated the ability of staphylococcal enterotoxin B (SEB) to modify the immediate hypersensitivity response induced in BALB/c mice following sensitization to ovalbumin (OVA), a response mediated by OVA-reactive Vβ8 T cells. Mice were sensitized by skin painting with OVA every second day over a period of 2 weeks. SEB, a potent activator of Vβ8⁺ T cells, was administered at the same site where OVA was applied (skin of the lower abdomen) following two different protocols. In protocol (A) SEB was injected intradermally 1 day before painting with OVA and on day 7; in protocol B, SEB was injected each time OVA was applied to the skin (eight times). SEB (but not SEA) altered the development of immediate hypersensitivity to OVA, as demonstrated by the reduction in allergen-specific IgE, decreased OVA-specific immediate skin test responsiveness, and prevented the development of increased airways responsiveness after bronchial challenge with OVA. Injections of SEB did not alter the proliferative responses of local draining lymph node cells or spleen mononuclear cells to OVA, indicating that administration of SEB did not inhibit the sensitization to OVA, but shifted the immune response away from an immediate type response (IgE/IgG₁) to IgG_2a, IgG_2b and IgG₃. Although both protocols of SEB treatment did not lead to a major deletion of the Vβ8 T cell population, they did reduce the proliferative response of Vβ8⁺ T cells to OVA. These data indicate that the bacterial toxin SEB is capable of modifying the immediate hypersensitivity response induced by OVA by altering the functional capacity of antigen-reactive Vβ8 T cells.     

Regulation of immune responses by CD1d-restricted natural killer T cells

Natural killer T (NKT) cells are a unique subset of T lymphocytes that share receptor structures and properties with conventional T lymphocytes and natural killer (NK) cells. NKT cells are specific for glycolipid antigens such as the marine sponge-derived agent α-galactosylceramide (α-GalCer) presented by the major histocompatibility complex (MHC) class I-like molcule CD1d. My laboratory has evaluated the function of NKT cells by generating and analyzing CD1d-deficient mice. These studies showed that CD1d expression is required for NKT cell development, but not absolutely necessary for the generation of polarized T helper (Th) cell responses. Further, we have studied the in vivo response of NKT cells toα-GalCer stimulation and the capacity of α-GalCer to modulate innate and adaptive immune responses. Our results revealed that, quickly following administration of α-GalCer, NKT cells expand and produce cytokines, trans-activate a variety of innate and adaptive immune cells, and promote Th2 responses that are capable of suppressing Th1-dominant autoimmunity. Our findings indicate that NKT cells play a regulatory role in the immune response and that specific activation of these cells may be exploited for therapeutic purposes.     

古草生机汤对H22荷瘤小鼠体内抑瘤及免疫调节作用的初步研究

目的：探究古草生机汤对H22荷瘤小鼠体内实体瘤的抑瘤功效和对机体的免疫调节作用机制。方法：本研究采用H22实体瘤动物模型,随机分为空白组、模型组、阳性药组、古草生机汤低、高剂量组,灌胃给药7 d,计算各组小鼠的体质量增长率、肿瘤抑制率、胸腺及脾指数,小鼠实体瘤病理切片苏木精-伊红（HE）染色观察瘤体组织和细胞形态,酶联免疫吸附试验法检测小鼠血清中γ干扰素（IFN-γ）、肿瘤坏死因子-α(TNF-α)、白细胞介素-2（IL-2）、Fas、Fas配体、天门冬氨酸氨基转移酶（AST）、丙氨酸氨基转移酶（ALT）的含量,实时荧光定量PCR测量小鼠瘤组织中Bax和Bcl-2mRNA的相对表达量。结果：古草生机汤有促进H22荷瘤小鼠体质量增长、抑制体内实体瘤增长和促进肿瘤凋亡坏死的作用,可提高小鼠的胸腺指数同时降低脾脏指数,可显著提高小鼠血清中IFN-γ、TNF-α、IL-2的含量并降低Fas、Fas配体、AST、ALT的含量（均P<0.01）,能够上调小鼠瘤组织中Bax mRNA的表达并下调Bcl-2mRNA的表达。结论：古草生机汤具有明显的体内抗肝肿瘤药效,在抑制肿瘤生长、促进肿瘤细胞凋亡、调节机体免疫方面有一定作用。     

胃癌免疫细胞浸润相关预后基因的共表达网络分析鉴定

背景与目的：肿瘤免疫细胞浸润在胃癌（GC）的进展和预后中起到关键作用,然而,目前影响GC预后的免疫细胞浸润调控基因还不清楚。本研究联合共表达网络分析研究和反卷积分法鉴定了GC免疫相关预后基因。  方法：从TCGA数据库中下载GC的mRNA表达数据并使用CIBERSORT反卷积算法来确定每个样品中免疫细胞的比例,并构建共表达网络,结合Kaplan-Meier法确定免疫相关预后基因。 结果：在纳入分析的22种免疫细胞中,有11种免疫细胞在肿瘤组织中显著高于正常组织（均P<0.05）。对上述11种免疫细胞进行生存分析,发现静止记忆性CD4 T细胞和调节型T细胞的浸润情况与GC患者生存率呈明显相关（均P<0.05）。基于上结果论进行共表达网络分析,得绿松石模块中的基因与上述两种类型的细胞相关性最显著,并鉴定得到了3个与记忆性CD4 T细胞相关（CGB5、LINC00106、LINC00392）和1个与调节型T细胞相关（UPK1B）的预后基因（均P<0.05)。 结论：本研究鉴定的4个胃癌免疫细胞浸润相关预后基因可能在GC的进展和预后中起到重要作用,这些基因可能会通过影响肿瘤免疫过程而作为潜在的胃癌免疫治疗靶点。      

Immunomodulating Effects of CKBM on the Cytokine Production in Peripheral Blood Mononuclear Cells (PBMCs) from Healthy Volunteers

The current study investigated the immunomodulating effect of CKBM on cytokine induction in peripheral blood mononuclear cells (PBMCs) isolated from 20 healthy volunteers. Cytometric Bead Analysis (CBA) was used to study IL‐2, IL‐4, IL‐6, IL‐10, TNF‐alpha and IFN‐gamma. TNF‐alpha and IL‐6 were significantly increased in a CKBM dose‐ and time‐dependent manner. Flow cytometry analysis showed an increased intracellular staining of IL‐6 but not of TNF‐alpha in CKBM treated PBMCs. In addition, MTT cell cytotoxicity assay showed that CKBM concentrations below 5% did not significantly affect the metabolic activities of PBMCs. The current study indicated that CKBM may modulate the immune response by inducing the secretions of TNF‐alpha and IL‐6, which are cytokine mediators of innate immunity and inflammation preparing or “priming” the body to combat diseases.