在人扁桃体细胞中诱生γ干扰素

本文初步研究了用人扁桃体细胞诱生γ干扰素及其影响因素。试验结果显示:细胞浓度在5×10~7/ml时产生的γ干扰素效价最高;PHA、PWM、ConA等T细胞促分裂剂诱生γ干扰素的适宜浓度范围分别为50～400μg/ml,2.5～40μg/ml,5～80μg/ml;诱生时间均以72小时为宜;2-ME对人γ干扰素的产生没有影响。本文还将人扁桃体细胞和外周血白细胞对4种γ干扰素诱生利(PHA、PWM、ConA、SEA)和3种α干扰素诱生剂(NDV、SPA菌体、CP)的反应进行了比较。     

牛磺酸对新生儿单个核细胞免疫功能的影响

本文以胎盘剥离前的脐血单个核细胞做体外试验,观察了牛磺酸对新生儿淋巴细胞增殖、γ-干扰素和抗体产生水平的影响,并与成人作对照。结果表明①牛磺酸可以促进淋巴细胞增殖,上升幅度以低出生体重儿最为显著。②牛磺酸可以提高ConA诱生干扰素和PWM刺激抗体生成的水平。③牛磺酸本身没有丝裂原和干扰素诱生剂的作用。提示牛磺酸有免疫促进作用。     

CMP促诱生IL—2,TNF—α,IL—6,IFNγ／α效应和产品中试

用羧甲基茯苓多糖（ＣＭＰ）预处理人外周血淋巴细胞（ＨＰＢＬ），在以ＰＨＡ和ＣｏｎＡ为主的复合诱生剂诱导培养２４、３６、４８、７２ｈ采样检测的促诱生组ＴＮＦ－α、ＩＬ－２、ＩＬ－６、ＩＦＮγ效价（ｘ±ｓ）分别为４５６．１±２４．２，４７５．６±１８５．７，７４３３±１１８９．９，５５８３．０±５３５．５ＩＵ／ｍｌ，分别比无ＣＭＰ的常规诱生组效价高１．５，３．８，１．８，１．４倍；在以ＮＤＶ－Ｆ病毒为诱生剂诱导培养２２ｈ采样，并经酸化和中性化后检测的ＩＦＮ－α效价可达２１２７４．２±５５２３．０ＩＵ／ｍｌ，比无ＣＭＰ的常规诱生组高１．２倍，均有显著差异（Ｐ＜０．０１），说明ＣＭＰ对上述五种细胞因子均有明显的促诱生效应。此外，还采用ＣＭＰ促诱生技术，对ＴＮＦ－α、ＩＬ－２、ＩＬ－６、ＩＦＮγ和ＩＦＮ－α细胞因子进行了中试，提出了低成本、高效益的中试工艺路线，有待于在制备天然细胞因子的细胞工程中推广应用     

S-O_2-1菌苗诱生干扰素的研究 Ⅰ小鼠体内诱生

本文研究了免疫增强剂——S-O_2-1菌苗在小鼠体内诱生干扰素的能力。结果表明,给(57BL/6小鼠注射菌苗1.5×1O~9菌/只后,血清中干扰素活性明显升高,于尾静脉注射后2小时,干扰素滴度达高峰(9.06±0.34log_2U/ml)。该干扰素具有小鼠I-型干扰素相同的理化特性。实验还表明,由S-O_2-1菌体提取的核糖体和脂多糖(LPS)亦均具有诱生干扰素的活性。因此我们推测,s-O_2-1菌苗诱生干扰素的能力可能对其抗肿瘤活性的发挥起一定的怍用。     

5种中药有效成分的干扰素促诱生效应

用100μg/ml的人参皂甙、云芝糖肽、人参多糖、羧甲基茯苓多糖及10μg/ml的刺五加多糖对人脐血白细胞进行了IFN-α和IFN-γ的促诱生试验,其促诱生组的IFN效价比常规诱生组(IFN     

慢性乙型肝炎病人单个核细胞γ干扰素的诱生和T淋巴细胞亚群的研究

本文对28例慢乙肝病人的外周血单个核细胞进行ConA诱生γ干扰素的测定,同时用T细胞单克隆抗体系统检测其T细胞亚群。结果发现,慢乙肝病人ConA诱生的γ干扰素效价为7.858±1.483log2U/ml,而正常对照组为7.839±0.621log2U/ml,两组间无显著性差异,但在慢乙肝病人组内个体间IFN-γ的效价差别显著大于对照组(P<0.05).慢乙肝病人组T细胞亚群的T_4/T_3比例与对照组无显著差异。但IFN-γ反应低下组的5人均有T_4/T_3比例异常。在28例病人中有14例T_4/T_3比例异常(高于或低于正常),且20例HBeAg阳性病人中有12例T_4/T_3比例异常;8例HBeAg阴性病人中仅2例T_4/T_3比例异常。     

S-O_2-1菌苗诱生干扰素的研究 Ⅱ小鼠脾脏细胞体外诱生

本文表明S-O_2-1菌苗具有较强的诱导小鼠脾细胞产生干扰素的能力。将事先用菌苗免疫的C57BL/6小鼠脾细胞体外培养时,即使不加菌苗也能产生较高滴度的干扰素活性,该活性在免疫后4天制备的脾细胞培养中开始出现,免疫后7天则达高峰(7.47±0.31 log_2U/ml),以后逐渐下降。若培养时添加菌苗,干扰素高峰出现稍晚些,于免疫后10天达高峰,但滴度明显上升(9.10±0.26 log_2U/ml)。另外,C57BL/6正常小鼠的脾细胞(5×10~6细胞/ml)添加菌苗(0.1亿菌/ml)体外培养12小时后可开始测出干扰活性,培养48小时后活性达高峰(8.23±0.30log_2U/ml)。该干扰素具有小鼠Ⅱ-型干扰素相同的特性。我们在实验中还发现,将S-O_2-1菌苗与刀豆素A(Con A)合用诱生的干扰素滴度明显高于两者单独使用时诱生滴度的累加值。     

干扰素的免疫调节作用

干扰素是由有关生物细胞在干扰素诱生剂作用下产生的一类诱生蛋白。国际干扰素命名委员会将其分为三种:α干扰素(IFN-α)、β干扰素(1FN-β)和γ-干扰素(1FN-γ)。根据氨基酸顺序的不同,又分为若干亚型:人α-干扰素至少有20个亚型,β干扰素有4个亚型,γ-干扰素可能也有4个亚型。     

P物质与生长抑素对美洲商陆诱导小鼠脾细胞产生…

本实验研究了Ｐ物质和生长抑素在体外对美洲商陆诱导小鼠脾细胞产生γ干扰素的调节作用。结果显示：０．１～１０μｍｏｌ／Ｌ ＳＰ对５，２．５及１．２５μｇ／ｍｌＰＷＭ诱生的ＩＦＮ－γ效价有明显增高作用，但０．００１～１μｍｏｌ／Ｌ ＳＰ对１０μｇ／ｍｌＰＷＭ诱生ＩＦＮ－γ没有影响。与ＳＰ相反，０．１～１０μｍｏｌ／ＬＳＯＭ可显著降低１，０．５及０．２５μｇ／ｍｌ ＰＷＭ诱生的ＩＦＮ－γ效价。用５、２．５     

正常人外伤脾多向混合淋巴细胞培养产生较高效价γ干扰素

三例正常人外伤猝死者,取其全脾细胞加无血清培养基作多向混合淋巴细胞培养,不加任何干扰素诱生剂,37℃5％CO_2中培育72h后,产生出γ干扰素(1FN-γ)的效价达10240IU/ml,比用一般诱生剂的产量高。这种多向混合淋巴细胞反应的转化率>48％。文中讨论了MLC产生IFN-Y的机理,多向MLC产生较高效价IFN-γ的原因以及与所谓混合淋巴细胸肿瘤细胞培养(MLTC)产生IFN的类型和过程的不同特点。     

尘螨诱导新生儿脐血单个核细胞ICOS与转录因子表达的研究

为了解尘螨(HDM)抗原对新生儿脐血单个核细胞(CBMC)及成人外周血单个核细胞(PBMC)CD3+ICOS+细胞阳性率、转录因子T-bet、GATA-3、Foxp3 mRNA表达水平以及培养上清中IL-4、IL-10、IFN-γ表达水平的影响。用流式细胞术,检测新生儿CBMC、成人PBMC体外经PHA和/或HDM抗原刺激前、后CD3+ICOS+细胞阳性率;用RT-PCR法检测细胞体外经PHA和/或HDM抗原刺激前、后T-bet、GATA-3以及Foxp3 mRNA表达水平;用ELISA法,检测细胞在体外经PHA和/或HDM刺激前、后培养上清液中IL-4、IL-10、IFN-γ表达水平。结果表明,高剂量HDM抗原显著下调CBMC经PHA刺激后的CD3+ICOS+阳性率(P<0.05),同时也显著上调PHA刺激前其T-bet mRNA表达(P<0.05),而显著下调该类细胞经PHA刺激后的Foxp3 mRNA的表达(P<0.05)。也显著增加其PHA刺激前的IFN-γ分泌(P<0.01),减少其PHA刺激后IL-10分泌(P<0.05)。高剂量HDM抗原对CBMC的作用强于PBMC,可下调CD3+ICOS+阳性率,显著上调PHA刺激前CBMC T-bet mRNA表达,增加PHA刺激前IFN-γ的分泌,减少PHA刺激后IL-10分泌,提示早期接触高剂量HDM抗原对机体免疫功能的影响较强,可促进新生儿Th1样反应,高剂量HDM抗原可能通过作用于下调Foxp3 mRNA的表达而下调CD4+CD25+调节性T细胞的功能。     

Macrophages from Nonobese Diabetic Mouse Have a Selective Defect in IFN-γ but Not IFN-α/β Receptor Pathway

Purpose

Aberrant regulation of innate immune cells such as macrophages has been implicated in the onset and progression of type 1 diabetes (T1D). Macrophages from nonobese diabetic (NOD) mouse, an animal model of T1D, entail developmental and functional defects that are often associated with hypo-responsiveness to interferon (IFN)-γ. We aimed to uncover a mechanism underlying this phenomenon.

Methods

We analyzed the receptor pathway along with the response of macrophages exposed to IFN-γ and the related IFNs such as IFN-α/β.

Results

We found that NOD macrophages failed to fully respond to IFN-γ but not to IFN-α for the production of inflammatory cytokines (e.g. TNF-α and IL-12). NOD macrophages were also resistant to apoptotic pathway induced by IFN-γ and LPS. Analyses of receptor pathway revealed that STAT1 pathway of intracellular signaling was selectively impaired in NOD macrophages exposed to IFN-γ but not to IFN-α/β. Further, these defects correlated with a low phosphorylation level of JAK2, and were related to impaired up-regulation of surface IFN-γ receptor 2 (IFN-γR2) by IFN-γ.

Conclusion

Taken together, our results suggest that NOD macrophages have a selective defect in IFN-γ but not IFN-α/β receptor pathway. As IFN-γ and IFN-α have been implicated in the development of autoimmunity towards β-cells, such an unanticipated selectivity in IFN responsiveness may provide a new insight into the pathogenesis of T1D.     

The anti-TNF-α antibody infliximab indirectly regulates PECAM-1 gene expression in two models of in vitro blood cell activation

Chronic inflammatory bowel diseases can be successfully treated with antibodies against the acute phase mediator TNF-α. The process of activation and of extravasation of inflammatory cells from the blood into the 'stressed' tissue site is controlled by cytokines and chemokines, which attract leukocytes and by adhesion molecules, which mediate their attachment and transmigration toward the affected cell(s). The changes in the gene expression of adhesion molecules taking place in those cells before attachment have been less investigated. Changes of PECAM-1, ICAM-1 and vascular cell adhesion molecule-1 (VCAM-1) gene expression were studied in phytohaemagglutinin (PHA)- and lipolysaccharide (LPS)-treated human peripheral blood leukocytes (PBLs), granulocytes and the human monocyte cell line U-937. Cells were treated either with PHA or with LPS in the presence or absence of infliximab and incubated with TNF-α, IFN-γ and/or transforming growth factor beta (TGF-β) and treated as above. Activation of PBLs by PHA or LPS treatment triggered a sharp upregulation of ICAM-1, VCAM-1 gene expression and a time-dependent downregulation of PECAM-1 gene expression reaching a minimum 4?h from start of the experiment. The anti-TNF-α antibody infliximab, by neutralizing TNF-α and IFN-γ production, completely reversed PECAM-1 mRNA downregulation and ICAM-1 and VCAM-1 upregulation. Immunostaining of PBLs cytospins with antibodies against PECAM-1 and ICAM-1 confirmed RT-PCR and western blot results. PBLs IFN-γ or TNF-α treatment downregulated PECAM-1 in parallel with the upregulation of ICAM-1 and VCAM-1 gene expression, whereas TGF-β upregulated PECAM-1- and downregulated ICAM-1 and VCAM-1 gene expression counteracting the effect of TNF-α or IFN-γ. Similar results were obtained in human U937 cells and in granulocyte cultures by TNF-α or IFN-γ treatment. Taken together, these results suggest that infliximab, blocking TNF-α and IFN-γ production, exerts its anti-inflammatory effect through inhibiting downregulation of PECAM-1 gene expression and upregulation of ICAM-1 and VCAM-1 expression in leukocytes of the peripheral blood. These results also suggest that TGF-β may thus be of therapeutic importance as an anti-inflammatory agent.     

Interferon Alpha Treatment of Patients with Impaired Interferon Gamma Signaling

Patients with deficiency in the interferon gamma receptor (IFN-γR) are unable to respond properly to IFN-γ and develop severe infections with nontuberculous mycobacteria (NTM). IFN-γ and IFN-α are known to signal through STAT1 and activate many downstream effector genes in common. Therefore, we added IFN-α for treatment of patients with disseminated mycobacterial disease in an effort to complement their IFN-γ signaling defect. We treated four patients with IFN-γR deficiency with adjunctive IFN-α therapy in addition to best available antimicrobial therapy, with or without IFN-γ, depending on the defect. During IFN-α treatment, ex vivo induction of IFN target genes was detected. In addition, IFN-α driven gene expression in patients’ cells and mycobacteria induced cytokine response were observed in vitro. Clinical responses varied in these patients. IFN-α therapy was associated with either improvement or stabilization of disease. In no case was disease exacerbated. In patients with profoundly impaired IFN-γ signaling who have refractory infections, IFN-α may have adjunctive anti-mycobacterial effects.     

IFN-γ-mediated suppression of coronavirus replication in glial-committed progenitor cells

The neurotropic JHM strain of mouse hepatitis virus (JHMV) replicates primarily within glial cells following intracranial inoculation of susceptible mice, with relative sparing of neurons. This study demonstrates that glial cells derived from neural progenitor cells are susceptible to JHMV infection and that treatment of infected cells with IFN-γ inhibits viral replication in a dose-dependent manner. Although type I IFN production is muted in JHMV-infected glial cultures, IFN-β is produced following IFN-γ-treatment of JHMV-infected cells. Also, direct treatment of infected glial cultures with recombinant mouse IFN-α or IFN-β inhibits viral replication. IFN-γ-mediated control of JHMV replication is dampened in glial cultures derived from the neural progenitor cells of type I receptor knock-out mice. These data indicate that JHMV is capable of infecting glial cells generated from neural progenitor cells and that IFN-γ-mediated control of viral replication is dependent, in part, on type I IFN secretion.     

Comparison of the effects of interleukin-1α, interleukin-lβ and interferon-γ-inducing factor on the production of interferon-γ by natural killer

Interferon-γ inducing factor (IGIF) is a recently identified cytokine which stimulates the production of interferon-γ (IFN-γ) by T cells and enhances natural killer (NK) cell cytolytic activity. Protein fold recognition, structure prediction and comparative modeling have revealed that IGIF is a member of the interleukin (IL)-1 cytokine family and has prompted the designation IL-1γ. Here we report functional similarities between members of the IL-1 family by comparing the effects of IL-1α, IL-1β and IGIF on NK cell production of IFN-γ. All three IL-1 types enhanced NK cell production of IFN-γ when induced by IL-2 or IL-12, although at high concentrations (>10 ng/ml), IGIF was five- to tenfold more potent than IL-1α or IL-1β. This effect correlated with enhanced levels of mRNA for IFN-γ when NK cells were stimulated with IGIF plus IL-12. In contrast to IL-12 and IL-2, the ability of IGIF to stimulate NK cell production of IFN-γ was not increased by IL-1α or IL-1β. The ability of IGIF to enhance IFN-γ production was independent of the type I and type II IL-1 receptors or the IL-1R accessory protein. Together, these results identify IGIF as a potent stimulator of NK cell production of IFN-γ and demonstrate that the effect of IGIF on NK cell production of IFN-γ is similar to that of IL-1α and IL-1β but distinct from that of IL-12.     

Increasing proliferation of murine adipose tissue-derived mesenchymal stem cells by TNF-α plus IFN-γ

The objective of the current study was to assess the effects of tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) along with their simultaneous application on proliferation and pluripotency genes of murine adipose tissue-derived mesenchymal stem cells (AT-MSCs). The proliferation, doubling time (DT), colony-forming unit–fibroblast (CFU-F), pluripotency genes expression, and proliferation-related immunomodulatory markers of MSCs were analyzed upon activation with TNF-α (10?ng/ml), IFN-γ (10?ng/ml) and both TNF-α and IFN-γ (5?ng/ml?+?5?ng/ml). Pluripotency genes including Oct-4, Sox-2, and Nanog as well as proliferation-associated immunomodulatory cytokines such as insulin-like growth factor 1 (IGF-1) and transforming growth factor-β (TGF-β) expression were evaluated using real-time PCR. Surface expression of Qa2 (HLA-G) was analyzed by flow cytometry. Pretreatment of MSCs with TNF-α plus IFN-γ led to significantly increased proliferation, DT and CFU-F as well as expression of pluripotency genes in AT-MSCs (p < 0.01). MSCs expressed more IGF-1, TGF-β, and Qa2 upon activation with TNF-α plus IFN-γ and IFN-γ. MSCs expressed significantly decreased amounts of TGF-β and Qa2 in presence of TNF-α. TNF-α combined with IFN-γ may be improved the proliferation of AT-MSCs. Conversely, expanded MSCs pointed out low levels of the immunomodulatory marker, s especially Qa2 in the presence of TNF-α. In conclusion, we showed that TNF-α together with IFN-γ increased the proliferation of MSCs and slightly enhanced the expression of pluripotency genes.