人疱疹病毒6型诱导单核细胞产生IL-10

目的探讨人疱疹病毒６型（ＨＨＶ－６）感染的免疫发病机理。方法用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）和双抗体夹心酶联免疫吸附试验（ＥＬＩＳＡ）检测白细胞介素－１０（ＩＬ－１０）等细胞因子的产生。结果ＨＨＶ－６诱导单核细胞表达和产生ＩＬ－１０，细胞因子ｍＲＮＡ动力学研究发现ＴＮＦ－α（肿瘤坏死因子）、ＩＬ－１β及ＩＬ－６随ＩＬ－１０ｍＲＮＡ积累而减少，用抗人ＩＬ－１０单克隆抗体阻断ＨＨＶ－６诱导的内源性ＩＬ－１０，ＴＮＦ－α、ＩＬ－１β和ＩＬ－６ｍＲＮＡ的表达和因子产生明显增加，表明ＨＨＶ－６诱导的内源性ＩＬ－１０在转录水平能抑制单核细胞因子产生。结论ＩＬ－１０具有抑制Ｉ类辅助性Ｔ细胞（Ｔｈ１）反应、下调单核巨噬细胞功能等多种生物作用，推测ＨＨＶ－６诱导产生的内源性ＩＬ－１０与该病毒长期潜伏感染及其参予的免疫功能紊乱有关。     

IL—6／IL—6R系统与重组IL—6—PE40融合蛋白

ＩＬ－６／ＩＬ－６Ｒ系统与重组ＩＬ－６－ＰＥ４０融合蛋白军事医学科学院附属医院免疫学研究室（北京１０００３９）奚永志ＩＬ－６／ＩＬ－６－ｒｅｃｅｐｔｏｒｓｙｓｔｅｍａｎｄｒｅｃｏｍｂｉｎａｎＩＬ６－ＰＥ４０ｅｘｏｔｏｘｉｎｆｕｓｉｏｎｐｒｏｔｅｉｎ...     

IFN—γ调节HHV—6诱导单核细胞产生IL—10和IL—12

为探讨人疱疹病毒６型（ＨＨＶ－６）感染对单核／巨噬细胞功能的影响，用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）和双抗体夹心ＥＬＩＳＡ法检测ＩＬ－１０及ＩＬ－１２的产生。ＨＨＶ－６诱导单核细胞、ＴＨＰ－１细胞表达及产生ＩＬ－１０和ＩＬ－１２。细胞因子ｍＲＮＡ动力学观察显示ＩＬ－１２随ＩＬ－１０ｍＲＮＡ积累而减少，用抗人ＩＬ－１０的单克隆抗体阻断ＨＨＶ－６诱导的内源性ＩＬ－１０，ＩＬ－１２ｍＲＮＡ表达和因子产生明显增加，提示内源性ＩＬ－１０抑制ＩＬ－１２产生。在ＨＨＶ－６感染培养中加入ＩＦＮ－γ，ＩＬ－１２基因表达随ＩＦＮ－γ浓度而增加，ＩＬ－１０ｍＲＮＡ测减少，ＩＬ－１２和ＩＬ－１０因子测定结果亦明显增加或降低，表明ＩＦＮ－γ在转录水平调节ＨＨＶ－６诱导的ＩＬ－１２和ＩＬ－１０产生，其机理可能主要与ＩＦＮ－γ抑制内源性ＩＬ－１０有关。     

中国人疱疹病毒6型（HHV—6）的分离与鉴定

４例婴儿玫瑰疹病人外周血淋巴细胞与脐血淋巴细胞共培养，３例病人标本见有细胞病，其中１株病毒经多次传代和冻存后，仍能产生ＣＰＥ。该病毒株感染细胞的超薄切片在电镜下可见疱疹病毒性颗粒，感染细胞涂片与抗ＨＨＶ－６单抗产生明显荧光，用该病毒株制备的单抗与ＨＨＶ－６ Ｚ２９株荧光试验呈阳性，表明分离的病毒为ＨＨＶ－６。     

人疱疹病毒—6型基因组及其编码蛋白的结构与功能研究进展

人疱疹病毒－６型（ＨＨＶ－６）是一种新发现的疱疹病毒科成员。本文介绍了近年来国外对ＨＨＢＶ－６基因组的结构，基因定位与功能，及其所编码蛋白的特征与功能方面的研究结果。     

重组人IL—6在E.coli中的高效表达

通过计算机分析设计,人工合成了寡核苷酸引物,并优化了翻译起始区,应用PCR及重组DNA技术,获得了重组人白细胞介素6(IL—6)高效表达工程菌。从全菌SDS—PAGE考马斯亮蓝染色后薄层密度扫描分析表达产率为38.6％,分子量为2.1×10~4。以IL—6依赖性的小鼠杂交瘤细胞株B9用MTT法测定表达重组人IL—6的HGF活性为2×10~7U/L菌液。不同菌株表达重组人IL—6研究表明,以E.coli DH5α/pBV IL—6表达产率最高。     

人疱疹病毒6型感染和儿科临床疾病

人疱疹病毒6型(HHV-6)是在1986年由Salahuddin SZ从艾滋病人和淋巴细胞增生性疾患病人的末梢血单核细胞中分离到的。以后根据这种新发现病毒的病毒学特征，归类于疱疹病毒，称疱疹病毒6型。1988年日本科学家YarrlanishiK证实HHV-6是幼儿急疹的病原，并且HHV-6在原发感染后潜伏于人体，当人体免疫低下时可以再活化。本文简述了HHV-6的病毒学特征、感染的发病机理，概述了幼儿、儿童感染HHV-6的临床方面的研究进展。     

IL—2,IL—6对地塞米松诱导小鼠胸腺细胞凋亡的调节作用

目的：以Ｄｅｘ诱导小鼠胸腺细胞凋亡为模型，研究细胞因子（ＩＬ－２、ＩＬ－６）对小鼠胸腺细胞凋亡的调节作用。方法：应用ＰＩ法检测亚二倍体细胞，二苯胺法测定胸腺细胞片段化ＤＮＡ含量（％），ＤＮＡ凝胶电泳，流式细胞计分析胸腺细胞表型、测定高钙细胞百分比。结果：发现地塞米松（Ｄｅｘ）与小鼠胸腺细胞共同培养引起细胞凋亡，胸腺细胞减少以ＣＤ４^ ＣＤ８^ 细胞最明显。细胞凋亡具有时间效应并与Ｄｅｘ浓度有关。３～４     

IL—2,IL—6对地塞米松诱导小鼠胸腺细胞凋亡的调节作用

目的：以Ｄｅｘ诱导小鼠胸腺细胞凋亡为模型,研究细胞因子（ＩＬ－２、ＩＬ－６）对小鼠胸腺细胞凋亡的调节作用。方法：应用ＰＩ法检测亚二倍体细胞,二苯胺法测定胸腺细胞片段化ＤＮＡ含量（％）,ＤＮＡ凝胶电泳,流式细胞计分析胸腺细胞表型、测定高钙细胞百分比。结果：发现地塞米松（Ｄｅｘ）与小鼠胸腺细胞共同培养引起细胞凋亡,胸腺细胞减少以ＣＤ４^ ＣＤ８^ 细胞最明显。细胞凋亡具有时间效应并与Ｄｅｘ浓度有关。３～４     

HIV-1 induces IL-10 production in human monocytes via a CD4-independent pathway

In HIV-infected patients, increased levels of IL-10, mainly produced by virally infected monocytes, were reported to be associated with impaired cell-mediated immune responses. In this study, we investigated how HIV-1 induces IL-10 production in human monocytes. We found that CD14(+) monocytes infected by either HIV-1(213) (X4) or HIV-1(BaL) (R5) produced IL-10, IL-6, tumor necrosis factor-alpha (TNF-alpha), and to a lesser extent, IFN-gamma. However, the capacity of HIV-1 to induce these cytokines was not dependent on virus replication since UV-inactivated HIV-1 induced similar levels of these cytokines. In addition, soluble HIV-1 gp160 could induce CD14(+) monocytes to produce IL-10 but at lower levels. Cross-linking CD4 molecules (XLCD4) with anti-CD4 mAbs and goat anti-mouse IgG (GAM) resulted in high levels of IL-6, TNF-alpha and IFN-gamma but no IL-10 production by CD14(+) monocytes. Interestingly, neither anti-CD4 mAbs nor recombinant soluble CD4 (sCD4) receptor could block IL-10 secretion induced by HIV-1(213), HIV-1(BaL) or HIV-1 gp160 in CD14(+) monocytes, whereas anti-CD4 mAb or sCD4 almost completely blocked the secretion of the other cytokines. Furthermore, HIV-1(213) could induce IL-10 mRNA expression in CD14(+) monocytes while XLCD4 by anti-CD4 mAb and GAM failed to do so. As with IL-10 protein levels, HIV-1(213)-induced IL-10 mRNA expression in CD14(+) monocytes could not be inhibited by anti-CD4 mAb or sCD4. Taken together, HIV-1 binding to CD14(+) monocytes can induce CD4-independent IL-10 production at both mRNA and protein levels. This finding suggests that HIV induces the immunosuppressive IL-10 production in monocytes and is not dependent on CD4 molecules and that interference with HIV entry through CD4 molecules may have no impact on counteracting the effects of IL-10 during HIV infection.     

Phenotypic alterations and survival of monocytes following infection by human herpesvirus-6

Summary. Freshly isolated monocytes rapidly undergo physiological changes in vitro, resulting in programmed cell death (apoptosis). Activation of monocytes, which promotes differentiation into macrophages, is known to inhibit apoptotic processes. In the present study, we report that human herpesvirus-6 (HHV-6) prevents monocytes from undergoing spontaneous apoptosis during the first 72 hours of culture. Furthermore, significant alterations in cell-surface phenotype were observed after 72 hours of infection with HHV-6. HHV-6-infected monocyte cultures have considerably reduced levels of CD14, CD64 (FcγRI) and HLA-DR antigen on their surface, while CD32 (FcγRII) expression is unaffected. On the basis of these results, we hypothesize that HHV-6 promotes monocytes survival and causes phenotypic modifications that could favor immune evasion and ensure its persistence within the infected host.     

Increased IL-10 production during spontaneous apoptosis of monocytes

Monocytes/macrophages undergo apoptosis and are in contact with apoptotic cells both in vitro and in vivo. The data show that monocytes undergoing spontaneous apoptosis in vitro change their cytokine production profile. We demonstrate that the lipopolysaccharide (LPS)-induced production of interleukin-10 (IL-10) is up-regulated, while production of pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) is either not affected or reduced. These differences seen both at the protein and mRNA level directly correlate with the appearance of apoptotic cells in the culture. Flow cytometry analysis using double staining, surface with annexin V and intracellular with anti-IL-10, suggested that annexin V-negative monocytes are the predominant source of IL-10. Analysis of sorted populations of monocytes indicated that the increase in IL-10 synthesis appears to result from direct interactions between non-apoptotic and apoptotic cells at the time of stimulation. Also non-apoptotic, freshly isolated monocytes produced more IL-10 upon stimulation with LPS, Staphylococcus aureus or zymosan when apoptotic neutrophils were added to the culture. In contrast, monocyte-derived macrophages did not produce more IL-10 in the presence of apoptotic neutrophils. Finally, we found that the presence of apoptotic monocytes in the culture may influence specific immune responses. The data show that in the presence of annexin V-positive monocytes CD4-positive memory T cells produce less IFN-gamma upon stimulation with purified protein derivative of tuberculin, which could be partially reversed by anti-IL-10 neutralizing antibodies. We conclude that these findings might illustrate the mechanisms operating within an inflammatory site and play an important immunoregulatory role during the resolution of inflammation and specific immune responses.     

IL-10 inhibits prostaglandin E2 production by lipopolysaccharide-stimulated monocytes

Since IL-10 has recently been shown to exhibit plelotroplc effectson human monocytes, It was of interest to determine the effectof this cytoklne on prostaglandin E2 (PGEJ production by monocytes.Recomblnant IL-10 (rlL-10) did not significantly affect PGE₂production by lipopolysaccharide (LPS)-unstimulated monocytes,but efficiently inhibited PGE₂ production by LPS-stlmulatedmonocytes. The inhibition by rlL-10 was achieved in a dose-dependentmanner. Recombinant IL-4 also inhibited PGE₂ production at thesame degree as rlL-10. Viral IL-10 Inhibited PGE2 productionby monocytes in a similar fashion as did human rlL-10. Endogenouslyproduced IL-10 was also shown to inhibit PGE₂ production byLPS-stlmulated monocytes. Kinetic studies showed that the inhibitionby rlL-10 on PGE₂ production was observed at least 3 h afterLPS stimulation. Taken together, these results indicate thatIL-10 may play an important role in modulating immunologlcalresponses via down-regulation of PGE₂ production by monocytes.     

Enhanced IL-10 production in vitro by monocytes in autoimmune haemolytic anaemia

Our previous studies on autoimmune haemolytic anaemia (AIHA) have shown a hyperactivation concerning cytokine production in T and B lymphocytes obtained from AIHA patients. In this study the production of interleukin (IL)-10, basal and stimulated by lipopolysaccharide (LPS), was determined in cultured monocytes obtained from patients affected by AIHA, either idiopathic or associated with other autoimmune diseases, e.g. idiopathic thrombocytopenic purpura (ITP), systemic lupus erythematosus (SLE), myastenia gravis (MG). In cultured AIHA monocytes the IL-10 basal production (mean+/-SD) was increased in all but one patient, compared to the controls, 125.96+/-76.10 pg/mL and 19.18+/-15.80 pg/mL respectively. Specifically, LPS stimulation was able to induce a higher IL-10 production by monocytes in two AIHA cases, whereas in five patients there was no difference and in two cases the LPS stimulated IL-10 production was even decreased compared to the levels observed in the controls. Interestingly in the latter two cases AIHA was associated with other autoimmune diseases (ITP, SLE). These results indicate a constitutively higher basal IL-10 production in monocytes from AIHA patients. The increased level of IL-10 could play an important role in the modified pathways of the immune response in AIHA.