Transforming growth factor-β stimulates IL-1β-induced monocyte chemoattractant protein-1 expression in human synovial cells via the ERK/AP-1 pathway

Objective and Design: Transforming growth factor- β (TGF-β) has not only a fibrogenic role, but also monocyte/ macrophage chemotactic properties in a synovial joint. However, little is known about the effects of TGF-β on monocyte chemoattractant protein-1 (MCP-1) expression in human synovial cells under inflammatory status. The aim of this study was to determine whether TGF-modulates MCP-1 production under the chronic inflammation, and to elucidate the cell signaling mechanism involved. Materials and methods: Human synovial cells were exposed to IL-1β, which mimics the environment of chronic inflammation. Production of MCP-1 protein and expression of MCP-1 mRNA were determined by ELISA and real-time PCR. Results: TGF-β upregulated the expression of MCP-1 mRNA and protein with or without IL-1β. TGF-β and IL-1β synergistically enhanced MCP-1 gene expression, and an AP-1 binding site was involved in the signal transduction. In addition, MEK inhibitor completely suppressed TGF-β-induced MCP-1 production. Conclusions: TGF-β and IL-1β synergistically enhance MCP-1 gene expression through the activation of the MEK/ERK1/2 pathways, which leads to AP-1 activation. The impairment of MCP-1 regulation by TGF-β in resident synovial cells might represent an important mechanism of chronic inflammation and tissue fibrosis in a synovial joint. MCP-1 should be considered a valid target for therapeutic intervention. Received 22 September 2005; returned for revision 3 February 2006; accepted by J. Skotnicki 29 May 2006     

八种粘附分子在急性淋巴细胞性白血病异常表达的研究

本文分析了３８名正常人和２５例急性淋巴细胞性白血病（ＡＬＬ）细胞表面ＣＤ１１ａ、ＣＤ１１ｂ、ＣＤ５４、ＣＤ４４等的表达。与正常人相比，肿瘤细胞上表达的ＣＤ１１ｂ、ＣＤ１８和ＣＤ５４降低，ＣＤ４４升高；ＣＤ１１ａ在Ｂ－ＡＬＬ和混合型ＡＬＬ表达减弱，在Ｔ－ＡＬＬ增强。３例急性淋巴细胞性白血病骨髓基质细胞表面ＣＤ５４、ＣＤｗ４９ｂ表达亦降低。认为粘附分子可能参与了急性淋巴细胞性白血病的发病机制。     

Monoclonal antibodies to the human mammary gland: III. Monoclonal antibody LICR-LON-M18 identifies impaired expression and excess sialylation of the I(Ma) cell-surface antigen by primary breast carcinoma cells

Monoclonal antibody LICR -LON- M18 identifies the immunodominant oligosaccharide sequence of the I(Ma) blood-group antigen: Gal beta 1----4GlcNAc beta 1----6--. In primary breast cancers this structure is almost totally cryptic, due to "masking" by sialic acid, but can be revealed by digestion with the specific glycosidase neuraminidase. Following desialylation, light microscopic immunohistochemical examination has revealed the epitope identified by LICR -LON- M18 to be heterogeneously distributed throughout the population of breast carcinoma cells. These tumor cells express the antigen as both a cytoplasmic and a surface membrane determinant. In the normal human breast, this structure is expressed exclusively along the luminal plasma membranes of the duct and alveolar littoral epithelial cells. Desialylation of tissue sections of normal resting and lactating breast epithelium with neuraminidase virtually abolishes the heterogeneous intercellular distribution of the I(Ma) determinant. In desialylated nonneoplastic breast tissues, the expression of this antigen is observed within the cytoplasm of some myoepithelial cells, but not in the littoral epithelial cells. The expression of the I(Ma) antigen by neoplastic and normal breast epithelial cells has also been compared with that of the oligosaccharide sequence Gal beta 1----3GalNAc. This structure, recognized by peanut agglutinin, forms the dominant portion of the Thomsen-Friedenreich antigen. With respect to normal and lactating breast epithelial cells, both oligosaccharide structures are sialylated and appear to be similarly misprocessed by breast carcinomas. The masking of surface carbohydrate determinants and the faulty processing of structures usually expressed on the surface of non-neoplastic breast epithelial cells may be important phenomena in the pathobiology of breast carcinomas.     

Detection of virus and cellular-determined antigens in situ using [125I]protein A and autoradiography.

The present paper describes the use of [125I]Protein A, isolated from Staphylococcus aureus, in detecting antigen-antibody complexes by autoradiography on single cells. the method is relatively quick, reproducible, potentially more sensitive than immunofluorescence, and should be useful in combination with conventional radioimmuno-assays. We have used it to detect the cellular expression of IgM, kappa, lambda, and beta 2-microglobulin, as well as the expression of Epstein-Barr Virus (EBV)-associated antigens expressed in human lymphoblastoid cell lines.     

H. pylori Escape Host Immunoreaction Through Inhibiting ILK Expression by VacA

Helicobacter pylori （H. pylori） persistently colonizes the gastric mucosa despite a vigorous immune response. Vacuolating cytotoxin secreted by H. pylori has turned out to be a potent immunomodulatory toxin, but the signal transduction pathways involved has not been studied in macrophages. We observed in this study that vacA-deficient H. pylori induced significantly higher expression of integrin-linked kinase （ILK） and endothelial nitric oxygen synthase （eNOS）, and significantly more production of reactive oxygen species （ROS） in monocyte/macrophage-like U937 cells, as compared with isogenic vacA^＋ H. pylori. The expression of eNOS mRNA in U937 cells overexpressing ILK was markedly increased compared with those transfected with empty vectors. Thus, vacA-deficient H. pylori appears to upregulate ILK expression, which modulates the expression of eNOS and as a result, stimulates the production of ROS. It is VacA that prevents such a process by inhibiting ILK expression, helping H. pylori escape host immunoreaction. This mechanism explains, at least in part, persistent infection of H. pylori in the stomach.     

小鼠肺腺癌细胞系四株克隆化亚系的转移潜能研究

本文对小鼠肺腺癌细胞系（ＬＡ－７９５）进行单细胞克隆化培养，分离；经筛选获得具有裸小鼠体内移植自发肺转移力相异的４株克隆化亚系，说明母系确实在存在转移潜在异质性，体外实验比较此四株亚系的结果显示：癌细胞的转移潜能不仅与其细胞增殖，分裂及胞浆骨架分布的不均紊乱程度有关（Ｐ〈０．０１），而且与其胞浆内架聚集于胞核－侧及内源性非特异性酯酶的活性表达有关（Ｐ〈０．０１）；同时，高转移潜能亚系比转移潜能亚系     

Cyclooxygenase-2 expression and its association with angiogenesis, Helicobacter pylori, and clinicopathologic characteristics of gastric carcinoma

Cyclooxygenase-2 (COX-2) is upregulated in gastric carcinoma, and its increased levels were found to have a prognostic significance in some studies. Both angiogenesis and Helicobacter pylori infection have been reported to be associated with COX-2 expression of gastric cancer in recent studies. In this study, COX-2 expression and its association with CD31 staining, H.-pylori infection, and well-known clinicopathological factors were investigated in 65 gastric cancer patients. COX-2 and CD31 expression assessment was done by immunohistochemical methods. Whartin Starry stain was performed for H.-pylori infection. Of 65 patients, 32 (49%) revealed intense COX-2 immunostaining. Among various clinicopathologic characteristics, COX-2 expression was inversely correlated with tumor size, TNM stage, and lymph node status. Thirty-two (49%) patients revealed intense CD31 immunostaining. Among various clinicopathologic characteristics, CD31 expression was associated only with lymph node metastasis. COX-2 expression was not correlated with CD31 staining and H.-pylori infection. Both COX-2 and CD31 staining had no prognostic significance. In conclusion, we found that COX-2 expression was significantly higher in earlier stages of gastric cancer. It can be suggested that COX-2 expression may be important in the initial development of gastric cancer but not in progression of the disease. Other factors which may be associated with COX-2 in gastric cancer, including angiogenesis and H.-pylori infection, should be investigated in further studies.     

鲨鱼软骨粉对小鼠移植性肿瘤的抑制作用与微血管的关系

观察鲨鱼软骨粉对小鼠移植性ＳＲＳ生长的影响与微血管密度的关系,方法肿瘤称重,切片用ｖＷＦ疫组化染色及微血管计算机扫描方法半定量。结果在已建立的小鼠ＳＲＳ实体瘤模型上应用口服鲨鱼软骨粉治疗,发现５４毫克／ｄ治疗１５ｄ后肿瘤重量及微血管密度较对照组明显减少。     

人白细胞介素4在大肠杆菌中的优化表达

目的：通过优化设计利用大肠杆菌表达人白细胞介素４（ＩＬ４） 并提高其表达量。方法：根据原核翻译起始序列的局部二级结构自由能,设计了ＡＵＧ 上下游序列,并在人ＩＬ４ 基因下游插入部分大肠杆菌ＬａｃＺ 序列以提高ｍＲＮＡ 的稳定性。结果：成功地构建了人ＩＬ４ 的高效表达克隆,命名为ｐＬＣＭ１８２ｈＩＬ４ 。ＳＤＳＰＡＧＥ 分析显示所表达的重组ＩＬ４ 蛋白质占细菌总蛋白的３０ ％ 。目的基因下游没有插入ＬａｃＺ序列所构建的表达克隆ｐＣＺＨｈＩＬ４ 在大肠杆菌中的表达量则占细菌总蛋白的２０ ％ 左右。结论：所设计的优化表达方法对人ＩＬ４ 基因的表达是成功的,在细胞因子的工程化表达中,优化设计对基因的表达量至关重要。     

Expression of FcgammaRs and mCD14 on polymorphonuclear neutrophils and monocytes may determine periodontal infection

Variance in expression of receptors for immunoglobulin G (FcgammaRs), complement (CR3) and lipopolysaccharide (mCD14) on polymorphonuclear neutrophils (PMNs) and monocytes might affect susceptibility for infection with certain pathogens in periodontitis, a chronic infectious disease of tooth-supportive tissues. Levels of FcgammaRI, IIa, III, CR3 and mCD14 on PMNs and monocytes were measured in 19 periodontitis patients and 18 healthy controls. Subgingival infection with Aggregatibacter actinomycetemcomitans (Aa) and Porphyromonas gingivalis (Pg) was determined. Activation of PMNs and monocytes in response to stimulation with Aa and Pg was assessed by means of change in mCD14 expression. Periodontitis is associated with an enrichment of the FcgammaRIII(+) monocytes (P = 0.015) with concomitant low mCD14 (P = 0.001). Unadjusted data showed that the subjects culture-positive for Aa (Aa(+)) had significantly lower expression of monocytic FcgammaRI (P = 0.005) and FcgammaRIIa (P = 0.015) than Pg(+) subjects. The FcgammaRI was still lower on monocytes from Aa(+) subjects after adjusting for the background factors (P = 0.037). PMNs from Aa(+) subjects responded in a hyper-reactive manner, in particular when stimulated with Aa (P = 0.011). Lower FcgammaRs expression by monocytes is related to a higher susceptibility of a subject to become infected with Aa. The higher proportion of FcgammaRIII(+) monocytes may be involved in the chronicity of this condition. Hyper-reactive PMNs in Aa(+) subjects may contribute to accelerated breakdown of tooth-supportive tissues.     

Perforin Expression in Plaque Psoriasis: An Immunohistochemical Study

Psoriasis (PsO) is T-cell-mediated disease resulting from aberrant activation of both innate and adaptive immunity. Perforin is a multi-domain, pore-forming protein. It is located within the cytoplasm of CD 8 cytotoxic T cells (CTLs) and natural killer cells (NK). The aim of this study was to evaluate the immunohistochemical (IHC) expression of perforin in lesional and perilesional skin of chronic plaque psoriatic patient and correlate its expression with the standard clinico-pathological variables. This prospective case–control study was conducted on 50 PsO patients and 30 age- and gender-matched healthy subjects as a control group. There were high-significant differences between lesional and perilesional skin of plaque PsO patients as regards to IHC perforin status and localization (p?p?p?=?0.02 and 0.03, respectively). Localization of IHC perforin positive lymphocytes in both epidermis and dermis was significantly associated with higher degree of acanthosis and higher degree of inflammatory infiltrates in comparison with positive cells located in dermis (p?=?0.001 for both). Perforin might have a putative signaling in early and late plaque PsO. Plaque psoriatic patients with positive perforin expression could be a candidate for a future target therapy to stop the proposed scenario and achieve a therapeutic response.     

In Vivo Expression of Classic PKC Isoforms in the Rat Dental Follicle as Related to Tooth Eruption

Activation of protein kinase C (PKC) can upregulate tooth eruption molecules such as osteoprotegerin (OPG) and vascular endothelial growth factor (VEGF). In this study, we examined the in vivo gene expression of classic isoforms of PKC in the dental follicle of postnatal rats. The expression level of PKC-α was significantly reduced at day 3 followed by a gradual return to day 1 level, a profile similar to OPG expression. The expression of PKC-β was the lowest at day 1 followed by elevated levels from day 3 to day 11. Expression of PKC-β is positively correlated with the expression of overall VEGF and VEGF120. The expression level of PKC-γ was relatively steady in the postnatal days. Injection of a PKC activator, phorbol 12-myristate 13-acetate (PMA), at late postnatal days, slightly accelerated first mandibular molar eruption. This study suggests that PKC isoforms may be involved in the regulation of tooth eruption.     

Functional characteristics and differential expression of class II DR,DS, and SB antigens on human melanoma cell lines

Although most cultured melanoma cell lines express DR Class II molecules, many of these do not also express the DS (MB) Class II molecules as detected by a monoclonal antibody specific for DS. Cells lacking either DR or DS molecules or both could only be induced to express DR antigens in rare cases by combined incubations with azacytidine and Interleukin-2 conditioned medium, although the expression of DR molecules on fibroblasts or U937 monocytes could more easily be induced under the same culture conditions. Melanoma cells expressing DR antigens could function in antigen presentation for the histocompatibility antigens themselves and for DR specific presentation of TNP determinants to allogeneic T-cells sensitized to TNP modified lymphocytes and showing restriction in their responses to the specificity of the DR molecules expressed on the original, autologous senzitizing cells. DR positive melanoma cells could not, however, be demonstrated to function in the presentation of any of the soluble antigens tested. All DR positive melanoma cells also expressed SB antigens, but these were not detected on DR negative melanoma cells. These studies collectively indicate that the expression of Class II histocompatibility antigens on diverse cell types is subject to differential regulatory control and is associated with differences in their functional activities.     

细胞繁殖及分化过程中核仁HMNA P24含量的变化

为探讨人恶性肿瘤相关核仁抗原P24的细胞生物学功能,本研究将Hela细胞进行同步化培养和分化的诱导,以免疫荧光显微分光光度技术测定在细胞周期和细胞分化过程中核仁P24含量的动态变化。结果表明:在G1早期到S早期,核仁P24含量大致恒定;S中期明显降低,随后迅速回升,至G2期达高峰。在用正丁酸钠诱导Hela细胞发生细胞分化过程中,核仁P24含量不断降低,72小时后接近消失。提示P24的表达与细胞的负分化平行,其在核仁中出现,尤其是在GO到G1期,对于癌细胞的快速增殖可能是必需的或重要的。     

Enhanced clearance of Candida albicans from the oral cavities of mice following oral administration of Lactobacillus acidophilus

Orally administered live Lactobacillus acidophilus was assessed for its capacity to enhance clearance from the oral cavity of DBA/2 mice shown previously to be 'infection prone'. L. acidophilus fed to DBA/2 mice significantly shortened the duration of colonization of the oral cavity compared to controls. Enhanced clearance of Candida albicans correlated with both early mRNA gene expression for interleukin (IL)-4 and interferon (IFN)-gamma and expression of their secreted products in cultures of cervical lymph nodes stimulated with Candida antigen. In addition rapid clearance correlated with higher levels of IFN-gamma and nitric oxide in saliva. Delayed clearance, less pronounced levels of the cytokine response, saliva IFN-gamma and nitric oxide, and later mRNA expression for IL-4 and IFN-gamma relative to feeding with the L. acidophilus isolate were noted in mice fed a different Lactobacillus isolate (L. fermentum). These observations indicate significant variations in individual isolates to activate the common mucosal system.     

Immunodeficiency diseases and expression of HLA antigens

The role of HLA antigens in lymphocyte differentiation is strongly suggested by the existence of a recently identified immunodeficiency associated with, and probably resulting from, the lack of expression of HLA-A, B, and C antigens as well as /gB₂ microglobulin on various cells of hematopoietic origin. This “bare lymphocyte syndrome” has been described in a family where the transmission appeared to be autosomal recessive, and the responsible gene was not borne by the sixth chromosome.Another infant with a severe combined immunodeficiency disease has been treated with fetal liver and thymus transplants (FLTT). A persisting chimerism has been documented: T cells derived from the donor and B cells from the host. Despite complete HLA-A, B, and DR mismatch, T and B cells did cooperate resulting in significant antibody production, and defense against viral infection has been normal. Such an observation may suggest that “allogeneic restriction” of T-cell effector functions can be circumvented.     

Xenotropic type C virus expression in murine thymomas induced by radiation or 3-methylcholanthrene

Thymic lymphoma incidence and thymic expression of MuLV with xenotropic infectivity was monitored in AKR, RF, and reciprocal F₁ mice of the AKR × RF cross after treatment with either γ radiation or the chemical carcinogen 3-methylcholanthrene (MCA). These two inbred strains and the F₁ hybrids developed similarly high incidences of thymoma, and lymphomatous cells from AKR mice and (AKR♀ × RF♂)F₁ mice were observed to be expressing MuLV with xenotropic host range. However, lymphoma cells from RF mice and (RF♀ × AKR♂)F₁ mice did not shed xenotropic MuLV. Thymic xenotropic virus expression was therefore not correlated with a high incidence of radiation or chemically induced thymoma, but rather appeared to be a phenotype genetically transmitted by AKR mice to F₁ mice of the AKR × RF cross as a dominant trait in induced thymomas. In addition, a maternal effect on thymic xenotropic virus expression in induced thymomas was observed by the comparison of reciprocal F₁ hybrids in this cross.     

Coordinate expression of the 48K host nuclear phosphoprotein and SV40 T ag upon primary infection of mouse cells

We have studied the expression of the 48K host phosphoprotein after primary infection of BALB/c 3T3 mouse cells with simian virus 40, using a monospecific antiserum against the purified 48K protein. The 48K host phosphoprotein, previously shown to complex with SV40 large T Ag, is shown to be expressed in the nuclei of BALB/c 3T3 cells 18–24 hr after primary infection by SV40, at the same time large T Ag is expressed. Expression of the 48K protein shows the same distribution and multiplicity dependence as that of large T Ag. We were able, by comparing percentages of cells stained with α94K, or α48K, or both antisera together, to show all cells expressing large T Ag and only those cells expressing large T Ag also express the 48K protein. The SV40 A locus governs the expression of the host 48K phosphoprotein, while deletions in the region 0.54 to 0.58 have no effect, suggesting it is large and not small T Ag which governs the expression of the host 48K phosphoprotein. The adenovirus-SV40 hybrid virus, AD2D2, also causes the expression of the host 48K protein, while adenovirus 2 alone does not, also indicating that it is the region 0.54-0.17 map units of the SV40 genome which governs the expression of the 48K protein.