Persistent Measles Virus Infection of Murine Neuroblastoma Cells Differentially Affects the Expression of PKC Individual Isoenzymes

Measles virus (MV) is among the infectious agents displaying a propensity for establishing persistent infections of the CNS. It is assumed that continuous presence of MV defective particles or viral genome in persistently infected cells may influence host cellular processes and perturb biochemical signal transduction pathways operating in linkage to various cell surface receptors. PKC expression in a MV persistently infected neuroblastoma cell line (NS20Y/MS) was investigated. The relative levels of PKC isoenzymes were determined by Western blot analysis. We found that protein levels of PKCα, ɛ and ζ, but not PKCδ, were increased in NS20Y/MS cells. PKCβ, γ and η were undetectable. Treatment of NS20Y/MS cells with anti-MV Abs, which downregulated MV protein synthesis, also reduced PKCα expression to the basal level observed in the uninfected NS20Y cells. Our results suggest that a persistent MV infection has specific effects on the expression of certain PKC isoenzymes. We postulate that the MV-associated neurologic changes may reflect virus induced changes in biochemical signaling pathways and that these effects are likely to be regulated by the host's anti-viral humoral immune response. This revised version was published online in July 2006 with corrections to the Cover Date.     

CD8+ T Cell Exhaustion During Persistent Viral Infection is Regulated Independently of the Virus-Specific T Cell Receptor

During chronic viral infections, responses by virus-specific CD8⁺ T cells become marginalized by the acquisition of functional defects and reduced cell numbers in a process defined as T cell exhaustion. Similarly, T cell tolerance to self-antigen is also characterized by impaired effector function and eventual deletion of self-reactive T cells. Induction of both tolerance and exhaustion involve many shared inhibitory mechanisms, thus similar therapeutic approaches have proven effective in these distinct environments. We previously demonstrated that tolerant self-reactive CD8⁺ T cells expressing dual-T cell receptors (i.e., dual-TCR) could be rescued by immunization through a second TCR specific for a foreign antigen. These data revealed that T cell tolerance was regulated at the level of the self-reactive TCR. Here, dual-TCR CD8⁺ T cells were used to examine if exhaustion during persistent viral infection could be rescued by an analogous strategy of immunization through a second TCR not involved in recognition of virus. In direct contrast to the rescue achievable in tolerant CD8⁺ T cells, exhausted T cells were equally impaired through both TCR. These findings suggest that exhaustion is maintained by defects downstream of the virus-specific TCR, and establish that exhaustion and tolerance are distinctly regulated states of T cell dysfunction.     

病毒感染与动脉粥样硬化相关性研究进展

动脉粥样硬化(atherosclerosis,AS)是心血管系统的常见疾病,严重威胁人类健康.AS发病机制复杂,有一个慢性的发病过程.在其发展过程中,血管内皮细胞的损伤、血小板和凝血因子的活化、体内促凝与抗凝的失衡、高血脂、高血黏度及血流动力学改变等因素均促进粥样斑块和血栓形成[1].近年来在该病的诊断、治疗上已取得很大进展,然而对于其原发因素和发病机理仍不清楚.     

Transcriptional regulatory regions of gap43 needed in developing and regenerating retinal ganglion cells

Mammals and fish differ in their ability to express axon growth‐associated genes in response to CNS injury, which contributes to the differences in their ability for CNS regeneration. Previously we demonstrated that for the axon growth‐associated gene, gap43, regions of the rat promoter that are sufficient to promote reporter gene expression in the developing zebrafish nervous system are not sufficient to promote expression in regenerating retinal ganglion cells in zebrafish. Recently, we identified a 3.6‐kb gap43 promoter fragment from the pufferfish, Takifugu rubripes (fugu), that can promote reporter gene expression during both development and regeneration. Using promoter deletion analysis, we have found regions of the 3.6‐kb fugu gap43 promoter that are necessary for expression in regenerating, but not developing, retinal ganglion cells. Within the 3.6‐kb promoter, we have identified elements that are highly conserved among fish, as well as elements conserved among fish, mammals, and birds. Developmental Dynamics 239:482–495, 2010. © 2009 Wiley‐Liss, Inc.     

细胞衰老与p16INK4a的转录调控

抑癌基因p16~(INK4a)可特异地抑制CDK4及CDK6,在抑制细胞生长、促进细胞衰老等方面发挥重要的生物学作用。由于p16~(INK4a)功能的重要性,近年来,针对p16~(INK4a)转录调控方面的研究取得了一系列进展,发现了一系列正性和负性调控元件和转录调控因子,如:E47、Id1、Jun B、Bmi-1、RREB等,为进一步认识细胞增殖规律以及衰老进程具有重要的理论意义。     

病毒感染与逃避免疫反应的研究进展

病毒和宿主共生过程中,病毒为了生存繁衍形成多种逃避宿主免疫反应的方式。其中病毒抗原变异逃逸抗体的中和和病毒干扰补体激活的关键环节来抑制宿主的抗病毒状态是病毒拮抗机体体液免疫反应的主要途径。此外,病毒还利用编码多种细胞因子的类似物干扰细胞因子的正常功能,造成有利于病毒生存复制的内环境,从而使得病毒能够持续感染机体。     

Regulation and Function of T-Cell-Mediated Immunity during Toxoplasma gondii Infection

The intracellular protozoan Toxoplasma gondii is a widespread opportunistic parasite of humans and animals. Normally, T. gondii establishes itself within brain and skeletal muscle tissues, persisting for the life of the host. Initiating and sustaining strong T-cell-mediated immunity is crucial in preventing the emergence of T. gondii as a serious pathogen. The parasite induces high levels of gamma interferon (IFN-γ) during initial infection as a result of early T-cell as well as natural killer (NK) cell activation. Induction of interleukin-12 by macrophages is a major mechanism driving early IFN-γ synthesis. The latter cytokine, in addition to promoting the differentiation of Th1 effectors, is important in macrophage activation and acquisition of microbicidal functions, such as nitric oxide release. During chronic infection, parasite-specific T lymphocytes release high levels of IFN-γ, which is required to prevent cyst reactivation. T-cell-mediated cytolytic activity against infected cells, while easily demonstrable, plays a secondary role to inflammatory cytokine production. While part of the clinical manifestations of toxoplasmosis results from direct tissue destruction by the parasite, inflammatory cytokine-mediated immunopathologic changes may also contribute to disease progression.     

Regulation of in vitro and in vivo T cell activation by CD43

Accessory molecule interactions can be critical in determining the outcome of a T cell's encounter with antigen. Cell adhesion proteins may augment T cell responses by facilitating TCR engagement of the antigen-MHC complex, while co-stimulatory molecules may deliver distinct signals that modulate T cell responsiveness. CD43 (leukosialin, sialophorin) has been suggested to influence cell activation by steric hindrance based upon the large size and glycosylation of the protein, as well as the relative abundance of the protein on the cell surface. In this paper we examine both in vitro and in vivo T cell-dependent responses in CD43-deficient mice. We demonstrate that T cells from CD43-deficient mice are hyper-responsive following both in vivo and in vitro activation, and that this is observed in response to not only TCR-CD3-mediated stimulation, but also following receptor-independent activation. This data suggests that mechanisms other than non-specific steric hindrance are important in the regulation of T cell activation by CD43.      

Persistent Superphosphorylation of Leukosialin (CD43) in Activated T Cells and in Tumour Cell Lines

CD43 (leukosialin) is a highly sialylated, single-chain molecule expressed on most human leucocytes. Regulatory signals appear to be transduced through the molecule as suggested by the ability of anti-CD43 antibodies to induce aggregation and proliferation of T cells and to enhance B-cell proliferation and natural killer cell activity. Activation of protein kinases is an essential event in signal transduction. We were therefore interested to study whether CD43 may function as a substrate for protein kinases during mitogenic activation of lymphocytes. We show that CD43 was rapidly superphosphorylated (within minutes) on serine residues following addition of phorbol ester (PMA) to peripheral blood lymphocytes. PMA treatment of the cells was not followed by rapid down-regulation of CD43. Activation of the lymphocytes by concanavalin A or anti-CD3 antibodies (OKT 3) also resulted in superphosphorylation of CD43. However, the phosphorylation was delayed as compared to that induced by PMA and was detected 3–4 h after the addition of the reagents. A plateau was reached after 24–48 h of Stimulation. Interestingly, the high level of phosphonrylation of CD43 was maintained in long-term cultures of T cells activated by various means. Furthermore. CD43 was found to be constitutively superphosphorylated (on serine and tyrosine) in continuously growing cell lines of T, B, and non-lymphoid origin. Taken together, the results suggest that CD43 has an important role during both early and late phases of T-cell activation and that modulation of its biochemical properties by protein kinases may be associated with progression through the cell cycle and with cellular growth.     

HTLV-III/LAV Viral Antigens in Lymph Nodes of Homosexual Men With Persistent Generalized Lymphadenopathy and AIDS

The presence of core antigens of retrovirus HTLV-III/LAV, referred to as "AIDS-related virus" (AV), has been sought in lymph node samples of patients with persistent generalized lymphadenopathy (PGL, 28 patients), prodromal AIDS (1 patient) and AIDS with Kaposi sarcoma (3 patients). In 30 patients the deposition of viral antigens, detected by monoclonal antibodies to HTLV-III and LAV, could be observed within the germinal centers (GCs) primarily within the extracellular network of immune complexes, and the two patients who were negative were atypical. No AV could be found in normal tonsil or in samples with follicular hyperplasia of unknown etiology (20 cases). These findings, taken together with the ultrastructural identification of typical retrovirus particles in all 9 PGL and 2 AIDS cases studied, indicates that the network of follicular dendritic (FD) cells is an important reservoir of AV virus antigen at this site. The persistence of this retrovirus inside the GCs helps explain how the follicular hyperplasia affecting FD cells and B blasts in PGL may in progressive cases be accompanied by destruction of FD cells and gradual development of T4+ lymphopenia. T4+ T cells may circulate through the GCs and become infected with AV there. In addition, the identification of retrovirus antigen in situ may be of diagnostic value.     

Expression of three oligosaccharide conjugates by neonatal rat dorsal root ganglion neurons: comparison with CGRP and GAP43 immunoreactivity

Adult dorsal root ganglion neurons express oligosaccharides conjugated to lipids that may be involved in cell–cell recognition, and consequently in the laminar organisation of their central terminations. This paper describes an immunohistochemical study of the developmental expression of 2 lactoseries (LA4 and LD2) and 1 globoseries (SSEA4) oligosaccharide conjugates in rats from embryonic d 19 to postnatal d 60. The expression of calcitonin gene related peptide and the growth associated protein GAP43 was also examined for comparative purposes. We found that these oligosaccharide conjugates begin to be expressed after birth, suggesting that they may be involved in maturation of the central or peripheral terminations, rather than axonal guidance.     

Ultrastructural double localization of B-50/GAP43 and synaptophysin (p38) in the neonatal and adult rat hippocampus

Summary B-50/GAP43, a neuron-specific phosphoprotein, is highly expressed in developing nervous tissue. Monospecific polyclonal affinity-purified B-50 antibodies were used to document the ultrastructural distribution of B-50 in the hippocampus of 90-day-old (P90) and 1-day-old (P1) rats. Double-labelling immunoprocedures were performed to compare the localization of B-50 and synaptophysin (p38), a protein specific for synaptic vesicles. By immunofluorescence light microscopy B-50 and p38 were similarly distributed in the CA1 neuropil of P90 rats. In contrast, in P1 rats B-50 was more widely distributed than p38. By electron microscopy of P90 rat hippocampus, B-50 was located at the plasma membranes of axon shafts and of p38-immunoreactive axon terminals. Some B-50 was found in the cytosol of axon terminals. B-50 was absent at the plasma membranes of apical dendrites and of pyramidal cells. In the P1 rat hippocampus, B-50 was detected at the plasma membrane of growth cones, axon terminals and axon shafts, but not in their cytosol. The plasma membranes of pyramidal cell bodies and their processes extending into the stratum radiatum were without B-50. B-50-immunoreactive organelles of the lysosomal family were found in the cytosol of pyramidal cells of the hippocampus of P1 and P90 rats. This ultrastructural study shows that during development of the stratum radiatum in the hippocampal field CA1, the localization of B-50 persists at the plasma membrane of axons and axon terminals in P1 and P90 rats. This localization of B-50 is consistent with the suggestion that B-50 acts as a regulator of neurotransmitter release and intracellular messengers.