Dissecting the autocrine and paracrine roles of the CCR2-CCL2 axis in tumor survival and angiogenesis

The CCL2 CCR2 axis is likely to contributes to the development and progression of cancer diseases by two major mechanisms; autocrine effect of CCL2 as a survival/growth factor for CCR2+ cancer cells and, the attraction of CCR2+ CX₃CR1+tumor associated macrophages that in the absence of CCR2 hardly migrate. Thus far no in vivo system has been set up to differentiate the selective contribution of each of these features to cancer development. Here we employed a chimera animal model in which all non-malignant cells are CCR2−/−, but all cancer cells are CCR2+, combined with an adoptive transfer system of bone marrow (BM) CX₃CR1+ cells from CCR2+ mice harboring a targeted replacement of the CX₃CR1gene by an enhanced green fluorescent protein (EGFP) reporter gene (cx₃cr1 ^gfp), together with the CD45.1 congene. Using this system we dissected the selective contribution of CX₃CR1+CCR2+ cells, which comprise only about 7% of CD11b+ BM cells, to tumor development and angiogenesis. Showing that aside for their direct pro-angiogenic effect they are essential for the recruitment of other CD11b+ cells to the tumor site. We further show that the administration of CCR2-Ig, that selectively and specifically neutralize CCL2, to mice in which CCR2 is expressed only on tumor cells, further suppressed tumor development, implicating for the key role of this chemokine supporting tumor survival in an autocrine manner. This further emphasizes the important role of CCL2 as a target for therapy of cancer diseases.     

Interaction of the TNF homologues BLyS and APRIL with the TNF receptor homologues BCMA and TACI

BLyS (also called TALL-1, THANK, or BAFF) [1] [2] [3] [4] is a member of the tumor necrosis factor (TNF) gene family that stimulates proliferation and immunoglobulin production by B cells. BLyS interacts with the TNF receptor (TNFR) homologue TACI (transmembrane activator and CAML-interactor) [5], and treatment of mice with a TACI-Fc fusion protein abolishes germinal center formation after antigenic challenge [6]. Here we report a novel interaction between BLyS and another TNFR homologue, BCMA (B cell maturation antigen) [7] [8]. Further, the TNF homologue APRIL [9], a close relative of BLyS, also bound to BCMA and TACI. BLyS or APRIL activated nuclear factor-kappaB (NF-kappaB) through TACI and BCMA, and each ligand stimulated immunoglobulin M (IgM) production by peripheral blood B cells. These results define a dual ligand-receptor system that may play an important role in humoral immunity.     

Structural and functional analysis of the interaction between the agonistic monoclonal antibody Apomab and the proapoptotic receptor DR5

Activation of the proapoptotic receptor death receptor5 (DR5) in various cancer cells triggers programmed cell death through the extrinsic pathway. We have generated a fully human monoclonal antibody (Apomab) that induces tumor cell apoptosis through DR5 and investigated the structural features of its interaction with DR5. Biochemical studies showed that Apomab binds DR5 tightly and selectively. X-ray crystallographic analysis of the complex between the Apomab Fab fragment and the DR5 ectodomain revealed an interaction epitope that partially overlaps with both regions of the Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand binding site. Apomab induced DR5 clustering at the cell surface and stimulated a death-inducing signaling complex containing the adaptor molecule Fas-associated death domain and the apoptosis-initiating protease caspase-8. Fc crosslinking further augmented Apomab's proapoptotic activity. In vitro, Apomab triggered apoptosis in cancer cells, while sparing normal hepatocytes even upon anti-Fc crosslinking. In vivo, Apomab exerted potent antitumor activity as a single agent or in combination with chemotherapy in xenograft models, including those based on colorectal, non-small cell lung and pancreatic cancer cell lines. These results provide structural and functional insight into the interaction of Apomab with DR5 and support further investigation of this antibody for cancer therapy.     

Targeting the extrinsic apoptosis pathway in cancer

Mutational inactivation of the p53 tumor-suppressor gene, which regulates apoptosis mainly via the cell-intrinsic pathway, reduces the sensitivity of many cancers to conventional treatments. Targeting the cell-extrinsic pathway, which triggers p53-independent apoptosis, offers a unique therapeutic strategy to induce apoptosis in cancer cells. This article focuses on two proapoptotic receptor agonists, recombinant human Apo2-ligand/TNF-related apoptosis-inducing ligand (rhApo2L/TRAIL) and Apomab, which activate death receptor (DR) 4 and/or DR5, thus stimulating the cell-extrinsic pathway. These agents are under investigation for the treatment of solid tumor and hematologic malignancies. Preclinical data indicate that both molecules cause significant regression or growth inhibition of malignant tumors without significant toxicity. Initial data on rhApo2L/TRAIL and Apomab from phase 1 safety trials also confirm that these agents are suitable for further clinical investigation.     

Native River Boats in ‘Irāq

The Maasai are cattle pastoralists, living on the grass savanna in southern Kenya and northern Tanzania. This article, which deals with Kenya, describes Maasai women as “heads of houses”. The house as a physical structure shelters and symbolically embodies a matrofocal unit of consumption and resource sharing which is the smallest unit of Maasai society. In terms of livestock property, such units are subordinated to larger, patriarchal units. According to the author, the relative autonomy of Maasai houses, and in particular women's control over food resources is significantly decreasing. The reason for this is the present commercialization of Maasai production.     

Foraging,roosting, and nesting habitats of the avian fauna of the Agmon wetland,northern Israel

The foraging, nesting and roosting habitats of the avian fauna of a newly created Agmon wetland and surrounding cultivated peat land (5 km2) in the Hula Valley, northern Israel, were evaluated (January 1996–February 1997) to assess the value as a habitat and for wildlife tourism. We recorded 180 bird species (herons, dabbling ducks, kingfishers, waders, wagtails and raptors) in different habitats (the lake, shores, cattail and reed-bed stands, trees, temporary inundated areas). The most heavily used habitat for foraging, breeding, and roosting was a large cattail stand in the southern third of the lake. The foraging habitat and diet data of 97 avian species were determined. The most intensively used foraging habitats were cultivated fields, lake shore reed-beds, shallow canals and the cattail stand. Forty six species nested in the wetland during March-October, with 2,040 colonial and solitary nests mostly in the cattail stand, near-shore reed-beds and young trees. Roosts, also mostly in the cattail and reed-bed stands and in trees, were used by 55 species. Based on this survey of available habitats, we identified several key habitats that are either missing or require further development (e.g., temporarily inundated mud flats, reed-beds and bare islands). We also provide suggestions for improving the delicate balance between requirements of this developing wetland ecosystem and of the adjacent agricultural areas.     

Localization of the gene for congenital dyserythropoietic anemia type I to a <1-cM interval on chromosome 15q15.1-15.3.

Congenital dyserythropoietic anemias (CDA) are a rare group of red-blood-cell disorders of unknown etiology that are characterized by ineffective erythropoiesis, pathognomonic cytopathology of the nucleated red blood cells in the bone marrow, and secondary hemochromatosis. In CDA type I, bone-marrow electron microscopy reveals characteristic findings in erythroid precursors, including spongy heterochromatin and enlarged nuclear pores. Since the genetic basis of CDA type I is not evident, we used homozygosity and linkage mapping to localize the genetic defect responsible for CDA type I in 25 Bedouins from four large consanguineous families. We report the linkage of this disease to markers on chromosome 15 located at q15. 1-q15.3. Fourteen markers within a 12-cM interval were typed in the relevant family members. Nine of the markers yielded maximum LOD scores of 1.625-12.928 at a recombination fraction of .00. Linkage disequilibrium was found only with marker D15S779. Haplotype analysis revealed eight different carrier haplotypes and highlighted the existence of a founder haplotype. Identification of historical crossover events further narrowed the gene location to between D15S779 and D15S778. The data suggest localization of the CDA type I gene within a 0.5-cM interval. The founder mutation probably occurred >/= 400 years ago. Sequence analysis of the coding region of protein 4.2, the only known erythroid-specific gene in the locus, did not reveal any change in the CDA type I patients. Future analysis of this locus may lead to the identification of a gene essential to normal erythropoiesis.     

Incorporation and uptake of thymidine and uridine by hela cells: Effect of DNA extracts prepared from normal human fibroblasts

Summary The incorporation and uptake of (³H) thymidine into HeLa cells markedly decreased in the presence of nuclear homogenates and DNA extracts that have been derived from normal diploid cell cultures. On the other hand, uridine uptake and incorporation were stimulated under the same conditions. The inhibition could be reversed immediately upon removal of the exogenous fraction from the culture medium. The inhibitory properties of the extracts are propagated by excreted cellular components as well as after DNAase treatment. The inhibitory factor is thermostable, resistant to pronase treatment, and seems to be related to nucleic acid. The material herein represents part of a dissertation presented by the senior author in partial fulfillment of the requirements for the degree of Doctor of Philosophy at the Tel Aviv University, Israel. This work was partially supported by a grant from the Israel Ministry of Health.