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 (cx3cr1gfp), 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

Gain-of-function human STAT1 mutations impair IL-17 immunity and underlie chronic mucocutaneous candidiasis

Chronic mucocutaneous candidiasis disease (CMCD) may be caused by autosomal dominant (AD) IL-17F deficiency or autosomal recessive (AR) IL-17RA deficiency. Here, using whole-exome sequencing, we identified heterozygous germline mutations in STAT1 in 47 patients from 20 kindreds with AD CMCD. Previously described heterozygous STAT1 mutant alleles are loss-of-function and cause AD predisposition to mycobacterial disease caused by impaired STAT1-dependent cellular responses to IFN-γ. Other loss-of-function STAT1 alleles cause AR predisposition to intracellular bacterial and viral diseases, caused by impaired STAT1-dependent responses to IFN-α/β, IFN-γ, IFN-λ, and IL-27. In contrast, the 12 AD CMCD-inducing STAT1 mutant alleles described here are gain-of-function and increase STAT1-dependent cellular responses to these cytokines, and to cytokines that predominantly activate STAT3, such as IL-6 and IL-21. All of these mutations affect the coiled-coil domain and impair the nuclear dephosphorylation of activated STAT1, accounting for their gain-of-function and dominance. Stronger cellular responses to the STAT1-dependent IL-17 inhibitors IFN-α/β, IFN-γ, and IL-27, and stronger STAT1 activation in response to the STAT3-dependent IL-17 inducers IL-6 and IL-21, hinder the development of T cells producing IL-17A, IL-17F, and IL-22. Gain-of-function STAT1 alleles therefore cause AD CMCD by impairing IL-17 immunity

Biocontrol potential of Microsphaeropsis ochracea on microsclerotia of Verticillium longisporum in environments differing in microbial complexity

The potential of the fungal antagonist Microsphaeropsis ochracea to control the soilborne pathogen Verticillium longisporum was investigated in environments with varying microbial complexity (in vitro vs. in vivo, sterile vs. unsterile, controlled conditions vs. field). A semi-quantitative PCR assay was developed for the detection of M. ochracea on unsterile plant debris. In vitro, M. ochracea caused high levels of mortality to V. longisporum microsclerotia (51-100 %) from 4 to 24 A degrees C, with a broad optimum between 16 and 24 A degrees C. In controlled conditions, M. ochracea significantly reduced the viability of V. longisporum microsclerotia grown on dead rapeseed stems in autoclaved sand, but not in unsterile soil. Likewise, in two experimental years, no significant reduction of V. longisporum inoculum was detectable on rapeseed straw buried in small plots in the field in any of the treatments (soil depths, exposure duration, doses of M. ochracea). Germination of M. ochracea pycnidiospores was inhibited by general soil fungistasis in unsterile soil from a field, botanical garden and grassland. Accordingly, V. longisporum infection of rapeseed plants in the greenhouse was reduced only at artificially high doses of M. ochracea inoculum and no biocontrol efficacy in disease control was recorded in field experiments conducted with winter oilseed rape during two subsequent seasons in an experimental field near Gottingen, with a soil homogenously infested with V. longisporum. The results demonstrate that M. ochracea, although having shown promising potential in controlling pathogens with melanised resting structures on leaf litter, evidently lacks microbial competitiveness to effectively control pathogens in the soil such as V. longisporum, even though the latter is effectively inhibited in vitro

Redox Mechanisms Influencing cGMP Signaling in Pulmonary Vascular Physiology and Pathophysiology

The soluble form of guanylate cyclase (sGC) and cGMP signaling are major regulators of pulmonary vasodilation and vascular remodeling that protect the pulmonary circulation from hypertension development. Nitric oxide, reactive oxygen species, thiol and heme redox, and heme biosynthesis control mechanisms regulating the production of cGMP by sGC. In addition, a cGMP-independent mechanism regulates protein kinase G through thiol oxidation in manner controlled by peroxide metabolism and NADPH redox. Multiple aspects of these regulatory processes contribute to physiological and pathophysiological regulation of the pulmonary circulation, and create potentially novel therapeutic targets for the treatment of pulmonary vascular disease

Inheritance of the Bantu/Benin haplotype causes less severe hemolytic and oxidative stress in sickle cell anemia patients treated with hydroxycarbamide

Beta S-globin gene cluster haplotypes (β S -haplotypes) can modulate the response to hydroxycarbamide (HC) treatment in sickle cell anemia (SCA) patients. In Brazil, the most common haplotypes are Bantu and Benin, and both confer a poor prognosis for patients when untreated with HC. We evaluated oxidative and hemolytic biomarkers in 48 SCA patients undergoing HC treatment separated in three subgroups: Bantu/Bantu, Bantu/Benin and Benin/Benin haplotype. On the basis of reduced haptoglobin (HP) levels, patients with Bantu/Bantu haplotypes had 3.0% higher hemolysis degree when compared with those with Bantu/Benin haplotypes (P=0.01). The Benin/Benin patients had 53.6% greater lipid peroxidation index than the Bantu/Bantu patients (P=0.01) because of evaluated thiobarbituric acid reactive species levels. The Bantu/Benin subgroup had intermediate levels of hemolytic and oxidative stress markers compared with the homozygous subgroups. Through strict inclusion criteria adopted, as well as consolidated and well-described hemolytic and the oxidative parameters evaluated, we suggest a haplotype-interaction response to HC treatment mediated by a 'balance' between the genetic factors of each haplotype studied.Department of Biology Hemoglobin and Hematological Genetic Diseases Laboratory Sao Paulo State University (UNESP), Cristovao Colombo street, no 2265Department of Molecular Biology Sao Jose do Rio Preto Medical School (FAMERP)Hematological State Institute 'Arthur de Siqueira Cavalcanti' (HEMORIO)Department of Biology Hemoglobin and Hematological Genetic Diseases Laboratory Sao Paulo State University (UNESP), Cristovao Colombo street, no 226