Inactivation of chemokine (C-C motif) receptor 1 (CCR1) suppresses colon cancer liver metastasis by blocking accumulation of immature myeloid cells in a mouse model

Recent reports have suggested critical roles of myeloid cells in tumor invasion and metastasis, although these findings have not led to therapeutics. Using a mouse model for liver dissemination, we show that mouse and human colon cancer cells secrete CC-chemokine ligands CCL9 and CCL15, respectively, and recruit CD34⁺ Gr-1⁻ immature myeloid cells (iMCs). They express CCL9/15 receptor CCR1 and produce matrix metalloproteinases MMP2 and MMP9. Lack of the Ccr1, Mmp2, or Mmp9 gene in the host dramatically suppresses outgrowths of disseminated tumors in the liver. Importantly, CCR1 antagonist BL5923 blocks the iMC accumulation and metastatic colonization and significantly prolongs the survival of tumor-bearing mice. These results suggest that CCR1 antagonists can provide antimetastatic therapies for patients with disseminated colon cancer in the liver.     

Compliance with seat belt use in Benin City, Nigeria

INTRODUCTION: Trauma is a major cause of death and disability worldwide. A quarter of all fatalities due to injury occur due to road traffic crashes with 90% of the fatalities occurring in low- and medium-income countries. Poor compliance with the use of seat belts is a problem in many developing countries. The aim of this study was to evaluate the level of seatbelt compliance in motor vehicles in Benin City, Nigeria. METHODS: A five-day, observational study was conducted in strategic locations in Benin City. The compliance rates of drivers, front seat passengers, and rear seat passengers in the various categories of vehicles were evaluated, and the data were subjected to statistical processing using the Program for Epidemiology. RESULTS: A total of 369 vehicles were observed. This consisted of 172 private cars, 64 taxis, 114 buses, 15 trucks, and four other vehicles. The seat belt compliance rate for drivers was 52.3%, front seat passengers 18.4%, and rear seat passengers 6.1%. Drivers of all categories of vehicles were more likely to use the seat belt compared to front seat passengers (p = 0.000) and rear seat passengers (p = 0.000). Drivers of private cars were more likely to use seat belts compared to taxi drivers (p = 0.000) and bus drivers (p = 0.000). Front seat passengers in private cars were more likely to use the seat belt compared to front seat passengers in taxis (p = 0.000) and buses (p = 0.000). Rear seat passengers in private cars also were more likely to use seat belts compared to rear seat passengers in taxis (p = 0.000) and buses (p = 0.000). CONCLUSIONS: Compliance with seat belt use in Benin City is low. Legislation, educational campaigns, and enforcement of seat belt use are needed.     

Factors associated with readiness to start antiretroviral therapy (ART) among young people (15–24 years) at four HIV clinics in Mulago Hospital,Uganda

IntroductionGlobally, the HIV burden continues to rise among young people despite the discovery of ART. This study assessed demographic and psycho-social factors among young people associated with readiness to be initiated on ART.MethodsA quantitative cross-sectional study was conducted among newly diagnosed HIV positive young people aged 15–24 years at 4 HIV clinics at Mulago Hospital. Readiness was measured as a self-report by the individual to the question, “How ready do you feel to start ART?ResultsOf the 231 young people enrolled, the mean age (SD) was 20.7years (+/-2.8) and most were female (66.2%). Majority were very ready (53.3%) and very motivated (51.1%) to start ART. Higher treatment readiness was associated with being female (95% CI [5.62, 8.31], p=0.003), thinking that ART cures HIV (95% CI [0.43, 0.86], p=0.005), history of having unprotected sex (95% CI [0.79, 0.87], p=<0.001), anticipating negative HIV results (95% CI [0.26, 0.88], p=0.017), internalized stigma (95% CI [0.83, 0.98], p=0.018) and knowledge of positive ART effects for others (95% CI [0.84, 0.93], p=<0.001).ConclusionsUnderstanding the underlying factors associated with ART readiness among young people can inform strategies to support and increase individuals'' readiness to initiate ART and early engagement in care.     

Characterization of binding and biological effects of monoclonal antibodies against a human tumor necrosis factor receptor

Three different antibodies against a human TNF receptor (htr-1, htr-5, and htr-9) have been examined for their binding pattern to U937 cells and ability to mimic TNF-alpha activity in U937 cells, Fs4 fibroblasts, and human endothelial cells. Flow cytometric analysis revealed that htr-5 and htr-9 bound specifically to a TNF receptor on U937 cells that could be blocked by pretreatment with rTNF-alpha. Pretreatment of U937 cells with rTNF-beta blocked the binding of htr-9, but to a lesser extent htr-5 binding. Pretreatment with htr-5 inhibited the binding of htr-9 to U937 cells while pretreatment with htr-9 did not inhibit htr-5 binding. These results indicate that htr-5 and htr-9 recognize distinct but overlapping epitopes of a human TNF receptor on U937 cells and that htr-5 may be close to a TNF-alpha-specific domain of the binding site. Pretreatment with htr-5 or htr-9 only minimally reduced binding of BrTNF-alpha to U937 cells; however, these antibodies were much more effective in inhibiting BrTNF-alpha binding to HL-60 cells. Furthermore, it was found that htr-1 and htr-9, but not htr-5, had TNF-alpha activity on U937 cells, Fs4 fibroblasts, and endothelial cells and that the TNF-alpha activity induced by htr-9 was completely inhibited by htr-5. However, the cytotoxic activity of TNF-alpha was only partially inhibited by htr-5 on U937 cells while htr-5 had no effect on TNF-alpha activity on Fs4 cells. The data suggest that a common epitope is involved in inducing TNF-alpha activity in three different cell systems.     

Inverse MCP-1/IL-8 ration in effluents of CAPD patients with peritonitis and in isolated cultured human peritoneal macrophages

An important event in intraperitoneal inflammation is the influx of leukocytes into the peritoneal cavity. Chemokines such as interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) play a major role in the recruitment of immune cells to the site of inflammation. We determined the concentrations of two members of the chemokine family, IL-8 and MCP-1, in the dialysate effluents of 18 continuous ambulatory peritoneal dialysis (CAPD) patients with peritonitis and of 18 non-infected CAPD patients by specific enzyme-linked immunosorbent assays (ELISA). Isolated peritoneal macrophages (PMs) from CAPD peritonitis patients were cultured and IL-8 and MCP-1 production was determined on protein (ELISA) and mRNA level (Northern blot) at designated timepoints over a 72-h culture period. PMs from non-infected patients served as controls. Much higher concentrations of IL-8 and MCP-1 were found in dialysate effluents of peritonitis patients than in effluents of non-infected patients: IL-8 2.39&plusnn;1.15 vs 0.05±0.01 ng/ml and MCP-1 release by cultured PMs from peritonitis patients and non-infected patients revealed significant differences: IL-8 40.3±2.2 ng/ml after 3 h and 194.2 ±34.9 ng/ml after 12 h compared to 21.02±6.15 ng/ml after 3 h and 89.64±30.28 ng/ml after 12 h, respectively; MCP-1 3.3±0.9 ng/ml after 3 h and 25.7±7.4 ng/ml after 12 h compared to 1.1±0.2 ng/ml and 1.8±0.2 ng/ml, respectively. Interestingly, the ratio of IL-8 to MCP-1 concentrations in the dialysate effluents (1:9.4) is reversed in the supernatants of cultured PMs. In the effluents and in the culture supernatants of PMs from CAPD peritonitis patients high amounts of IL-8 and MCP-1 are detectable, suggesting that PMs are an important source for these chemokines during peritonitis. Because of the inverse ratio of IL-8 and MCP-1 in the effluents and culture supernatants it can be assumed that PMs are responsible for the MCP-1 concentration to a lesser extent than for the IL-8 concentration in the effluents.     

A skin-selective homing mechanism for human immune surveillance T cells

Effective immune surveillance is essential for maintaining protection and homeostasis of peripheral tissues. However, mechanisms controlling memory T cell migration to peripheral tissues such as the skin are poorly understood. Here, we show that the majority of human T cells in healthy skin express the chemokine receptor CCR8 and respond to its selective ligand I-309/CCL1. These CCR8(+) T cells are absent in small intestine and colon tissue, and are extremely rare in peripheral blood, suggesting healthy skin as their physiological target site. Cutaneous CCR8(+) T cells are preactivated and secrete proinflammatory cytokines such as tumor necrosis factor-alpha and interferon-gamma, but lack markers of cytolytic T cells. Secretion of interleukin (IL)-4, IL-10, and transforming growth factor-beta was low to undetectable, arguing against a strict association of CCR8 expression with either T helper cell 2 or regulatory T cell subsets. Potential precursors of skin surveillance T cells in peripheral blood may correspond to the minor subset of CCR8(+)CD25(-) T cells. Importantly, CCL1 is constitutively expressed at strategic cutaneous locations, including dermal microvessels and epidermal antigen-presenting cells. For the first time, these findings define a chemokine system for homeostatic T cell traffic in normal human skin.     

Different strategies of mixed chimerism induction may determine stem/progenitor cell populations in recipient mice

Mixed chimerism has been suggested to induce tolerance to transplanted alloantigens. As the precise influence of mixed chimerism induction on the host organism has still not been fully elucidated, the aim of the present study was to explore this phenomenon in relation to the stem cell compartment.The experiment was performed on B6.SJL-Ptprc^aPep3^b mice. Mixed chimerism induction protocols involved 3 Gy TBI (Day − 1 of the experiment), injection of 20-30 × 10⁶ Balb C bone marrow cells (Day 0), and administration of blocking antibodies against CD40L (Day 0 and Day 4), anti-CD8 (Day − 2) with/without anti-NK1.1 (Day − 3). Selected groups of mice were also treated with cyclophosphamid (175 mg/kg) on Day 2. The presence of mixed chimerism was assessed in peripheral blood, bone marrow, and spleen, as well as in various subpopulations of leukocytes (CD4⁺, CD8⁺, CD45/B220⁺, Gr-1⁺, lin⁻/Sca-1⁺/c-kit⁻, lin⁻/Sca-1⁺/c-kit⁺, lin⁻/Sca-1⁻/c-kit⁺). Furthermore, the percentage of stem/progenitor cells (lin⁻/Sca-1⁺/c-kit⁻, lin⁻/Sca-1⁺/c-kit⁺, lin⁻/Sca-1⁻/c-kit⁺, VSEL, HSC) was analysed for the first time in bone marrow and peripheral blood of chimeric mice.The range of mixed chimerism differed significantly among various cell populations: it was lowest in CD8-positive cells and lin⁻/Sca-1⁺/c-kit⁻ cells, and highest in granulocytes. The induction of mixed chimerism revealed a significant impact on the stem/progenitor cell frequency in recipient mice, providing potential therapeutic insights into the long-term immunologic tolerance observed in chimeric mice. Collectively, these findings contribute to further optimization of mixed chimerism induction protocols and might help in the introduction of this phenomenon into clinical practice.