Efficient activation of T cells by human monocyte-derived dendritic cells (HMDCs) pulsed with Coxiella burnetii outer membrane protein Com1 but not by HspB-pulsed HMDCs

<p>Abstract</p> <p>Background</p> <p><it>Coxiella burnetii </it>is an obligate intracellular bacterium and the etiologic agent of Q fever; both coxiella outer membrane protein 1 (Com1) and heat shock protein B (HspB) are its major immunodominant antigens. It is not clear whether Com1 and HspB have the ability to mount immune responses against <it>C. burnetii </it>infection.</p> <p>Results</p> <p>The recombinant proteins Com1 and HspB were applied to pulse human monocyte-derived dendritic cells (HMDCs), and the pulsed HMDCs were used to stimulate isogenic T cells. Com1-pulsed HMDCs expressed substantially higher levels of surface molecules (CD83, CD40, CD80, CD86, CD54, and CD58) and a higher level of interleukin-12 than HspB-pulsed HMDCs. Moreover, Com1-pulsed HMDCs induced high-level proliferation and activation of CD4⁺and CD8⁺cells, which expressed high levels of T-cell activation marker CD69 and inflammatory cytokines IFN-γ and TNF-α. In contrast, HspB-pulsed HMDCs were unable to induce efficient T-cell proliferation and activation.</p> <p>Conclusions</p> <p>Our results demonstrate that Com1-pulsed HMDCs are able to induce efficient T-cell proliferation and drive T cells toward Th1 and Tc1 polarization; however, HspB-pulsed HMDCs are unable to do so. Unlike HspB, Com1 is a protective antigen, which was demonstrated by the adoptive transfer of Com1-pulsed bone marrow dendritic cells into naive BALB/c mice.</p

HSPB1, HSPB6, HSPB7 and HSPB8 Protect against RhoA GTPase-Induced Remodeling in Tachypaced Atrial Myocytes

BACKGROUND: We previously demonstrated the small heat shock protein, HSPB1, to prevent tachycardia remodeling in in vitro and in vivo models for Atrial Fibrillation (AF). To gain insight into its mechanism of action, we examined the protective effect of all 10 members of the HSPB family on tachycardia remodeling. Furthermore, modulating effects of HSPB on RhoA GTPase activity and F-actin stress fiber formation were examined, as this pathway was found of prime importance in tachycardia remodeling events and the initiation of AF. METHODS AND RESULTS: Tachypacing (4 Hz) of HL-1 atrial myocytes significantly and progressively reduced the amplitude of Ca²⁺ transients (CaT). In addition to HSPB1, also overexpression of HSPB6, HSPB7 and HSPB8 protected against tachypacing-induced CaT reduction. The protective effect was independent of HSPB1. Moreover, tachypacing induced RhoA GTPase activity and caused F-actin stress fiber formation. The ROCK inhibitor Y27632 significantly prevented tachypacing-induced F-actin formation and CaT reductions, showing that RhoA activation is required for remodeling. Although all protective HSPB members prevented the formation of F-actin stress fibers, their mode of action differs. Whilst HSPB1, HSPB6 and HSPB7 acted via direct prevention of F-actin formation, HSPB8-protection was mediated via inhibition of RhoA GTPase activity. CONCLUSION: Overexpression of HSPB1, as well as HSPB6, HSPB7 and HSPB8 independently protect against tachycardia remodeling by attenuation of the RhoA GTPase pathway at different levels. The cardioprotective role for multiple HSPB members indicate a possible therapeutic benefit of compounds able to boost the expression of single or multiple members of the HSPB family

Thermal Control of Engineered T-cells

Genetically engineered T-cells are being developed to perform a variety of therapeutic functions. However, no robust mechanisms exist to externally control the activity of T-cells at specific locations within the body. Such spatiotemporal control could help mitigate potential off-target toxicity due to incomplete molecular specificity in applications such as T-cell immunotherapy against solid tumors. Temperature is a versatile external control signal that can be delivered to target tissues in vivo using techniques such as focused ultrasound and magnetic hyperthermia. Here, we test the ability of heat shock promoters to mediate thermal actuation of genetic circuits in primary human T-cells in the well-tolerated temperature range of 37–42 °C, and introduce genetic architectures enabling the tuning of the amplitude and duration of thermal activation. We demonstrate the use of these circuits to control the expression of chimeric antigen receptors and cytokines, and the killing of target tumor cells. This technology provides a critical tool to direct the activity of T-cells after they are deployed inside the body

Spoilage indicator bacteria in farmed Atlantic salmon (Salmo salar) stored on ice for 10 days

peer-reviewedThis study investigated the growth of indicator and spoilage bacteria on whole Atlantic salmon (Salmo salar) stored aerobically at 2 °C. On days 0, 2, 3, 6, 8 and 10 microbiological analysis was carried out on inner flesh and outer skin samples as well as outer skin swabs (25 cm2 surface areas). Mesophilic total viable counts (TVCm) on skin, flesh and swab samples increased from 1.9, 1.1 and 2.7 log10 CFUcm2 to 6.0, 5.1 and 5.7 log10 CFU/cm2 after 10 days, respectively. Psychrotrophic counts (TVCp), increased from 2.2, 1.8 and 3.1 log10 CFU/cm2 to 6.2, 5.3 and 5.9 log10 CFU/cm2, for skin, flesh and swab samples respectively. Hydrogen sulphide producing bacteria (HSPB), lactic acid bacteria (LAB), Pseudomonas spp., Brochothrix thermosphacta and Photobacterium spp. grew well with similar growth rates (mean generation times of 17.2–26 h). It was concluded that the shelf-life of salmon at 2 °C was approximately 10 days and that HSPB, LAB, Pseudomonas spp., Br. thermosphacta and Photobacterium spp. may be a better indicator of fish spoilage rather than TVC growth, with a count of 5–6 log10 CFU/cm2 indicating the end of shelf-life

The Effect of Organic Acid, Trisodium Phosphate and Essential Oil Component Immersion Treatments on the Microbiology of Cod (Gadus morhua) during Chilled Storage

peer-reviewedSpoilage is a major issue for the seafood sector with the sale and exportation of fish limited by their short shelf-life. The immediate and storage effects of immersion (30 s at 20 °C) with 5% (w/v) citric acid (CA), 5% (v/v) lactic acid (LA), 5% (w/v) capric acid (CP) and 12% trisodium phosphate (TSP) (experiment 1) and essential oil components (EOC) (1% (v/v) citral (CIT), 1% (v/v) carvacrol (CAR), 1% (w/v) thymol (THY) and 1% (v/v) eugenol (EUG)) (experiment 2) on the concentrations of indicator (total viable counts (TVC) (mesophilic and psychrophilic) and total Enterobacteriaceae counts (TEC)), and spoilage organisms (Pseudomonas spp., lactic acid bacteria (LAB), Brochothrix thermosphacta, Photobacterium spp. and hydrogen sulphide producing bacteria (HSPB)) on cod (Gadus morhua) (stored aerobically at 2 °C) was investigated. There was no significant reduction for most treatment-bacteria combinations, with the following exceptions; TSP and TVCm (time t = 6), TSP and TVCp (t = 6), CP and LAB (t = 6, 8 and 10), CP and Br. thermosphacta (t = 4, 6, 8, 10, 14 and 16), TSP and Photobacterium spp. (t = 4), CAR and Br. thermosphacta (t = 6) and CAR and HSPB (t = 3, 6, 9, 12, 15 and 18). Although the majority of treatments did not significantly (P > 0.05) reduce bacterial counts, the limited success with CP and CAR warrants further investigation