Dense granule protein 3 of Toxoplasma gondii plays a crucial role in the capability of the tissue cysts of the parasite to persist in the presence of anti-cyst CD8+ T cells during the chronic stage of infection.

Toxoplasma gondii establishes chronic infection by forming tissue cysts, and this chronic infection is one of the most common parasitic infections in humans. Our recent studies revealed that whereas CD8+ T cells of genetically resistant BALB/c mice have the capability to remove the tissue cysts of the parasite through their perforin-mediated activities, small portions of the cysts are capable of persisting in the presence of the anti-cyst CD8+ T cells. It is currently unknown how those small portions of the cysts resist or escape the T-cell immunity and persist in the hosts. In the present study, we discovered that the cysts, which persisted in the presence of the perforin-mediated CD8+ T-cell immunity, have significantly greater mRNA levels for four dense granule proteins, GRA1, GRA2, GRA3, and GRA7, and one rhoptry protein, ROP35, than the total population of the cysts present in the absence of the T cells. In addition, increased levels of mRNA for GRA1, GRA3, and ROP35 in the cysts significantly correlated with their successful persistence through the condition in which greater degrees of reduction of the cyst burden occurred through anti-cyst CD8+ T cells. In addition, GRA3-deficient T. gondii displayed significantly enhanced elimination of the cysts by anti-cyst CD8+ T cells when compared to the wild-type parasite. These results indicate that GRA3 is a key molecule that mediates in the capability of T. gondii cysts to persist by resisting or evading the anti-cyst activity of CD8+ T cells during the later stage of infection.

IFN-γ production by brain-resident cells activates cerebral mRNA expression of a wide spectrum of molecules critical for both innate and T cell-mediated protective immunity to control reactivation of chronic infection with Toxoplasma gondii.

We previously demonstrated that brain-resident cells produce IFN-γ in response to reactivation of cerebral infection with Toxoplasma gondii. To obtain an overall landscape view of the effects of IFN-γ from brain-resident cells on the cerebral protective immunity, in the present study we employed NanoString nCounter assay and quantified mRNA levels for 734 genes in myeloid immunity in the brains of T and B cell-deficient, bone marrow chimeric mice with and without IFN-γ production by brain-resident cells in response to reactivation of cerebral T. gondii infection. Our study revealed that IFN-γ produced by brain-resident cells amplified mRNA expression for the molecules to activate the protective innate immunity including 1) chemokines for recruitment of microglia and macrophages (CCL8 and CXCL12) and 2) the molecules for activating those phagocytes (IL-18, TLRs, NOD1, and CD40) for killing tachyzoites. Importantly, IFN-γ produced by brain-resident cells also upregulated cerebral expression of molecules for facilitating the protective T cell immunity, which include the molecules for 1) recruiting effector T cells (CXCL9, CXCL10, and CXCL11), 2) antigen processing (PA28αß, LMP2, and LMP7), transporting the processed peptides (TAP1 and TAP2), assembling the transported peptides to the MHC class I molecules (Tapasin), and the MHC class I (H2-K1 and H2-D1) and Ib molecules (H2-Q1, H-2Q2, and H2-M3) for presenting antigens to activate the recruited CD8+ T cells, 3) MHC class II molecules (H2-Aa, H2-Ab1, H2-Eb1, H2-Ea-ps, H2-DMa, H2-Ob, and CD74) to present antigens for CD4+ T cell activation, 4) co-stimulatory molecules (ICOSL) for T cell activation, and 5) cytokines (IL-12, IL-15, and IL-18) facilitating IFN-γ production by NK and T cells. Notably, the present study also revealed that IFN-γ production by brain-resident cells also upregulates cerebral expressions of mRNA for the downregulatory molecules (IL-10, STAT3, SOCS1, CD274 [PD-L1], IL-27, and CD36), which can prevent overly stimulated IFN-γ-mediated pro-inflammatory responses and tissue damages. Thus, the present study uncovered the previously unrecognized the capability of IFN-γ production by brain-resident cells to upregulate expressions of a wide spectrum of molecules for coordinating both innate and T cell-mediated protective immunity with a fine-tuning regulation system to effectively control cerebral infection with T. gondii.

Effect of Si, Mn, Be and Sr Addition on the Tensile Properties of 6061 Type Alloys: Role of Aging Treatment.

The present study was performed on a 6061-type alloy to examine the effects of minor additions (Si, Mn, Be, Sr) of the type of precipitated Fe-based intermetallics, in terms of Fe/Si ratios. All alloys were grain refined (0.15%Ti in the form of Al-5%Ti-1%B) to minimize hot tearing during casting. The effect of these intermetallics on the alloy tensile properties was also investigated. Tensile test bars were solutionized at 520 °C followed by quenching in warm water at 60 °C to avoid cracking. The quenched bars were aged at 175 °C for periods up to 100 h. Characterization of the formed intermetallics as well as phase precipitation were carried out using field emission scanning electron microscopy. In Be-treated alloys, α-Al8Fe2SiBe phase may precipitate along with α-Al15(Fe, Mn)3Si2 phase. In addition, Be results in fragmentation of the α-Fe phase when the alloy was Sr-modified, leading to better tensile properties, compared to those obtained from the base alloy under same conditions. It should be noted that this study does not promote the use of Be as it is a toxic element.

The Role of Mg Content and Aging Treatment on the Tensile and Fatigue Properties of Die-Cast 380 Alloy.

The main objective of this contribution was to determine the impact of magnesium (Mg) concentration and solidification rate (about 800 °C/s) on the mechanical properties of commercial A380.1 die-cast alloy. Respective amounts of 0.10%, 0.30%, and 0.50% Mg were used to establish their influence on the main tensile properties, namely, the ultimate limit, the elastic limit, and the percentage of elongation to fracture. The study also focused on the effect of magnesium on the fatigue behavior of A380.1 alloy where the role of surface defects and internal defects (porosity, oxide films, and inclusions) on the alloy fatigue life was also determined. The tensile properties were analyzed in order to optimize the heat treatments of T6 (under-aging) and T7 (over-aging). Consequently, the influence of several parameters was evaluated using tensile testing and optical and scanning electron micrography. Fatigue strength was investigated by performing rotational bending tests. The results show that the alloy tensile strength parameters improve with up to 0.3% Mg. Further addition of Mg, i.e., 0.5%, does not produce any significant improvement with respect to either traction or fatigue. It is observed that the tensile properties fluctuate according to the Guinier-Preston zones which occur during heat treatment, while the fatigue properties decrease as the Mg content increases. In contrast to a mechanical fatigue failure mechanism, in the present study, cracks were initiated at the sample's outer surface and then propagated toward the center.

Human MHC class I molecule, HLA-A2.1, mediates activation of CD8+ T cell IFN-γ production and the T cell-dependent protection against reactivation of cerebral Toxoplasma infection.

To examine whether the HLA-A2.1, one of the most common MHC class I molecules in humans, activates the protective immunity against reactivation of cerebral infection with Toxoplasma gondii, HLA-A2.1-transgenic and wild-type (WT) mice were infected and treated with sulfadiazine to establish chronic infection in their brains. One month after discontinuation of sulfadiazine, which initiates reactivation of the infection, mRNA levels for tachyzoite (the acute stage form)-specific SAG1 and numbers of the foci associated tachyzoites were significantly less in the brains of the HLA-A2.1-transgenic than WT mice. Greater numbers of IFN-γ-producing CD8+ T cells were detected in the spleens of infected transgenic than WT mice, and CD8+ T cells from the former produced markedly greater amounts of IFN-γ than the T cells from the latter in response to tachyzoite antigens in vitro. When their CD8+ T cells were systemically transferred to infected immunodeficient NSG mice expressing the HLA-A2.1, the CD8+ T cells from HLA-A2.1-transgenic mice inhibited reactivation of the cerebral infection in the recipients more efficiently than did the WT T cells. Furthermore, the inhibition of reactivation of the infection by CD8+ T cells from the transgenic mice was associated with increased cerebral expression of IFN-γ and effector molecules against tachyzoites in the recipients when compared to the WT CD8+ T cell recipients. Thus, the human HLA-A2.1 is able to effectively activate IFN-γ production of CD8+ T cells against T. gondii tachyzoites and confer a potent protection against reactivation of cerebral infection with this parasite through the CD8+ T cells activation.

Mechanical Performance and Precipitation Behavior in Al-Si-Cu-Mg Cast Alloys: Effect of Prolonged Thermal Exposure.

Al-Si-Cu-Mg cast (354) alloys are used in the automotive sector owing to their remarkable properties which are achievable after applying appropriate thermal treatments. Zirconium, Nickel, and Manganese were added to this category of Al-alloys to preserve good mechanical properties while being exposed to elevated temperatures for long times. The ultimate and yield strength values obtained at room temperature for the stabilized (thermally-exposed) T5-treated condition were comparable to those of the stabilized T6-treated condition, whereas the same properties for T5-treated alloys were higher than those of T6-treated ones for elevated-temperature tensile testing. Interestingly, the results showed that the addition of 0.75 wt.% Mn was competitive with the addition of 2 and 4 wt.% Ni with respect to the elevated-temperature and ambient temperature strength values, respectively. In addition, the Mn-containing alloy M3S exhibited improved ductility values at ambient temperature and at 250 °C, compared to the Ni-containing alloys. Examination of the fracture surface of tested samples revealed the advantageous role of sludge particles in enhancing the performance of Mn-containing alloys through their resistance to the propagation of cracks that developed in many intermetallic phases. This finding is considered to be economically significant in view of the lower price of manganese compared to that of nickel.

Deficiency in indoleamine-2, 3-dioxygenase induces upregulation of guanylate binding protein 1 and inducible nitric oxide synthase expression in the brain during cerebral infection with Toxoplasma gondii in genetically resistant BALB/c mice but not in genetically susceptible C57BL/6 mice.

We examined the roles of indoleamine-2, 3-dioxygenase 1 (IDO1) in controlling cerebral Toxoplasma gondii infection in both genetically resistant and susceptible strains of mice. In susceptible C57BL/6 mice, IDO expression was immunohistochemically detected only in a minority (22.5%) of tachyzoite-infected cells in their brains during the later stage of infection. When C57BL-6-background IDO1-deficient (IDO1-/-) mice were infected, their cerebral tachyzoite burden was equivalent to those of wild-type (WT) animals. In contrast, in resistant BALB/c mice, IDO expression was detected in a majority (84.0%) of tachyzoite-infected cerebral cells. However, tachyzoite burden in BALB/c-background IDO1-/- mice remained as low as that of WT mice, which was 78 times less than those of C57BL/6 mice. Of interest, IDO1-/- mice of only resistant BALB/c-background had markedly greater cerebral expressions of two other IFN-γ-mediated effector molecules, guanylate binding protein 1 (Gbp1) and nitric oxide synthase 2 (NOS2), than their WT mice. Therefore, it would be possible that IDO1 deficiency was effectively compensated by the upregulated expression of Gbp1 and NOS2 to control cerebral tachyzoite growth in genetically resistant BALB/c mice, whereas IDO1 did not significantly contribute to controlling cerebral tachyzoite growth in genetically susceptible C57BL/6 mice because of its suppressed expression in infected cells.