The impact of a native hemiparasite on a major invasive shrub is affected by host size at time of infection.

Few studies have examined how parasite impact is affected by host size. In a glasshouse experiment, we investigated the impact of the Australian native hemiparasitic vine, Cassytha pubescens, on a major invasive shrub, Ulex europaeus, of different sizes. Infected plants had significantly lower total, shoot and root biomass, but the parasite's impact was more severe on small than large hosts. When infected small but not large hosts had significantly lower nodule biomass. Irrespective of size, infection significantly decreased host shoot/root ratio, predawn and midday quantum yields, maximum electron transport rates and carbon isotope composition, and host nodule biomass g-1 root biomass significantly increased in response to infection. Infection did not affect host foliar nitrogen concentration or midday shoot water potential. Parasite biomass was significantly lower on small relative to large hosts, but was similar g-1 host total biomass. Parasite stem nitrogen, phosphorous and potassium concentration were significantly greater when C. pubescens was growing on small than large hosts. Our results clearly show that C. pubescens strongly decreases performance of this major invasive shrub, especially when hosts are small. This suggests that C. pubescens could be used most effectively as a native biocontrol when deployed on smaller hosts

High water availability increases the negative impact of a native hemiparasite on its non-native host

© 2015 The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology. Environmental factors alter the impacts of parasitic plants on their hosts. However, there have been no controlled studies on how water availability modulates stem hemiparasites' effects on hosts. A glasshouse experiment was conducted to investigate the association between the Australian native stem hemiparasite Cassytha pubescens and the introduced host Ulex europaeus under high (HW) and low (LW) water supply. Cassytha pubescens had a significant, negative effect on the total biomass of U. europaeus, which was more severe in HW than LW. Regardless of watering treatment, infection significantly decreased shoot and root biomass, nodule biomass, nodule biomass per unit root biomass, F v/F m, and nitrogen concentration of U. europaeus. Host spine sodium concentration significantly increased in response to infection in LW but not HW conditions. Host water potential was significantly higher in HW than in LW, which may have allowed the parasite to maintain higher stomatal conductances in HW. In support of this, the δ13C of the parasite was significantly lower in HW than in LW (and significantly higher than the host). C. pubescens also had significantly higher F v/F m and 66% higher biomass per unit host in the HW compared with the LW treatment. The data suggest that the enhanced performance of C. pubescens in HW resulted in higher parasite growth rates and thus a larger demand for resources from the host, leading to poorer host performance in HW compared with LW. C. pubescens should more negatively affect U. europaeus growth under wet conditions rather than under dry conditions in the field

A native parasitic plant affects the performance of an introduced host regardless of environmental variation across field sites

Increasing evidence from glasshouse studies shows that native hemiparasitic plants can significantly impact the performance and growth of introduced host plants. We investigated the effect of the native Australian hemiparasite Cassytha pubescens R.Br. on the introduced shrub Ulex europaeus L. at three field sites in South Australia. Parasite infection significantly decreased midday PSII efficiency (ΦPSII) and the maximum electron transport rates (ETRmax) of U. europaeus across sites. The impact of C. pubescens on the photosynthetic performance of U. europaeus may have been caused by infected plants having significantly lower N and K, but higher Fe and Al than uninfected plants at all sites. Significant Al and Fe enrichment in infected plants may be possibly due to the parasite indirectly inducing rhizosphere acidification. At two sites, C. pubescens significantly affected host Fv/Fm, indicating chronic photoinhibition in response to infection. The impact of infection on Fv/Fmwas greatest at the wettest site, in line with an experiment where C. pubescens had more impact under high water availability. At this site, infected plants also had the highest foliar Fe and Al. The C isotope (δ13C) of infected plants was significantly lower than that of uninfected plants at only one site. Unusually, the δ13C of the parasite was the same as or significantly higher than that of the hosts. There were no site effects on parasite Fv/Fmor ΦPSII; however, ETRmaxand δ13C varied across sites. The results suggest that this native parasite has negative effects on U. europaeus in the field, as was found for glasshouse studies. The abundance of this introduced weed in Australia could be negatively affected by C. pubescens infection

Differentiation of regulatory myeloid and T-cells from adult human hematopoietic stem cells after allogeneic stimulation

IntroductionDonor hematopoietic stem cell (DHSC) infusions are increasingly being studied in transplant patients for tolerance induction.MethodsTo analyze the fate of infused DHSCs in patients, we developed an in vitro culture system utilizing CD34+DHSCs stimulated with irradiated allogeneic cells in cytokine supplemented medium long-term.ResultsFlow cytometric analyses revealed loss of the CD34 marker and an increase in CD33+ myeloid and CD3+ T-cell proportion by 10.4% and 72.7%, respectively, after 21 days in culture. T-cells primarily expressed TcR-αβ and were of both CD4+ and CD8+ subsets. Approximately 80% of CD3+ T cells lacked expression of the co-stimulatory receptor CD28. The CD4+ compartment was predominated by CD4+CD25+CD127-FOXP3+ Tregs (>50% CD4+CD127- compartment) with <1% of all leukocytes exhibiting a CD4+CD127+ phenotype. Molecular analyses for T-cell receptor excision circles showed recent and increased numbers of TcR rearrangements in generated T cells over time suggesting de novo differentiation from DHSCs. CD33+ myeloid cells mostly expressed HLA-DR, but lacked expression of co-stimulatory receptors CD80 and CD83. When studied as modulators in primary mixed lymphocyte reactions where the cells used to stimulate the DHSC were used as responders, the DHSC-lines and their purified CD8+, CD4+, CD33+ and linage negative subsets inhibited the responses in a dose-dependent and non-specific fashion. The CD8+ cell-mediated inhibition was due to direct lysis of responder cells.DiscussionExtrapolation of these results into the clinical situation would suggest that DHSC infusions into transplant recipients may generate multiple subsets of donor “chimeric” cells and promote recipient Treg development that could regulate the anti-donor immune response in the periphery. These studies have also indicated that T cell maturation can occur in vitro in response to allogeneic stimulation without the pre-requisite of a thymic-like environment or NOTCH signaling stimulatory cell line

Does nitrogen affect the interaction between a native hemiparasite and its native or introduced leguminous hosts?

© 2016 The Authors. New Phytologist © 2016 New Phytologist Trust Associations between plants and nitrogen (N)-fixing rhizobia intensify with decreasing N supply and come at a carbon cost to the host. However, what additional impact parasitic plants have on their leguminous hosts’ carbon budget in terms of effects on host physiology and growth is unknown. Under glasshouse conditions, Ulex europaeus and Acacia paradoxa either uninfected or infected with the hemiparasite Cassytha pubescens were supplied (high nitrogen (HN)) or not (low nitrogen (LN)) with extra N. The photosynthetic performance and growth of the association were measured. Cassytha pubescens significantly reduced the maximum electron transport rates and total biomass of U. europaeus but not those of A. paradoxa, regardless of N. Infection significantly decreased the root biomass of A. paradoxa only at LN, while the significant negative effect of infection on roots of U. europaeus was less severe at LN. Infection had a significant negative impact on host nodule biomass. Ulex europaeus supported significantly greater parasite biomass (also per unit host biomass) than A. paradoxa, regardless of N. We concluded that rhizobia do not influence the effect of a native parasite on overall growth of leguminous hosts. Our results suggest that C. pubescens will have a strong impact on U. europaeus but not A. paradoxa, regardless of N in the field

Immune responses and their regulation by donor bone marrow cells in clinical organ transplantation

Infusions of donor bone marrow derived cells (DBMC) continue to be tested in clinical protocols intended to induce specific immunologic tolerance of solid organ transplants based on the observations that donor-specific tolerance is induced this way in animal models. We studied the immunological effects of human DBMC infusions in renal transplantation using modifications in lymphoproliferation (MLR) and cytotoxicity (CML) assays. The salient observations and tentative conclusions are summarized in this review. Among many types of organs transplanted using DBMC at this center, it was found that the cadaver renal recipients (CAD) had significantly decreased chronic rejection and higher graft survival when compared to equivalent non-infused controls. DBMC infusion was also associated with a marginal and non-specific immune depression. It was also observed that the number of chimeric donor cells gradually increased in the iliac crest bone marrow compartment with a concomitant decrease in the peripheral blood and that the increase was more rapid in living-related donor (LRD)-kidney/DBMC recipients in spite of a lower number of DBMC infused (<25%) than in the CAD-kidney/DBMC group. In the LRD recipients with residual anti-donor responses, purified chimeric cells of either donor or recipient inhibited recipient immune responses to the donor significantly more strongly than the freshly obtained bone marrow from the specific donor or volunteer suggesting an active regulatory role for chimeric cells. A number of (non-chimeric) subpopulations of bone marrow cells including CD34 + stem cells and the CD34 − early progeny like CD38 +, CD2 +, CD5 + and CD1 + lymphoid cells as well as CD33 + (but CD15 −) myeloid cells down-regulated the MLR and CML responses of allogeneic PBMC stimulated with (autologous) donor spleen cells. These regulatory effects appeared to be refractory to the action of commonly used immunosuppressive drugs and occurred during the early phase of the immune response through cell–cell interactions. Most of these DBMC sub-populations had stimulatory capabilities, albeit markedly lower than donor spleen cells, but only through the indirect antigen presentation pathway. When co-cultured with allogeneic stimulators, purified CD34 + cells were found to give rise both to CD3 − TCR αβ +, as well as CD3 + TCR αβ + cells and, thereby, responded in MLR to allogeneic stimulation (but did not generate cytotoxic effector cells). Also, a number of DBMC subpopulations inhibited the CML and to a lesser extent the MLR, of autologous post-thymic responding T cells stimulated with allogeneic irradiated cells, mediated through soluble factors. Finally, non-chimeric DBMC also inhibited the proliferative and cytotoxic responses of autologous T cells to EBV antigens, inducing T suppressor cells, which in turn could inhibit autologous anti-EBV CTL generation and B cell anti-CMV antibody production. These studies all suggested a strong inhibitory property of a number of DBMC sub-populations in vitro and in vivo with the notion that they promote unresponsiveness

Inhibition of NF-κB during human dendritic cell differentiation generates anergy and regulatory T-cell activity for one but not two human leukocyte antigen DR mismatches

We examined the in vitro inhibition of human monocyte-derived dendritic cells (DC) maturation via NF-κB blockade on T-cell allostimulation, cytokine production, and regulatory T-cell generation. DC were generated from CD14 + monocytes isolated from peripheral blood using GM-CSF and IL-4 for differentiation and TNF-α, IL-1β, and PGE 2 as maturational stimuli with or without the NF-κB inhibitors, BAY 11-7082 (BAY-DC) or Aspirin (ASA-DC). Stimulator and responder cells were one versus two HLA-DR mismatched in direct versus indirect presentation assays. Both BAY-DC and ASA-DC expressed high levels of HLA-DR and CD86 but always expressed less CD40 compared with controls. Some experiments showed slightly lower levels of CD80. Both BAY- and ASA- allogeneic DC and autologous alloantigen pulsed DC were weaker stimulators of T cells (by MLR) compared with controls, and there was reduced IL-2 and IFN-γ production by T cells stimulated with BAY-DC or ASA-DC (by ELISPOT) (more marked results were always observed with ASA-treated DC). In addition, NF-κB blockade of DC maturation caused the generation of T cells with regulatory function (T regs) but only when T cells were stimulated by either allogeneic (direct presentation) or alloantigen pulsed autologous DC (indirect presentation) with one HLA-DR mismatch and not with two HLA-DR mismatches (either direct or indirect presentation). However, the T regs generated from these ASA-DC showed similar FoxP3 mRNA expression to those from nontreated DC. Extension of this study to human organ transplantation suggests potential therapies using one DR-matched NF-κB blocked DC to help generate clinical tolerance