The aims of this study were
to assess the role of Nrf2 in rosuvastatin-mediated antioxidant effects in endothelial cells and to further elucidate the molecular mechanisms of renoprotective effect of rosuvastatin treatment. Methods: Wild type (WT) and Akita diabetic mice (AKITA) were treated with RSV for 4 weeks. Urinary albumin Cilomilast clinical trial excretion and renal histology were examined. Nrf2-antioxidant response element (ARE) activity was measured in human umbilical vein endothelial cell (HUVEC) with luciferase assay after transfection of reporter plasmids containing AREs. The expression of Nrf2-regulated genes was also examined. Results: Increased urinary albumin excretion in AKITA mice was significantly reduced by RSV treatment. The amount of lectin-stained glomerular endothelial surface layer, important for permselectivity in the vascular wall, was significantly reduced in AKITA mice and preserved with RSV treatment. RSV significantly increased the transcriptional activity of the AREs and Stem Cell Compound Library subsequent expression of Nrf2-regulated genes in HUVEC. Additional experiments with cycloheximide and actinomycin D indicated that RSV extended the half-life of Nrf2 protein. Furthermore, RSV increased p21cip1 expression and thereby inhibited degradation of Nrf2 through direct binding of Nrf2 with p21cip1. Conclusion: These data indicate that rosuvastatin has anti-oxidative effects through activation of Nrf2, thereby restoring glomerular
endothelial function and preventing development of albuminuria in diabetes. FAN QIULING, PU SHI, LIU NAN, LV XIAOMENG, JIANG YI, WANG LINING Department of Nephrology, The First Hospital, China Medical University, Shenyang, China Introduction: To explore the pathogenesis and the biomarkers for early detection of diabetic nephropathy (DN), the circulating microRNA expression profile of DN patients was analyzed by AB Taqman human miRNA array. Methods: We
obtained serum samples from 5 diabetic nephropathy patients proven by renal biopsy as nodular diabetic glomerulosclerosis, 5 diabetic patients without microalbuminuria (DM) and 5 healthy BCKDHA controls (N). Serum miRNAs were analyzed with the TaqMan Low Density Array and then validated with a quantitative reverse-transcription PCR assay with 30 individual samples. Results: The urinary microalbumin/creatinine ratio and serum creatinine in diabetic nephropathy patients were higher than that of diabetic patients and healthy control (p < 0.05). 20 miRNAs were upregulated and 22 miRNAs were downregulated in serum of diabetic patients compared with that of healthy controls. 42 miRNAs were upregulated and 19 miRNAs were downregulated in serum of diabetic nephropathy patients compared with that of diabetic patients. Among them, along with the progression of diabetes and diabetic nephropathy, miR-1179 was gradually increased (2.03 times in DM/N and 2.14 times in DN/DM), miR-148b, miR-150 were gradually reduced (2.04 times in DM/N, 2.
 Serotonergic drugs, such as selective serotonin reuptake inhibitors (SSRIs) and serotonin noradrenaline reuptake inhibitors (SNRIs), are widely used to treat panic disorder and depression, and ameliorated OAB in selected patients. These drugs are thought to act on both efferent and afferent fibers from the bladder. On the other hand, brain corticotropin-releasing factor (CRF) has anxiogenic effects and increases
bladder sensation. Irritable bowel syndrome is highly prevalent in anxiety and mood disorders, and CRF receptor antagonists could ameliorate increased bowel sensation in those patients. selleck chemicals These findings suggest that increased bladder sensation can be a reflection of biological changes in both the emotion and micturition circuits within the brain. In contrast, the emotional mechanism
underlying the underactive/acontractile detrusor is not well understood. Neurogenic cases such as brain tumor and stroke[57, 58] and functional imaging studies[15, 16] have suggested that the cingulate cortex and insular cortex are the key areas for the generation of micturition impulses, which are sent to the brainstem structures. Therefore, functional changes in these areas might also occur in depressive/anxious patients with bladder Y-27632 nmr dysfunction. In somatoform disorders other than autonomic, functional neuroimaging studies have shown a decrease in the activity of frontal and subcortical circuits involved in motor control, and increases in the activities of supplementary motor area and midline regions for hysterical
motor paralysis.[59-61] However, in somatoform disorder of the bladder, no functional neuroimaging Ceramide glucosyltransferase studies are available. Serotonergic and GABAergic drugs are the mainstay in the treatment of depression/anxiety. What is the effect of these drugs on the bladder function? Central serotonergic neurons participate in a variety of physiological functions. Recent evidence has shown that centrally administered serotonin has modulatory effects on bladder function, the main actions of which are facilitation of urine storage.[52, 62] While inhibiting the bladder, serotonin facilitates sacral anterior horn cells innervating the urethra, presumably via inhibitory interneurons, leading to urethral contraction.[52, 63] Most central serotonin is physiologically released from nerve terminals of the brainstem raphe nucleus. There is a variety of micturition-related neuronal activity in the raphe nucleus, and microstimulation has been shown to elicit inhibition of the bladder. This effect might be due to activation of the raphe-spinal descending pathways, which in turn suppresses the sacral preganglionic neurons via inhibitory serotonin 1A receptors; it might also be due to suppression of the sensory afferent in the spinal posterior horn.
Background: CVD is the leading cause of mortality worldwide and cardiac troponins have been the cornerstone in the risk stratification of individuals with and without CVD. In a community-based population study, hsTropI may identify high-risk Erlotinib ic50 individuals several years prior to CVD-related mortality but this association using this newly established troponin assay has not been
validated in other population cohorts and it remains unclear whether this association is modified by baseline kidney function. Methods: This was a prospective observational study of 1,235 women over the age of 70 from the Calcium Intake Fracture Outcome Study. Baseline hsTropI was measured by immunoassay with level of detection of 4 ng/L. Association between hsTropI and 10-year risk of CVD hospitalisation/mortality was examined using Cox regression analysis. Results: Mean ± SD of CKD-EPI estimated glomerular filtration rate (eGFR) and hsTropI were 66.6.3 ± 13.3 mL/min/1.73 m2
and 6.8 ± 11.5 ng/L respectively. Less than 2% of participants had prevalent selleck products kidney disease. Above-median hsTropI was associated with a greater risk of CVD hospitalisation/mortality in the model adjusted for age, baseline eGFR, prevalent vascular and renal disease, diabetes and hypertension
(hazard ratio [HR] 1.56, 95%CI 1.17–2.09, P = 0.003). Baseline eGFR was an effect modifier between hsTropI and CVD hospitalisation/mortality (p-value for interaction 0.03). When stratified by eGFR < or ≥60 mL/min/1.73 m2, the association between above-median hsTropI and CVD hospitalisation/mortality was present only for participants with eGFR ≥60 mL/min/1.73 m2 (HR 1.73, 95%CI 1.16, 2.59, P = 0.007). Conclusions: The association between the newly established hsTropI and CVD hospitalisation/mortality may not be as robust in of elderly women with reduced kidney function but this finding requires confirmation in larger studies. 182 THE IMPACT OF ADVANCE CARE PLANNING FOR RENAL PATIENTS D MAWREN1, K DETERING1, D CHAFFERS1, S FRASER1, D POWER2, W SILVESTER1 1Respecting Patient Choices, Austin Health, Melbourne; 2Department of Nephrology, Austin Health, Melbourne, Australia Aim: To evaluate the impact of the introduction of ACP to the Austin Hospital renal unit. Background: Research indicates that renal patients are uninformed about care options and have limited knowledge about illness prognosis and trajectories.
In this case, downregulation of Drosha using the siRNA technique should increase titers of miRNA-encoding retroviral particles. To test this hypothesis, we first determined the amount of Drosha-specific siRNA required to efficiently downregulate the enzyme by transfecting Phoenix cells via the calcium phosphate method with synthetic siRNA against Drosha. In western blots, the signal for Drosha was already reduced with 50 pmol and barely detectable with 800 pmol siRNA (Supporting Information Fig. 2). Next, we co-transfected Phoenix cells with a retroviral expression vector encoding miR-106b and selleck chemicals llc with
200 pmol of Drosha siRNA or a control siRNA against luciferase. As expected, western blot analysis verified the successful downregulation of Drosha only in cultures that were co-transfected with siRNA against Drosha (Fig. 1B). As revealed by flow cytometry, frequencies
of GFP-positive cells were quite GPCR Compound Library cost similar in all transfected Phoenix cultures, ranging from 87 to 98% (Fig. 1C). Downregulation of Drosha did not lead to altered abundance of Dicer, the second RNaseIII enzyme needed to release mature miRNAs from the hairpin precursors. siRNA-mediated downregulation of Drosha in Phoenix cells should increase the amount of viral particles in the culture medium of cells transfected with retroviral constructs. As summarized in Supporting Information Fig. 3, this was indeed the case. More importantly, flow cytometry detected approximately Fossariinae 80% GFP-positive
cells in NIH3T3 cultures that were infected with retroviral supernatants of Phoenix cells co-transfected with pCLEP-106b and 200 pmol Drosha siRNA (Fig. 1C), similar to the frequency of GFP-positive cells in NIH3T3 cultures infected with the empty control virus. In contrast, only 32 and 47% of NIH3T3 cells could be infected with miR-106b virus from Phoenix cells transfected without Drosha siRNAs or a control siRNA against luciferase, respectively. Transfection of Phoenix cells with 800 pmol of Drosha siRNA yielded a very similar picture (data not shown). We next confirmed the effect of Drosha siRNA with pCLEP-30c. Addition of siRNA against Drosha in the Phoenix transfection cocktail led to a three- to four-fold increase in GFP-positive cells in infected NIH3T3 cultures (data not shown). Therefore, titers of miRNA-encoding retroviral particles were increased by co-transfecting the packaging line with the retroviral expression vector and an siRNA against Drosha. To test whether the addition of Drosha siRNA also improves the transduction efficiency in primary B-cell cultures, we infected pre-activated primary splenic (CD43−) B cells with supernatants from Phoenix cells transfected either with the control vector pCLEP, with pCLEP-106b or with pCLEP-106b together with 200 pmol of Drosha siRNA (Fig. 1D).
Spleens from DENV-2-infected mice were surgically removed at different time-points and single cell suspensions were prepared. Approximately 0·5 × 106 to 1 × 106 spleen cells were incubated with either 10 μg/ml of the indicated peptide, RPMI-1640 medium (Gibco) or PMA (0·1 μg/ml) + ionomycin (1 μg/ml). Golgi plug (BD Biosciences, San Jose, CA) was added to each of the selleck compound above samples and incubated at 37° for 6 hr. Cells
were washed with FACS buffer, blocked with Fc block (2.4G2) for 10 min and then surface stained with peridinin chlorophyll protein-Cy5.5-hCD45 (clone 2D1), phycoerythrin-Cy7-mCD45 (clone 30-F11), FITC-hCD3 (clone UCHT1), phycoerythrin-hCD8 (clone H1T8a) and Pacific Blue-hCD4 (clone RPAT4) antibodies for 20 min at room temperature. Cells were washed with FACS buffer, then permeabilized using Cytofix/Cytoperm buffer (BD Biosciences) and stained with allophycocyanin-hIFN-γ (clone B27) and Alexa700-TNF-α for 20 min at room temperature. AUY-922 In all experiments the viability marker LIVE/DEAD® Aqua (Molecular Probes, Eugene, OR) was added to exclude dead cells. All cell preparations were fixed with Cytofix (BD Biosciences). Cytokine levels were also assessed by ELISA; 0·5 × 106 to 1 × 106 spleen cells from DENV-2-infected mice
were incubated with either 10 μg/ml of the indicated peptide, RPMI-1640 medium (Gibco) or PMA (0·1 μg/ml) + ionomycin (1 μg/ml) and incubated at 37° for 96 hr. Culture supernatants were collected and IFN-γ level was determined by IFN-γ ELISA (R&D Systems, Minneapolis, MN). Levels of DENV-2 envelope (E) protein-specific antibody in the serum of DENV-infected
engrafted, uninfected engrafted and non-engrafted mice were determined using a standard ELISA. Ninety-six-well microplates were coated overnight with 100 ng/well of DENV-2 E protein (Hawaii Biotech, Aiea, HI) or 1 : 40 dilution of DENV-2-infected Vero cell lysate. The plates were blocked with 1% bovine serum albumin for 90 min and a 1 : 20 dilution of sera diluted with PBS was added to the wells for 1 hr. Plates were washed with PBS containing 0·1% Tween-20. Horseradish peroxidase-labelled goat anti-human IgM or IgG (Bethyl Laboratories Inc., Montgomery, Sucrase TX) was added as the secondary antibody. TMB Solution (Sigma-Aldrich Inc., St Louis, MO) was used as the substrate. The enzyme reaction was stopped by addition of 1 m HCl and the plates were read at 450 nm. Positive controls included sera from a known DENV-positive human. Sera from uninfected engrafted and non-engrafted mice were used as negative controls. All assays were carried out in duplicate or triplicate. Splenocytes from DENV-2 S16803-infected or naive mice were stimulated in vitro with CpG (2·5 μg/ml) + interleukin-2 (1 μg/ml) and Epstein–Barr virus (50 μl/ml). Supernatants of stimulated splenocytes collected 14 days after in vitro stimulation were tested for DENV-specific IgM antibodies and for DENV neutralization activity.
C57BL/6 mice (2 months old) were i.n. infected with 5 HAU of influenza virus. After 3 days, lung mononuclear cells were isolated from infected mice or uninfected mice, and then the cell suspensions were layered on a Histopaque-1083 gradient (Sigma-Aldrich), and centrifuge at 400 × g for 30 min at room temperature. Subsequently NK cells were purified using a negative selection mouse NK cell enrichment kit (StemCell Technologies), and labeled by CellTrace™
Violet (Invitrogen Corporation). As described previously [52, 53], 2 × 106 NK cells in 0.25 mL PBS were injected i.v. into recipient mice via the tail vein. On the same Selleck BGJ398 day, the mice were i.n. infected with 5 HAU of influenza A/PR8 virus. After infection, NK cells from lung and spleen were analyzed by flow cytometry 15 h later. The survival rate and body weight of
infected mice were monitored daily. Two months check details old C57BL/6 mice were i.n. infected with 5 HAU of influenza virus or normal egg allantoic fluid on day 0. At days 2, 4, and 6 after infection, mice were euthanized and lungs were isolated and fixed in 10% buffered formalin, then embedded in paraffin and sectioned. Specimens were stained with H&E and examined using a Zeiss Axio Imager M1 microscope equipped with an AxioCam HRc camera under control of AxioVision 4 software (Carl Zeiss Canada Ltd.). GraphPad Prism 4.00 (GraphPad Software, Inc., San Diego, CA, USA) was used for all analyses. Differences among experimental groups were assessed by one-way ANOVA followed by Tukey multiple comparison test. Unpaired t-test (two-tailed) was used to compare pairs of groups. Survival curves were assessed by survival analysis in Prism. Values were reported as the mean ± SEM. This work was supported by operating grants from the Canadian Institutes for Health Research (to K.P.K.). We thank Suellen Lamb, Dr. L. Tyrrell Laboratory, University of Alberta for making histologic sections
and performing hematoxylin and eosin staining. We thank Donger Gong for her technical support. The authors declare no financial or commercial conflict of interest. “
“The description of highly Wilson disease protein exposed individuals who remain seronegative (HESN) despite repeated exposure to human immunodeficiency virus (HIV)-1 has heightened interest in identifying potential mechanisms of HIV-1 resistance. HIV-specific humoral and T cell-mediated responses have been identified routinely in HESN subjects, although it remains unknown if these responses are a definitive cause of protection or merely a marker for exposure. Approximately half of HESN lack any detectible HIV-specific adaptive immune responses, suggesting that other mechanisms of protection from HIV-1 infection also probably exist.
In comparison with adult cattle, we have previously demonstrated that the immune response of calves involves early IL-12 expression with consequent IFN-γ production, a nitric oxide burst and modulation selleck products by IL-10 (6–9). This age-related immunity is dependent upon cellular events within the spleen as splenectomy of calves renders them equally susceptible (5,10). Our studies have utilized a technique to
marsupialize the spleen of calves (11) so that cells could be acquired for ex vivo analysis (microplate assays and flow cytometry) (12–16). Such analyses have proven valuable in determining the function of various splenic cell phenotypes but lack the ability to place these cell populations within their anatomical context which include the marginal zone, red and white pulp (17). Amongst many factors that comprise an effective immune response to haemoparasitic infection, trafficking and interaction of cells within such domains are central (18). Intravital imaging techniques have been used to dynamically study such factors within
superficial lymphoid organs (19,20) and, to a limited extent, also within deeper structures including Selleckchem Epigenetics Compound Library the spleen of mice (21). But current techniques are not well suited to study the spleen of large mammals because of the limits on depth resolution (22). An approach readily applied to the spleen of large mammals is the serial analysis of the distribution of phenotyped cells in tissue sections. Similar to a recent study on the acute immune response of naïve mice to haemoparasitic infection (23), we have applied this technique to the spleen of naïve calves infected with Babesia bovis. The results document acute change in the distribution of several cells thought to be important to the spleen-dependent response of naïve calves to B. bovis
and serve to underscore common themes in the acute response to haemoparasitic infections. In addition, this is the first documented use of magnetic resonance imagery to measure spleen volume in calves. Twelve Holstein–Friesian steer calves were obtained at 8 weeks of age, vaccinated against pathogenic Clostridium species, castrated and dehorned. All animals were cELISA seronegative for Anaplasma marginale (VMRD, Pullman WA, USA) and B. bovis and B. bigemina (24–26). pheromone The care and use of these calves were approved by the Institutional Animal Care and Use Committee at Washington State University (Pullman, WA, USA). At 12 weeks of age, all calves underwent a surgical procedure to marsupialize the spleen (11). When necessary, spleen cell aspirates were obtained under local lidocaine anaesthesia into 60cc syringes containing ACD and prepared for in vitro studies as previously described (14,27). Ten of the twelve calves were inoculated intravenously with 1 × 105 erythrocytes infected with the T2Bo virulent isolate of B. bovis (7).
However, its judicious use helps in the assessment
of selected patients with PI associated with chronic infective or inflammatory disease. SCIG is becoming well established as a viable alternative to IVIG for patients with primary antibody deficiency. SCIG is as efficacious as IVIG in infection prophylaxis and in achieving satisfactory serum IgG levels as has been demonstrated in several recent key clinical studies of a 16% SCIG versus IVIG formulation. A total of 158 patients with PI were assessed in three different studies and no difference in mean infection scores and in duration of infections was observed for SCIG versus IVIG [3,24,25]. Of particular interest is that for European-based studies, the Vivaglobin® dose given Selleck MK2206 is equivalent when switching patients from IVIG to SCIG, whereas in North American studies the United States
Food and Drug Administration (US FDA) requires the initial SCIG dose at switching to be 1·37 times the previous IVIG dose, in order to achieve a similar area under the IgG concentration–time curve. Despite this, no difference between the rate of SBIs was observed in these European versus North American studies. check details There were, however, differences in the overall infection rate, an observation which should generate further evaluation. The European Hizentra trial showed that an increase in IgG dose upon switching from IVIG to SCIG is not necessary
to maintain a low frequency of SBIs, but is beneficial in reduction of the rate of non-serious infections and the associated rates of hospitalization and antibiotic use . As SCIG is given more frequently in smaller doses compared with IVIG, it allows increasing the total monthly dose more easily without risk of compromising tolerability. Additionally, SCIG has a very favourable AE profile. In contrast to IVIG, there have been no reports of associated renal impairment, aseptic meningitis or anaphylaxis. Moreover, SCIG has been used successfully in cases of IVIG-induced anaphylaxis associated with anti-IgA antibodies. In a recent US study, 49 patients previously on IVIG were switched to IgPro20, a 20% liquid SCIG stabilized Liothyronine Sodium with l-proline . No SBIs (defined as per US FDA criteria) were observed and the rate of non-serious infections was low (2·76 infections/patient/year). Subcutaneous administration allows infusion of up to 1·2 g/kg/month and a 20% SCIG formulation enables administration of even higher doses . Furthermore, SCIG therapy results in more stable serum IgG levels over time, as smaller doses are given more frequently compared to the larger IVIG boluses given every 3–4 weeks . A maintenance of serum IgG levels can be achieved with SCIG even with a reduction in total monthly dose compared to the previously IVIG administered dose .
Despite the apparent importance of inherited susceptibility in the development of T1D, genetics alone cannot account for the disease’s entire aetiological spectrum. One of the most important indications
is the rapid increase in T1D incidence since the 1950s, particularly within the age group younger than 5 years [21–24]: a too-rapid growth to be explained reasonably Cilomilast mouse by genetic changes. In addition, twin studies have identified concordance rates that do not exceed 40% . Such arguments lend support to the notion that one, or perhaps multiple, environmental event(s) should be factored in to explain disease development, and particularly the onset of clinical hyperglycaemia in predisposed individuals. Humans – like all other organisms on the planet – respond to environmental
influences. Until somewhat recently, humans had only a dim notion of, and so generally neglected, the concept of hygiene. Consequently, exposure to faecal–oral transmitted microorganisms and viruses was high from birth onwards. One disease that is spread by a faecal–oral-transmitted HEV and which was rare in our collective past, but became horrifically important to human health in the 20th century, is poliovirus (PV)-induced poliomyelitis. It is of interest that T1D, also rare in the past but common today, has also been linked closely to HEV infections (reviewed in ; recent meta-analysis in ). In the case of PV, immunity acquired by a combination of passive immune transfer through nursing from and environmental exposure to infectious PV resulted in poliomyelitis being CHIR-99021 datasheet rarely manifested. Could a similar effect with other viruses, such as species B HEV , have resulted in maintaining T1D at a low level in our human past? Experimental data showing that autoimmune T1D is suppressed in NOD mice following inoculation with HEV  and that such exposure can promote expansion of a protective regulatory T cell (Treg)
population  support this hypothesis. In a modern society, in which common exposure to faecal pathogens such as HEV has been greatly minimized, failure to become immune to one or more specific HEV by a certain age leaves one open to an HEV infection and a potentially aggressive attack on the pancreas, which may lead in turn to T1D onset . Observational data of T1D incidence indifferent countries and societies  generally support the concept that more rural and/or less developed populations have a lower T1D risk than do populations in highly developed societies, in which it might be expected that higher hygiene standards are more widespread. What could be the pathological mechanisms that link viral infection to the onset of islet autoimmunity and eventually development of T1D? Several models have been postulated in attempts to answer this question.
4, right-hand graph and Fig. 1C). We also studied the IFN-γ production by tumour-infiltrating CD4+ and CD8+ T cells Copanlisib research buy and found this to be correlated with the suppression of tumour growth after ITADT in each of the experimental groups (data not shown). These results suggest that not only the injected syngeneic DC but also host-derived in situ APC functioned well as pAPC in ITADT, resulting in an efficient antitumour effect. Therefore, ITADT using MHC-incompatible allogeneic DC resulted in an efficient
antitumour effect if there was no rejection of the injected DC, and these effects were likely mediated indirectly via the MHC-compatible in situ pAPC of the host. Taken together, all three factors — [(1) survival of injected DC, (2) MHC compatibility of the injected DC and (3) function of host-derived pAPC] – affected the antitumour response induced by ITADT, but the survival time of the injected DC was the most important factor when using allogeneic DC. As described earlier, the host-derived pAPC functioned so well in ITADT that semi-allogeneic DC showed efficient antitumour INCB024360 concentration effects. Even fully allogeneic DC had significant antitumour effects if the alloresponse of the host was abrogated. We next investigated whether semi-allogeneic DC or fully allogeneic DC would have an antitumour effect if injected subcutaneously at sites distant from
the tumour (we refer to this as SCDT). In this case, tumour-associated host-derived pAPC could not contribute to any antitumour effect by priming TAA-specific T cells. In addition, we also investigated the antitumour effects of SCDT using syngeneic DC and compared the results with those of ITADT using syngeneic DC. For SCDT, we used DC that had been pulsed with tumour lysate. ITADT using syngeneic BL6 DC showed an efficient antitumour clonidine effect, resulting in significant suppression
of tumour growth (4/5 tumours eradicated) and significantly improved survival rates compared with PBS-treated controls (Fig. 5A,B, P < 0.01). SCDT using syngeneic BL6 DC also showed a significant antitumour effect compared with controls (Fig. 5A,B, P < 0.05). However, the antitumour effect observed in SCDT using BL6 DC was significantly weaker than that of ITADT using BL6 DC, and no tumour eradication was observed. Additionally, the survival rates in the SCDT group using BL6 DC were significantly worse compared with those in the ITADT group using BL6 DC (Fig 5A,B, P < 0.01). It was noted that SCDT using either semi-allogeneic BDF1 DC or fully allogeneic DBA/2 DC did not show a significant antitumour effect (Fig. 5A,B). We also investigated the effects of SCDT and ITADT against CT26 tumours. SCDT using syngeneic B/c DC had a significant antitumour effect in terms of tumour growth suppression and prolonged survival times relative to PBS controls (Fig. 5C,D, P < 0.01).