Neither the withholding of nor withdrawing from dialysis is euthanasia. No physician-assisted suicide (PAS) is entirely different to the ceasing of a treatment.

PAS is a positive act done by a patient to cease life and where a physician has assisted in its execution (usually by prescribing medications used in the suicide). The KPT-330 supplier withdrawing of treatment, including dialysis, is an entirely different act where the death, when it results is due to the underlying disease and not due to the action taken by the patient. Lisa Phipps and Robert Walker With variable availability of renal supportive care (RSC) programmes available throughout Australia and New Zealand, there is a need for provision of training in this area to be available to all medical and paramedical staff Online resources may be a potential source of training material for staff and information for patients and families. The possibility of exchange programmes between renal medicine and palliative care should be explored as a way of enhancing education in both fields. The ANZSN and the ANZ Society of Palliative Cobimetinib chemical structure Care both have special interest groups in RSC. The potential for

bringing these two groups together to facilitate cross-specialty training should be explored. The incidence of end-stage kidney disease (ESKD) in Australia and New Zealand is increasing (ANZDATA 2011). Patients with ESKD both on dialysis and conservative care pathways are sicker and more debilitated than in the past.[1] Patients with chronic kidney disease (CKD) and ESKD are amongst the most Tau-protein kinase symptomatic of any chronic disease group.[2, 3] With increasing evidence that patients with multiple co-morbidities may not benefit from dialysis,[4-6] it is essential that nephrologists are trained in the conservative management of ESKD. The current curricula for Australian and New Zealand Nephrology advanced

trainees (http://www.rpctraining.com.au) recognizes this under learning objective 2.3.8 ‘plan and manage the non-dialysis pathway’. Manage common ESKD problems – pruritus, fatigue, xerostomia, depression, constipation, insomnia, nausea, vomiting, dyspnoea and pain Adjust drug doses according to reduced GFR Liaise with allied health staff Describe reduced life expectancy to a patient with respect, empathy and dignity. However with only a small number of conservative care clinics in Australia and New Zealand, trainees and nephrologists may receive very limited exposure to symptom control and conservative management. This has been the experience overseas, with a survey of nephrology trainees in the US revealing their training resulted in them feeling least prepared to manage a patient at the end of life.

parvum antigens on dendritic cells, we generated an enriched population of immature DCs by culturing whole BM cells in GM-CSF. We assessed the differentiation status of the loosely adherent cells by day 14. On the day of the BM harvest, <5% of whole BM cells were expressing the myeloid DC markers. By the time the cells were harvested

from the plates, at day 14, >90% of the cells were expressing CD11c and CD11b and a subset expressed other DC markers, such as CD86, CD80, CD40 and MHCII (Figure 1). These unstimulated DCs were then used for subsequent in vitro studies. The same time frame and format was used for the DCs generated from the whole BM Opaganib datasheet of the MyD88 KO mice (data not shown). In order to identify the differentiation/maturation status of the BMDC, we examined the expression levels of DC-SIGN (CD209) as well as CD86, CD80, CD40, MHCI and MHCII as shown in Figure 1. CD86 and CD80 were already high in the unstimulated cells, whereas marked increases were observed with CD40, MHCII and CD209 when DCs were treated with either sporozoites

or cryptosporidial antigen-treated cultures. In order to investigate the role DCs play in eliciting responses to different C. parvum antigen presentation/maturation, we incubated DCs with either freshly excysted intact sporozoites or solubilized sporozoite lysate. We also looked at the responses to several recombinant antigens, such as Cp23, Cp40, Cp17 and P2 (18,22,24). All antigen preparations as well as conditioned media preparations were tested for endotoxin and were below 0·03 EU. Lipopolysaccharide was used as a positive Epigenetics Compound Library molecular weight control and was also tested at different concentrations and yielded consistent results, indicating that MoDCs were biologically active. As shown in Figure 2(a), solubilized sporozoite antigen was able to induce significant increases in the expression of IL-12p70

from DCs as compared to Thymidylate synthase media alone (>200-fold increase), whereas freshly excysted sporozoites induced much lower-level IL-12 responses. In contrast, expression levels of IL-12p70 from DCs isolated from MyD88 KO mice were at or below background levels (Figure 2a, b). Recombinant antigens Cp40 and Cp23 were also able to significantly increase IL-12p70 expression, as observed in Figure 2(b). This finding indicates that the solubilized as well as recombinant antigens can induce the maturation of the DCs and subsequently initiate an innate immune response. Treatment of dendritic cells with cryptosporidial antigens induced increased expression levels of the Th1 cytokine, IL-2 (Figure 3 a, b). Again, significantly reduced expression levels of IL-2 were observed in the BMDCs of MyD88 KO mice in responses to C. parvum antigen, with the exception of LPS that has been shown to induce the maturation of MyD88-deficient dendritic cells (25).

9,15–18 Further studies are needed to increase our understanding of the roles of eosinophils and IL-5 in inflammatory responses and other diseases in which hypereosinophilia occurs. The differential migration of eosinophils versus neutrophils to thyroids of IFN-γ−/− and WT mice during the development of G-EAT offers a unique opportunity to examine the role of eosinophil trafficking to sites of inflammation and to investigate the potential role of these

cells in the induction and resolution of inflammation. Neutralization of IL-5 markedly inhibited migration of eosinophils to thyroids of IFN-γ−/− mice during development of G-EAT. However, IL-5 neutralization had no effect on the severity or rate of resolution of inflammation in G-EAT, suggesting that eosinophil migration has no apparent pathogenic role in G-EAT. WT and IFN-γ−/− DBA/1 mice were produced Everolimus in our animal facilities at the University of Missouri as previously described.6–8 Both male and female mice (6–10 weeks old) were used. G-EAT was induced as previously described.1,5 Briefly, mice were injected intravenously

(i.v.) twice at 10-day intervals with 150 μg of MTg3 and 15 μg of lipopolysaccharide (LPS) (Escherichia coli 011:B4; Sigma Chemical Co., St Louis, MO). Seven days later, donor spleen cells were re-stimulated in vitro selleck products with 25 μg/ml MTg and 5 ng/ml IL-12.1 Cells were harvested after 72 hr and washed twice, and 3·5 × 107 cells were transferred i.v. to 500-Rad irradiated

syngeneic recipients. Anti-IL-5 was purified from culture supernatants of the anti-IL-5-producing hybridoma TRFK-5 (provided by Dr Robert Coffman, DNAX Research Institute, Palo Alto, CA, USA) using protein G. IFN-γ−/− recipients of IFN-γ−/− donor cells were given 300 μg of anti-IL-5 intraperitoneally (i.p.) or rat immunoglobulin G (control IgG) every 4 days beginning on the Rebamipide day of cell transfer until euthanasia. WT recipients of WT donor cells were used for comparison. Thyroids were removed from groups of five or six recipient mice 20 days (peak of disease) or 40–50 days (fibrosis versus resolution) after cell transfer.1–6 Thyroids were fixed in formalin, sectioned and stained with haematoxylin and eosin (H&E), and scored quantitatively for G-EAT severity (the extent of inflammatory cell infiltration and thyroid follicle destruction) using a scale of 1+ to 5+, as described previously.6 1+ thyroiditis is defined as an infiltrate of at least 125 cells in one or several foci; 2+ is 10–20 foci of cellular infiltration involving up to 25% of the gland; 3+ indicates that 25–50% of the gland is infiltrated; 4+ indicates that > 50% of the gland is destroyed by infiltrating inflammatory cells; and 5+ indicates virtually complete destruction of the thyroid with few or no remaining follicles. Thyroid lesions were also evaluated qualitatively.

No serum miRNA was regulated exclusively in aUC compared with iUC patients. Four miRNAs were higher and three miRNAs

were lower in the mucosa of aCD than iCD. Two miRNAs were higher and three miRNAs were lower in the mucosa of aUC than iUC. No serum miRNAs coincided with tissue miRNAs in aCD and aUC patients. Our results suggest Decitabine in vivo the existence of specific miRNA expression patterns associated with IBD and their different stages and support the utility of miRNA as possible biomarkers. This pilot study needs to be validated in a large prospective cohort. Inflammatory bowel disease (IBD), including Crohn’s disease (CD) and ulcerative colitis (UC), is a chronic inflammatory gastrointestinal disorder, the pathophysiology of which remains unclear. The theory accepted most commonly is that IBD

and the associated gastrointestinal inflammation are likely to be the result of the interaction between a defective immune response to a luminal factor (probably intestinal flora), epigenetic and environmental factors (e.g. smoking) and its influence in genetically predisposed subjects [1-3]. Genetic factors involved in inflammation and immune functions are known to play a very important role in IBD physiopathology. Micro-RNAs (miRNAs) are a class of small non-coding RNAs, involved in the control of gene expression at the post-transcriptional level [4]. Following the discovery of miRNAs, the number of publications regarding their biogenesis and functions has been increasing exponentially and the miRNA sequence database, miRBase, is growing continuously [5, 6].

BVD-523 mouse MiRNAs are involved in the regulation of many biological processes such as the cell cycle, differentiation, Exoribonuclease proliferation, apoptosis, fibrosis and immune function [7]. Emerging evidence has demonstrated that miRNAs can also play an important role in the development of cancer as well as in the induction of chronic inflammatory and autoimmune diseases [8, 9]. miRNAs have been found in tissues, serum, plasma and other body fluids. It has been demonstrated that the levels of miRNAs in serum are stable, reproducible and consistent among individuals of the same species [10]; for this reason, such levels are now being used as a non-invasive biomarker for different pathologies (i.e. cancer, autoimmune disease, inflammation) [10, 11]. Previous studies, focused particularly on cancer, have discovered that circulating miRNA profiles can be correlated with tissue miRNA profiles [12, 13]. In most cases, those changes in circulating miRNA profiles can precede the standard blood biomarkers and possess prognostic value [12, 14, 15]. These properties mean that miRNAs are attractive, blood-based, non-invasive biomarkers. Recently, several papers have focused investigation on the altered expression of miRNAs in IBD and their important role as regulators and possible diagnostic biomarkers in IBD [8, 16-18].

© 2012 Wiley Periodicals, Inc. Microsurgery, 2012. “
“Pulsed acoustic cellular expression (PACE) is a treatment that applies focused acoustic shock waves to promote tissue healing. The aim of this study was to assess the effect of PACE treatment on inflammatory responses in a cremaster muscle ischemia/reperfusion injury model. Seventeen cremaster muscle flaps were evaluated

in four groups: nonischemic controls (n = 5), 5-hour www.selleckchem.com/products/Temsirolimus.html ischemia controls (n = 4), preischemic (5-hour) PACE conditioning (n = 4), and postischemic (5-hour) PACE conditioning (n = 4). The expression of proinflammatory cytokines (TNFα, IL-6, IL-1α, IL-1β, GM-CSF) and chemokines (CCL3, CCL4, CXCL4) was assessed using TaqMan® real-time PCR. Expression of ELAM-1, VCAM-1, and ICAM-1 was assessed by immunostaining. Preischemic PACE conditioning upregulated expression of IL-6, CCL3, CCL4, and CXCL4, and downregulated expression of TNFα, GM-CSF, and IL-1α. Postischemic PACE conditioning significantly decreased expression of all evaluated genes. Pre- and postischemic PACE conditioning decreased expression of ELAM-1 and ICAM-1. Results of the study indicate

that application Ruxolitinib clinical trial of PACE conditioning may have a beneficial effect on the recovery of tissues subjected to the ischemia/reperfusion injury. Postischemic PACE conditioning revealed anti-inflammatory effect as confirmed by decreased expression of inflammatory cytokines, chemokines, and cell adhesion molecules (ELAM-1 and Rho ICAM-1) that are responsible for leukocyte

recruitment into ischemic tissues. Hence, PACE therapy may be used effectively in clinical practice as a convenient therapeutic strategy to protect tissues against ischemia/reperfusion related injury after microsurgical procedures of free tissue transfers. © 2012 Wiley Periodicals, Inc. Microsurgery, 2013. “
“The reconstruction of complex hand injury such as multifinger soft tissue defect remains a challenging problem. Two cases of repair of multifinger injury with exposed bones using the free chimeric flaps based on the dorsalis pedis vessels are presented. Two male patients, 46 years old and 36 years old, suffered from a thermocompression injury to the dorsum of fingers resulting in soft tissue defects of multiple fingers. The chimeric free flap was designed and applied to cover the defects. The donor sites were covered by skin grafts. The postoperative courses were uneventful. Both patients were followed up for 10–12 months. The maximal flexion angle of the distal interphalangeal, proximal interphalangeal, and metacarpophalangeal joints were 40°–85° at the end of the follow-up. The protective sensation was achieved on the dorsal fingers. The report suggests that the free chimeric flaps based on the dorsalis pedis artery may be an alternative for the reconstruction of the multifinger dorsal soft tissue defects. © 2013 Wiley Periodicals, Inc. Microsurgery 33:660–666, 2013.

For intracellular staining LY294002 of GM-CSF, isolated leukocytes were incubated with 50 ng/mL PMA, 500 ng/mL ionomycin, Golgi-Plug (1 μL/mL) containing brefeldin A in RPMI-1640 at 37°C for 4 h. Thereafter,

cells were stained with rat antimouse CD4-FITC, rat antimouse CD45-V450, fixed and permeabilized with Cytofix/Cytoperm (BD), and stained with rat antimouse GM-CSF-PE (BD). Apoptotic and dead CD4+ T cells were detected by staining with 7-AAD and CD4-allophycocyanin. Fas expression on CD4+ T cells was analyzed by staining with hamster antimouse Fas-PE and CD4-FITC. Controls were stained with isotype-matched control antibodies. All antibodies and reagents were obtained from BD Biosciences (Heidelberg, Germany) unless otherwise mentioned. Flow cytometry was performed on a FACScan (BD Biosciences), and the data were analyzed with WinMDI or Cell Quest software. Primary astrocytes Poziotinib clinical trial were isolated from 1- to 2-day-old newborn mice and cultured as published before [43]. To obtain pure astrocytes, cells were harvested from astrocyte cultures and stained with rat antimouse CD11b-PE. Pure astrocytes (CD11b−) were then separated from CD11b+ microglia with a FACSVantage cell sorter (BD). Neuronal cultures were obtained according to Lenz et al. [44]

with slight modifications. Briefly, pregnant female mice were sacrificed by cervical dislocation at gestational day 18.5, and dissociated cells of each embryonic brain were cultivated in flasks coated with poly-L-lysine in Neurobasal medium supplemented with B27 (Invitrogen) and 500 μM L-glutamine (Gibco). The purity of cultures for neurons was ≥98%, as determined by immunofluorescence

staining for Janus kinase (JAK) neuron-specific class III β-tubulin. DNA was isolated from sorted astrocytes and microglia, respectively, as well as from cultured neurons using a DNA isolation kit (Qiagen, Germany). For the detection of FasL expressed on the surface of astrocytes, mixed astrocyte/microglia cultures were stained with mouse antimouse FasL-PE and CD11b-FITC. Controls were stained with isotype-matched control antibodies. For histology on paraffin sections, mice anesthetized with methoxyflurane were perfused with 0.1 M PBS followed by 4% paraformaldehyde in PBS, spinal cords were processed and stained with hemalum and eosin, cresyl violet, and luxol fast blue. In addition, paraffin sections were used for immunohistochemical demonstration of GFAP, neurofilament, Mac3, and CD3 (Serotec, Düsseldorf, Germany) by an ABC protocol as described [45]. Total mRNA was isolated from the spinal cords of nonimmunized and MOG35–55- immunized mice (RNeasy kit, Qiagen, Germany) at day 15 and day 22 p.i., respectively. SuperScript reverse transcriptase kit with oligo (dT) primers (Invitrogen, Germany) was used to generate cDNA from total mRNA.

64±10.87×106 and WT: 31.54±15.52×106 for B220+; Hax1−/−: 3.71±0.77×106 and WT: 2.55±1.05×106 for T1; Hax1−/−: 6.91±3.61×106 and WT: 4.73±2.23×106 for T2; Hax1−/−: 5.89±2.89×106 and WT: 4.53±2.39×106 for mature B cells; Hax1−/−: 2.92±1.84×106 and WT: 2.34±1.16×106 for MZ B cells). Our data clearly demonstrate that Hax1−/− LSK cells in a Hax1+/+ environment were able to fully reconstitute the lethally irradiated hosts. To further investigate the reason for the massive B lymphocyte deficiency, we investigated www.selleckchem.com/products/pd-0332991-palbociclib-isethionate.html the expression of CXCR4 and BAFFR on splenic B cells. CXCR4 is expressed on hematopoietic precursors 22 as well as on centroblasts within the germinal centre

18. CXCR4-expressing cells migrate towards CXCL12, expressed by stromal cells and germinal center dark zone compartments. Thus, an impaired CXCR4 expression would severely impede normal B-cell development. Alternatively, signals through the BAFFR have a significant role in promoting B-cell survival and homeostatic proliferation 23. For real time analysis, we isolated total splenocytes of four 10-wk-old WT and Hax1−/− mice and enriched for B lymphocytes using magnetic cell sorting. Both the CXCR4 and the BAFFR DNA Damage inhibitor amplification showed prominent amplification products. Most interestingly, CXCR4 expression

in HAX1-deficient B cells was decreased by around 70% compared to WT cells. BAFFR expression was slightly, but not significantly, decreased in HAX1-deficient B cells (Fig. 7A). However, the decreased expression had no effect on the formation of follicular structures. No differences in the distribution of B- Teicoplanin and T-cell areas, as stained by CD3 and B220, were detectable (Fig. 7B). Because of the fact that the transfer of Hax1−/− bone marrow cells into a HAX1+ environment gave rise to normal levels of B220+ cells and functional B-cell subsets, we conclude that the severely decreased

CXCR4 expression on HAX1-deficient B cells is not solely responsible for the described B-cell loss in Hax−/− mice. Previously, we described HAX1 as interaction partner of membrane bound IgE (mIgE) 24. From that point of view, it would have been of most interest to analyse IgE responses on a Hax1-deficient background. However, the short lifespan of Hax1−/− mice impeded a direct analysis. Therefore, we focused on the detailed investigation of the biological function of HAX1 during lymphocyte development. Hax1−/− mice are characterized by a severely diminished cellularity of lymphoid tissues accompanied by a significant reduction of B and Tlymphocytes. Recently, Chao et al. 25 reported on the role of HAX1 with a similar approach. Our results demonstrate that the developmental impairment is not restricted to specific developmental stages. We observed reduced numbers of B cells from the pro-pre B-cell stage in the bone marrow to mature stages in the spleen. The analysis of splenic subpopulations clearly demonstrated a continuation of the developmental defects for T1 and T2 B cells 26, 27.

044). Group one showed two good, two satisfactory, six moderate, and one bad results while the second group showed five good, six satisfactory, one bad and no moderate results (P = 0.026). The first time to show clinical response in group one was the third month while in the second group it was at 1.5 month (P < 0.001). In addition, the first time to show electromyographic response in group one was at the sixth month while in group two it was at the third month Vein wrapping is a simple technique that could be used reliably to augment primary neurorrhaphy particularly in cases with associated vascular or tendon injuries GSK126 to prevent scarring and enhance functional and electrophysiological

recovery. © 2013 Wiley Periodicals, Inc. Microsurgery 34:361–366, 2014. “
“A 19-year-old male patient with type 1 von Willebrand’s disease underwent two separate superficial inferior epigastric artery free flap tissue transfers and three revision procedures for reconstruction of a postextirpative mid-facial

defect. Intravenous 1-desamino-8-D-arginine vasopressin (DDAVP) was administered as bleeding prophylaxis prior to incision for free tissue transfer. For each debulking procedure, DDAVP was administered by intranasal sprays in minutes prior to incision and redosed 12 and 24 hours postoperatively. There were no incidents of either thrombosis or bleeding. This outcome indicates that 0.3 μg/kg intravenous DDAVP may be effective as bleeding prophylaxis for patients with mild and quantitative defects in von Willebrand factor undergoing microvascular reconstruction. © 2011 Wiley-Liss, Inc.

Microsurgery, 2011. “
“Postoperative Selleckchem APO866 vascular compromise is a common but critical complication requiring emergent re-exploration, and remains a chief cause of free flap failure. This study investigated the relationship between postanesthetic shivering (PAS) and the development of postoperative complications associated with free flap reconstruction. One hundred thirty-six patients who underwent head and neck cancer resection selleck and free flap reconstruction were retrospectively enrolled. Fifteen patients were assigned to the PAS group, while the others were assigned to the non-PAS (NPAS) group. The odds ratios of acute re-exploration or total failure of the free flap in the PAS group was 3.5 and 14.9, respectively. The dose of meperidine was positively correlated with PAS prevention in our statistical ROC curve analysis. The minimum effective dose of meperidine for PAS prevention was 0.35 mg/kg with 75% sensitivity and 60% specificity. These findings indicate that an optimal dose of meperidine could prevent PAS, which is shown to be associated with a decrease in the incidence of the early post-surgical re-exploration rate of these free flaps related to circulatory compromise. © 2013 Wiley Periodicals, Inc. Microsurgery 34:106–111, 2014. “
“Several authors have reported the usefulness and benefits of lymphoscintigraphy.

To examine the involvement of IL-12 from DCs in the activation of NK cells, we co-cultured NK cells with AFP-DCs or Alb-DCs with or without the presence of neutralizing antibody for IL-12. The cytolytic activity of NK cells co-cultured with Alb-DCs decreased to the level of that with AFP-DCs on addition of anti-IL-12 neutralizing antibody. Moreover, adding IL-12 to the co-culture of AFP-DCs and NK cells resulted in enhancement of the cytolytic activity of NK cells to the levels

of Alb-DCs and NK cells. Taken together, these data demonstrated Midostaurin cell line that IL-12 derived from AFP-DCs plays essential roles in the impairment of NK cytotoxicity, which is consistent with the results of the production of IL-12 from AFP-DCs and Alb-DCs. Serum AFP is often high in patients with cirrhosis without HCC [8]. Oka et al. reported that the incidence of HCC development is significantly high in cirrhosis patients who had elevated serum levels of AFP [17], which suggests that high production of AFP in cirrhosis patients might also impair innate immunity, leading to HCC development. Our results might offer support for the hypothesis that elevation of AFP in cirrhosis patients impairs innate immunity which plays an essential role in the deletion of micro HCC, and thus results in promotion of HCC development. Although the expression of antigen-presenting related molecules on AFP-DCs was not altered,

the maturation of AFP-DCs was inhibited compared with Alb-DCs. This is consistent with a previous report [13] suggesting that the EPZ-6438 molecular weight presence of AFP impairs DC maturation. DCs have been implicated in the activation of NK cells

[19]. However, activated NK cells have been shown to recognize and lyse DCs in vitro and in vivo, but maturation of DCs results in resistance to NK lysis [19]. In HCC patients, it has been shown that impairment of DCs is associated with increased tumour progression [20] and that the levels of activated DCs are significantly low in HCC tissues [21]. High levels of AFP produced by HCC tissues may impair DC maturation, which would enhance HCC progression by removing DCs from HCC tumour areas. IL-12 exhibits a number of immunologically important activities, including the ability Bay 11-7085 to enhance NK cell and CTL functionality, to polarize CD4+ T cell responses by preferentially supporting the T helper type 1/cytotoxic T cell (Th1/Tc1)-type and to suppress Th2-type immunity [22]. We have demonstrated that the production of IL-12 protein from LPS-stimulated or Poly(I:C)-stimulated AFP-DCs was impaired significantly compared with that from Alb-DCs, which might affect immunosuppression in AFP-elevated patients. The expression of mRNA of both IL-12p35 and IL-12p40 were also inhibited significantly in AFP-DCs compared with Alb-DCs but not those of TLR-4, LPS receptor and TLR-3, Poly(I:C) receptor.

Interestingly, R788 the avidity of response to the OVA was similar (1.7×10−9 M) to the response to TRP2 (1.3×10−9 M) suggesting that there is no deletion of the

repertoire to this self Ag. However responses to both epitopes could be increased over 100-fold, by using an Ab–DNA vaccine compared to peptide immunization. These results suggest that at the each peptide MHC complex interacts with a defined number of TCR within the repertoire playing an important role in determining the original avidity 28 but this can then be further modulated at the clonal level. The range of avidities observed in the mice analyzed spans five logs, yet within individual experiments this variation is less. This probably reflects the plasticity of the avidity to any given TCR:MHC/peptide combination with optimal immunization leading to a high avidity. The avidity with DNA vaccination depends upon the degree of direct v cross presentation, GSK-3 beta pathway which may vary between experiments. However this does not explain the reduced variability within one

experiment. Our explanation is that despite careful operating procedures, this is related to the efficacy of immunization/monitoring of the response. We are aware that timing for harvesting the splenocytes to plating into an assay is a key parameter and endeavor to keep this constant. Finally experiments were performed over a 2-year period and factors such as subtle changes in mice, environment and batches of DNA have to be considered. Within the small groups these factors would be more consistent. The avidity of the responses to the TRP2/HepB human IgG1 DNA vaccine varied from 5×10−13 M to 5×10−8 M in different mice but was on average

5×10−10 M. Is this avidity sufficient to result in effective immune response? An elegant study by Dutoit et al. demonstrated that T cells cloned from cancer patients exhibited an exponential increase in killing with T-cell avidity greater than 10−9 M Ureohydrolase 2. A similar study with T-cell clones showed that only high-avidity clones adoptively transferred caused tumor rejection in mice 1. The avidity resulting in tumor killing will depend upon the expression level of the Ag/MHC. Our study is in agreement with these demonstrating that selective vaccination can increase avidity to a level sufficient for therapy. The frequency and avidity of the responses from human IgG1 DNA immunization was significantly higher than that observed from peptide immunization. Initially unlinked peptides were used but due to lack of T-cell help, these gave very weak responses (results not shown). To give a more reasonable comparison, the CTL epitopes were linked to a well known helper epitope which still gave poor responses. This was perhaps not surprising as even linked helper-CTL peptides have a very short half life and are poor immunogens in vivo29.