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Purpose: Colon antigen-1 (COA-1) was recently identified as a novel antigen of colorectal cancer encoded by the UBXD5 gene. Here, we evaluated whether a specific T-cell-mediated response directed against this molecule can occur in colorectal cancer patients.Experimental Design: Antigen- and tumor-specific immunologic responses of peripheral blood mononuclear cells (PBMC) stimulated in vitro with the MHC class II-associated immunogenic epitope of COA-1 (FSTFPPTLYQDDTLTLQAAG) were analyzed by IFN-γ ELISPOT assay.Results: COA-1-specific and tumor-reactive T lymphocytes were isolated from all (n = 7) HLA-DRβ1*0402+ or *1301+ colorectal cancer patients with progressive disease (Dukes' C and D) but not in patients (n = 4) with early-stage tumor (Dukes' A and B) and in healthy donors (n = 5), suggesting that the immune response against this antigen is associated with the progression of colorectal cancer. COA-1- and tumor-specific T lymphocytes displayed a CD3+CD4+CD69+CD45RA+ phenotype, compatible with the activated effector-type T-cell subset, and most of them exerted cytotoxic activity against HLA-matched and COA-1+ tumor cells. COA-1-specific T cells could also be isolated by in vitro stimulation of peripheral blood mononuclear cells with autologous dendritic cells loaded with tumor lysate, suggesting that this antigen can generate a dominant immunologic response against colorectal cancer cells. Notably, we could identify also COA-1-derived epitopes binding to HLA-A*0201 molecules that elicited antigen- and tumor-specific CD8+ T-cell-mediated responses in colorectal cancer patients.Conclusions: Both CD4+ and CD8+ T-cell responses against COA-1 can occur in colorectal cancer patients with metastatic disease, suggesting that this antigen is suitable for immunotherapeutic protocols of these patients.

Supplementary Data from Induction of Both CD8+ and CD4+ T-Cell–Mediated Responses in Colorectal Cancer Patients by Colon Antigen-1

CT-PET scan showing clinical response of Pt#2.

Supplementary Table I: IFN-DC release criteria and median values registered among patient-derived IFN-DC (min-max range); Supplementary Table 2: Patients MHC-I and MHC-II alleles and IGHV epitope sequences

Purpose:This study was aimed at evaluating the feasibility, safety, immunologic and clinical responses in patients with follicular lymphoma treated with monocyte-derived dendritic cells generated in the presence of IFNα and GM-CSF (IFN-DC) in combination with low doses of rituximab.Patients and Methods:Firstly, we analyzed in vitro and in vivo the immunologic properties of IFN-DC against follicular lymphoma. Thus, we performed a phase I trial in 8 patients with refractory and relapsed follicular lymphoma based on sequential intranodal injections of low-dose of rituximab and unloaded IFN-DC and report the safety, clinical, and immunologic results of the enrolled patients.Results:Preclinical studies indicated that IFN-DC can synergize with rituximab leading to increased cytotoxicity and T-cell tumor infiltration. The clinical evaluation showed that the combined treatment was totally safe. The overall response rate was 50%, PET-negative complete response rate 37%, and remission is still ongoing in 2/4 of responding patients (median follow-up 26 months, range 11–47). Notably, following the combined therapy all patients showed induction/enhancement of T-cell responses by CD107 degranulation or IFNγ ELISPOT assay against patient-specific tumor IGHV sequences.Conclusions:These results represent the proof-of-principle on the effectiveness of unloaded IFN-DC in inducing durable clinical responses and promoting induction of tumor-specific peripheral T cells, thus suggesting the occurrence of an effective endogenous antitumor vaccination. The overall findings indicate that some unique properties of IFN-DC can be successfully exploited to induce/enhance antitumor responses, thus representing a valuable antitumor strategy for novel and more effective combination therapies in patients with cancer.

CT-PET scan showing clinical response of Pt#3

Chimeric antigen receptor T cell therapies are revolutionizing the clinical practice of hematological tumors, whereas minimal progresses have been achieved in the solid tumor arena. Multiple reasons have been ascribed to this slower pace: The higher heterogeneity, the hurdles of defining reliable tumor antigens to target, and the broad repertoire of immune escape strategies developed by solid tumors are considered among the major ones. Currently, several CAR therapies are being investigated in preclinical and early clinical trials against solid tumors differing in the type of construct, the cells that are engineered, and the additional signals included with the CAR constructs to overcome solid tumor barriers. Additionally, novel approaches in development aim at overcoming some of the limitations that emerged with the approved therapies, such as large-scale manufacturing, duration of manufacturing, and logistical issues. In this review, we analyze the advantages and challenges of the different approaches under development, balancing the scientific evidences supporting specific choices with the manufacturing and regulatory issues that are essential for their further clinical development.

Purpose: This study was aimed at evaluating the feasibility, safety, immunologic and clinical responses in patients with follicular lymphoma treated with monocyte-derived dendritic cells generated in the presence of IFNα and GM-CSF (IFN-DC) in combination with low doses of rituximab. Patients and Methods: Firstly, we analyzed in vitro and in vivo the immunologic properties of IFN-DC against follicular lymphoma. Thus, we performed a phase I trial in 8 patients with refractory and relapsed follicular lymphoma based on sequential intranodal injections of low-dose of rituximab and unloaded IFN-DC and report the safety, clinical, and immunologic results of the enrolled patients. Results: Preclinical studies indicated that IFN-DC can synergize with rituximab leading to increased cytotoxicity and T-cell tumor infiltration. The clinical evaluation showed that the combined treatment was totally safe. The overall response rate was 50%, PET-negative complete response rate 37%, and remission is still ongoing in 2/4 of responding patients (median follow-up 26 months, range 11–47). Notably, following the combined therapy all patients showed induction/enhancement of T-cell responses by CD107 degranulation or IFNγ ELISPOT assay against patient-specific tumor IGHV sequences. Conclusions: These results represent the proof-of-principle on the effectiveness of unloaded IFN-DC in inducing durable clinical responses and promoting induction of tumor-specific peripheral T cells, thus suggesting the occurrence of an effective endogenous antitumor vaccination. The overall findings indicate that some unique properties of IFN-DC can be successfully exploited to induce/enhance antitumor responses, thus representing a valuable antitumor strategy for novel and more effective combination therapies in patients with cancer.

For more than 25 years, dendritic cell (DC) based vaccination has flashily held promises to represent a therapeutic approach for cancer treatment. While the vast majority of studies has focused on the use of antigen loaded DC, the intratumoral delivery of unloaded DC aiming at in situ vaccination has gained much less attention. Such approach grounds on the ability of inoculated DC to internalize and process antigens directly released by tumor (usually in combination with cell-death-inducing agents) to activate broad patient-specific antitumor T cell response. In this review, we highlight the recent studies in both solid and hematological tumors showing promising clinical results and discuss the main pitfalls and advantages of this approach for endogenous cancer vaccination. Lastly, we discuss how in situ vaccination by DC inoculation may fit with current immunotherapy approaches to expand and prolong patient response.

Human gammaherpesviruses such as Epstein-Barr virus (EBV) cause lifelong infections and associated diseases, by virtue of their ability to establish latent infection. Many studies performed in the past years in murine herpesvirus 68 (MHV-68) model of infection suggested that the limited immunity generated against isolated viral components by subunit vaccines cannot counteract the multiple immune evasion strategies operated by gammaherpesviruses. Indeed, a significant inhibition of long-term latency establishment could be observed in mice vaccinated with strains of genetically modified MHV-68 defective in reactivation or establishment of latency. In this study, we focused on the effects of interferon-α (IFN-α) on both the lytic and latent phase of MHV-68 infection, as exerted by the constitutive release of IFN-α1 by a clone of MHV-68 genetically modified to produce this cytokine (MHV-68mIFNα1). Although the MHV-68mIFNα1 recombinant virus exhibited in vitro replication features indistinguishable from those of the wild type MHV-68, its pathological properties were severely attenuated in vivo in immunocompetent mice and not in mice rendered genetically unresponsive to type I IFN, suggesting that a stronger immune response was primed in the presence of the cytokine. Notably, MHV-68mIFNα1 attenuation did not result in a reduced level of long-term spleen latency establishment. These results prompted us to evaluate the efficacy of MHV-68mIFNα1 in a prophylactic vaccination regimen aimed at inhibiting the symptoms of acute virus infection and the establishment of long-term latency after MHV-68 challenge. Our results show that mice vaccinated with MHV-68mIFNα1, administered as a live-attenuated or partially inactivated (by Psoralen and UV treatment) vaccine, were protected against the challenge with wt MHV-68 from all phases of infection. The ability of MHV-68mIFNα1 to produce IFN-α at the site of the infection, thus efficiently stimulating the immune system in case of virus reactivation from latency, makes this recombinant virus a safer live-attenuated vaccine as compared to the previously reported latency-deficient clones.

Purpose: We have developed a novel DC population differentiated from human monocytes in the presence of GM-CSF and IFN-alpha (IFN-DC), which is highly efficient in internalizing tumour cell antigens and in the cross-priming of CD8+T cells, promoting efficient anti-lymphoma response. Preclinical results prompted us to start a clinical trial of IFN-DC combined with low-dose Rituximab(RTX) in patients with Follicular lymphoma (FL), which is an indolent, immune-responsive, but still incurable cancer. Patients and Methods: In October 2014 we started a phase I clinical trial (EudraCT: 2013-003158-25) testing safety, immunogenicity and systemic clinical responses of low-dose RTX combined with autologous IFN-DC, administered by intra-nodal injections. Patients features: age 18-75y, indolent FL, relapsed or refractory (R/R), stage III-IV low tumour burden, with at least 1 superficial pathologic lymph node. One leukapheresis was scheduled at study entry for harvesting mononuclear cells. Eight cycles of RTX (5-20mg) and IFN-DC (30±10x106) were planned: the first 4 every two weeks, followed by other 4 cycles administered monthly. All injections were delivered with the aid of an ultrasound guide. Blood test for immune-responses and autoimmunity markers were scheduled at 0,+72d, +132 days ; +9; +18;+24 months. Results: as of the 31st July 2018, 8 patients have completed treatment and have been evaluated for response (median follow-up=24 months; range=8-45 mo). Median age of the study population was 60 years (range 27-76). All subjects had disseminated disease and enlarged lymph nodes or lesions greater than 1.5 cm at multiple sites available for local injections and monitoring for distant response. All patients were R/R after previous systemic therapy (median lines=2, range1-4) and 3/8 (37.5%) after auto-transplantation. The treatment was very well tolerated and manageable. No relevant adverse events were observed. Four out of 8 (50%) patients had objective response (OR) with 3/8(37.5%) attaining complete remission (CR); 2/8 (25%) are in stable disease at +8 and +25 months; 2/8 (25%), who did not achieve OR, progressed at +8 and +14 respectively. One patient progressed 4 months after achieving CR (Figure1). In all patients who achieved OR or who are in stable disease, induction of a tumour specific immune response was observed. Conclusions: local RTX and IFN-DC combined therapy was effective in eliciting in 50% of R/R FL with low tumour burden systemic and durable responses, which were associated with induction of specific anti-FL T cell responses. Disclosures No relevant conflicts of interest to declare.

Background Advanced melanoma patients have an extremely poor long term prognosis and are in strong need of new therapies. The recently developed targeted therapies have resulted in a marked antitumor effect, but most responses are partial and some degree of toxicity remain the major concerns. Dendritic cells play a key role in the activation of the immune system and have been typically used as ex vivo antigen-loaded cell drugs for cancer immunotherapy. Another approach consists in intratumoral injection of unloaded DCs that can exploit the uptake of a wider array of tumor-specific and individual unique antigens. However, intratumoral immunization requires DCs endowed at the same time with properties typically belonging to both immature and mature DCs (i.e. antigen uptake and T cell priming). DCs generated in presence of interferon-alpha (IFN-DCs), due to their features of partially mature DCs, capable of efficiently up-taking, processing and cross-presenting antigens to T cells, could successfully carry out this task. Combining intratumoral immunization with tumor-destructing therapies can induce antigen release in situ, facilitating the injected DCs in triggering an antitumor immune response. Methods We tested in a phase I clinical study in advanced melanoma a chemo-immunotherapy approach based on unloaded IFN-DCs injected intratumorally one day after administration of dacarbazine. Primary endpoint of the study was treatment safety and tolerability. Secondary endpoints were immune and clinical responses of patients. Results Six patients were enrolled, and only three completed the treatment. The chemo-immunotherapy was well tolerated with no major side effects. Three patients showed temporary disease stabilization and two of them showed induction of T cells specific for tyrosinase, NY-ESO-1 and gp100. Of interest, one patient showing a remarkable long-term disease stabilization kept showing presence of tyrosinase specific T cells in PBMC and high infiltration of memory T cells in the tumor lesion at 21 months. Conclusion We tested a chemo-immunotherapeutic approach based on IFN-DCs injected intratumorally one day after DTIC in advanced melanoma. The treatment was well tolerated, and clinical and immunological responses, including development of vitiligo, were observed, therefore warranting additional clinical studies aimed at evaluating efficacy of this approach. Trial registration Trial Registration Number not publicly available due to EudraCT regulations: https://www.clinicaltrialsregister.eu/doc/EU_CTR_FAQ.pdf

INTRODUCTION Follicular lymphoma (FL) is considered a chronic, immune responsive and still incurable tumour. Recently two papers have shown the efficacy of active immunotherapy by repeated administration of autologous dendritic cells (DC) in Follicular Lymphoma (FL). Notably, in both studies a third of patients achieved long lasting remission. A novel DC population (IFN-DC), differentiated from human monocytes in the presence of GM-CSF and IFN-alpha has been developed in the laboratory of the Istituto Superiore di Sanità (ISS). IFN-DC have been demonstrated highly efficient in internalizing tumour-cell antigens, recovering "in vitro" T-cell responses from FL patients, as well as in mediating NK cell activation and enhanced cytotoxic effector function toward autologous FL cells. We have recently started a phase I study (approved by EC in 2014, EudraCT: 2013-003158-25) of intranodal immunotherapy based on sequential injection of low-dose anti-CD20 antibody (Ab) and IFN-D, in patients with advanced FL. Here we report early results of the ongoing trial. STUDY DESIGN IFN-DC Trial (EudraCT: 2013-003158-25) is a phase I trial aiming at evaluating safety and tolerability as well as immune and clinical responses of a IFN-DC based therapy in combination with rituximab for the treatment of patients with advanced FL. · The Primary endpoints are: I) Evaluation of safety and tolerability of treatment II) Evaluation of tumor-specific immune responses. by determining: i) Tumor-specific immune response in peripheral blood; ii) Intratumoral infiltration of immune cells; iii) DTH test. · The secondary endpoint is clinical response. The study population consists of patients with relapsed/refractory FL, aged 18-75 years, not needing immediate retreatment basing on the GELF Criteria. The · Treatment: the regimen foresees eight injection cycles, each consisting of the combined administration of rituximab and IFN-DC. The first four administered every two-weeks and the remaining four administered monthly. Rituximab (5-10mg) and low or high doses of IFN-DC (20±5 x106 or 40±5x106cells respectively), are administered by intranodal direct injection (IDI). RESULTS Five patients have been enrolled so far. The treatment proved to be safe, feasible and well tolerated. Three patients have completed treatment and two experienced complete remission with disappearance of both proximal and distal lesions, as confirmed by PET scan. Notably in both patients the distal lymph nodes, which had not been directly treated by intranodal injections, reduced to normal size earlier than those treated by IDI (abscopal effect). One of these two is still in durable complete clinical and molecular remission after 17 months while the latter patient, who had progressed after completing eight cycles of treatment, experienced a complete clinical remission after a second course of immunotherapy ( Fig.1). Two out of three patients performed the DHT test and, despite one of the two achieved remission, they both resulted non-responsive. The characterization of IFN-DC and lymphoma nodes in FL patients have shown high expression of PD-1 and PD-L1 in FL microenvironment and on IFN-DC respectively. Immunomonitoring is currently underway. However, quantification of NF-kB (p65) nuclear translocation by multispectral image flow cytometry, as a marker of NK cell activation, showed the persistence of higher percentages of activated NK cells in the two responding patients (Fig.2). CONCLUSION Currently no definitive data prove the benefit of IDI treatment, however the trial is ongoing and other patients are under screening. Notably both the responding patients showed the abscopal effect. The possible set-up of an effective and comparatively economic treatment, that exploits patients' immune system against this chronic and immune responsive tumour, is worth of exploring. Furthermore it can be envisaged that the combination of immune checkpoint inhibitor antibodies to DC-based therapies, might improve clinical activity of DC-based strategies. Disclosures No relevant conflicts of interest to declare.

The Nm23 protein is a nucleoside diphosphate kinase (NDPK) and is thought to play a critical role in metastatic behavior. It has been reported that a NDPK activity is present in microtubules assembled in vitro. Since microtubule assembly is determinant in cell growth and differentiation, we investigated whether Nm23-M1 forms molecular complexes with beta-tubulin in murine cells either actively proliferating or differentiating. For this purpose a polyclonal antibody against the GST-Nm23-M1 fusion protein was generated and employed to detect Nm23-M1/beta-tubulin complexes in murine tumor cells derived from the Lewis lung carcinoma (3LL) and in undifferentiated and differentiated myogenic cells (C2C12). Immunoblotting and immunoprecipitation experiments performed using the anti-fusion protein antibody demonstrated that the Nm23-M1 protein is detectable in in vitro tumor cell lines and in in vivo primary tumors but not in spontaneous lung metastases. These data are in good agreement with data previously reported. Immunoprecipitation experiments demonstrated that the Nm23-M1 protein forms complexes with beta-tubulin in in vitro tumor cell lines, but not in primary tumors. Furthermore, the Nm23-M1 protein forms complexes with beta-tubulin in myogenic cells prior to and after differentiation. Interestingly, however, the level of the Nm23-M1/beta-tubulin complexes is remarkably increased in differentiated myotubes. In conclusion, the results indicate that the Nm23-M1 protein forms molecular complexes with beta-tubulin and that the number of complexes increases during the differentiation process of murine cells.

One of the underlying justifications of the patent system is to encourage dissemination of scientific knowledge and promote innovation. Yet, the patent system is not a green card to innovation. Indeed, given our progress in science and the increasing rate of technological developments it is ironic that there is arguably a declining rate of innovation. From this perspective, it might be contended that the patent system is not meeting its said objectives. But the patent system should be seen in a wider context. While well-intended, it is far too simplistic to argue that patents are good or bad for innovation, or that they ‘freeze or spur’ innovation. The challenges to the patent system are complex and cannot solely be attributed to its laws. That is, the patent system’s constituting laws do not tell the full story. Often more telling are the institutional frameworks and socio-cultural and economic practices at the pre-patent inventive stage and the post-patent commercialization stage and beyond. And so when attempting to predict future developments, and ultimately, craft solutions, understanding and ensuring that the wider context is effective is paramount.

Guppies pretreated with 1.1 μmol/L diazinon (0.5 times the LC50 with the guppy) for 7 d were less susceptible to the toxicity of the compound. The LC50 in these fish was about five times higher than that in nonpretreated animals. The lethal body burden and the slope of the concentration-response curve of diazinon in pretreated guppies were similar to those in zebra fish. The 7-d pretreatment of zebra fish with 12 μmol/L diazinon (0.5 times the LC50 with the zebra fish) did not affect the susceptibility of this species to diazinon. The pretreatment of guppies resulted in a strong inhibition of the formation of diazoxon and pyrimidinol during incubations of diazinon with the hepatic postmitochondrial supernatant. During exposure of guppies and zebra fish to diazinon, the tissue concentration of the metabolite, pyrimidinol, initially increased and then declined to very low levels. In the guppy, concurrently with the decrease of the metabolite concentration, the parent compound began accumulating again after a temporary steady state. The hypotheses of this paper are that the toxicity of diazinon in the guppy is due to its metabolism to a highly toxic metabolite, likely diazoxon; and in zebra fish or pretreated guppies having low rates of diazinon metabolism toxicity is due to the accumulation of the parent compound.

We have investigated the molecular mechanisms underlying the peculiar cross-presentation efficiency of human dendritic cells (DCs) differentiated from monocytes in the presence of Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) and Interferon (IFN)-α (IFN-DCs). To this end, we evaluated the capability of IFN-DCs to present and cross-present epitopes derived from Epstein–Barr Virus (EBV) or human melanoma-associated antigens after exposure to cell lysates or apoptotic cells. In an autologous setting, IFN-DCs loaded with Lymphoblastoid Cell Lines (LCL) lysates or apoptotic LCL were highly efficient in expanding, respectively, EBV-specific class II- or class I-restricted memory T cell responses. Of note, IFN-DCs loaded with apoptotic LCL were more potent than fully mature DCs in triggering the cytotoxicity of CD8 + T lymphocytes recognizing a subdominant HLA-A*0201-restricted epitope derived from EBV latent membrane protein 2 (LMP2). In addition, IFN-DCs loaded with apoptotic human melanoma cells were highly efficient in cross-presenting the MART-1 27–35 epitope to a specific CD8 + cytotoxic T cell clone, and this functional activity was proteasome-dependent. These IFN-DC properties were associated with an enhanced expression of all the immunoproteasome subunits as well as of TAP-1, TAP-2 and tapasin, and, interestingly, to a higher proteasome proteolytic activity as compared to immature or mature DCs. Altogether, these results highlight new mechanisms by which IFN-α influences antigen processing and cross-presentation ability of monocyte-derived DCs, with potentially important implications for the development of DC-based therapeutic vaccines.

The 96-h LC 50 value of the organophosphorus pesticide diazinon ( O , O -Diethyl- O -2-isopropyl-4-methyl-6-pyrimidyl thiophosphate) was about 0.8 mg l for the guppy and about 8 mg l for the zebra fish. Both the guppy and the zebra fish metabolized diazinon. The detected metabolite was 2-isopropyl-6-methyl-4-pyrimidinol (pyrimidinol). Diazoxon ( O , O -diethyl- O -2-isopropyl-4-methyl-6-pyrimidylphos-phate) could not be found as a metabolite. After short exposures (up to 48 h) to low sublethal diazinon concentrations the body concentration of pyrimidinol was linearly dependent on the body concentration of diazinon. The ratio of pyrimidinol to diazinon body concentrations was about 5 times higher in the guppy than in the zebra fish. In both species however, the pyrimidinol level reached a maximum at concentrations corresponding to 1 4 to 1 2 of the LC 50 value and these levels were remarkably similar. Kinetic experiments at diazinon concentrations of 0.1 and 0.4 ppm pesticide in water showed that an apparent steady state was reached after 24 to 48 h. After longer exposures an increased accumulation of diazinon was observed concomitantly with the inhibition of pyrimidinol formation. The inhibition of the brain acetylcholinesterase reached 73% in the guppy and 83% in the zebra fish. The maximum of the inhibition was already reached when the concentration of diazinon in water was 25% of the LC 50 value. The factor of bioconcentration (BCF) was dependent on the level of the pesticide in the water and varied between 39 (in the guppy, at a water concentration of 0.1 ppm) and more than 300 (in the zebra fish, at a water concentration of 4 ppm). The results indicate that the metabolism plays a key role in the bioaccumulation and in the species specific toxicity of diazinon in fish.

Dendritic cells (DCs) play a crucial role in linking innate and adaptive immunity, by virtue of their unique ability to take up and process antigens in the peripheral blood and tissues and, upon migration to draining lymph nodes, to present antigen to resting lymphocytes. Notably, these DC functions are modulated by cytokines and chemokines controlling the activation and maturation of these cells, thus shaping the response towards either immunity or tolerance.An ensemble of recent studies have emphasized an important role of type I IFNs in the DC differentiation/activation, suggesting the existence of a natural alliance between these cytokines and DCs in linking innate and adaptive immunity. Herein, we will review how type I IFNs can promote the ex vivo differentiation of human DCs and orient DC functions towards the priming and expansion of protective antitumor immune responses. We will also discuss how the knowledge on type I IFN-DC interactions could be exploited for the design of more selective and effective strategies of cancer immunotherapy.

Purpose: Colon antigen-1 (COA-1) was recently identified as a novel antigen of colorectal cancer encoded by the UBXD5 gene. Here, we evaluated whether a specific T-cell-mediated response directed against this molecule can occur in colorectal cancer patients. Experimental Design: Antigen- and tumor-specific immunologic responses of peripheral blood mononuclear cells (PBMC) stimulated in vitro with the MHC class II-associated immunogenic epitope of COA-1 (FSTFPPTLYQDDTLTLQAAG) were analyzed by IFN-γ ELISPOT assay. Results: COA-1-specific and tumor-reactive T lymphocytes were isolated from all (n = 7) HLA-DRβ1*0402+ or *1301+ colorectal cancer patients with progressive disease (Dukes' C and D) but not in patients (n = 4) with early-stage tumor (Dukes' A and B) and in healthy donors (n = 5), suggesting that the immune response against this antigen is associated with the progression of colorectal cancer. COA-1- and tumor-specific T lymphocytes displayed a CD3+CD4+CD69+CD45RA+ phenotype, compatible with the activated effector-type T-cell subset, and most of them exerted cytotoxic activity against HLA-matched and COA-1+ tumor cells. COA-1-specific T cells could also be isolated by in vitro stimulation of peripheral blood mononuclear cells with autologous dendritic cells loaded with tumor lysate, suggesting that this antigen can generate a dominant immunologic response against colorectal cancer cells. Notably, we could identify also COA-1-derived epitopes binding to HLA-A*0201 molecules that elicited antigen- and tumor-specific CD8+ T-cell-mediated responses in colorectal cancer patients. Conclusions: Both CD4+ and CD8+ T-cell responses against COA-1 can occur in colorectal cancer patients with metastatic disease, suggesting that this antigen is suitable for immunotherapeutic protocols of these patients.

An integrated health record (IHR) that enables clinical data to be shared at a national level has profound implications for medical research. Data that have been useful primarily within a single clinic will instead be free to move rapidly around a national network infrastructure. This raises challenges for technologists, clinical practice, and for the governance of these data. This article considers one specific issue that is currently poorly understood: how intellectual property (IP) relates to the sharing of medical data for research on large-scale electronic networks. Based on an understanding of current practices, this article presents recommendations for the governance of IP in an integrated health record.

Canada's Supreme Court ruled that publishers need permission from freelance authors before reproducing their works in online databases. But in contrast to the approach taken in Europe and the US, publishers are now entitled to republish articles on CD-ROM without asking or compensating the freelance authors. Ultimately, irrespective of the publishing medium, the Court defers to private ordering to clarify ambiguities in new use clauses that will continue to persist as technologies evolve to the detriment of freelancers. Thus this decision will have little, if any, impact in practice.