Irradiated Autologous Tumor Cells Research Articles

Abstract 6800: Unique autologous cancer vaccine comprised of irradiated whole tumor cells and MBTA (rWTC-MBTA) triggers antitumor immune response to prevent metastasis

Abstract Despite the advances made in cancer treatment over the past decades, one statistic has remained unchanged: metastatic cancer accounts for 90 percent of annual cancer deaths in the United States. Our group previously demonstrated that an autologous whole tumor cell vaccine (rWTC-MBTA: irradiated autologous whole tumor cells pulsed with Mannan-BAM, TLR ligands, and anti-CD40 antibody) had a potent anti-tumor immune response and prolonged survival in a mouse colon carcinoma model. To investigate whether rWTC-MBTA would work in “cold” tumor models, we evaluated the vaccine’s effect on preventing and treating tumor metastasis in 4T1 breast tumors. Our data showed that vaccinated mice could significantly prevent lung metastasis in both intravenous injection and mammary pad subcutaneous implantation animal models. Further, we used another metastasis model that more closely mimics clinical practice by resecting the primary tumor after metastasis development. The data showed that vaccinated mice could prevent tumor recurrence, increase T-cell infiltration in the metastatic tumor, and prolong the survival curve. The mechanistic investigation by immunophenotyping revealed that rWTC-MBTA vaccination induced both effector memory (CD44+CD62L−) and center memory (CD44+CD62L+) T cells as well as increased overall CD4+ and CD8+ T-cell count while depletion experiments demonstrated that CD8+ T cells were required for vaccine efficacy. Isolated splenocytes co-cultured with 4T1 cells showed rWTC-MBTA significantly increased T-cell mediated cytotoxicity through TNF-a and IFN-γ in CD107a+CD4+ T cells and Granzyme B and IFN-γ in CD107a+CD8+ T cells. In all experiments, vaccination exerted negligible systemic toxicity. Collectively, our study demonstrates that the rWTC-MBTA vaccine is a safe and promising therapeutic option to prevent and treat tumor metastasis by triggering an antitumor immune response. Citation Format: Juan Ye, Herui Wang, Mitchell Sun, Ondrej Uher, Mark R. Gilbert, Karel Pacak, Zhengping Zhuang, Rogelio Medina, Samik Chakraborty, Jan Zenka. Unique autologous cancer vaccine comprised of irradiated whole tumor cells and MBTA (rWTC-MBTA) triggers antitumor immune response to prevent metastasis. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6800.

762P Personalized immunotherapy with encapsulated cell technology: Feasibility, safety and efficacy results from the first-in-human clinical trial in advanced relapsing solid tumors

Preclinical data shows that sustained, local delivery of low doses of GM-CSF by irradiated, genetically engineered tumor cells at the immunization site leads to specific, long-lasting anti-tumor immunity in several tumor types. Providing sustained levels of GM-CSF SC in a clinical setting remains challenging. Encapsulated Cell Technology enables the sustained and controlled delivery of GM-CSF by allogeneic cells. MVX-ONCO is an active, personalized cancer immunotherapy combining irradiated autologous tumor cells and encapsulated, genetically engineered allogeneic cells producing GM-CSF.

MVX-ONCO-1 in advanced refractory cancers: Safety, feasibility, and preliminary efficacy results from all HNSCC patients treated in two ongoing clinical trials.

e18005 Background: MVX-ONCO-1 is an active, personalized cancer immunotherapy combining irradiated autologous tumor cells and encapsulated, genetically engineered allogeneic cells producing GM-CSF. The immunostimulant cytokine is released locally in a sustained manner over one week at the immunization site. All cancer types are potential candidates and more than 40 patients have been treated in two clinical trials. Here we report the data on all HNSCC patients treated, as of 31st January 2021. Methods: Eleven (11) patients with locally advanced/metastatic HNSCC relapsing after at least one line of systemic therapy were enrolled in 2 open-label, single-arm clinical trials (NCT02193503 and NCT02999646) evaluating the safety, feasibility and efficacy of MVX-ONCO-1. All patients were treated with at least 5 administrations of MVX-ONCO-1 over 8 weeks (W1,2,3,4,6,8). Each administration consists of the subcutaneous implantation in healthy skin of 2 capsules containing each 8x105 cells, producing > 20ng/24h of GM-CSF, and an injection of 4x106 irradiated autologous tumor cells between the 2 capsules. Results: Eleven (11) patients are evaluable for safety and feasibility. Ten patients (91%) with at least 6 months, respectively 9 patients (82%) with 12 months follow-up are evaluable for efficacy analysis. Eight patients (80%) failed both Cisplatin based chemotherapy and anti-PD1 therapies prior to enrolment. The most common treatment-related adverse event (AE) was local hematoma at implantation site in 3 patients (27%). None experienced > grade 2 treatment-related AE. Overall Survival is 70% (7/10pts) at 6 months, and 56% (5/9pts) at 12 months. Eight patients (80%) presented some degree of disease control with 4SDs, 2PRs and 2CRs. Both CRs are longstanding, with both patients not any longer on anticancer therapy for 24 and 6 months respectively. While 1 PR was observed on MVX-ONCO-1, 1PR and 1CR were observed on subsequent Nivolumab therapy (both CPS0), and 1 CR was observed after carboplatin-cetuximab. Two patients had progressive disease as best overall response. Conclusions: Treatment with MVX-ONCO-1 is feasible, safe and well tolerated. Preliminary efficacy data shows an encouraging rate of tumor control including prolonged CR in patients subsequently treated with nivolumab or chemotherapy. Efficacy of the concurrent use of anti-PD-1 and MVX-ONCO will be assessed in a planned clinical trial. Clinical trial information: NCT02999646.

Autologous tumor cell vaccination combined with systemic CpG-B and IFN-\u03b1 promotes immune activation and induces clinical responses in patients with metastatic renal cell carcinoma: a phase II trial

In this study the toxicity and efficacy of an irradiated autologous tumor cell vaccine (ATV) co-injected with a class-B CpG oligodeoxynucleotide (CpG-B) and GM-CSF, followed by systemic CpG-B and IFN-α administration, were examined in patients with metastatic renal cell carcinoma (mRCC). A single-arm Phase II trial was conducted, in which patients with mRCC were intradermally injected with a minimum of three whole-cell vaccines containing 0.7–1.3 × 107 irradiated autologous tumor cells (ATC), admixed with 1 mg CpG-B and 100 µg GM-CSF, followed by bi-weekly s.c. injections with 8 mg CpG-B and s.c. injections with 6 MU IFN-α three times per week. Fifteen patients were treated according to the protocol. Treatment was well tolerated. Objective clinical responses occurred in three patients, including one long-term complete response. Disease stabilization occurred in another three patients. Positive delayed type hypersensitivity (DTH) responses to ATC were absent before treatment but present in 13 out of 15 patients during treatment. Immune monitoring revealed activation of plasmacytoid dendritic cells, non-classical monocytes and up-regulation of both PD-1 and CTLA4 on effector T cells upon treatment. Moreover, a pre-existing ex vivo IFN-γ response to ATC was associated with clinical response. ATV combined with systemic CpG-B and IFN-α is tolerable, safe, immunogenic and able to elicit anti-tumor responses in patients with mRCC. Immune activation and treatment-induced up-regulation of PD-1 and CTLA4 on circulating T cells further suggest an added benefit of combining this approach with immune checkpoint blockade [added]

Abstract A021: Vaccination with autologous, nonattenuated, live glioblastoma cells induces potent peripheral and intratumoral anti-tumoral responses: A first-in-human study

Abstract Glioblastoma (GBM), which grows uninhibited in the brain, almost never metastasizes outside of it. The rare occurrence (<0.5%) of overt GBM extracranial metastasis is paradoxically associated with significantly enhanced patient survival. We previously reported that the highly malignant F98 Fischer-rat GBM model, which grows aggressively in the brain, spontaneously regresses when injected subcutaneously as live unattenuated cells. The peripheral growth and regression of live glioma cells markedly enhanced the survival of all rats, and cured (survival >9 months) approximately 50% of rats challenged with F98 cells intracranially. The results in the F98 model were corroborated in the CNS1 astrocytoma model in Lewis rats. The fraction of cured rats in both models were the highest reported in published literature. We found that the spontaneous rejection of live glioma cells inoculated in peripheral sites was immune-mediated. This location-biased immune response was termed “Split Immunity”—a tumor that thrives in an immune-privileged site (e.g., the brain) may be inhibited by injecting live unmodified tumor cells in a site that is not privileged, generating protective immunity that spreads back to the privileged site.To evaluate the applicability of the “Split Immunity” approach in humans, we ran a first-in-human (FIH) study on two recurrent GBM patients. The patients were vaccinated subcutaneously, initially with autologous irradiated tumor cells, and then with autologous, non-attenuated, live tumor cells. The treatment caused no serious adverse events. The injected live tumor cells did not grow at the inoculation site, nor did they spread metastatically, as evaluated by choline 18F PET-CT. Standard biochemical and hematologic blood tests showed no treatment-related adverse effects. Both patients’ quality of life (QoL) questionnaires demonstrated subjective improvements in self-evaluated “global health status.” The treatment had demonstrated several signs indicative of efficacy. MRI evaluation using “treatment response assessment maps” (TRAM) demonstrated strong increases in the “treatment-response” component following live-cell vaccination, compatible with “pseudo-progression-like” responses in both patients. Using elaborate 8-color flow cytometric panels, we detected potent tumor-cell specific polyfunctional activation in 2.5% and 5% of peripheral cytotoxic T-cell (CTL) and T helper cells (Th), respectively, 3 days following live-cell vaccination. Flow cytometric immune mapping of all intratumoral immune cell subsets showed major changes from baseline that had occurred shortly after live-cell vaccinations. We observed major increases (8 to 200 fold) in both patients of all monitored intratumoral dendritic cell (DC) subsets (CD1c+ and CD141+ myeloid DC, CD123+ plasmacytoid DC). Patient-1 showed the highest frequency of intratumoral CD56brt natural killer (NK) cells from all previously fully mapped brain tumor patients. Patient-2’s intratumoral CTL and Th were increase by 6- and 17-fold, respectively. Patient-2’s CD141+ and CD1c+ myeloid DC had exhibited upregulated CD86 maturation marker following treatment. Overall, the immune infiltrate following treatment in Patient-2 exceeded the immune infiltrates found in 42 previously fully-mapped brain tumor patients. The flow cytometric results were corroborated by the patients’ immunohistochemistry data. Patient-2, who had been treated by a tighter vaccination protocol than Patient-1, had survived 19 months after GBM recurrence, which is considerably longer than the median 6- to 8-month survival of recurrent GBM patients. Taken together, the accumulated FIH results are suggestive of safety and efficacy of the “Split Immunity” approach. Based on the these results, we have recently received an ethical approval for a phase IIa clinical trial on 8 recurrent- and 4 newly-diagnosed GBM patients. Citation Format: Ilan Volovitz, Nati Shapira, Rachel Grossman, Zvi Ram. Vaccination with autologous, nonattenuated, live glioblastoma cells induces potent peripheral and intratumoral anti-tumoral responses: A first-in-human study [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr A021.

Randomized phase II trial of autologous dendritic cell vaccines versus autologous tumor cell vaccines in metastatic melanoma: 5-year follow up and additional analyses

BackgroundDespite improved survival following checkpoint inhibitors, there is still a potential role for anti-cancer therapeutic vaccines. Because of biological heterogeneity and neoantigens resulting from each patient’s mutanome, autologous tumor may be the best source of tumor-associated antigens (TAA) for vaccines. Ex vivo loading of autologous dendritic cells with TAA may be associated with superior clinical outcome compared to injecting irradiated autologous tumor cells. We conducted a randomized phase II trial to compare autologous tumor cell vaccines (TCV) and autologous dendritic cell vaccines (DCV) loaded with autologous TAA.MethodsShort-term autologous tumor cell lines were established from metastatic tumor. Vaccines were admixed with 500 micrograms of GM-CSF and injected weekly for 3 weeks, then at weeks 8, 12,16, 20, and 24. The primary endpoint was overall survival. Secondary objectives were identification of adverse events, and results of delayed type hypersensitivity (DTH) reactions to intradermal tumor cell injections.ResultsForty-two patients were randomized. All were followed from randomization until death or for five years; none were lost to follow-up. DCV was associated with longer survival: median 43.4 versus 20.5 months (95% CI, 18.6 to > 60 versus 9.3 to 32.3 months) and a 70% reduction in the risk of death (hazard ratio = 0.304, p = 0.0053, 95% CI, 0.131 to 0.702). Tumor DTH reactions were neither prognostic nor predictive. The most common treatment-related adverse events were mild to moderate local injection site reactions and flu-like symptoms; but grade 2 treatment-related adverse events were more frequent with TCV. Serum marker analyses at week-0 and week-4 showed that serum markers were similar at baseline in each arm, but differed after vaccination.ConclusionsThis is the only human clinical trial comparing DCV and TCV as platforms for autologous TAA presentation. DCV was associated with minimal toxicity and long-term survival in patients with metastatic melanoma. DTH to autologous tumor cells was neither prognostic for survival nor predictive of benefit for either vaccine.Trial registrationClinical trials.gov NCT00948480 retrospectively registered 28 July 2009.

Vaccination with Irradiated Autologous Tumor Cells Mixed with Irradiated GM-K562 Cells Stimulates Antitumor Immunity and T Lymphocyte Activation in Patients with Recurrent Malignant Glioma

Recurrent malignant glioma carries a dismal prognosis, and novel therapies are needed. We examined the feasibility and safety of vaccination with irradiated autologous glioma cells mixed with irradiated GM-K562 cells in patients undergoing craniotomy for recurrent malignant glioma. We initiated a phase I study examining the safety of 2 doses of GM-K562 cells mixed with autologous cells. Primary endpoints were feasibility and safety. Feasibility was defined as the ability for 60% of enrolled subjects to initiate vaccination. Dose-limiting toxicity was assessed via a 3+3 dose-escalation format, examining irradiated tumor cells mixed with 5 × 10(6) GM-K562 cells or 1 × 10(7) GM-K562 cells. Eligibility required a priori indication for resection of a recurrent high-grade glioma. We measured biological activity by measuring delayed type hypersensitivity (DTH) responses, humoral immunity against tumor-associated antigens, and T-lymphocyte activation. Eleven patients were enrolled. Sufficient numbers of autologous tumor cells were harvested in 10 patients, all of whom went on to receive vaccine. There were no dose-limiting toxicities. Vaccination strengthened DTH responses to irradiated autologous tumor cells in most patients, and vigorous humoral responses to tumor-associated angiogenic cytokines were seen as well. T-lymphocyte activation was seen with significantly increased expression of CTLA-4, PD-1, 4-1BB, and OX40 by CD4(+) cells and PD-1 and 4-1BB by CD8(+) cells. Activation was coupled with vaccine-associated increase in the frequency of regulatory CD4(+) T lymphocytes. Vaccination with irradiated autologous tumor cells mixed with GM-K562 cells is feasible, well tolerated, and active in patients with recurrent malignant glioma. Clin Cancer Res; 22(12); 2885-96. ©2016 AACR.

5-year follow up for 72 metastatic melanoma patients treated with eltrapuldencel-T (CLBS20) vaccine.

3090 Background: CLBS20 is a patient-specific therapeutic vaccine consisting of autologous dendritic cells loaded with antigens from irradiated cells from an autologous tumor cell line. It potentially presents the entire repertoire of unique antigens resulting from nonsynonymous mutations in a patient’s self-renewing melanoma cells. In previously reported single arm and randomized phase 2 trials, CLBS was associated with a 72% 2-year overall survival (OS). In the randomized trial median OS for CLBS20-treated patients was more than twice that for a vaccine consisting of irradiated autologous tumor cells. Mild local injection site reactions was the most common toxicity. Methods: During 2001 to 2011 72 patients with stage 4 or recurrent stage 3 melanoma were treated with CLBS20 (Clinicaltrials.gov NCT00948480 & NCT00436930). Each had a metastatic melanoma lesion surgically resected, from which a tumor cell line was established, cells from which were incubated with autologous dendritic cells to produce the vaccine that was injected s.c. in 500 micrograms sargramostim (GM-CSF) weekly x 3 weeks and monthly x 5 months (mos). This report summarizes long-term survival data for all CLBS20-treated patients from date of first injection. Results: There were 45 men and 27 women. Median age is 52 years (17 to 83). Tumor sources were 37-lymph node, 15-soft tissue, 20-viscera. Prior therapies included 72-surgery, 41-chemotherapy, 33-Interleukin-2, 30-interferon alpha, 24-GM-CSF, 19-radiation therapy (RT), and 15-RT for brain metastases. All 33 surviving patients were followed 5 years; none were lost to follow up. Median OS is 45.6 mos; 5-year OS is 46%. Median progression free survival (PFS) is 4.4 mos, 5-year PFS is 19%. The 20 patients with recurrent stage 3, have a 70% 5-year OS, median PFS 19.8 mos, and 35% 5-year PFS. For the 52 patients with stage 4, median OS is 41.0 mos, 5-year OS is 36%, and median PFS is 3.8 mos. One patient with refractory progressive measurable disease experienced a delayed complete response that was ongoing at 5 years. Conclusions: CLBS20 is associated with encouraging 5-year overall survival. Because of its unique mechanism of action and absence of toxicity, it should be evaluated further in similar patients. Clinical trial information: NCT00948480. Clinical trial information: NCT00436930.

Colony stimulating factors secreted by irradiated autologous tumor cell vaccines inhibit immunity

Meeting abstracts Autologous tumor cell-based vaccines (ATCVs) have a number of potential advantages including multivalency and patient specificity. ATCVs contain many potential antigens, both known and unknown which potentiate polyclonal responses capable of responding to a more diverse population

5PD - Cell-based immunotherapy combining encapsulation cell technology and irradiated autologous tumor cells: A novel technology platform that is both safe and feasible. Results from first in man trial

5PD - Cell-based immunotherapy combining encapsulation cell technology and irradiated autologous tumor cells: A novel technology platform that is both safe and feasible. Results from first in man trial

531 MVX-ONCO-1: First in man, Phase I clinical trial combining encapsulation cell technology and irradiated autologous tumor cells for personalized cell-based immunotherapy. Safety, feasibility and clinical outcome results

Reference EPFL-CONF-214198View record in Web of Science Record created on 2015-12-02, modified on 2016-08-09

Phase III multicenter trial of eltrapuldencel-T: Autologous dendritic cells loaded with irradiated autologous tumor cells (DC-TC) in granulocyte-macrophage colony stimulating factor (GM-CSF) in patients with metastatic melanoma (INTUS trial).

TPS9082 Background: Melanomas harbor non-synonymous mutations that can result in unique, immunogenic tumor associated antigens (TAA). Recognition of TAA can be induced or enhanced by vaccination. T...

Phase III randomized, double-blinded, placebo-controlled multicenter trial of Eltrapuldencel-T: autologous dendritic cells loaded with irradiated autologous tumor cells (DC-TC) in GM-CSF in patients with metastatic melanoma

Phase III randomized, double-blinded, placebo-controlled multicenter trial of Eltrapuldencel-T: autologous dendritic cells loaded with irradiated autologous tumor cells (DC-TC) in GM-CSF in patients with metastatic melanoma

Seven Year Followup for Bystander Vaccine Therapy in Mantle Cell Lymphoma (MCL)

Background:A Phase I study revealed that vaccination of cancer patients with irradiated autologous tumor cells and GM.CD40L bystander cells (engineered to secrete GM-CSF and express CD40L) is safe, recruits/activates dendritic cells, and elicits tumor-specific T cell responses. We report final results using this vaccine strategy in patients with MCL, an aggressive and incurable B-cell malignancy.Methods:After lymph node resection for autologous tumor harvest, 4-6 cycles of chemotherapy, and restaging (CT, endoscopy, bone marrow biopsy), patients with usable vaccine who achieved a PR or CR lasting 1 month were vaccinated x4 at 28-day intervals with IL-2 (0.5 x 106 U SC BID x 14 d/cycle). Patients were monitored for toxicity, tumor response, tumor-specific immune responses, and median EFS/OS.Results:43 were enrolled, including 21 with relapsed MCL, 20 with int/high MIPI, and 6 with blastoid MCL. Twenty never received vaccine: 2 withdrew, 1 progressed prior to nodal harvest, 7 had insufficient or contaminated specimens, and 10 progressed/died during chemotherapy. The unvaccinated were older (68.4y vs 62.8y; p=.026) but otherwise did not differ significantly by stage, LDH, MIPI, de novo status, or number of prior treatments.Among 23 treated, 10 had relapsed disease (median of 3 prior therapies), 10 had an int/high MIPI, and 2 had blastoid MCL. Pre-vaccination response following chemotherapy included 7 CR, 15 PR, and 1 SD. At 6 months after vaccination, 2 pts in PR had resolution of MRD within the bone marrow, 10 progressed (including 3 who progressed after 1-2 vaccines), and 11 had no change. Grade 3 toxicities were observed in three patients, and consisted of leukocytosis, neutropenia, and cough, none of which were attributed to vaccination. One patient remains in CR at 84 mo. A second who relapsed at 64mo remains without need for salvage therapy at 82 mo, suggestive of more indolent relapse post vaccination. Median EFS and OS are calculated from receipt of first vaccine. Median EFS is 9 mo, and median OS is 101 mo (median follow-up 81 mo).DTH was not observed. Biopsy of vaccine sites revealed lymphocytic response in 62%, however, this was not correlated with EFS or OS. Increased IFN gamma post vaccination was observed in 76%, and accompanied by an increase in EFS (p=.06). Increased GMCSF post vaccination was observed in 89%, with a trend for improved survival among those with higher peak levels.Conclusions:Extended follow up continues to show a sustained benefit in regards to overall survival for patients with de novo (85% alive at 81 months) and relapsed MCL (median OS 25 months), and support therefore the need for further studies of vaccination strategies in the context of new immunotherapeutic agents that are improving the outcomes of MCL patients. [Display omitted] DisclosuresShah:Seattle Genetics, Inc.: Research Funding; NCCN: Consultancy; Celgene: Consultancy, Speakers Bureau; SWOG: Consultancy; Pharmacyclics: Consultancy; Janssen: Consultancy. Off Label Use: SGN-CD19A is an investigational agent being studied in patients with B-cell malignancies. SGN-CD19A is not approved for use. .

Glioblastoma exosomes and IGF-1R/AS-ODN are immunogenic stimuli in a translational research immunotherapy paradigm.

Glioblastomas are primary intracranial tumors for which there is no cure. Patients receiving standard of care, chemotherapy and irradiation, survive approximately 15 months prompting studies of alternative therapies including vaccination. In a pilot study, a vaccine consisting of Lucite diffusion chambers containing irradiated autologous tumor cells pre-treated with an antisense oligodeoxynucleotide (AS-ODN) directed against the insulin-like growth factor type 1 receptor was found to elicit positive clinical responses in 8/12 patients when implanted in the rectus sheath for 24 h. Our preliminary observations supported an immune response, and we have since reopened a second Phase 1 trial to assess this possibility among other exploratory objectives. The current study makes use of a murine glioma model and samples from glioblastoma patients in this second Phase 1 trial to investigate this novel therapeutic intervention more thoroughly. Implantation of the chamber-based vaccine protected mice from tumor challenge, and we posit this occurred through the release of immunostimulatory AS-ODN and antigen-bearing exosomes. Exosomes secreted by glioblastoma cultures are immunogenic, eliciting and binding antibodies present in the sera of immunized mice. Similarly, exosomes released by human glioblastoma cells bear antigens recognized by the sera of 6/12 patients with recurrent glioblastomas. These results suggest that the release of AS-ODN together with selective release of exosomes from glioblastoma cells implanted in chambers may drive the therapeutic effect seen in the pilot vaccine trial.

Phase III randomized, double-blinded, placebo-controlled multicenter trial of melapuldencil-t: autologous dendritic cells loaded with irradiated autologous tumor cells (dc-tc) in gm-csf in patients with metastatic melanoma

Meeting abstracts Genomic analyses have shown that melanoma patients have hundreds to thousands of non-synonymous mutations including unique tumor associated antigens (TAA) that can be recognized by their immune systems. The efficacy of anti-checkpoint monoclonal antibodies provides further proof

Phase I trial of active specific immunotherapy with autologous dendritic cells pulsed with autologous irradiated tumor stem cells in hepatitis B positive patients with hepatocellular carcinoma

Background Active or chronic infections may be barriers to successful vaccines, and patients with hepatitis typically are excluded from clinical trials testing therapeutic anticancer vaccines. One concern is vaccines may exacerbate the underlying infection. Hepatocellular carcinoma (HCC) often arises in the context of viral hepatitis B (HBV) or hepatitis C (HBC), with or without cirrhosis. HCC is seldom cured by standard therapy because of undetectable micrometastatic disease that is present at diagnosis. Effective nontoxic adjunctive treatments might improve outcomes for HCC patients who are at high risk for recurrence and death despite surgical resection of their primary hepatoma. Active specific immunotherapy (ASI) with autologous dendritic cells loaded with antigens from autologous tumor stem cell lines has been associated with promising long-term survival in metastatic melanoma, but patients with HBV or HCV were ineligible. Therefore a Phase I safety trial in HCC patients with HBV and/or HIV was warranted. Patients and methods Patients with previously untreated HCC who were candidates for surgical resection, but not curable by resection or liver transplant were enrolled in Shanghai, China. Patients had a solitary lesion >5 cm in diameter, or 3 lesions with at least one > 3 cm, but no regional or distant metastatic disease. Patients had a good performance status, adequate blood counts and organ function. An autologous tumor cell (TC) line was established from the resected tumor. Irradiated autologous TC were incubated with autologous dendritic cells (DC) to create a patient-specific DC-TC product which was cryopreserved. Approximately eight weeks after one course of transarterial chemoembolization therapy (TACE), patients received three weekly subcutaneous injections of DC-TC suspended in 500 micrograms of sargramostim. Patients were monitored for toxicity for 8 weeks from the start of treatment.

Vaccination with Irradiated Autologous Tumor Cells Mixed with Irradiated GM-K562 Cells Provokes Lymphocyte Activation and Specific Antitumor Immunity in Patients with Recurrent Glioblastoma

We have previously reported the safety and feasibility of vaccinating recurrent malignant glioma patients with irradiated autologous cells mixed with irradiated GM-K562 cells. Here, we report the impact of vaccination on the expression of T-lymphocyte co-stimulatory and co-inbhibitory molecules as well as on regulatory T-lymphocytes. Furthermore, we explored the impact of vaccination on patient humoral immune responses with particular attention to antibody titers against molecules associated with tumor angiogenesis. Blood was collected from 9 of 10 treated patients at regular intervals and submitted for immune analysis. 5-color flow cytometry was employed after staining of whole blood by an immunophenotyping antibody panel. Antibody titers to tumor-associated antigens were measured by ELISA of patient plasma - all assays were performed in duplicate. Treatment-associated change in percent expression of various markers was analyzed via the non-parametric sign-rank test. p < 0.05 was considered statistically significant. In patients undergoing craniotomy for recurrent malignant glioma, vaccination with irradiated autologous tumor cells and irradiated GM-K562 cells was associated with statistically significant increases in CD4+ T-lymphocyte expression of costimulatory molecules OX40 and 4-1BB (CD137). CD8+ T-lymphocytes saw increased expression of 4-1BB as well. Likewise, there were increases in CD4+ T-lymphocyte expression of CTLA-4, PD1, and FOXP3 - each of which is associated with negative immune regulation and are targetable with existing monoclonal antibodies. 7 of 9 patients had marked treatment-driven increases in antibody titers to Angiopoietin 2, which may represent a novel glioma vasculature-associated antigen. Vaccination with irradiated autologous tumor cells mixed with irradiated GM-K562 cells in patients with recurrent malignant glioma is biologically active and activates both T-lymphoctes and humoral antitumor immunity. Vaccine-induced expression of costimulatory molecules may provide the rationale for combination therapies with either co-stimulatory receptor agonist or co-inhibitory receptor antagonist molecules.

Autologous CLL cell vaccination early after transplant induces leukemia-specific T cells

Patients with advanced hematologic malignancies remain at risk for relapse following reduced-intensity conditioning (RIC) allogeneic hematopoietic stem cell transplantation (allo-HSCT). We conducted a prospective clinical trial to test whether vaccination with whole leukemia cells early after transplantation facilitates the expansion of leukemia-reactive T cells and thereby enhances antitumor immunity. We enrolled 22 patients with advanced chronic lymphocytic leukemia (CLL), 18 of whom received up to 6 vaccines initiated between days 30 and 45 after transplantation. Each vaccine consisted of irradiated autologous tumor cells admixed with GM-CSF-secreting bystander cells. Serial patient PBMC samples following transplantation were collected, and the impact of vaccination on T cell activity was evaluated. At a median follow-up of 2.9 (range, 1-4) years, the estimated 2-year progression-free and overall survival rates of vaccinated subjects were 82% (95% CI, 54%-94%) and 88% (95% CI, 59%-97%), respectively. Although vaccination only had a modest impact on recovering T cell numbers, CD8+ T cells from vaccinated patients consistently reacted against autologous tumor, but not alloantigen-bearing recipient cells with increased secretion of the effector cytokine IFN-γ, unlike T cells from nonvaccinated CLL patients undergoing allo-HSCT. Further analysis confirmed that 17% (range, 13%-33%) of CD8+ T cell clones isolated from 4 vaccinated patients by limiting dilution of bulk tumor-reactive T cells solely reacted against CLL-associated antigens. Our studies suggest that autologous tumor cell vaccination is an effective strategy to advance long-term leukemia control following allo-HSCT. Clinicaltrials.gov NCT00442130. NCI (5R21CA115043-2), NHLBI (5R01HL103532-03), and Leukemia and Lymphoma Society Translational Research Program.

Autologous Whole Tumor Cell Vaccination Early After Allogeneic Stem Cell Transplantation Elicits Anti-Tumor T Cell Responses in Patients with Advanced Chronic Lymphocytic Leukemia (CLL)

Abstract 1892▪▪This icon denotes a clinically relevant abstractPatients with advanced hematological malignancies remain at high risk for eventual disease progression following reduced intensity conditioning (RIC) allogeneic hematopoietic stem cell transplantation (allo-HSCT). We hypothesized that vaccination with whole leukemia cells during the critical period of immune reconstitution early after transplant may enhance antitumor immunity and facilitate expansion of leukemia-reactive T cell responses. We tested this hypothesis in a prospective clinical trial, in which patients with advanced chronic lymphocytic leukemia (CLL) received up to 6 vaccine doses initiated between day 30–45 following RIC allo-HSCT. Each vaccine consisted of 1×107 irradiated autologous tumor cells admixed with 1×107 irradiated K562 bystander cells secreting GM-CSF (GM-K562). All patients received tacrolimus and mini-methotrexate as graft-versus-disease (GvHD) prophylaxis. Tacrolimus was maintained at therapeutic levels during the vaccination period without taper.Twenty-two patients were enrolled, all with advanced disease (median number of prior therapies 3; range 2–11). Many of the leukemias expressed markers associated with aggressive disease (e.g. unmutated IgVH - 68%) and displayed high-risk cytogenetic abnormalities (sole del(11q) - 41%; sole del(17p) - 23%; del(11q and 17p) - 18%). Greater than 50% (n=13) of patients had persistent marrow involvement (≥10%) at time of allo-HSCT. Eighteen of 22 subjects were vaccinated after allo-HSCT and received a median of 6 (range 1–6) vaccines. The remaining 4 patients were precluded from vaccination due to development of acute GvHD before day 45. Vaccines were generally well tolerated, but mild, transient injection site erythema was common. Only one grade 4 event (neutropenia) with a possible attribution to treatment occurred. We observed a similar incidence of grade II-IV aGvHD at 1 year in the 18 vaccinated patients (39%; 95% CI: 17–61%) and 42 control CLL patients that underwent RIC allo-HSCT at our institution from 2004–2009 (31%; 95%CI: 18–46%). At a median follow-up of 2.9 (range 1–4) years, the estimated 2-year rates of progression-free survival and overall survival of vaccinated study participants were 80% (95% CI: 54–92%) and 84% (95% CI: 58–95%).With these promising clinical results, we next focused on gaining insight into the mechanism that generated the observed clinical graft-versus-leukemia (GvL) responses. To delineate the specific contribution of vaccination to the overall GvL effect, we performed T cell assays to detect CLL-specific reactivity in serial pre- and post-HSCT samples obtained from vaccinated patients (n=9) who received median of 6 vaccines (range 3–6). In comparison, we examined T cell responses in study subjects (n=4) that developed aGvHD at a median of 44.5 days (range 26–56) after HSCT; and control CLL patients (n=4; no vaccine, no GvHD in the early post-transplant period) that were not enrolled in the study. Although early post-transplant vaccination had no impact on recovering absolute T cell numbers, reactivity of CD8+ T cells from the vaccinated patients was consistently directed against autologous tumor cells but not alloantigen bearing-recipient cells (PHA T cell blasts and fibroblasts) in IFNγ ELISpot assays. A peak response against autologous tumor cells was reached at day 60 after allo-HSCT (average 221 SFC/5×105 cells vs. 29 and 33 average SFC/5×105cells for PHA blasts and fibroblasts, respectively). CD8+ T cell clones were isolated from 4 vaccinated study subjects by limiting dilution and 17% (range 13–33%) reacted solely against CLL-associated antigens. In contrast, broad CD8+ T cell reactivity indicating an alloantigen response was observed in GvHD patients, while no increase in T cell reactivity against tumor-associated or alloantigens was seen in control patients. Tumor-reactive CD8+ T cells isolated from vaccinated patients secreted a broad profile of effector cytokines (GM-CSF, TNFα and IP10). Moreover, the amount of cytokines secreted by these CLL-specific CD8+ T cells steadily increased following early post-transplant vaccination, but not after allo-HSCT alone or in relation to GvHD.Our studies reveal that vaccination with autologous whole CLL/GM-K562 cells between days 30–100 after allo-HSCT is associated with induction of immunity against recipient CLL cells, and suggest that this is an effective strategy for promoting GvL following RIC allo-HSCT. Disclosures:Brown:Genzyme, Celgene: Research Funding; Calistoga, Celgene, Genentech, Pharmacyclics, Novartis, Avila: Consultancy. Cutler:Pfizer, inc: Research Funding; Astellas, Inc: Consultancy, Research Funding.