Immunotherapy Products Research Articles

Promoter usage regulating the surface density of CAR molecules may modulate the kinetics of CAR-T cells in vivo

Although chimeric antigen receptor T (CAR-T) cell therapy achieves high remission rates, challenges (e.g., toxicity management and relapse prevention) remain. The major risks are cytokine release syndrome and related neurological toxicity. The influence of the CAR surface density on the efficacy/safety of CAR-T cell therapy and the factors determining CAR density were not elucidated comprehensively. Here, we discovered that the use of the MND promoter increased the transduction rate and reduced the CAR surface density. Additionally, MND-driven CAR-T cells had prolonged antileukemia activity in a mouse model. In an initial dual-armed anti-CD19 CAR-T cell pilot study (ClinicalTrials.gov: NCT03840317), eight and six subjects were infused with MND and EF1α promoter-driven autologous CAR-T cells (3 × 105 CAR-T cells/kg), respectively. MND subjects developed mild fever and lower interferon gamma (IFN-γ) concentrations than in the EF1A19 group. All but one subject in each cohort reached minimal residual disease (MRD)-negative complete remission after the first month of evaluation. These results represent the first comprehensive study on the promoter-driven modulation of CAR-T cell functionality. These findings encourage further evaluation of the potential of the MND promoter to drive CAR-T cells as a broadly applicable cellular product for anticancer immunotherapy.

Emerging developments in the forefront of peanut oral immunotherapy.

Despite the COVID-19 pandemic, progress continued in the field of peanut oral immunotherapy over the past 12 to 18 months. Of importance, the first oral immunotherapy product for the treatment of peanut allergy was approved by the US Food and Drug Administration in January 2020. Suggested modifications to the practice of oral immunotherapy, some of which may have lasting impacts, were circulated as a result of the pandemic. New advances in pathophysiology, sustained unresponsiveness, quality of life, safety, and cost effectiveness were also published. During 2020, COVID-19 influenced the daily practice of allergy and immunology, with peanut oral immunotherapy being no exception. However, clinicians now have a FDA-approved treatment option for peanut allergy in children, a welcome development for a difficult disease. Future research is needed to clarify several knowledge deficits surrounding the best use of peanut OIT.

In silico identification of natural products from Traditional Chinese Medicine for cancer immunotherapy

Advances in immunotherapy have revolutionized treatments in many types of cancer. Traditional Chinese Medicine (TCM), which has a long history of clinical adjuvant application against cancer, is emerging as an important medical resource for developing innovative cancer treatments, including immunotherapy. In this study, we developed a quantitative and systems pharmacology-based framework to identify TCM-derived natural products for cancer immunotherapy. Specifically, we integrated 381 cancer immune response-related genes and a compound-target interaction network connecting 3273 proteins and 766 natural products from 66 cancer-related herbs based on literature-mining. Via systems pharmacology-based prediction, we uncovered 182 TCM-derived natural products having potential anti-tumor immune responses effect. Importantly, 32 of the 49 most promising natural products (success rate = 65.31%) are validated by multiple evidence, including published experimental data from clinical studies, in vitro and in vivo assays. We further identified the mechanism-of-action of TCM in cancer immunotherapy using network-based functional enrichment analysis. We showcased that three typical natural products (baicalin, wogonin, and oroxylin A) in Huangqin (Scutellaria baicalensis Georgi) potentially overcome resistance of known oncology agents by regulating tumor immunosuppressive microenvironments. In summary, this study offers a novel and effective systems pharmacology infrastructure for potential cancer immunotherapeutic development by exploiting the medical wealth of natural products in TCM.

An Exploratory Field Study With A Subcutaneously Administered Tyrosine Adsorbed Modified Grass Allergen + MPL

This exploratory field study explores the safety and efficacy of an optimized dose of a modified grass allergen subcutaneous immunotherapy (SCIT) product (1.0 ml) with microcrystalline tyrosine (MCT) and monophosphoryl lipid A (MPL) adjuvant system for the treatment of allergic rhinoconjunctivitis (ARC) due to grass pollen.

Allergen Immunotherapy: Biomarkers and Clinical Outcome Measures.

Clinical trials for allergen immunotherapy products’ development and approval are conducted, aiming to monitor safety and efficacy of them. Symptom scores and the use of rescue medication are the primary clinical endpoints used in the conducted clinical trials, while Quality of Life scores and symptom-free days are measurements also used as secondary endpoints. Although the use of in vitro biomarkers might have been more practical and objective, there are yet no broadly used reliable ones accurately reflecting the clinical effects of allergen immunotherapy. On the contrary, in vivo biomarkers, such as the nasal allergy provocation test, are reliable and successfully used. The aim of this review is to describe how to adapt and use biomarkers and clinical outcomes in the everyday practice of Allergists who perform allergen immunotherapy.

No Severe Anaphylaxis in Clinical Trials of House Dust Mite Sublingual Immunotherapy Tablet Using a Standardized Identification Algorithm

A standardized anaphylaxis definition is needed for clinical trials of allergy immunotherapy products. An algorithm in the Standardized MedDRA Query (SMQ) was used to identify anaphylaxis in house dust mite (HDM) sublingual immunotherapy (SLIT)-tablet clinical trials.

Patients' Perspectives and Needs on Novel Food Allergy Treatments in the United States.

PurposeFood allergy management places a daily psychosocial burden on patients and their caregivers. New food allergy treatments may positively impact their lives, but also introduce new stressors. The purpose of this paper is to provide an overview of the current state of the literature regarding patients’ and caregivers’ food allergy experiences and needs within the United States as well as a set of recommendations regarding how best to proceed with patient-centered development and evaluation of new food allergy treatments.Recent findingsThe first pharmaceutical-grade product for peanut oral immunotherapy was approved in the United States for children aged 4–17 years following a successful international Phase 3 trial. This new treatment is only the first of several food allergy treatments currently under development. Patients will soon be presented with multiple options for food allergy treatment and will need to make decisions about what treatment is best for them.SummaryAllergy researchers and providers are encouraged to consider patients’ perspectives and needs when developing and evaluating new food allergy treatments. Recommendations regarding next steps include the development of new patient-reported outcome tools, focus on psychosocial support, health disparities, and financial implications, and research harmonization and interdisciplinary collaboration.

Kinetic Measurement of Apoptosis and Immune Cell Killing Using Live-Cell Imaging and Analysis.

The rapid, efficient detection of cell death is critical for characterizing the underlying biology of in vitro disease models and, in particular, immunotherapy products used for preclinical therapeutic research. Traditional endpoint assays are laborious to perform for mass screening of therapeutic candidates and may fail to fully capture the kinetics of events surrounding the initiation, duration, and mechanisms of cell death-important events that may affect translational relevance and impact therapeutic decision-making during development. Here, we describe simple, efficient methods to measure apoptosis and immune cell killing in both adherent and nonadherent cell populations using the Incucyte® Live-Cell Analysis system and associated nonperturbing reagents, cells, and protocols. Assays are performed in the user's own incubator with minimal disturbance and may be readily incorporated into existing workflows. Users may multiplex to maximize data collection from each sample. The integrated, user-friendly software does not require advanced technical training, enabling rapid analysis. Taken together, this method provides essential kinetic insight for greater understanding of cell death and the dynamic interactions between immune cells and their targets.

DDRE-20. GENERATION OF A HIGH-AFFINITY EGFR/EGFRvIII-TARGETED D2C7 CAR-T CELL

Abstract BACKGROUND The intent of this investigation is to validate a novel, high-affinity epidermal growth factor receptor (EGFR) chimeric antigen receptor (CAR) T-cell product for adoptive immunotherapy of EGFR-expressing malignancies of the central nervous system (CNS). Wild type EGFR is ubiquitously expressed on glial tumors in both children and adults, and the majority of solid tumors that metastasize to the CNS but is not expressed on healthy CNS tissues. EGFR and its isoform mutant, EGFRvIII, are hyperactivated or overexpressed receptor tyrosine kinases described in many human cancers. D2C7 is a recombinant monoclonal antibody short chain variable fragment (scFv) that binds to both wild type EGFR and EGFRvIII. We hypothesize that D2C7 adapted as a CAR on primary human T cells will generate potent, cytotoxic activity against EGFR-expressing tumors in vivo. METHODS In vitro interferon gamma release assays comparing the D2C7 CAR to the EGFRvIII CAR have proven that D2C7 CAR-T cells have high specificity and potent cytotoxicity against established murine glioblastoma (GBM) tumor cell lines, expressing both EGFR and EGFRvIII. Next steps include pre-clinical in vivo testing of the D2C7 CAR-T cells in murine models of CNS tumors expressing EGFR/EGFRvIII such as GBM, non-small cell lung carcinoma (NSCLC), breast carcinoma, and melanoma. CONCLUSIONS To improve and validate the effectiveness of CAR T-cell therapy for EGFR-expressing CNS tumors, our team has designed a high-affinity CAR-T cell that targets both EGFRvIII and wild-type EGFR, known as a D2C7 CAR-T cell. This investigation has established pre-clinical anti-tumor activity of D2C7-CAR T cells in vitro and we plan to present more mature data regarding efficacy in orthotopic murine models of GBM, NSCLC, breast carcinoma, and melanoma at the meeting this Fall.

PRAME mRNA Expression in AML/MDS and HLA Genotype Analysis: Impact on Population Coverage and Design of TCR-Based Immunotherapies

The cancer-testis antigen PRAME (PReferentially expressed Antigen in MElanoma) is expressed at high levels in several oncological indications, including leukemias and solid tumors, as confirmed by mRNA or protein expression studies. Its low to absent expression on normal tissues allows suitable discrimination of tumors for targeted immunotherapy. As an intracellular protein, PRAME-derived peptides can be presented by several HLA class I allotypes, including HLA-A*02:01, and specific T cell recognition has been reported for different PRAME peptide-HLA (pHLA) ligands. Thus, developing a small set of TCRs covering different HLAs could increase the number of PRAME-positive patients eligible for treatment with T cell receptor (TCR)-based immunotherapy approaches. We determined PRAME expression in a group of Caucasian patients with acute myeloid leukemia (AML) and high-risk myelodysplastic syndrome (MDS). In addition, we analyzed the HLA-A genotype distribution in a set of healthy blood donors to determine how many and which different TCRs would be needed to optimally cover a Caucasian population in Germany. For the analysis of PRAME expression, bone marrow (BM) and peripheral blood (PB) samples from 165 patients (AML=133, MDS=32), obtained from multiple clinical sites across Germany, were studied. Ninety-six samples were from newly diagnosed patients (58%) while 68 samples were from relapsed/refractory patients (41%), irrespective of percentage of leukemic blasts. Disease status was not known for one patient. PRAME mRNA expression was measured by quantitative reverse transcriptase polymerase chain reaction (RT-qPCR). A threshold for PRAME-negativity was derived by measuring average PRAME mRNA expression in 22 healthy individuals. Detection of 99 or more copies of PRAME, as measured in 12.5 ng RNA, was considered positive. Of the 165 AML/MDS patients, 79 (48%; AML= 69, MDS=10) were positive for PRAME mRNA. Among relapsed/refractory patients, 52% (35/68) were classified as PRAME-positive, versus 45% (43/96) of first-diagnosis AML/MDS patients. NGS high-resolution HLA-A genotyping showed that 55% of all patients had an HLA-A*02:01 allotype, which is higher than the approximately 45% expected for a random Caucasian population and accounted for by the fact that some patients were included with known HLA-A2 status due to prior assessment for transplant eligibility. In combination, 43 (26%) AML/MDS patients were positive for PRAME above the threshold level as well as for HLA-A*02:01, thereby being potential candidates for treatment with PRAME-specific, HLA-A*02:01-restricted TCR immunotherapy. In a group of 141 healthy local blood donors, NGS high-resolution HLA-A genotyping showed an expected distribution of HLA-A*02:01, HLA-A*01:01, HLA-A*03:01, HLA-A*24:02 and HLA-A*11:01 of 43%, 26%, 25%, 19% and 8% respectively, with some donors heterozygous for two of these particular allotypes. To determine the smallest number of TCRs needed to address the largest number of patients, we assessed how the non-overlapping distribution for each potential HLA-A allotype would contribute to patient coverage. Starting with the 43% of patients who would be covered by an HLA-A*02:01-restricted TCR immunotherapy product, additional coverage of 21%, 14%, 9%, and 1% of patients could be achieved with TCR products using HLA-A*01:01-, HLA-A*03:01-, HLA-A*24:02- and HLA-A*11:01-restricted TCRs, respectively. In conclusion, approximately half of AML/MDS patients are PRAME-positive and could be addressed with PRAME-specific TCR immunotherapy approaches. An HLA-A*02:01-restricted TCR addresses the most patients in a population in Germany at 43%, while coverage could be increased to 87% with addition of three TCRs restricted by the next most common HLA-A allotypes, contingent upon suitable TCR discovery. Disclosures Addo: Medigene Immunotherapies GmbH: Current Employment. Davari:Medigene Immunotherapies: Current Employment. Raffegerst:Medigene Immunotherapies GmbH: Current Employment, Current equity holder in publicly-traded company. Tippmer:Medigene Immunotherapies: Current Employment. Schendel:Medigene AG: Current Employment, Current equity holder in publicly-traded company. Pinkernell:Medigene Immunotherapies GmbH: Current Employment, Current equity holder in publicly-traded company.

Oral Immunotherapy for Food Allergy-a US Regulatory Perspective.

The recent approval of Palforzia for treatment of peanut allergy by the US Food and Drug Administration (USFDA) predicts that additional products for oral immunotherapy (OIT) are on the horizon. In this article, the authors review the legal framework by which the USFDA regulates products for OIT of food allergy and the clinical data that demonstrated that the safety and effectiveness profile of Palforzia supported approval and conclude with a discussion of oral food challenge (OFC) as a clinical endpoint to demonstrate safety and effectiveness of OIT products.

The Impact of a New Interleukin-2-Based Immunotherapy Candidate on Urothelial Cells to Support Use for Intravesical Drug Delivery.

(1) Background: The intravesical instillation of interleukin-2 (IL-2) has been shown to be very well tolerated and promising in patients with bladder malignancies. This study aims to confirm the use of a new IL-2 containing immunotherapy candidate as safe for intravesical application. IL-2, produced in mammalian cells, is glycosylated, because of its unique solubility and stability optimized for intravesical use. (2) Materials and Methods: Urothelial cells and fibroblasts were generated out of porcine bladder and cultured until they reached second passage. Afterwards, they were cultivated in renal epithelial medium (REM) and Dulbecco’s modified Eagles medium (DMEM) with the IL-2 candidate (IMS-Research) and three more types of human interleukin-2 immunotherapy products (IMS-Pure, Natural IL-2, Aldesleukin) in four different concentrations (100, 250, 500, 1000 IU/mL). Cell proliferation was analyzed by water soluble tetrazolium (WST) proliferation assay after 0, 3, and 6 days for single cell culture and co-culture. (3) Results: Proliferation assays showed that all IL-2 products induced very similar cultivation results and none of the IL-2 variants had a negative impact on the proliferation of urothelial cells and fibroblast in either concentration. (4) Conclusion: Human recombinant glycosylated IL-2 as well as human non-glycosylated IL-2 have no negative influence on the tissue cell proliferation of porcine urothelial cells and fibroblasts in vitro and represent a promising and innovative potential intravesical therapy candidate for patients in high need.

The potential markers of NK-92 associated to cytotoxicity against K562 cells

Markers associated to NK cytolytic activity are in a great need to regulate NK cell immunotherapy products. We assume that biomarkers which response to cytolysis will change their transcription, expression or secretion. To find NK-92 indicator to cytolytic activity, we have evaluated the potential markers by quantifying the expression of well-known cytotoxicity functional molecules (cytokine IFN-γ, Granzyme B, perforin, CD69 and CD107a), and explored candidate markers by a sweeping transcription picture of NK-92 using a direct cytolysis model (incubation with K562). We found that IFN-γ secretion was highly correlated to cytotoxicity of NK-92, neither Granzyme B, perforin secretion, nor CD69, CD107a positive population were upregulated by K562 stimulation. RNAseq revealed 432 genes expression changed during cytolysis, several genes (BIRC3, CSF2, VCAM1 and TNFRSF9) mRNA expression were validated by real time RT-PCR under K562 being killed or protected from being killed conditions. Results suggested IFN-γ secretion, BIRC3 and TNFRSF9 transcription in NK-92 were responsive to K562 cytolysis. In a word, our results confirmed one marker and reveal an array of novel candidate markers associated with NK-92 cytotoxicity. Further studies are greatly needed to determine the roles these new makers play in NK-92 cytolysis process.

A smart dual-drug nanosystem based on co-assembly of plant and food-derived natural products for synergistic HCC immunotherapy

Nanotechnology has emerged as an ideal approach for achieving the efficient chemo agent delivery. However, the potential toxicity and unclear internal metabolism of most nano-carriers was still a major obstacle for the clinical application. Herein, a novel “core‒shell” co-assembly carrier-free nanosystem was constructed based on natural sources of ursolic acid (UA) and polyphenol (EGCG) with the EpCAM-aptamer modification for hepatocellular carcinoma (HCC) synergistic treatment. As the nature products derived from food-plant, UA and EGCG had good anticancer activities and low toxicity. With the simple and “green” method, the nanodrugs had the advantages of good stability, pH-responsive and strong penetration of tumor tissues, which was expected to increase tumor cellular uptake, long circulation and effectively avoid the potential defects of traditional carriers. The nanocomplex exhibited the low cytotoxicity in the normal cells in vitro, good biosafety of organic tissues and efficient tumor accumulation in vivo. Importantly, UA combined with EGCG showed the immunotherapy by activating the innate immunity and acquired immunity resulting in significant synergistic therapeutic effect. The research could provide new ideas for the research and development of self-assembly delivery system in the future, and offer effective intervention strategies for clinical HCC treatment.

New guidance on the regulation of allergen products: key aspects and outcomes.

Key aspects and outcomes from the recently published guidance on the regulation of allergen products are summarized. A new regulatory guideline has been published to enhance harmonized national approaches on the regulation of allergen products and thereby strengthen the availability of high-quality products across the European Union (EU). As the guideline was developed, critical aspects for allergen products regulation were identified and are discussed in the document, including recommendations on the regulatory procedures to be applied for diagnostics, allergen immunotherapy products and named-patient products. The new guidance is expected to provide clarifications on and support harmonization of the regulation of allergen products in the EU.

Maintaining excellence in a high volume academic cell therapy production facility through optimization of space utilization, operations, scheduling and staffing

Background & Aim The Clinical Cell and Vaccine Production Facility (CVPF) at UPenn is a FACT accredited resource with a 25 year record of novel first-in-human cell and gene therapy clinical trials. CVPF is integrated with research, process development, and correlative studies laboratories, QC/QA, clinical operations and monitoring, apheresis facilities and infusion sites. CVPF has two clean room facilities in which cell immunotherapy products are currently manufactured in support of >15 investigational new drug (IND) protocols. Methods, Results & Conclusion CVPF recently developed a clean room space utilization model to increase production capacity. Previously, a product occupied a single processing suite for the length of manufacture. Improvements to the processing room layout and implementation of a “bioreactor nursery” allowed for expansion of multiple products in a single space. Other benefits included fewer equipment pieces required and built-in redundancy. Additionally, we developed a standardized production slot model to reduce the rate of unfilled manufacturing slots and maximize facility efficiency. A standardized schedule along with the application of scheduling software has improved resource management, effort tracking, production task scheduling and capacity projections. Standardized scheduling is more efficient in scheduling manufacturing staff, sample transport, environmental monitoring, facility cleaning, equipment maintenance, QC testing, batch record preparation and review and product release. We developed an accelerated training program that reduced time-to-proficiency (TTP) for new hires through resource assignment, dedicated training space and re-designed curricula. A mock processing suite was established outside of the clean room space to allow for non-GMP materials to be used during training, greatly reducing cost. Still, staff recruitment, retention and training continue to be a challenge for academic manufacturing facilities as staff renewal limits production capacity and results in fewer patients enrolled in clinical trials. CVPF joined forces with University leadership to develop a novel metrics based career advancement track for cell therapy technologists. These techniques for increased productivity coupled with improved training methods and staff retention initiatives can be adopted by academic cell therapy facilities to optimize staffing, reduce costs and increase efficiencies, thereby increasing production capacity and patient clinical trial enrollment.

EMA Review of Axicabtagene Ciloleucel (Yescarta) for the Treatment of Diffuse Large B-Cell Lymphoma.

On June 28, 2018, the Committee for Advanced Therapies and the Committee for Medicinal Products for Human Use adopted a positive opinion, recommending the granting of a marketing authorization for the medicinal product Yescarta for the treatment of adult patients with relapsed or refractory diffuse large B-cell lymphoma and primary mediastinal large B-cell lymphoma, after two or more lines of systemic therapy. Yescarta, which was designated as an orphan medicinal product and included in the European Medicines Agency's Priority Medicines scheme, was granted an accelerated review timetable. The active substance of Yescarta is axicabtagene ciloleucel, an engineered autologous T-cell immunotherapy product whereby a patient's own T cells are harvested and genetically modified ex vivo by retroviral transduction using a retroviral vector to express a chimeric antigen receptor (CAR) comprising an anti-CD19 single chain variable fragment linked to CD28 costimulatory domain and CD3-zeta signaling domain. The transduced anti-CD19 CAR T cells are expanded ex vivo and infused back into the patient, where they can recognize and eliminate CD19-expressing cells. The benefits of Yescarta as studied in ZUMA-1 phase II (NCT02348216) were an overall response rate per central review of 66% (95% confidence interval, 56%-75%) at a median follow-up of 15.1 months in the intention to treat population and a complete response rate of 47% with a significant duration. The most common adverse events were cytokine release syndrome, neurological adverse events, infections, pyrexia, diarrhea, nausea, hypotension, and fatigue. IMPLICATIONS FOR PRACTICE: Yescarta (axicabtagene ciloleucel) was the first chimeric antigen receptor T-cell therapy to be submitted for evaluation to the European Medicines Agency and admitted into the "priority medicine" scheme; it was granted accelerated assessment on the basis of anticipated clinical benefit in relapsed/refractory diffuse large B-cell lymphoma, a condition of unmet medical need. Indeed, Yescarta showed an overall response rate of 66% and a complete response rate of 47% with a significant duration and a manageable toxicity that compared very favorably with historical controls. Here the analysis of benefits and risks is presented, and specific challenges with this important novel product are highlighted, providing further insights and reflections for future medical research.

Collection of PBMCs From Healthy Subjects for the Development of Cellular Immunotherapy Product

Collection of PBMCs From Healthy Subjects for the Development of Cellular Immunotherapy Product

Identification and Characterization of Tropomyosin in a Potential Shrimp Oral Immunotherapy Product

Identification and Characterization of Tropomyosin in a Potential Shrimp Oral Immunotherapy Product

Allogeneic Chimeric Antigen Receptor-Invariant Natural Killer T Cells Exert Both Direct and Indirect Antitumor Effects through Host CD8 T Cell Cross-Priming

Introduction Chimeric antigen receptor (CAR) T cells have shown impressive results in B-cell malignancies but, unfortunately, their use is still limited by the logistical and financial burdens related to the need of generating autologous cell products. The development of universal allogeneic CAR T cells to be used off-the-shelf has faced major limitations, namely the risk of Graft-versus-Host-Disease (GvHD) induction and the rejection of the cells by the host immune system. Invariant Natural Killer-T (iNKT) cells are innate lymphocytes deprived of GvHD induction potential and displaying antitumor effects, both directly and indirectly, through enhancement of CD8 T cell responses. Preclinical studies in xenogeneic mouse models demonstrated the feasibility of using iNKT cells as a platform for CAR-based therapies, and two clinical trials are currently ongoing. Objective To assess the immunoadjuvant effect of allogeneic CD19-specific iNKT CAR on the host immune system. Methods We transduced murine iNKT cells from FVB/N (H-2Kq) mice with a CD19-specific CD28/CD3ζ CAR and assessed their antitumor effect in vitro and in vivo. Results CD19-iNKT CAR had a direct cytotoxic effect in vitro against the CD19+ A20 lymphoma cells and significantly improved survival of mice after administration to major histocompatibility complex (MHC)–mismatched, immunodeficient BALB/c (H-2Kd) Rag2−/− gamma-chain−/− mice receiving A20 cells (2 × 10e4; Figure 1A) without inducing any signs of GvHD. To test the efficacy of iNKT CAR cells in the presence of host immune cells, we employed BALB/c mice receiving sublethal irradiation (4.4 Gy), resulting in only a partial and transient lymphopenia. The antitumor effect of allogeneic iNKT CAR cells was greatly enhanced (Figure 1B) suggesting the participation of host cells in the antitumor effect. Interestingly, the iNKT CAR effect was partially abrogated when we employed as recipients BALB/c BATF3−/− mice, in which CD8 T cell cross-priming is impaired as a result of the absence of BATF3-dependent CD103+ CD8a+ dendritic cells known to play a role in iNKT cell interactions with other immune effector cells (Figure 1C). Co-administration of allogeneic FVB/N iNKT CAR with autologous BALB/c CD8 T cells at the time of transfer of T-cell-depleted autologous bone marrow cells and A20 lymphoma cells into lethally irradiated BALB/c recipients resulted in a synergistic effect (Figure 1D). Due to their immuno-adjuvant effect on host CD8 T cells, low numbers of allogeneic iNKT CAR outperformed allogeneic conventional CAR T (Tcon CAR) when administered to sublethally irradiated BALB/c mice receiving A20 cells (Figure 1E-F). Conclusion Collectively, these results demonstrate the potent immunoadjuvant effect exerted by allogeneic iNKT CAR cells toward the host immune system suggesting that iNKT CAR cells are an attractive off the shelf product for adoptive immunotherapy.