BackgroundImmunotherapy based on the adoptive transfer of gene modified T cells is an emerging approach for the induction of tumor-specific immune responses. Memory stem T cells, due to their enhanced antitumor and self-renewal capacity, have become potential candidate for adoptive T cell therapy of cancer. Methods to generate memory stem T cells ex vivo rely on CD3/CD28 costimulation and the use of cytokines such as IL-7 and IL-15 during the entire culture period. However, a strong costimulation may induce differentiation of memory stem T cells to effector memory T cells. Here we show that manipulation of the length of the costimulation and addition of IL-21 enhance the ex vivo expansion of memory stem T cells.MethodsPurified naïve T cells from healthy donors were cultured in the presence of anti-CD3/CD28 coated beads, IL-7, IL-15 and/or IL-21 (25 ng/ml). T cells phenotype from the different memory and effector subpopulations were analyzed by multiparametric flow cytometry.ResultsA short anti-CD3/CD28 costimulation of naïve T cells, combined with IL-7 and IL-15 significantly increased the frequencies of CD4+ and CD8+ memory stem T cells ex vivo, compared to a prolonged costimulation (34.6 ± 4.4 % vs 15.6 ± 4.24 % in CD4+; p = 0.008, and 20.5 ± 4.00 % vs 7.7 ± 2.53 % in CD8+; p = 0.02). Moreover, the addition of IL-21 to this condition further enhanced the enrichment and expansion of CD4+ and CD8+ memory stem T cells with an increase in the absolute numbers (0.7 × 106 ± 0.1 vs 0.26 × 106 ± 0.1 cells for CD4+; p = 0.002 and 1.1 × 106 ± 0.1 vs 0.27 × 106 ± 0.1 cells for CD8+; p = 0.0002; short + IL-21 vs long).ConclusionsThese new in vitro conditions increase the frequencies and expansion of memory stem T cells and may have relevant clinical implications for the generation of this memory T cell subset for adoptive cell therapy of patients with cancer.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-016-0973-y) contains supplementary material, which is available to authorized users.
Objectives. Adoptive cell therapy (ACT) with mature T cells modified with a chimeric antigen receptor has demonstrated improved outcome for B-cell malignancies. However, its application for others such as Hodgkin lymphoma remains a clinical challenge. CD30 antigen, expressed in Hodgkin lymphoma cells, is absent in most healthy tissues, representing an ideal target of ACT for this disease. Despite that, efficacy of CD30-chimeric antigen receptor (CAR) T cells for Hodgkin lymphoma remains modest. Here, we have developed and tested a novel CD30-CAR T to improve efficacy of CD30-CAR therapy, using a targeting epitope within the non-cleavable part of CD30 receptor, and memory stem T cells (T SCM ) to improve engraftment, persistence and antitumor activity. Methods. T SCM-like cultures were generated and expanded ex vivo and transduced at day 1 or 2 with a lentiviral vector encoding the CD30-CAR. Therapeutic in vivo experiments were performed using NSG mice injected with L540 (sc) or L428 (iv) and treated with CD30-CAR T cells when the tumor was established. Results. CD30-CAR T SCM-like cells generated and expanded ex vivo, despite CD30 expression and fratricide killing of CD30 + CAR T cells, were not impaired by soluble CD30 and completely eradicated Hodgkin lymphoma in vivo, showing high persistence and long-lasting immunity. In addition, highly enriched CD30-CAR T SCM-like products confer a survival advantage in vivo, in contrast to more differentiated CAR T cells, with higher tumor infiltration and enhanced antitumor effect. Conclusion. This study supports the use of a refined CD30-CAR T cells with highly enriched T SCM-like products to improve clinical efficacy of CAR T for Hodgkin lymphoma.
BackgroundThe generation of new immunogens able to elicit strong specific immune responses remains a major challenge in the attempts to obtain a prophylactic or therapeutic vaccine against HIV/AIDS. We designed and constructed a defective recombinant virus based on the HIV-1 genome generating infective but non-replicative virions able to elicit broad and strong cellular immune responses in HIV-1 seropositive individuals.ResultsViral particles were generated through transient transfection in producer cells (293-T) of a full length HIV-1 DNA carrying a deletion of 892 base pairs (bp) in the pol gene encompassing the sequence that codes for the reverse transcriptase (NL4-3/ΔRT clone). The viral particles generated were able to enter target cells, but due to the absence of reverse transcriptase no replication was detected. The immunogenic capacity of these particles was assessed by ELISPOT to determine γ-interferon production in a cohort of 69 chronic asymptomatic HIV-1 seropositive individuals. Surprisingly, defective particles produced from NL4-3/ΔRT triggered stronger cellular responses than wild-type HIV-1 viruses inactivated with Aldrithiol-2 (AT-2) and in a larger proportion of individuals (55% versus 23% seropositive individuals tested). Electron microscopy showed that NL4-3/ΔRT virions display immature morphology. Interestingly, wild-type viruses treated with Amprenavir (APV) to induce defective core maturation also induced stronger responses than the same viral particles generated in the absence of protease inhibitors.ConclusionsWe propose that immature HIV-1 virions generated from NL4-3/ΔRT viral clones may represent new prototypes of immunogens with a safer profile and stronger capacity to induce cellular immune responses than wild-type inactivated viral particles.
BackgroundInvariant natural killer T (iNKT) cells are a small population of lymphocytes with unique specificity for glycolipid antigens presented by non-polymorphic CD1d receptor on dendritic cells (DCs). iNKT cells play a central role in tumor immunology since they are implicated in the coordination of innate and adaptive immune responses. These cells can be activated with the prototypic lipid α-galactosylceramide (α-GalCer), stimulating interferon gamma (IFN-γ) production and cytokine secretion, which contribute to the enhancement of T cell activation.MethodsWe evaluated the antitumor effect of a combination of dendritic cells (DCs) and tumor cells with the iNKT cell agonist α-GalCer in a therapeutic model of B cell lymphoma. iNKT, NK and T cell phenotype was determined by flow cytometry. Serum cytokines were analyzed by Luminex technology. Significant differences between survival curves were assessed by the log-rank test. For all other data, Mann–Whitney test was used to analyze the differences between groups.ResultsThis vaccine induced a potent (100% survival), long-lasting and tumor-specific antitumor immune response, that was associated with an increase of both Th1 cytokines and IFN-γ secreting iNKT cells (4.59 ± 0.41% vs. 0.92 ± 0.12% in control group; p = 0.01) and T cells (CD4 IFN-γ+: 3.75 ± 0.59% vs. 0.66 ± 0.18% p = 0.02; CD8 IFN-γ+: 10.61 ± 0.84% vs. 0.47 ± 0.03% p = 0.002). Importantly, natural killer (NK) cells played a critical role in the antitumor effect observed after vaccination.ConclusionsThis study provides clinically relevant data for the development of iNKT-cell based immunotherapy treatments for patients with B cell malignancies.
2019) The novel agonistic iNKT-cell antibody NKT14m induces a therapeutic antitumor response against B-cell lymphoma, OncoImmunology, 8:2, e1546543, ABSTRACT Invariant natural killer T (iNKT) cells are a small population of T lymphocytes that expresses an invariant T cell receptor with a unique specificity for glycolipid antigens. Their activation using the glycolipid αgalactosylceramide (α-GalCer) triggers innate and adaptive immune responses. The use of α-GalCer in preclinical models as a single antitumor treatment showed moderate effect, but its efficacy in cancer patients was less effective. In addition, this glycolipid induces long-term iNKT-cell anergy precluding the possibility of retreatment. Recently, the first murine iNKT-cell agonistic antibody, NKT14m, has been developed. Here, we analyzed, for the first time, the antitumor efficacy of NKT14m in a B-cell lymphoma model. In a therapeutic setting, a single dose of NKT14m had a moderate antitumor efficacy that was associated with an increase of IFN-γ producing iNKT cells even after a second dose of the NKT14m antibody. Importantly, the combination of a single dose of NKT14m with cyclophosphamide had a potent antitumor efficacy and long-lasting immunity in vivo. Our findings provide the first evidence of the in vivo antitumor efficacy of NKT14m antibody, showing that, either alone or in combination with chemotherapy, induces an effective antitumor response. These results open new opportunities for iNKTcell mediated immunotherapy to treat B-cell lymphoma. ARTICLE HISTORY
Immunotherapy with T cells genetically modified with chimeric antigen receptors (CARs) has shown significant clinical efficacy in patients with relapsed/refractory B-cell lymphoma. Nevertheless, more than 50% of treated patients do not benefit from such therapy due to either absence of response or further relapse. Elucidation of clinical and biological features that would predict clinical response to CART19 therapy is of paramount importance and eventually may allow for selection of those patients with greater chances of response. In the last 5 years, significant clinical experience has been obtained in the treatment of diffuse large B-cell lymphoma (DLBCL) patients with CAR19 T cells, and major advances have been made on the understanding of CART19 efficacy mechanisms. In this review, we discuss clinical and tumor features associated with response to CART19 in DLBCL patients as well as the impact of biological features of the infusion CART19 product on the clinical response. Prognosis of DLBCL patients that fail CART19 is poor and therapeutic approaches with new drugs are also discussed.
Introduction: Adoptive transfer of gene modified T cells (ACT) with chimeric antigen receptors (CARs) is becoming a clinically relevant immunotherapy approach for cancer treatment. One important question involves the selection of the T cell subpopulation for gene-modified ACT. Recent preclinical studies have shown that memory stem T cells (TSCM) (with a phenotype CD45RO-, CD45RA+, CCR7+, CD62L+, CD95+) have higher in vivo persistence, self-renewal, proliferative and antitumor capacities compared to other memory and effector T cell subpopulations. For these reasons, TSCM may represent an ideal candidate for ACT with CARs. Ex vivo generation of TSCM rely on CD3/CD28 costimulation and the use of cytokines such as IL-7 and IL-15 during the entire culture period. However, a strong costimulation may induce differentiation of TSCM to effector memory T cells. In this study, we have analyzed the impact of the costimulation length and the addition of the IL-21 on the ex vivo generation of TSCM cells. Methods: Purified naïve T cells from healthy donors were cultured in the presence of anti-CD3/CD28 coated beads, IL-7, IL-15 and/or IL-21 (25ng/ml). T cells phenotype from the different memory (TCM, TSCM and TEM) and effector subpopulations were analyzed by multiparametric flow cytometry. Particularly, TSCM were identified by a specific plotting strategy. Briefly, CD4+ and CD8+ cells were gated and a CCR7 against CD45RO plot were used to select CCR7+ CD45RO- cells in each subpopulation. Within these cells, a CCR7+ CD45RO- CD45RA+ cell population was identified by plotting CCR7 against CD45RA. Then, CD27 was used to detect CCR7+ CD45RO- CD45RA+ CD27+ cells. Finally, a plot of CD95 against CCR7 was used to identify the CCR7+ CD45RO- CD45RA+ CD27+ CD95+ cell population. Cellular proliferation was analyzed by flow cytometry using CFSE. Results: A short (48 hours) anti-CD3/CD28 costimulation of naïve T cells combined with IL-7 and IL-15 significantly increased the frequencies of CD4+ and CD8+ TSCMex vivo, compared to a long (10 days) costimulation (34.6%±4.4% vs 15.6%±4.24% for CD4+; p=0.008, and 20.5%±4.00% vs 7.7%±2.53% for CD8+; p=0.02). Moreover, the addition of IL-21 to this condition further enhanced the enrichment (47.9%±4.1% vs 34.6%±4.4% for CD4+; p=0.006, and 34.1%±3.5% vs 20.5%±4% for CD8+; p<0.0005) and expansion of CD4+ and CD8+ TSCM, with an increase in absolute numbers (311.3±39 vs 192.8±58.6 fold expansion in CD4+; p=0.04, and 728.1±53 vs 442.7±122 fold expansion in CD8+; p=0.04). In addition, IL-21 did not preferentially enriched for the CD8+ population since the CD4+ TSCM were maintained over the cell culture period in all the conditions tested (CD4+/CD8+ TSCM ratio:1.03±0.28 vs 1.34±0.19 for long costimulation versus long + IL-21, and 0.68±0.14 vs 0.72±0.002 for short vs short + IL-21; p=0.23). Conclusion: We show that these refined in vitro conditions significantly increase the frequencies and expansion of TSCM. Since the reagents used are available in a clinical grade setting, these data may have relevant clinical implications for the generation of this memory T cell subset for adoptive cell therapy with CAR-T cells of patients with cancer. Disclosures No relevant conflicts of interest to declare.
Background: Background:Up to 30% of Hodgkin lymphoma (HL) patients are refractory or relapse (R/R) after first treatment and their prognosis is poor. We have developed a refined CD30-CAR-T (HSP-CAR30) targeting a proximal epitope within the CD30 molecule to overcome soluble CD30 and generated products enriched in memory T-cells to ensure efficient engraftment, persistence and enhancement of antitumor efficacy (Alvarez-Fernandez et al, 2021). Here, we report the results of our Phase 1 study evaluating HSP-CAR30 for the treatment of R/R HL and CD30 + T-cell non-Hodgkin lymphoma (T-NHL) (NCT04653649).
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.