Anita Kremer

Hyperactive mTOR pathway promotes lymphoproliferation and abnormal differentiation in autoimmune lymphoproliferative syndrome.

Autoimmune lymphoproliferative syndrome (ALPS) is a human disorder characterized by defective Fas signaling, resulting in chronic benign lymphoproliferation and accumulation of TCRαβ(+) CD4(-) CD8(-) double-negative T (DNT) cells. Although their phenotype resembles that of terminally differentiated or exhausted T cells, lack of KLRG1, high eomesodermin, and marginal T-bet expression point instead to a long-lived memory state with potent proliferative capacity. Here we show that despite their terminally differentiated phenotype, human ALPS DNT cells exhibit substantial mitotic activity in vivo. Notably, hyperproliferation of ALPS DNT cells is associated with increased basal and activation-induced phosphorylation of serine-threonine kinases Akt and mechanistic target of rapamycin (mTOR). The mTOR inhibitor rapamycin abrogated survival and proliferation of ALPS DNT cells, but not of CD4(+) or CD8(+) T cells in vitro. In vivo, mTOR inhibition reduced proliferation and abnormal differentiation by DNT cells. Importantly, increased mitotic activity and hyperactive mTOR signaling was also observed in recently defined CD4(+) or CD8(+) precursor DNT cells, and mTOR inhibition specifically reduced these cells in vivo, indicating abnormal programming of Fas-deficient T cells before the DNT stage. Thus, our results identify the mTOR pathway as a major regulator of lymphoproliferation and aberrant differentiation in ALPS.

Human Leukocyte Antigen–DO Regulates Surface Presentation of Human Leukocyte Antigen Class II–Restricted Antigens on B Cell Malignancies

Hematological malignancies often express surface HLA class II, making them attractive targets for CD4+ T cell therapy. We previously demonstrated that HLA class II ligands can be divided into DM-resistant and DM-sensitive antigens. In contrast to presentation of DM-resistant antigens, presentation of DM-sensitive antigens is suppressed by HLA-DM but can be rescued by HLA-DO. We also showed that HLA-DO expression remains low in nonhematopoietic cells under inflammatory conditions, suggesting that DM-sensitive antigens may be ideal T cell targets with a low risk for graft-versus-host disease. Here, we demonstrated that B cell malignancies often express HLA-DO and that levels are in particular high in chronic lymphocytic leukemia. Moreover, we showed that surface presentation of DM-sensitive antigens is regulated by HLA-DO, and that DM-sensitive antigens are relevant T cell targets for B cell malignancies and, especially, chronic lymphocytic leukemia. These data open the perspective to target HLA class II ligands with specific processing and presentation behavior for CD4+ T cell therapy of hematological malignancies.

Advances in cellular therapy: 6th international symposium on the clinical use of cellular products, March 24 and 25, 2011, Erlangen, Germany

‘‘Cellular Therapy 2011’’: The 6th International Symposium on the Clinical Use of Cellular Products was held in Erlangen, Germany, on March 24–25, 2011. Twelve years ago, the conference took place for the first time in Regensburg and was originally initiated by Andreas Mackensen. Over the past years, it gained reputation and attracted leading researchers in the field from all over the world. This year’s conference was jointly organized at the University of Erlangen by the Department of Hematology & Oncology (Andreas Mackensen), the Department of Dermatology (Gerold Schuler), Erlangen, and the Department of Hematology & Oncology, Regensburg (Reinhard Andreesen). In principal, the ‘‘Cellular Therapy Meeting’’ aims at providing a multidisciplinary forum for basic and clinical researchers to communicate and to learn about the broad scope of cell therapy from different perspectives. The two-day meeting was attended by approximately 400 international participants and included sessions in the field of immune effector cells (T cells and NK cells), regulatory cells (Treg and MSC), antigen-presenting cells, cancer vaccination, gene-based cellular therapy, metabolism as well as regulatory issues and clinical applications of cellular therapy. These topics were presented by talks from 23 invited international experts. In addition, 20 short talks selected from abstracts of highest scientific quality and almost 130 posters were presented at the meeting. In this report, we summarize the most important research findings and highlight the main discussion topics of the plenary sessions.

Natural T-cell ligands that are created by genetic variants can be transferred between cells by extracellular vesicles

CD4 T cells play a central role as helper cells in adaptive immunity. Presentation of exogenous antigens in MHC class II by professional antigen‐presenting cells is a crucial step in induction of specific CD4 T cells in adaptive immune responses. For efficient induction of immunity against intracellular threats such as viruses or malignant transformations, antigens from HLA class II‐negative infected or transformed cells need to be transferred to surrounding antigen‐presenting cells to allow efficient priming of naive CD4 T cells. Here we show indirect antigen presentation for a subset of natural HLA class II ligands that are created by genetic variants and demonstrated that (neo)antigens can be transferred between cells by extracellular vesicles. Intercellular transfer by extracellular vesicles was not dependent on the T‐cell epitope, but rather on characteristics of the full‐length protein. This mechanism of (neo)antigen transfer from HLA class II‐negative cells to surrounding antigen‐presenting cells may play a crucial role in induction of anti‐tumor immunity.

Abstract CT028: Adoptive transfer of CMV- and EBV- specific peptide-stimulated T cells after allogeneic stem cell transplantation: A Phase I/IIa clinical trial

Allogeneic stem cell transplantation (ASCT) to date is the only permanent curative treatment for many hematological cancers. Besides the development of Graft versus Host disease (GvHD), infections are the major adverse effect of ASCT. Specifically, reactivation of viruses is highly problematic in the aftermath of ASCT. Reactivation of human CMV and EBV negatively impacts on outcome after ASCT. 40-50% of patients reactivate CMV following ASCT, while the only CMV specific antiviral therapy available is ganciclovir, with other drugs being used off-label. For the 20-30% of patients reactivating EBV, only rituximab is available to control EBV. Rituximab leads to long term B-cell depletion requiring frequent administration of immunoglobulins. To cover this unmet medical need of CMV and EBV control after ASCT, we investigate a somatic cell therapy approach by adoptive transfer of CMV- and EBV-specific peptide-stimulated T cells. We specifically are scrutinizing the effect of application of the cells to prevent virus reactivation before its onset or preemptively at the early stages of viral infection. We set up a prospective randomized controlled phase I/IIa multi-center clinical trial to evaluate the safety and efficacy of preventive and preemptive adoptive transfer of this ATMP in patients after ASCT (EudraCT number 2012-004240-30). The design of the trial allows to applicate low numbers of activated T cells, thereby reducing the potential risks of GvHD for the recipient of adoptively transferred T cells. The multi-center trial is currently recruiting, so far, 13 patients have been randomized. ASCT patients are randomly assigned to the intervention or control group. Subjects of the control group receive standard of care. For all subjects of the intervention group, a personalized cell product (ATMP) containing a standardized number of virus specific T cells is manufactured from an aliquot of the leukapheresis product previously used for ASCT and cryopreserved until being administered. Subjects receive the cell product in intervals of at least 30 days, starting with the first adoptive transfer 30 days after ASCT at the earliest. Cells are transferred as preventive, preemptive, or also as therapeutic treatment. Subjects are monitored for occurrence of GvHD, for viral load, as well as for immune reconstitution and composition of the TCR pool, especially of virus-specific T cells. Citation Format: Michael Aigner, Regina Gary, Andreas Moosmann, Stefanie Maas, Julian Strobl, Robert Zimmermann, Jurgen Zingsem, Anita Kremer, Andreas Mackensen, Armin Gerbitz. Adoptive transfer of CMV- and EBV- specific peptide-stimulated T cells after allogeneic stem cell transplantation: A Phase I/IIa clinical trial [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr CT028. doi:10.1158/1538-7445.AM2017-CT028