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The concept of immunological surveillance. Prog Exp Tumor Res ; Burnet M.

Tumor Immunology and Immunotherapy

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Tumor Immunology

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Castro 8. Methods to edit T cells for cancer immunotherapy Laurie Menger 9. Generating stem-like memory T cells with antioxidants for adoptive cell transfer immunotherapy of cancer Enrico Lugli Reverse immunology: from a peptide sequence to a tumor-killing T-cell clone Pierre van der Bruggen Rapid isolation and enrichment of mouse NK cells for experimental purposes Ignacio Melero Rapid, high-yield isolation of human NK cells from the peripheral blood Jitka Fucikova Dyanmic two-dimensional evaluation of NK cell mediated lysis Anne Caignard Identification of innate lymphoid cells in mouse models Pedro Romero Applications of microfluidic devices in advancing NK-cell migration studies Sam K.

Kung Berzofsky Mixed lymphocytes reaction assay in drug discovery for immune checkpoint blockades Rogze O. Lu Speiser A flow cytometry-based method to screen for modulators of tumor-specific T cell cytotoxicity Santos Manes Tumor Immunology and Immunotherapy — Cellular Methods Part A, Volume , the latest release in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field.

Experts in the field who may want to expand their technical horizons and to newcomers who need detailed introductions to basic techniques.

Lorenzo Galluzzi is best known for major experimental and conceptual contributions to the fields of cell death, autophagy, tumor metabolism and tumor immunology. In particular, he provided profound insights into the links between adaptive stress responses in cancer cells and the activation of a clinically relevant tumor-targeting immune response in the context of chemotherapy and radiation therapy.

Lorenzo Galluzzi has published more than scientific articles in international peer-reviewed journals. One of the now well known regulatory mechanisms which serve to dampen or shut down T-cell responses are immune checkpoint molecules expressed on the T-cell surface, including CTLA-4 and PD An area of active investigation is the dynamic ability of tumor cells to up-regulate or express ligands for these checkpoints such as PD-L1 or PD-L2.

Clearly a combinatorial approach will be needed to overcome the multiple layers of negative regulation, which could include combined checkpoint blockade [ 42 ], adoptive T-cell transfer with lymphablation, or incorporating other modalities such as radiotherapy, which has been shown to upregulate MHC expression and increase susceptibility to immune mediated cell death [ 43 ].

Cancer vaccines, like the conventional vaccines used to prevent infectious diseases, generally involve inoculating a patient with a reagent designed to induce an antigen specific immune response. Infectious disease vaccines, though, are preventative vaccines which rely upon priming the adaptive immune response to generate a memory response which can more rapidly expand upon pathogen exposure and prevent full-blown infection.

In the setting of cancer, oncogenic viruses are an ideal target for preventative cancer vaccines, and the HPV vaccine has been shown in large clinical trials to drastically reduce the chances of developing cervical cancer [ 44 ]. Importantly, multiple other tissues such as oral and anal mucosa are susceptible to HPV mediated transformation, and thus the HPV vaccine has the potential to reduce development of multiple different types of cancer. Other examples of preventative cancer vaccines include the HBV vaccine which can significantly reduce the incidence of hepatocellular carcinoma [ 45 ].

David C. Linehan, M.D.

Therapeutic cancer vaccines, on the other hand ,aim to treat cancer after diagnosis. This task can be much more difficult given the development of immune tolerance mechanisms and the obstacles to immune function as described above. In general, several broad categories of therapeutic cancer vaccines include peptide based vaccine, cell-based vaccines, virus-based vaccine and vaccines based on ex-vivo generated dendritic cells.

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This is a broad topic, which has been well covered in several recent reviews [ 46 , 47 ]. Probably due primarily to recent clinical success [ 48 - 50 ] a great deal of excitement in immunotherapy has surrounded further understanding and modulating immune checkpoints for cancer immunotherapy.

Costimulation classically involves the interaction of B7 with CD28, and disruption of this interaction by the presence of CTLA-4 on the surface of T cells is one example of coinhibition [ 51 , 52 ]. Early work in the field led by Allison and colleagues showed that in preclinical models blockade of CTLA-4 induces an anti-tumor immune response [ 53 ]. This initial body of work culminated in a phase III trial in which CTLA-4 blockade with an anti-CTLA-4 mAb improved overall survival in patients with metastatic melanoma compared to patients receiving a tumor vaccine [ 54 ], and to subsequent approval of the anti-CTLA-4 antibody ipilimumab for metastatic melanoma [ 55 ].

As discussed above, PD-1 inhibition appears to have clinical activity in a variety of cancers, showing durable responses in a proportion of patients, many of whom failed other therapies [ 48 , 49 ]. These checkpoint inhibitors are also being tested in a variety of tumor types including non-small cell lung carcinoma NSCLC , small cell lung carcinoma SCLC , renal cell carcinoma RCC , prostate cancer, and hematological malignancies.

Further emphasizing interest in combined checkpoint blockade, a phase I trial combining PD-1 and LAG-3 inhibition has recently opened. In particular, combining radiation therapy with immunotherapy is an area of intense clinical and pre-clinical research activity, incited to some degree by a recent case report of a potential abscopal effect in a patient with metastatic melanoma [ 60 ].

Continued delineation of the most effective combinatorial approaches for patients is important, as optimal combinations will likely be different for various tumor types. Immune monitoring can define immune correlates of clinical responses and delineate the specificity of anti-tumor immune responses induced by various forms of immunotherapy.

More broadly, monitoring can encompass multiple fields including immunology, pathology, genomics, proteomics, and imaging. At a more cellular level, techniques include ELISPOT assays, flow cytometric analyses, tetramer staining, and intracellular staining for cytokines. Multiplexed assays, sequencing, and array based technologies can also be used to screen more broadly for potential immune responses.

These higher throughput methods include phenotyping of populations, multiplex cytokine arrays, immunogenomics of T-cells and B-cells, sequencing of TCR and BCR, seromics assays to profile antibodies, and immunohistochemistry and imaging of T-cells and B-cells. When selecting assays to monitor immune responses one must address the question of whether the immune response is readily detectable. Another question is whether to draw samples for immune monitoring from the peripheral blood versus locally in-situ. Given the specificity and compartmentalization of the immune system the source of sample for immune monitoring can be critical for proper readout and interpretation of the assay.

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Correlating specific immune responses with clinical outcomes is an aspirational goal for immune monitoring. Such successful correlative studies are relatively rare, but include an association between IFN-gamma response to vaccine and survival in a trial of an autologous DC lysate vaccine in GBM [ 62 ]. In a series of therapeutic vaccine trials for renal cell cancer, patients who received single dose cyclophosphamide and manifested a multipeptide immune response to the IMA vaccine had a longer survival than those with no detectable immune responses or single peptide immune responses [ 63 ].

These studies highlight the potential relevance of immune monitoring when using immunotherapy, as the clinical data gained from analyzing patient responses could be critical to guiding scheduling, dosing, and optimal incorporation of immunotherapy into current treatment paradigms. Cancer immunotherapies, despite being attractive and potentially curative treatment strategies, have not demonstrated sufficient clinical activity to justify routine use in most malignancies. Many challenges exist in the field including the difficulty assessing clinical response due to delayed response kinetics, the lack of biomarkers, questions regarding optimal dosing and scheduling of various therapies, and potentially inappropriate patient selection.

Patent selection may be of particular importance in early Phase I studies, which typically enroll heavily pre-treated patients who might be less likely to benefit from immunotherapy. One approach to overcoming these challenges is to perform phase IA and IIA pre-surgical trials — which are often called neo-adjuvant studies.

The primary goal of a pre-surgical clinical trial is to collect tissue and blood for biomarker analysis in order to provide mechanistic insights into the immunotherapy utilized in that trial. These trials are hypothesis-generating endeavors that use a discovery-driven approach in order to bring clinical questions to the lab and then back again to the clinic [ 64 ].

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For example, at the MD Anderson Cancer Center, pre-surgical trials in bladder and prostate cancer have provided insights into the mechanism of action of anti-CTLA-4 therapy. This was the first study reporting immunological changes in both tumor tissues and peripheral blood after treatment with anti-CTLA-4 therapy. Thus, a preoperative clinical trial model is a powerful tool that can help identify biomarkers and other molecular pathways that can be targeted in combination with currently approved agents.

Detailed knowledge of the function of the immune system has been increasing dramatically over the past decade. Though these relationships may play a role in suppressing the formation or progression of certain tumors, there are clearly scenarios in which endogenous anti-tumor immune responses are inhibited through a variety of mechanisms. Thus, a continued exploration of the workings of the innate and adaptive immune systems is paramount to utilizing therapeutic techniques for cancer immunotherapy.

There have been a number of recent advancements in therapeutic approaches utilizing dendritic cells, cancer vaccines, anti-tumor antibodies, adoptive T cell therapy, immune checkpoint blockade, and combinations of these strategies with other modalities such as chemotherapy or radiation therapy. In addition, there are still many challenges and obstacles to overcome in fully realizing the potential of immunotherapy, and there are also important implications for the future of clinical trial design as well in the era of personalized medicine. Some of the approaches reviewed here have led to groundbreaking medical advancements for many cancer patients.

This shared goal of ultimately improving outcomes for patients is the impetus behind educating researchers and clinicians through the SITC Primer. Indeed, a more detailed understanding of the mechanisms of the immune system and its interaction with the tumor microenvironment is central to developing effective therapeutic strategies for cancer immunotherapy. No potential competing interests were disclosed by the other authors. RR, AS, and AW performed the literature search; drafted and edited the manuscript; and RR incorporated all edits from all authors into the manuscript.

PS and CD supervised the preparation of the manuscript and contributed background research and writing for the manuscript.