Immunologic approaches to antigen discovery for cancer vaccines.

Since the early 1990s, scientists have identified an ever-expanding number of antigens to serve as targets for experimental cancer treatments, based on the stimulation of a patient's immune system. Using both immune cells and serum to screen potential candidates, several promising antigens are currently components of vaccines directed against a wide range of tumour types. These antigens vary in their tumour- and tissue-specificity. Their utility as a single reagent or as part of a multi-dimensional approach is as varied as the genes themselves. However, there are already reports indicating that the promise of evoking a clinically beneficial immune response, toward human tumours, is being fulfilled. In this review, we provide a summary of the current status of immunologic approaches to antigen discovery. We also discuss the need for additional, supportive data from non-immunologic techniques, as well as the progression of the preclinical process towards target validation.

Prostate cancer immunotherapy at the dawn of the new millennium.

Standard treatments for adenocarcinoma of the prostate, such as surgery, hormones, radiation and chemotherapy, often achieve a clinical response, but this is usually short-lived. Prostate cancer frequently recurs and second-line therapies have a poor response rate. Many clinicians seem comfortable in limiting their philosophy of treating advanced recurrent disease merely to new regimens of failed therapies, such as combination chemotherapy. However, other medical researchers have chosen to pursue novel approaches, including immunotherapy, several of which are summarised in this review. Although ranging widely in antigen specificity, all attempt to exploit the body's natural antitumour immunity. Furthermore, all aim to stimulate immunity above a threshold level necessary for tumour regression or to induce stability in the face of progression. The goal of in vivo or ex vivo gene therapy is the modification of gene expression within an antigen-presented cell by the introduction of a vector, DNA, or RNA. Within that field, much progress has been made and is ongoing currently concerning gene delivery systems, target identification and characterisation. Comparatively, monoclonal antibodies are an established type of cancer immunotherapy. However, the more recent development of humanized or fully human antibodies, as well as novel moieties they can be coupled to, renews their prospects for clinical impact. Lastly, various cell-based therapies are the focus of several recent clinical studies demonstrating tumour regression or stabilisation. Immune cells, for example, T-lymphocytes and dendritic cells, have already demonstrated treatment benefit, as well as the ability to maintain an excellent quality of life for participants. Overall, there is a multitude of approaches being considered for the treatment of prostate cancer. The following review concentrates on those approaches that are currently in human or animal studies and have a specific emphasis on prostate cancer.

Progress in active-specific immunotherapy of brain malignancies.

Despite the significant advances in neurosurgical techniques and oncology treatment regimens, the prognosis of patients with brain malignancies remains dismal. Brain tumours remain as lethal in the beginning of this new millennium as they were 30 years ago. Among the promising treatment modalities being tested are various immunotherapeutic approaches. Development of cancer vaccines, also known as active-specific immunotherapy, for malignant brain tumours is summarised in this review. Understanding the mechanisms behind vaccinations and the initiation of immune response have helped the design and improvement of the efficacy of clinical vaccines. The emergence of the antigen-presenting properties of dendritic cells brings the cancer vaccine field into a new generation. Preclinical work on the use of dendritic cell-based vaccine for malignant brain tumours are encouraging. The move from these preliminary studies to the clinic is anticipated with high hope.

Dendritic cell-based immunotherapy of prostate cancer: immune monitoring of a phase II clinical trial.

We assessed both non- and peptide-specific immune responses in prostate cancer patients before and after immunotherapy with dendritic cells exogenously pulsed with the prostate-specific membrane antigen-derived peptides, PSM-P1 and PSM-P2. For all subjects, we observed that clinical responses were strongly associated with two indicators of immunocompetence: skin test responses to recall antigens and cytokine secretion by T cells after nonspecific stimulation. In a subset of responders, we observed cytokine secretion or cytotoxicity against the immunizing peptides or an immunodominant epitope from an influenza recall antigen. The clinical results support the use of monitoring for overall immunocompetence to help determine why a patient has or has not responded to therapy. Moreover, it could be useful as an inclusion criterion to select those more likely to benefit from treatment.

American Association for Cancer Research: dendritic cells: strategies and vaccines. April 1-5, 2000, San Francisco, CA, USA.

Several major sessions on dendritic cells (DCs) reflected the importance of these members of the immune system within cancer research. Technical hurdles to their routine propagation were largely overcome within the past decade, so the study of precursors, as well as the numerous types of dendritic cells, is the focus of intense research. In vitro studies were presented that described further refinements in dendritic cells are collected and culture. Cytokines and other growth factors have been analysed to assess their contribution to the ability of dendritic cells to intake, process and present antigen. In some instances, the dendritic cells themselves are genetically altered to improve their natural biological activity. Many studies have advanced from in vitro work and demonstrated utility in various animal models. Ultimately, there is the hopeful, steady progress into the increasing numbers of experimental immunotherapies against human cancer.

American Association for Cancer Research: Clinical trials of immunotherapeutics and immunologic monitoring. April 1-5, 2000, San Francisco, CA, USA.

A major session at this annual gathering of the cancer researchers from around the globe dealt with the current state of immunotherapy for cancer. Immunotherapy is a form of cancer treatment that enhances its scientific promise and legitimacy with each passing year. As a result, this topic has become one of the most highly attended and anticipated of all the sessions during the AACR. This year's session included further progress from the laboratory to the clinic involving an ever-increasing number of cancers. For example, brain, lung and prostate cancer are now as well-represented as melanoma and lymphoma at such forums. This year's session continued the trend of impressive biosafety of both cell- and antibody-based vaccines. Therefore, these cancer vaccines offer optimism in treatment benefit as well as a minimal impact on quality of life. Lastly, the increasing number of clinical responses allows for true immunological monitoring, as scientists strive to unlock the mysteries behind what makes one patient respond to treatment and another progress. Several groups discussed in vitro immune parameters that were studied in concert with their clinical trials. Others discussed ways in which those in the immunotherapy community can work towards more reliable immune monitoring and ultimately a surrogate marker for response.

Immune monitoring of cancer patients undergoing experimental immunotherapy.

Advancements in the understanding of cellular immunity within the last decade, along with the characterization of tumor antigens, have led to immunotherapeutic approaches for cancer therapy. This mode of treatment is expected to provide more tumor-specific activity, thereby being less toxic to normal cells than standard modalities. Clinical trials are underway throughout the world to determine whether immunotherapy is a practical and viable alternative to conventional cancer therapies. Unlike radiotherapy and chemotherapy, wherein tumor regression is the standard for determining efficacy of the regimens, immunotherapy has to be evaluated by the examination of several immunological parameters within patients. The purpose of this article is to review the methods currently utilized to evaluate the induction, maintenance, and duration of antitumor immune reactivity in cancer patients undergoing immunotherapy.

AACR - 91st meeting. Dendritic cell vaccine strategies.

Inevitably, the sessions on dendritic cells at the AACR Annual Meeting were some of the most consistently well attended. Interest has been intense for several years, largely since the technical obstacles to the routine culture of these cells and their precursors, both from animal and human sources, were removed in the early 1990s. Several important advances were presented towards further optimizing dendritic cell-based immunotherapy in general. Groups reported on improved culture conditions, as well as more efficient means of obtaining larger quantities of dendritic cells or their precursors. As expected, there were several strong reports on the beneficial bioactivity of cytokines, such as IL-12, GM-CSF, and IL-2. In addition, enticing work continues with Flt3-ligand and has begun with progenipoietin, a more recently identified hematopoietic growth factor. As shown in this year's sessions, the clinical promise of tumor lysate and apoptotic bodies continues to move steadily from the bench to the clinic. Finally, although dendritic cells are excellent antigen-presenters, work to determine if they could be engineered to be even more effective continues. As a result, several reports were given on gene-modifying this type of immune cell.

Expression and purification of prostate-specific membrane antigen in the baculovirus expression system and recognition by prostate-specific membrane antigen-specific T cells.

Antigen-specific immunotherapy of cancer depends on a consistent source of well-defined protein antigen. Production of recombinant protein offers the obvious solution to this problem but few comparisons of recombinant and native proteins in cellular immune assays have been reported. We report expression of a putative immunotherapy antigen, prostate-specific membrane antigen (PSMA), in insect cells using a baculovirus vector. T cells stimulated with recombinant PSMA or native PSMA derived from the LNCaP cell line recognized both native PSMA and recombinant, baculoviral PSMA. These data indicate that PSMA produced in Sf9 cells is immunologically cross-reactive with native PSMA and therefore suitable for immunotherapy as it is recognized by both cellular and humoral immune responses.

Construction and binding analysis of recombinant single-chain TCR derived from tumor-infiltrating lymphocytes and a cytotoxic T lymphocyte clone directed against MAGE-1.

The TCR is responsible for the specificity of cytotoxic T lymphocytes (CTL) by recognizing peptides presented in the context of MHC. By producing recombinant soluble TCR, it is possible to study this interaction at the molecular level. We generated single-chain TCR (scTCR) from tumor infiltrating lymphocytes (TIL) and one CTL clone directed against melanoma-associated antigen (MAGE)-1. Sixty-eight day anti-MAGE-1 TIL and one cloned anti-MAGE-1 CTL were analyzed by PCR for their Valpha and Vbeta gene usage. The TIL population showed a restriction in Valpha and Vbeta usage with only Valpha4 and Valpha9 and Vbeta2 and Vbeta7 expressed. The anti-MAGE-1 CTL clone demonstrated absolute restriction with only Valpha12 and Vbeta1 expressed. DNA sequence analysis was performed on all V regions. For the TIL, each possible Valpha-Vbeta combination (i.e. Valpha4-Vbeta2, Valpha9-Vbeta2, Valpha4-Vbeta7 and Valpha9-Vbeta7) was constructed as a distinct scTCR and the recombinant proteins expressed in bacteria. From the anti-MAGE-1 TIL, Valpha4-Vbeta2 scTCR demonstrated binding activity to HLA-A1(+) cells pulsed with MAGE-1 peptide. Results obtained from screening a panel of our scTCR constructs on HLA-A1(+) cells pulsed with MAGE-1 peptide or irrelevant peptide demonstrated that Vbeta2 plays a significant role in binding to the MAGE-1 peptide. Amino acid alignment analysis showed that each Vbeta sequence is distinctly different from the others. These findings demonstrate that soluble TCR in single-chain format have binding activity. Furthermore, the results indicate that in TCR, like antibodies, one chain may contribute a dominant portion of the binding activity.

Phase II prostate cancer vaccine trial: report of a study involving 37 patients with disease recurrence following primary treatment.

BACKGROUND: A phase II trial was conducted to assess the efficacy of infusions of dendritic cells (DC) and two HLA-A2-specific prostate-specific membrane antigen (PSMA) peptides (PSM-P1 and -P2). This report describes the evaluation of 37 subjects admitted with presumed local recurrence of prostate cancer after primary treatment failure. METHODS: All subjects received six infusions of DC pulsed with PSM-P1 and -P2 at 6-week intervals. Clinical monitoring was conducted pre-, during, and post-phase II study. Data included: complete blood count, bone and total alkaline phosphatase, prostate markers, physical examination, performance status, bone scan, ProstaScint scan, and chest X-ray, as well as other assays to monitor cellular and humoral immune responses. RESULTS: One complete and 10 partial responders were identified from this group based on National Prostate Cancer Project criteria, or on a 50% reduction of prostate-specific antigen (PSA), or on a significant resolution in lesions (biopsy-proven when possible) on ProstaScint scan. CONCLUSIONS: About 30% of study participants in this group showed a positive response at the conclusion of the trial. This study suggests that DC-based cancer vaccines may provide an alternative therapy for prostate cancer patients whose primary treatment failed.

Infusion of dendritic cells pulsed with HLA-A2-specific prostate-specific membrane antigen peptides: a phase II prostate cancer vaccine trial involving patients with hormone-refractory metastatic disease.

BACKGROUND: A phase II trial was conducted to assess the efficacy of infusions of dendritic cells (DC) and two HLA-A2-specific PSMA peptides (PSM-P1 and -P2). This report describes thirty three subjects with hormone-refractory metastatic prostate cancer without prior vaccine therapy history who were evaluated and reported as a group. METHODS: All subjects received six infusions of DC pulsed with PSM-P1 and -P2 at six week intervals. Clinical monitoring was conducted pre-, during, and post- phase II study. Data collected include: complete blood count, bone and total alkaline phosphatase, prostate markers, physical examination, performance status, bone scan, ProstaScint scan, chest x-ray, as well as assays to monitor cellular immune responses. RESULTS: Six partial and two complete responders were identified in the phase II study based on NPCP criteria, plus 50% reduction of prostate-specific antigen (PSA), or resolution in previously measurable lesions on ProstaScint scan. CONCLUSIONS: Over 30% of study participants in this group showed a positive response at the conclusion of the trial. This study suggested that DC-based cancer vaccines may provide an alternative therapy for prostate cancer patients whose disease no longer responds to hormone therapy.

Dendritic cell-based immunotherapy for prostate cancer.

Dendritic cells are unique in their ability to stimulate naive T cells. These investigators have developed a prostate cancer vaccine using autologous dendritic cells as a vehicle to present prostate antigens to T cells in vivo.

Viral vectors for gene transfer into antigen presenting cells.

Crucial insights for vaccine development have come from examining how the immune system responds to antimicrobial vaccines, as well as to viral vectors employed for gene therapy. The effectiveness of a vaccine depends upon both the method of antigen delivery and the presentation of antigen to lymphocytes. Much focus has turned to delivering antigens to dendritic cells, to promote clinically beneficial T- and B-cell responses. Recombinant viral vectors represent a powerful vehicle to deliver genes encoding microbial- or tumor-derived antigens to generate clinically beneficial immunity. Dendritic cell-based and viral vector-based vaccines are currently being evaluated in clinical trials as a means of inducing antitumor immunity.