Frank P. Holladay

Autologous tumor cell vaccination combined with adoptive cellular immunotherapy in patients with Grade III/IV astrocytoma

SummaryBrain tumors are highly resistant to treatment. Their diffuse infiltrative nature and the relative inaccessibility of the brain to blood and lymph are barriers to surgical and cytotoxic treatments alike. Preclinical animal studies demonstrated that intravenously administered tumor antigen-specific T lymphocytes will reject tumors growing in the brain. Specifically activated effector T lymphocytes may be generated by in vivo immunization followed by restimulation of antigen-primed T cells with autologous tumor cells in vitro. In order to apply these findings to humans, feasibility studies of combined active immunization and specific adoptive cellular immunotherapy were performed on fifteen patients with recurrent astrocytoma. The objective was to determine whether; 1) T cells could be grown from peripheral blood of patients immunized with autologous tumor cells, and 2) whether stimulated cells could be safely readministered to patients. Patients were immunized with a combination of their own irradiated tumor cells and Bacillus of Calmette and Guerin. Two weeks later, a mononuclear cell-rich fraction of blood was obtained by leukapheresis. Mononuclear cells were cultured with irradiated autologous tumor cells and interleukin-2. Selective expansion of CD4+ and CD8+ T lymphocytes occurred. Intravenous transfer of stimulated cells to the fifteen patients on twenty-four separate occasions with or without systemic administration of interleukin-2 was tolerated with limited toxicity. The studies established the feasibility of conducting controlled studies of the anti-tumor effects of tumor antigen-specific cellular immunotherapy.

Successful treatment of a malignant rat glioma with cytotoxic T lymphocytes.

Brain tumors are highly resistant to therapy. Their diffuse infiltrative nature and the relative inaccessibility of brain tissue to blood and lymph are barriers to surgical and cytotoxic treatments alike. The purpose of this study was to produce immune cells specifically reactive with an anaplastic rat glioma (RT2) and determine whether those cells could affect tumor progression in the brain. RT2-specific cytotoxic cells were prepared by priming rats in vivo with RT2 tumor cells and Corynebacterium parvum and stimulating the primed lymphocytes in vitro with irradiated RT2 tumor cells and interleukin-2 (IL-2). Cultured cells exhibited a high level of cytotoxicity against RT2, but not C6 (an allogeneic glioma), 3M2N (a syngeneic mammary tumor), or CSE (a syngeneic fibrosarcoma) tumor cells. To generate a model for therapy, rats were injected intracerebrally with RT2, generating progressing brain tumors, which killed untreated rats in approximately 2 weeks. To test the therapeutic potential of the effector cells, tumor-bearing rats were treated by intravenous injection of lymphocytes on Day 5 of tumor growth. Treated rats also received a 5-day course of systemic IL-2 beginning on Day 5. Treatment with IL-2 alone, RT2-primed spleen cells, or RT2-primed spleen cells stimulated in vitro with C6 did not affect rat survival. However, tumor-bearing rats treated with RT2-stimulated lymphocytes exhibited increased survival or were cured. Systemic IL-2 was an essential adjunct, because survival was not affected by treatment with effector cells alone. Therapy initiated on Day 8 of tumor progression lacked effect on survival.(ABSTRACT TRUNCATED AT 250 WORDS)

Generation of cytotoxic immune responses against a rat glioma by in vivo priming and secondary in vitro stimulation with tumor cells.

Cytotoxic T lymphocyte (CTL) responses to most antigens are generated by in vivo priming and secondary stimulation with antigen in vitro. The present studies were designed to determine whether that strategy could be used to stimulate development of CTL against brain tumors. Rats were primed with one of two tumors, RT2, an astrocytoma, or 9L, a gliosarcoma, and Corynebacterium parvum. Spleen cells from primed rats were stimulated with tumor cells and interleukin-2 in vitro to generate CTL. CTL generated against RT2 killed RT2 and 9L, but not allogeneic or histopathologically unrelated tumor cells, suggesting that the killing was brain tumor-specific and major histocompatibility complex gene product-restricted. Similar results were obtained with rats primed and secondarily stimulated with 9L. Specific cytotoxic cells only developed when syngeneic brain tumor cells were used for both priming and secondary stimulation. The cytotoxic cell populations were composed of OX-19+ T cells with a mixed CD4/CD8 phenotype. Controls consisting of spleen cells from unprimed or primed rats tested before culture exhibited low levels of cytotoxicity against brain tumor targets. Culturing unprimed or primed cells with interleukin-2 alone stimulated cell proliferation, but the cells that grew out exhibited only low levels of cytotoxicity for brain tumor cells. Cell populations exhibited consistent cytotoxicity against natural killer cell targets. None of the cell populations killed lymphokine-activated killer cell targets. The results demonstrated that brain tumor-specific CTL could be produced by priming in vivo followed by secondary stimulation with brain tumor cells in vitro. The results further demonstrated that RT2 and 9L share antigens that both induce and serve as target structures for specific cytotoxic cells.

Primary Meningeal Extraosseous Ewing??s Sarcoma: Case Report

A 25-year-old man presented with a suspected right-sided subdural hematoma after a skiing accident. A large hemorrhagic mass was found and was evacuated. Histological studies demonstrated a highly cellular neoplasm with extensive hemorrhage. Further histological, immunohistochemical (including staining for Ewings sarcoma cell surface antigen), and ultrastructural analyses of the tumor were consistent with Ewings sarcoma. Search for other foci of this neoplasm by bone scan, full body computed tomographic scans, magnetic resonance imaging scans of the spine, and a bone marrow aspiration with biopsy failed to detect any soft tissue or bony involvement outside the cranium. This case appears to represent the first report of a primary extraosseous Ewings sarcoma occupying the cranial subdural area.

Primary Meningeal Extraosseous Ewing's Sarcoma

ABSTRACTA 25-YEAR-OLD man presented with a suspected right-sided subdural hematoma after a skiing accident. A large hemorrhagic mass was found and was evacuated. Histological studies demonstrated a highly cellular neoplasm with extensive hemorrhage. Further histological, immunohistochemical (includi

Generation of cellular immune responses against a glioma-associated antigens(s)

The study demonstrated that RT2, a highly malignant anaplastic glioma, expresses antigens that make it susceptible to in vivo adoptive immunotherapy with cytotoxic T lymphocyte-containing immune cell populations. Rats were immunized with irradiated RT2 tumor cells and the adjuvant C. parvum. Lymphocytes from immunized rats were restimulated in vitro with irradiated RT2 tumor cells plus interleukin-2 (IL-2). The cells that proliferated and differentiated in vitro effectively killed RT2, but only low levels of cytotoxicity were observed against other histopathologically related and unrelated, syngeneic and allogeneic target cells. Adoptive transfer of immune cells combined with a 5-day course of systemic IL-2 produced specific regression of brain tumors growing as lung microfoci.

Immune rejection of intracerebral gliomas using lymphocytes from glioma-bearing rats

Naturally occurring malignancies do not induce immune responses against cancer antigens. Is the lack of an immune response caused by an antigen presentation defect or by induced antigen-specific immune suppression? The current study was performed to determine whether a progressing intracerebral malignancy affects production of peripheral autologous glioma antigen-specific immune responses. Peripheral immunization of both glioma-bearing and non-glioma-bearing animals with cancer cells and adjuvant generated similar levels of glioma antigen-specific cytotoxic T lymphocyte activity. However, immune cell populations from glioma bearers were significantly less efficient than immune cell populations from non-cancer bearers in their ability to reject progressing intracerebral tumors. A variety of manipulations designed to reduce nonspecific immune suppression in vivo and in vitro had no effect on the in vivo efficacy of the activated T-lymphocyte populations. The presence of progressing tumors appeared to augment rather than suppress cancer antigen-specific responses, leading to the speculation that reduced efficacy was caused not by generalized immune suppression but rather by a reduction in the number of immune effector cells by either clonal anergy or clonal deletion. Most importantly, the data demonstrated that, despite decreased in vivo efficacy, immune effector cells capable of rejecting an intracerebral malignancy could be generated from cancerous hosts.

cerebellar oligodendroglioma in a child

Oligodendrogliomas occur primarily in the cerebral hemispheres of adults. A rare case of an oligodendroglioma in the cerebellum of a child is presented. The tendency for oligodendroglioma to metastasize through the cerebrospinal fluid (CSF) is reviewed and emphasized. A recommendation for CSF cytology and possible spinal axis irradiation in the treatment of oligodendrogliomas is made.

Autologous Vaccine and Adoptive Cellular Immunotherapy as Treatment for Brain Tumors

This chapter discusses a process that combines vaccination to induce an immune response (IR) against autologous tumor-associated antigens (TAAs) with adoptive transfer of autologous cancer Ag-specific effector T-lymphocytes to treat individuals with progressing cancer. The use of this approach for the treatment of brain malignancy is described. The rationale is that the multiple genetic defects accumulating in cells during malignant transformation and subsequent tumor growth lead to production of altered protein molecules that confer immunogenicity. The host immune system can recognize malignant cells as nonself. Vaccination with autologous cancer cells and an immunologic adjuvant primes T-lymphocytes against cancer Ags, and overcomes the Ag presentation defect that prevents malignancies from being recognized during their natural progression. Cancer Ags and nonspecific Ag receptor stimuli (such as anti-CD3) can activate primed T-lymphocytes to differentiate into Ag-specific effector cells in vitro, and interleukin-2 (IL-2) stimulates these cells to proliferate. Activated effector T-lymphocytes are able to travel to sites of tumor growth, enter and reject the tumor, and potentially cure tumor-bearing animals after being infused into the blood stream. This strategy has been used to cure several types of experimental gliomas, demonstrating that immune privilege, immune suppression, the blood-brain barrier (BBB), Ag presentation defects, and other theoretical barriers to successful IT can be overcome. Phase I/II clinical trials using patients with recurrent malignant astrocytoma have demonstrated that cancer Ag and adoptive transfer (AT) IT is feasible, minimally toxic, and potentially efficacious. The data justify further clinical trials of cancer Ag and AT in humans with central nervous system malignancies.