Form Of Adoptive Immunotherapy Research Articles

Safety and Adverse Reactions of CAR T Therapy for the Treatment of Multiple Myeloma: A Systematic Review

Introduction: Despite multiple treatment options, multiple myeloma (MM) remains an incurable disease. Chimeric antigen receptor T-cell (CAR T) therapy, a form of adoptive immunotherapy, has shown favorable outcomes in the treatment of hematological malignancies. The B cell maturation antigen (BCMA) which is present on myeloma cells, serves as a valid target site for CAR T cells. Early studies have shown promising outcomes in patients (pts) with relapsed and refractory MM (RRMM). However, CAR T therapy is also associated with multiple adverse effects. The main objective of this review is to evaluate the all grades of CRS and other commonly occurring adverse events (AE) associated with CAR T therapy in pts with RRMM. Methods: A systematic search of databases (following PRISMA guidelines) PubMed, Cochrane, Embase and, clinicaltrials.gov was performed with no restrictions of language and time period. A total of 249 articles were identified initially and after a detailed screening, we finalized 16 studies including 12 phase I and 3 phase Ib/II clinical trials and 1 retrospective analysis for data extraction. Results: Total included trials were 15 and trial-level data collected from 392 pts is summarized in Table 1 and Table 2. Commonly used CAR T regimens for the treatment of MM were bb2121 BCMA, P BCMA 101 CAR, LCAR B38 M, and anti CD19 CAR. Single Target; Anti BCMA CAR T (Table 1) BCMA CAR T, LCAR-B38M: In a phase I trial, Cohen et al. (2019) reported G1&2 CRS in 56% pts (n=25) and G3 CRS in 32% pts when treated with BCMA specific CAR T. Leukopenia, neutropenia, and lymphopenia were seen in 44%, 44% and 36% of pts, respectively. In a phase I trial, Wang et al. (2019) reported G1 & G2 CRS in 47% and 35% pts (n=57), respectively and G3 CRS in 7% pts when treated with LCAR-B38M CAR T. Pyrexia, thrombocytopenia, and leukopenia were seen in 90%, 49% and 30% pts, respectively. In a phase I trial, Chen et al. (2019) reported G1/2 CRS in 58% pts (n=17) and G3 and G5 CRS in 35% and 5% pts respectively when treated with LCAR-B38M. Hepatotoxicity and cytopenia were seen in 100% and 82% pts, respectively. In a phase 1 trial, Brudno et al. (2018) reported G1/2 in 56% pts (n=16) and G3 in 37% pts when treated with BCMA CAR T. Anemia, leukopenia and thrombocytopenia were observed in 68%, 62% and 43% pts, respectively. JNJ-4528, bb2121 In a phase Ib/II trial, Berdeja et al. (2020) reported G1/2 CRS in 86% pts (n=29), G3 and G4 CRS in 3.5% pts each when treated with JNJ-4528 (an LCAR-B38M) CAR T. Neutropenia, thrombocytopenia and leukopenia were seen in 100%, 69% and 59% pts, respectively. In a phase I trial, Berdeja et al. (2019) reported G1, G2 and G3 CRS in 22%, 32% and 5% pts (n=22), respectively when treated with bb2121 CAR T. Neurotoxicity was observed in 22% pts. In a phase I trial, Raje et al. (2017) reported CRS of any grade in 76% pts (n=33) and G3 CRS in 6% pts when treated with bb2121 CAR T. Neurotoxicity, hypotension and hyponatremia were observed in 42%, 15% and 15% pts, respectively. Dual Target CAR T (Table 2) Anti-CD19/BCMA, PF-3135, Dual Target BM38: In a phase Ib trial, Shi et al. (2019) reported G1/2 CRS in 97% pts (n=32) while G3/4 CRS in 3% pts when treated with anti-CD19/BCMA CAR T. Other AE were slight and reversible. In a phase I/II trial, Yan et al. (2019) reported G1/G2 CRS in 86% pts (n=21) and G3/4 CRS in 5% pts when treated with humanized anti-CD19/BCMA CAR T. Neutropenia, Anemia and thrombocytopenia were observed in 86%, 62% and 62% pts, respectively. In a phase I trial, Raje et al. (2019) reported G1/2 CRS in 24% pts (n=17) and G3 CRS in 5% pts when treated with PF-3515 (anti-CD3-BCMA) CAR T. Hepatotoxicity, thrombocytopenia, Neutropenia and pyrexia were observed in 18%, 24%, 18% and 18% pts, respectively. In a phase I trial, Li et al. (2019) reported G1/2 CRS in 62% pts (n=16) and G3 CRS in 25% pts when treated with Dual-target BM38 CAR T. No dose-limiting (>/G3) toxicities were observed at a median follow up of 36 months. Conclusion: CAR T therapy is emerging as a treatment modality for RRMM. With promising initial results, there is evidence for multiple side effects, the most concerning and common one is CRS. The most frequently occurring CRS grade is G 1&2 (requiring symptomatic treatment), but G3 CRS is also reported which can be dose-limiting and life-threatening. Neurotoxicity, hematological abnormalities, hepatotoxicity and pyrexia are the other common side effects Disclosures Anwer: Incyte, Seattle Genetics, Acetylon Pharmaceuticals, AbbVie Pharma, Astellas Pharma, Celegene, Millennium Pharmaceuticals.: Honoraria, Research Funding, Speakers Bureau.

Immunotherapy in Pediatric B-Cell Acute Lymphoblastic Leukemia: Advances and Ongoing Challenges.

Leukemia, most commonly B-cell acute lymphoblastic leukemia (B-ALL), accounts for about 30% of childhood cancer diagnoses. While there have been dramatic improvements in childhood ALL outcomes, certain subgroups-particularly those who relapse-fare poorly. In addition, cure is associated with significant short- and long-term side effects. Given these challenges, there is great interest in novel, targeted approaches to therapy. A number of new immunotherapeutic agents have proven to be efficacious in relapsed or refractory disease and are now being investigated in frontline treatment regimens. Blinatumomab (a bispecific T-cell engager that targets cluster of differentiation [CD]-19) and inotuzumab ozogamicin (a humanized antibody-drug conjugate to CD22) have shown the most promise. Chimeric antigen receptor T (CAR-T) cells, a form of adoptive immunotherapy, rely on the transfer of genetically modified effector T cells that have the potential to persist in vivo for years, providing ongoing long-term disease control. In this article, we discuss the clinical biology and treatment of B-ALL with an emphasis on the role of immunotherapy in overcoming the challenges of conventional cytotoxic therapy. As immunotherapy continues to move into the frontline of pediatric B-ALL therapy, we also discuss strategies to address unique side effects associated with these agents and efforts to overcome mechanisms of resistance to immunotherapy.

Donor Lymphocyte-Derived Natural Killer Cells Control MHC Class I-Negative Melanoma.

Natural killer (NK) cells provide a natural defense against MHC-I-negative tumors, such as melanoma. Donor lymphocyte infusion (DLI) containing NK cells, a form of adoptive immunotherapy used after allogenic bone marrow transplantation (allo-BMT), promotes antitumor immune responses but is often associated with life-threatening complications such as graft-versus-host disease (GvHD). Here, we showed that without prior allo-BMT, DLI provoked melanoma control associated with the infiltration and persistence of the transferred NK cells. This allograft acceptance did not correlate with an increase of GvHD; instead it correlated with the expansion and activation of tumor-infiltrating NK cells that expressed the cytotoxic molecules (e.g., IFNγ and granzyme B) and maturation signatures (e.g., CD11bhiCD27lo and KLRGhi/CD43hi). The development of beneficial tumor-infiltrating NK cells of DLI origin required host CD4+ T-cell help in part by producing IL2, as well as by limiting regulatory CD4+ T cells (Treg). IL2 blockade impaired the NK-dependent melanoma control, which could not be rescued by IL2 administration beyond CD4+ T-cell help. Our findings linked NK allograft acceptance-CD4+ T-cell help crosstalk to melanoma development without the need of allo-BMT. We thereby helped define that tumor-infiltrating NK cells of DLI origin may serve as effective therapeutic targets for controlling melanoma.

Is Initial CD3+ T Cell Dose in Donor Lymphocyte Infusion for Relapsed Hematological Malignancies a Prognostic Determinant?

▪Introduction: Patients after relapsing allogeneic hematopoietic stem cell transplantation (AHSCT) have a treatment option of donor lymphocyte infusion (DLI). DLI is a form of adoptive immunotherapy which induces graft versus leukemia effect. Severe forms of graft versus host disease (GVHD) and marrow aplasia are known complications. In this study, we aimed to detect the effect of the infused CD3+ T cell dose on response, GVHD and overall survival (OS) of hematological malignancies after AHSCT.Methods: We retrospectively evaluated 78 DLI procedures in 57 patients with different hematological malignancies for relapse after AHSCT from June 2000 through June 2015. Initial DLI CD3+ cell dose/kg recipient body weight was ≤ 1x10^7 (n=20; Group 1), >1.0 to ≤ 5 x10^7 (n=29; Group 2), 5-10 x10^7 (n=29; Group 3). Chi-square test was used in comparison between groups. P<.05 was considered statistically significant.Results: The median age during DLI was 33,8 ± 13,2 (range 16-67 years) in 57 patients (38M/19F). The median time interval from HCT to DLI was 8.08 (2-69) months. Ten patients (18%) had bone marrow while 47 patients (82%) had peripheral blood as stem cell source. All patients received transplants from HLA-matched related siblings. There was no difference found between the three groups according to age, diagnosis, stem cell source and conditioning regimen. Achieving complete remission after DLI was detected in thirty two (56%) patients. Disease status after DLI, acute and chronic GVHD rates were not statistically different between the groups. Overall survival which was not statistically significant according to initial DLI cell dose at 2 years were 25%, 38% and 24%, respectively.Conclusion: An initial DLI CD3+ cell dose of 10 x10e7 or higher has been shown improve overall survival but increase GVHD rates with no effect on risk of relapse in previous reports. Response rate, GVHD and survival outcomes were found to be similar in comparison of three different DLI dose groups in our study. DisclosuresNo relevant conflicts of interest to declare.

Donor CD3+ lymphocyte infusion after reduced intensity conditioning allogeneic stem cell transplantation: Single-center experience

Donor lymphocyte infusions (DLI) can induce remission in patients with hematologic malignancies who relapse after allogeneic stem cell transplantation. However, graft-vs-host disease (GVHD) remains a major complication of this strategy. We have used escalating doses of DLI for many years, and wanted to assess the risk factors for GVHD and transplant-related mortality as well as disease outcomes according to the reason for DLI. We analyzed 65 patients who received a total of 111 DLI for different reasons and at different intervals after transplantation. Median number of DLI was 2 (range, 1-4), median interval between transplantation and DLI was 9 months (range, 1-41 months) and median number of infused CD3(+) cells/kg recipient body weight was 2.5 × 10(7) (range, 1 × 10(6)-11.8 × 10(7)). Reasons for DLI were relapse or progression in 37 patients (57%), residual disease in 15 patients (23%), and persistence of mixed chimerism in 13 patients (20%). Seven patients (11%) developed acute GVHD grade II to IV and 5 patients (8%) developed extensive chronic GVHD. In univariate analysis, we could identify a transplantation-DLI interval ≤6 months, the dose of DLI (≥1 × 10(7)), and DLI number as predictive factors of GVHD. In multivariate analysis, these results were confirmed only for the transplantation-DLI interval (hazard ratio = 19.48; 2.23-170.34; p = 0.007). Our findings indicate that this form of adoptive immunotherapy is well tolerated and induces a low incidence of GVHD and transplant-related mortality, supporting further investigation as an upfront modality to enhance the graft-vs-tumor response in high-risk patients.

Adoptive Immunotherapy with Cytokine-Induced Killer Cells for Patients with Relapsed Hematologic Malignancies after Allogeneic Hematopoietic Cell Transplantation

Donor leukocyte infusions induce remissions in some patients with hematologic malignancies who relapse after allogeneic hematopoietic cell transplantation (HCT); however, graft-versus-host disease (GVHD) remains the major complication of this strategy. Cytokine-induced killer (CIK) cells are a unique population of cytotoxic T lymphocytes that express the CD3(+)CD56(+) phenotype and show marked up-regulation of the natural killer cell receptor NKG2D (CD314). CIK cells are non-major histocompatibility complex-restricted and NKG2D-dependent in target recognition and cytotoxicity. We explored the feasibility of exvivo expansion of allogeneic CIK cells in patients with relapsed hematologic malignancies after allogeneic HCT. Eighteen patients (median age, 53 years; range, 20-69 years) received CIK cell infusions at escalating doses of 1 × 10(7) CD3(+) cells/kg (n = 4), 5 × 10(7) CD3(+) cells/kg (n = 6), and 1 × 10(8) CD3(+) cells/kg (n = 8). The median expansion of CD3(+) cells was 12-fold (range, 4- to 91-fold). CD3(+)CD56(+) cells represented a median of 11% (range, 4%-44%) of the harvested cells, with a median 31-fold (range, 7- to 515-fold) expansion. Median CD3(+)CD314(+) cell expression was 53% (range, 32%-78%) of harvested cells. Significant cytotoxicity was demonstrated invitro against a panel of human tumor cell lines. Acute GVHD grade I-II was seen in 2 patients, and 1 patient had limited chronic GVHD. After a median follow-up of 20 months (range, 1-69 months) from CIK infusion, the median overall survival was 28 months, and the median event-free survival was 4 months. All deaths were due to relapsed disease; however, 5 patients had longer remissions after infusion of CIK cells than from allogeneic HCT to relapse. Our findings indicate that this form of adoptive immunotherapy is well tolerated and induces a low incidence of GVHD, supporting further investigation as an upfront modality to enhance graft-versus-tumor responses in high-risk patient populations.

Cytokine Induced Killer (CIK) Cells as Post-Transplant Immunotherapy Following Allogeneic Hematopoietic Cell Transplantation.

Donor leukocyte infusions induce remissions in some patients(pts) with hematologic malignancies who relapse after allogeneic hematopoietic cell transplantation(AHCT). Response rates are disease-specific and acute graft vs host disease(GVHD) remains the major complication of this strategy. CIK cells are a unique population of cytotoxic T lymphocytes that express the CD3+CD56+ phenotype and show marked upregulation of the NK cell receptor, NKG2D. CIK cells are non-MHC restricted, NKG2D dependent in target recognition and killing and demonstrate cytotoxicity against a broad array of tumor cell lines including leukemia targets. In preclinical studies, CIK cells have exhibited superior antitumor activity compared to IL-2 activated NK cells with a markedly reduced capacity for acute GVHD. The primary objective of this trial was to determine the feasibility of ex vivo expansion of allogeneic CIK cells suitable for clinical application for pts with relapsed hematologic malignancies after AHCT(myeloablative or non-myeloablative) and to determine the maximum tolerated dose of CIK cells based on CD3+ cell dose. CIK cells were generated from CD3+ precursors by culturing unmobilized peripheral blood mononuclear cells (PBMC) from the patient's matched sibling donor. The PBMCs were cultured for 21–28 days in the Aastrom Replicell® biochamber in the presence of anti-CD3+ monoclonal antibody, IL-2 and IFN-γ. Ten patients with a median age of 50 yo(range 29–63 yo) received CIK cell infusions based on CD3+cells/kg at a dose of 1x107 (n=3), 5x107 (n=6) and 1x108 (n=1). The diagnoses of these pts included acute myeloid leukemia(n=4), non-Hodgkins lymphoma(n=2), multiple myeloma(n=3) and Hodgkins disease(n=1). Prior to CIK infusion, all pts underwent cytoreduction for tumor debulking. One pt experienced infusional toxicities consisting of ventricular arrhythmias and transient hypotension. After infusion, grade 3–4 toxicities were seen at the 2nd dose level and included symptomatic ventricular arrhythmias in two patients with 1 of these pts also experiencing transient elevation of transaminases. These dose limiting toxicities(DLTs) resulted in expansion of the cohort at this dose level. No further DLTs were seen in subsequent pts allowing escalation to the 3rd and current dose level. Grade 1 skin acute GVHD was seen in 1 pt and 2 pts had limited chronic GVHD. After a median follow-up time of 432 days (range 2–649) from CIK infusion, the 1 year event free survival and overall survival was 20% and 76%, respectively. The median time to progression was 90 days(range 9–577). Analysis of cell cultures showed that most recovered cells were CD3+ (median 98%, range 90–99%) with a median viability of 87%( range 73–95%) with the median expansion of CD3+ cells being 16 fold(range 9–91 fold). CD3+CD56+ cells represented a median of 12% (range 8–32%) of the harvested cultures with a median 96 fold (range 26–515 fold) expansion. CD8+NKG2D+ expression ranged from 17–61%(median 38%) of harvested cells. Upregulation of the activation markers, CD25+ and CD69+ was also seen. Significant tumor cell killing was demonstrated in vitro by cytotoxicity assays against a panel of tumor cell lines. This data demonstrates successful ex vivo expansion of allogeneic CIK cells in the clinical setting. This form of adoptive immunotherapy is well tolerated by pts, induces a low incidence of GVHD and shows evidence for clinical efficacy.

Pilot Clinical Trial of Adoptive Immunotherapy with Chimeric, Co-Stimulated Tumor-Infiltrating Lymphocytes after Allogeneic Hematopoietic Stem Cell Transplantation.

Treatment of refractory or recurrent malignancy with donor lymphocyte infusion (DLI) after allogeneic hematopoietic stem cell transplantation (alloHSCT) is often not curative, with graft-vs-tumor (GVT) effects frequently accompanied by graft-versus-host disease (GVHD), requiring immune suppression that compromises efficacy. After alloHSCT, chimeric T lymphocytes infiltrating residual tumor (chimeric TIL) may provide enhanced antigen specificity and maintain tumor-specific homing. Compared with DLI, they may have a better GVT effect with less GVHD. Based on the success of autologous TIL therapy for melanoma, we tested the hypothesis that enhanced GVT with limited GVHD could be achieved through administration of ex-vivo activated chimeric TIL after alloHSCT. Preclinical TIL production carried out on several tumor types from non-transplanted patients demonstrated effective T cell isolation, expansion and activation using anti-CD3/CD28 bead co-stimulation, yielding a 10- to 30-fold expansion of CD3+ cells. Clinical evaluation of chimeric TIL therapy was initiated with a 51 year-old woman for metastatic breast cancer whose disease progressed after a T cell-depleted reduced-intensity alloHSCT with delayed DLI from a 6/6 HLA-matched sibling donor and subsequent conventional therapy plus DLI. Two weeks after administration of unmanipulated DLI, two thoracic metastases were surgically removed. T cells were liberated from 9.4 cm of tumor using enzymatic digestion and mechanical dispersion, lymphocyte-enriched by density gradient separation, and expanded for 14 days through co-stimulation with anti-CD3/CD28-coated magnetic beads (3:1 bead-to-total nucleated cell ratio) and media containing IL-2 (100 or 1000 IU/mL). This process yielded 42.5 x 106 cells, 33% expressing CD3, and generated 14.7 x 109 chimeric TIL, 85% expressing CD3 (a 3.1-log T cell expansion). There was no tumor contamination of the T cell product by immunohistochemistry. Flow cytometry demonstrated that the CD4/CD8 T cell ratio increased from 1.3 to 1.9 after expansion. Three infusions of the chimeric TIL product were given in a dose-escalating manner (5, 25 and 100 x 106 CD3+ cells/kg). A fourth infusion was given in conjunction with low-dose IL-2 (6mu SQ per day x 3D). CT scan performed after each infusion monitored disease response, with progressive disease the indication for the next dose administration. No infusion-related or delayed toxicities were observed, except for rigors following IL-2 administration. The patient shows no evidence of GVHD, even after the highest dose of 108 allogeneic T cells. Evaluation of the remaining thoracic lesion demonstrated progressive disease after the first two chimeric TIL dose-levels, transient disease stability one month after the third dose-level, and stable disease one month after the fourth dose with IL-2. This is the first clinical report of the application of chimeric TIL and represents a novel approach for developing new forms of adoptive immunotherapy in the setting of alloHSCT.

Therapeutic effects of adoptive splenocyte transfer following in situ AdIL-12 gene therapy in a mouse prostate cancer model

We developed a preclinical prostate cancer model to study the feasibility of adoptive immunotherapy for residual tumor following neo-adjuvant in situ adenoviral-vector-mediated interleukin 12 (AdIL-12) gene therapy. Splenocytes were obtained from mice with orthotopic 178-2 BMA metastatic mouse prostate cancers treated previously with AdIL-12, or a vector with the IL-12 genes plus the costimulatory gene B7-1 (AdIL-12/B7), or a control gene (Adbetagal). The splenocytes were subsequently injected intravenously into syngeneic mice bearing orthotopic 178-2 BMA tumors generated 3 days previously. Significant orthotopic tumor growth suppression was achieved with splenocytes derived from mice whose tumors had been injected with AdIL-12 compared to splenocytes from control Adbetagal mice (P = 0.0005) and splenocytes from AdIL-12/B7-treated mice significantly suppressed spontaneous lung metastases compared to splenocytes from control mice (P = 0.0356). Adoptive transfer of splenocytes from either AdIL-12 (P = 0.004) or AdIL-12/B7 (P = 0.009)-treated mice significantly prolonged survival relative to controls. Transfer of NK and tumor-specific CTL activities was detected and depletion of CD4+ and CD8+ T cells by in vitro antibody-mediated complement lysis of the splenocytes prior to injection abrogated the effects. Systemic IL-12 administration delivered by intramuscular AdIL-12 injection enhanced the antitumor effects of adoptive splenocyte transfer and boosted the CTL response. Our data provide evidence that this form of adoptive immunotherapy can enhance the effectiveness of neo-adjuvant in situ IL-12 gene therapy in cases of persistent malignancy.

Graft-versus-myeloma effect in two cases

Summary Background Allogeneic bone-marrow transplantation (BMT) is associated with the graft-versus-leukaemia effect because of the antileukaemic action of donor lymphocytes. We describe a graft-versus-myeloma effect after BMT in multiple myeloma. Methods Two patients with recurrent multiple myeloma after allogeneic BMT (T cells partly depleted, 105 T cells infused per kg) received leucocyte infusions obtained by leukapheresis from their original marrow donors. The patients were a 48-year-old woman and a 49-year-old man. Findings Both patients developed graft-versus-host disease and achieved complete remission of myeloma. Chimerism was complete in both patients in that all peripheral blood cells were of donor origin. Interpretation We see our results as evidence for a graft-versus-myeloma effect. Using this form of adoptive immunotherapy, we could administer 1000-3000 times more T cells than with the earlier BMT.

Treatment of metastatic renal cell carcinoma with autolymphocyte therapy: Low Toxicity Outpatient Approach to Adoptive Immunotherapy Without Use of In Vivo Interleukin-2

Thirty-six patients with Stage IV renal cell carcinoma were treated with autolymphocyte therapy (ALT). This new form of adoptive immunotherapy is based on the infusion of relatively small numbers of autologous lymphocytes that are depleted of suppressor cells and immunized in vitro by a method designed for antigen-specific activation using a 3M KCl extract of autologous tumor and an autologous lymphokine mixture. Patients received six monthly infusions of immunized lymphocytes, all on an outpatient basis. The majority of patients experienced no toxicicity The few reactions that occurred were minor and self-limiting; none required any medical intervention or subsequent delay in therapy. Patients also received oral cimetidine to reduce in vivo suppressor cell function. Survival at twenty-four months is 36 percent. Median survival is fifteen months, a significant improvement over the natural history of this disease. A multi-site, randomized, controlled trial of ALT in renal cell carcinoma has been initiated to confirm that this treatment causes a significant prolongation of survival with virtually no toxicity in these patients.

The use of microcapsules for high density growth of human tumor infiltrating lymphocytes and other immune reactive T cells

The use of microcapsules to achieve high density growth of tumor infiltrating lymphocytes (TIL) and other antigen-specific human T cells is described. Whereas human T cells in suspension cultures usually do not exceed 1–2 × 10 6 cells/ml, densities approaching that found in living tissues (> 10 8 cells/ml) have been observed for microcapsule cultures. TIL and human peripheral blood-derived T cells can be routinely recovered from microcapsules with viabilities ⩾90%. The recovered cells retain their antigen reactivities and bear cell surface phenotypes identical to their counterparts grown in suspension culture. These findings suggest that microcapsule technology could prove valuable in generating the vast numbers of cells required for TIL therapy and other forms of adoptive immunotherapy with T cells.

Preliminary trial of specific targeting therapy against malignant glioma

The results of specific targeting therapy by use of lymphokine-activated killer (LAK) cells treated with bispecific antibody in 10 patients with malignant glioma were compared with the results of therapy with untreated LAK cells in 10 other patients. Both treated and untreated cells were given locally. The bispecific antibody was an anti-CD3 monoclonal antibody (mAb) chemically conjugated to anti-glioma mAb. Of the 10 patients given specific targeting therapy, 4 showed regression of tumour, and in another 4 patients computed tomography and histology suggested eradication of the glioma cells left behind after surgery. No recurrence was detected in the 10 to 18 months of follow-up. Patients receiving untreated LAK cells had recurrences within 1 year except in 1 case, and 8 patients died within 4 years. So far specific targeting therapy seems to be a new useful form of adoptive immunotherapy.

Adoptive immunotherapy for stage IV renal cell carcinoma: A novel protocol utilizing periodate and interleukin-2-activated autologous leukocytes and continuous infusions of low-dose interleukin-2

In a pilot study involving 13 patients with advanced stage IV renal cell carcinoma, anti-tumor effects and toxicity of a novel form of adoptive immunotherapy were determined. The protocol utilizes infusions of autologous mononuclear leukocytes treated with the oxidizing mitogen sodium periodate (IO 4 −) and cultured in medium containing human recombinant interleukin-2 (IL-2), and continuous infusions of low-dose IL-2 (mean ± SD dose = 39.5 ± 8.6 × 10 3 U/kg/24 hours). Leukocytes (5 to 10 × 10 9) were removed by leukapheresis three times per week, mononuclear cells were separated, activated with IO 4 − and cultured in medium containing IL-2 (500 U/ml) for 48 to 72 hours. The cells were re-infused following the next leukapheresis procedure. IL-2 was administered five days per week. Treatment was continued for two three-week cycles. An increase in peripheral blood mononuclear cells bearing the natural killer cell (NK) surface marker, Leu 11, an increase in NK- and antibody-dependent cell-mediated cytotoxicity, and a slight increase in spontaneous cytotoxicity for non-NK targets were noted. Regressions (more than 50 percent decrease in tumor mass) of pulmonary, liver, bone, or soft tissue metastases were induced in six patients. Severe fluid retention did not develop in any patient and no patient required treatment in the intensive care unit. Five of the patients who showed a response have experienced a relapse at 5.2 ± 1.0 (mean ± SD) months. These observations indicate that IO 4 −/IL-2-activated killer cells plus continuous infusions of low-dose IL-2 can result in regression of metastatic renal cell carcinoma.

LOW-TOXICITY OUTPATIENT ADOPTIVE IMMUNOTHERAPY OF METASTATIC CANCER WITH AUTOLOGOUS LYMPHOCYTES GENERATED BY IN VITRO IMMUNIZATION (IVI)

We have previously described results with a novel form of adoptive immunotherapy using in vitro immunized (IVI) autologous lymphocytes. We now report results from an on-going study using an improved version of that method. Patient lymphocytes are depleted of suppressor T-cells and in vitro immunized for 60 hours in medium containing autologous serum, mixed lymphocyte culture supernatant and a 3M KCl autologous tumor extract. The resultant IVI cells are washed, depleted again of suppressor cells and infused into the patient. All patients also receive cimetidine to block in vivo suppressor cells. Patients will be treated monthly for 6 months. To date, 30 patients have received a total of 113 cell infusions, all delivered on an outpatient basis, (average of 109 IVI cells/infusion). Toxicity has been minimal, consisting only of mild, transient fever and chills accompanying 42 of the infusions (37%). Twenty-one patients have reached the first evaluation timepoint at 3 mqnths.This approach to immunotherapy is safe and appears to be biologically active, in that 7 of 21 evaluable patients (33%) have evidence of stable disease or objective response after only 3 months of treatment.