bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lenalidomide and pomalidomide strongly enhance tumor cell killing in vitro during antibody-dependent cellular cytotoxicity (ADCC) mediated by trastuzumab, cetuximab and rituximab
Authors: Bartlett J. B. , Wu L. , Adams M. , Schafer P. , Muller G. , Stirling D. 
Published: March 2020

3023 Background: Potential mechanisms of action of lenalidomide and pomalidomide (CC-4047) include anti-angiogenic, anti- proliferative and immunomodulatory activities, e.g., enhancement of T cell and NK cell function. Both drugs appear to enhance T cell activation and Th1-type cytokines in cancer patients. Also, both drugs have been shown to enhance rituximab-mediated protection in a mouse lymphoma model. Methods: We have utilized an in vitro ADCC system to assess the ability of these drugs to enhance human NK cell function in response to the approved therapeutic antibodies trastuzumab, cetuximab and rituximab. Results: Pre-treatment of NK cells with either pomalidomide or lenalidomide greatly enhanced IFN-γ production by NK cells in response to IgG in the presence of either IL-2 or IL- 12. In a series of functional ADCC assays, Her2/neu overexpressing breast cancer cells (SKBR3 & MCF-7) pre-coated with trastuzumab, EGFR positive colorectal cancer cells (HCT-116) pre-coated with cetuximab and NHL cell lines (Namalwa, Farage & Raji) pre-coated with rituximab, were exposed to NK cells pre-treated with pomalidomide, lenalidomide or thalidomide. Both pomalidomide and lenalidomide synergistically increased (up to 6-fold) NK cell-mediated killing of antibody coated tumor cells in a dose-dependent manner. Thalidomide had no effect in this system. There was minimal tumor cell killing by antibody alone or in the absence of antibody. The presence of IL-2 or IL-12 was required to see enhanced killing by either drug. Enhanced ADCC was associated with increased signaling in NK cells, specifically with inhibition of the negative regulator SHIP-1. Other downstream effects on signaling pathways are also being investigated. Monocyte-mediated tumor cell ADCC was also enhanced by both drugs. Conclusions: We have shown that pomalidomide and lenalidomide (but not thalidomide) strongly enhance the ability of therapeutic antibodies to induce ADCC via NK cell/monocyte-mediated killing of tumor cells in vitro. These results provide a strong rationale for combination of either lenalidomide or pomalidomide with antibodies to tumor-specific surface antigens in cancer patients. No significant financial relationships to disclose.
A phase II trial of trastuzumab and low dose interleukin-2 in patients with metastatic breast cancer who have previously failed trastuzumab
Authors: Mani A. , Roda J. , Caligiuri M. , Fleming G. , Kaufman P. , Brufsky A. , Carson W. , Shapiro C. 
Published: March 2020

3028 Background: Trastuzumab (TZB) mediates lysis of Her2-pos. breast cancer cells by interleukin-2 (IL2) primed natural killer (NK) cells. We hypothesized that IL2 would augment the effects of TZB. The objective of this study was to determine the response rate to and toxicity of low dose IL2 plus TZB in patients with Her2-pos. metastatic breast cancer who had progressed within 12 months of receiving TZB. Also, we measured the ability of patient (pt) peripheral blood mononuclear cells (PBMC) to produce cytokines and conduct antibody-directed cellular cytotoxicity (ADCC) against Her2-pos. target cells. Methods: Pts received low (1 million IU/m2) and intermediate dose (12 million IU/m2) IL2 plus TZB (4 mg/kg) in each cycle. Low dose IL2 was given on days 2–7 and days 12–21 of cycle 1, and days 4–14 of later cycles. Intermediate dose IL2 was given on days 9–11 of cycle 1, and on days 1–3 of later cycles. TZB was given on day 1 and 8 of cycle 1, and on day 1 of later cycles. Pt plasma and PBMCs were analyzed for levels of serum cytokines and ADCC against a TZB-coated cell line, respectively. Results: Thirteen pts with median age of 52 (range 30–71), and a median of 1 (range 1–2) prior TZB-containing regimens were enrolled. The median number of cycles completed was 4. Five pts had grade 3 or greater toxicities, including fever, nausea, vomiting, diarrhea, dyspnea, and hypercalcemia. Twelve pts had progression of disease, and 1 pt withdrew consent. ADCC of pt PBMCs against a TZB-coated cell line was enhanced in only 1 pt. Two pts had elevated plasma levels of interferon-gamma (IFNg), and one of these pts had a 20-fold increase in IFNg transcript levels. The antiangiogenic chemokines MIG and IP-10 rose significantly over baseline in 11 pts. Conclusions: TZB given with low and intermediate pulse-dose IL2 did not produce a robust immune or clinical response in this pt population. A clinical trial in a TZB-naive population may help to determine the immune effects of this combination of IL2 plus TZB. No significant financial relationships to disclose.
Correlation between NK function and response to trastuzumab in metastatic breast cancer
Authors: Beano A. , Signorino E. , Polimeni M. A. , Mistrangelo M. , Ardine M. , Spadi R. , Donadio M. , Ciuffreda L. , Matera L. 
Published: March 2020

3036 Background: Trastuzumab is a monoclonal antibody selectively directed against Her2 approved for the treatment of Her2 overexpressing breast cancer patients. Its proposed mechanisms of action include also a role in mediating antibody-dependent cellular cytotoxicity (ADCC), through the triggering of the FcγRIII on natural killer (NK) cells. This study addressed the correlation between overall NK function and clinical trastuzumab activity. Methods: Between March and September 2006 22 metastatic patients in treatment with trastuzumab alone (8 mg/kg load and then 6 mg/kg every 3 weeks until disease progression) as maintaining therapy after chemotherapy were analyzed for clinical and immunological responses. According to RECIST criteria, 14 patients obtained a response to trastuzumab, while 8 patients had a disease progression. Patient’s peripheral blood mononuclear cells were tested for cytotoxic activity against standard NK target (the MHC class I-negative K562 cell line) and trastuzumab-coated MCF7 (Her2-negative) and SKBR3 (Her2-positive) human cell lines in a 4-h 51Cr-release cytotoxicity assay in the presence of grading concentrations of effector cells. Results: NK activity was significantly (p<0.05) higher in responder compared to non responder patients at all the four effector:target (50:1 to 6:1) ratios tested. NK activity of non responder patients was significantly lower than that of 25 sex and age matched controls (p<0.02) and this was not merely due to chemotherapy- or tumor-associated immunosuppression, since the values of responder patients did not significantly differ from those of controls. ADCC activity against Her2-positive SKBR3 cells was also significantly higher in responder compared to non responder patients (p<0.05) and markedly so when compared to controls (p<0.001). Conclusions: The fact that, as shown here, normal levels of FcγR- independent and higher than normal levels of FcγR-dependent cytotoxicity are required for trastuzumab response, lends support to a paramount role of NK cells in the mechanism of action of this drug. No significant financial relationships to disclose.
Antitumor effects of lenalidomide in combination with IDEC114 (anti CD80) in lymphoma bearing severe combined immunodeficiency (SCID) mouse model
Authors: Reddy N. M. , Hernandez-Ilizalituri F. , Knight J. , Czuczman M. S. 
Published: March 2020

3037 Background: Lenalidomide is a thalidomide analogue with immunomodulatory effects. We previously demonstrated that lenalidomide enhances the biological activity of rituximab. In our current work we further studied the effects of combining lenalidomide with IDEC114, a primatized anti-CD80 monoclonal antibody, which is undergoing clinical testing in B-cell lymphoma. Methods: Raji cells were exposed in vitro to lenalidomide (10μg/ml) or DMSO over five days. Changes in DNA synthesis were determined by [3H]-thymidine uptake. For ADCC and CMC assays, lenalidomide or control exposed Raji cells were labeled to 51Cr and then exposed to IDEC114 or isotype control and PBMC’s or human serum. For in vivo studies, 6–8 week old SCID mice were inoculated with 1×106 Raji cells via tail vein injection and after a period of 72 hours animals were divided into four cohorts. Lenalidomide was administered intraperitoneally (i.p) at 0.5mg/kg/dose on days +3, +4, +8, +9, +13, +14, +18 and +19. IDEC114 was administered via tail vein injection at 10mg/kg/dose on days +5, +10, +15 and +20. Difference in survival between treatment groups was performed by Kaplan-Meier analysis. Results: In vitro exposure of Raji to lenalidomide for five consecutive days enhanced the anti-proliferative effects of IDEC114 when compared to control. In addition, an improvement in IDEC114-associated ADCC was observed in lenalidomide-exposed Raji cells. In vivo treatment of SCID mice with lenalidomide in combination with IDEC114 led to prolongation of survival (44 days) compared to either biological agent alone (p<0.01). Conclusions: Our current research demonstrates that Lenalidomide when added to IDEC114 has augmented in vitro antitumor activity (i.e, antiproliferation and ADCC) and synergistic effects in vivo (i.e., prolongation of survival). We hypothesize and currently are evaluating whether the improvement in in vivo antitumor activity of IDEC114, when combined with Lenalidomide, is secondary to potential changes in the tumor microenvironment and/or IMiD-primed upregulation of NK cells and ADCC. This promising unique combination of biologics warrants evaluation as a clinical trial. (Supported by USPHS grant PO1-CA103985 from the National Cancer Institute.) No significant financial relationships to disclose.
In vitro expansion of CMV-specific CD4+ T-cells by artificial antigen presentation for adoptive immunotherapy in the post- transplant setting
Authors: Ricafort R. J. , Stephan M. , O’Reilly R. , Sadelain M. 
Published: March 2020

3049 Background: CMV continues to be a major cause of morbidity and mortality in marrow allograft recipients. The importance of CD4+ T-cells in maintaining immunity and therapeutic efficacy in adoptive cellular strategies has been appreciated. With a novel artificial antigen presenting cell (AAPC) system, we induced the expansion of CMV-specific CD4+ T-cells. Methods: AAPCs were generated by using a standardizable line of murine 3T3 cells sequentially transduced to express human ICAM-1, LFA-1, B7.1, HLA A*0201 and DRB1*1101 alleles. Antigen was provided either by peptide pulsing with the HLA-DRB1*1101-restricted, CMV- pp65-derived P92 peptide, or the influenza matrix peptide (IMP) as a control, or by transgene expression of pp65. PBMCs were isolated from a CMV+, HLA*A02, DRB1*11-positive healthy donor after informed consent, and enriched for T-cells via positive selection using sheep RBCs. Cocultures of T-cells with AAPCs were established at a ratio of 10:1. 20 IU/ml of IL-2 was added on d7 and every third day thereafter. On d12- 14 of co-culture, the T-cells were restimulated on a fresh monolayer of AAPCs with antigen. Cells were counted in triplicate via hemocytometer with trypan blue exclusion. Immunophenotyping of expanded T-cells was done via flow cytometric analysis for cell surface expression of CD3, CD8, CD4, and soluble Class II (HLA-DRB1*1101) tetramer specific for the P92 peptide of CMV-pp65. Results: Following 3 rounds of stimulation, antigen-specific fold expansion, defined by DR11,P92-tetramer-positive, CD4+ T-cells, was 313-fold when stimulated on AAPC.A2.DR11.PP65, compared to 5–35-fold using the other AAPC controls. Conclusion: We are able to expand clinically relevant numbers of CMV-specific CD4+ T-cells using a rapid, practicable, and broadly applicable approach. These preclinical studies show the feasibility of generating virus-specific T-cells of desired specificity and HLA restriction for adoptive immunotherapy of severe CMV infections. [Table: see text] No significant financial relationships to disclose.
Accumulation of lymphocytes following adoptive transfer of allogeneic T lymphocytes equipped with an HLA-A2 restricted, MART-1 reactive T Cell Receptor
Authors: Duval L. , Schmidt H. , Steiniche T. , Kaltoft K. , Fode K. , Jensen J. J. , Soerensen S. M. , Nishimura M. I. , von der Maase H. 
Published: March 2020

3050 Background: Adoptive transfer of cytotoxic, antigen specific lymphocytes represents an immunotherapeutic approach currently investigated for the ability to reinforce immunologic reactivity against tumor cells in cancer patients. The present study aimed to elucidate immunological reactions during a phase I immunotherapeutic trial with receptor modified T lymphocytes in stage IV melanoma. The aim was in particular to assess if treatment induced a local immunologic response in terms of accumulation of immune cells in metastases used for injection of cells, and if such a response propagated to metastases not used for injection of cells. Treatment consisted of intra-tumoral injections of allogeneic, cytotoxic T lymphocytes equipped with a MART-1 antigen recognizing T cell receptor. Expression of the T cell receptor was obtained through transduction with the receptor-encoding gene. Methods: Core needle were sampled both from metastases used for injection of cells, and from metastases not used for injection of cells. Biopsies were sampled at day 1, before any administration of cells, and at day 14 following the first 4 injections administered at day 1, 4, 7 and 10. Hereafter, biopsies were formalin fixed, paraffin embedded and immunohistochemically stained by a two layer detection technique with antibodies against; CD4, CD8 and CD57, before enumeration by a stereological counting technique.Wilcoxon matched pairs signed rank-sum test was used for comparison. Results: A total of forty six biopsies were sampled from twelve patients without complications. The number of CD8 positive T lymphocytes and NK cells (CD57+) was shown to increase significantly from day 1 to 14 in metastases used for injection. Analyses of the number of cells in metastases which were not directly used for injection of cells showed differences in some patients but overall, changes were not significant. Conclusions: The current study showed a statistically significant accumulation of immune cells in metastases following intra-tumoral injections of allogeneic, antigen specific, cytotoxic T lymphocytes in stage IV melanoma patients. The number of immune cells in metastases not used for injection did not change significantly during treatment. No significant financial relationships to disclose.
Combination of gemcitabine and adoptive cell therapy of autologous anti-tumor CTL induces clinical activities in patients with refractory lung cancer
Authors: Toh U. , Fujii T. , Takamori S. , Fukunaga M. , Ogo E. , Seki N. , Yamana H. , Shirouzu K. 
Published: March 2020

3053 Background: Cytotoxic T cells selectively kill autologous tumor cells is powerful for adoptive cell therapy of cancer. Gemcitabine (GEM) is able to induce molecular changes in cancer cells that make them to induce an antigen specific CTL response. This study was to evaluate the anti-tumor and the immunological activity using the CTL in combination with GEM. Methods: The 50Gy irradiated autologous tumor cells were cocultured with PBMCs and CTLs was developed with RPMI 1640 and rIL-2 (50 u/ml) for 7–14 days. Nine pts with non small cell lung cancer failed their prior chemotherapy were enrolled this pilot study. GEM regimen [intravenous (iv.) GEM (1000mg/m2) - day 1, 8 and 15 ] was started without CTL administration for 1st cycle. GEM was administered at least 2 cycles with a 1-week interval and combined with iv. CTL therapy (0.9 x 108 - 4.6 x 108 cells/injection + IL-2 0.4 MIU; biweekly for 6 to 12 injections) from 2nd cycle. The mean total administered T cells were reached to 3.9 x 109 - 5.6 x 109. PBMCs were analyzed their surface markers by Flow Cytometry and the cytokine productions of IFN-γ etc. in the serum were measured by ELISA before and after 1st cycle of GEM administration and 3rd cycles of CTL injection. Results: After finishing 2 cycles of GEM and 3 injections of CTL, the ratio of CD4/CD8 in PBMCs increased in 7/9 pts. In contrast, CD3/CD19 decreased in 6/9 pts. The cytokine production of IFN-γ in the serum revealed an increase after treatment, the levels of TGF-β were decreased simultaneously. There was no remarkable change in the levels of NKG2D in the PBMCs and MIG, IP10 in the serum. The clinical response showed PR/SD/PD was 2/5/2. The tumor marker proteins (CEA) were also decreased significantly in 4 of 9 pts. The adverse effects were tolerable with grade <2 fever, nausea and fatigue and no bone marrow suppression was observed. Conclusions: These results suggested the synergistic enhancement of antitumor effect might be induced between CTLs and anti-cancer agent GEM. Marked clinical responses were observed in two pts after the treatment. Thus this chemo-immunotherapy will be applicable for the patients with refractory lung cancer. No significant financial relationships to disclose.
Efficacy of gabapentin on pruritus induced by interleukin-2 (IL-2) treatment in patients with metastatic renal cell carcinoma and melanoma
Authors: Lee S. , Rusciano V. , Dutcher J. P. 
Published: March 2020

3056 Background: IL-2 treatment causes pruritus, but the pathogenic mechanism is not well understood. Its relation to histamine is controversial, and it does not seem to correlate with the eosinophilia induced by IL-2. Direct stimulation of pain nerve fibers, such as A-delta and C fibers by the cytokines induced by IL-2 treatment has been proposed as a possible cause of pruritus. Stabilizing the nerve membrane or nerve synapses by pharmacologic means may improve pruritus, and we have used gabapentin for this purpose. Here we evaluate its efficacy. Methods: 54 patients, 34 Male, 20 female, were treated with IL-2 from January, 2005 to December, 2006. Among 30 patients who complained of pruritus, 17 patients were given gabapentin. Concurrent follow up notes of these 17 patients were reviewed. They were interviewed immediately following IL-2 treatment and retrospectively using a specific IRB approved questionaire, which quantified pruritus according to CTCAE 3.0 criteria, before and after the administration of gabapentin. The CTCAE grading scale is 0 to 3, from none, localized, widespread, to widespread pruritus interfering with daily activities. Data was analyzed with paired student T-test using SPSS 13.0 for data analysis. Results: 15 patients had renal cell carcinoma and 2 had melanoma. Mean age was 56 years. 12 patients were male and 5 were female. Gabapentin was started in hospital in 9 patients and outpatient in 8. Mean initial dose of gabapentin was 865 mg/day, and the mean time for gabapentin to provide symptom relief was 1.088 hours after each dose. According to CTCAE scale, the mean pruritus before gabapentin was 2.41 and it decreased to 0.65 after gabapentin treatment. Paired student T-test for the difference of means before and after the treatment was statistically significant (p < 0.0005). Conclusions: Pruritus induced by IL-2 infusion responds well to gabapentin, which stabilizes nerve fibers and synapses. Although other confounding causes of pruritus cannot be excluded, the major mechanism is likely due to direct or indirect effect of IL-2, which then stimulates the nerve fibers. Thus, we propose gabapentin as an effective treatment against pruritus induced by IL-2. No significant financial relationships to disclose.
HLA-A0201-positive, IL-7/B7.1-cotransfected allogeneic tumor cells as a vaccine in metastatic renal cell cancer—A clinical phase-I trial
Authors: Floercken A. , Kopp J. , Schabath R. , Joehrens-Leder K. , Pohla H. , Schendel D. , Blankenstein T. , Dörken B. , Westermann J. , Pezzutto A. 
Published: March 2020

3072 Background: Tumor vaccination remains a promising experimental approach in RCC. HLA-A0201-restricted T cell immunity against RCC is well established. RCC26 is an allogeneic HLA-A0201+ human RCC cell line, a T cell clone specifically recognising RCC26 and other RCC in the context of HLA-A0201 has previously been generated, the TCR of this T cell clone was characterised. Furthermore, IL-7/B7.1 cotransfected tumor cells are a potent vaccine in animal models. Methods: RCC26 was transfected with pKEx-IL-7-IR-B7 coding for human IL-7 and B7.1 (CD80). 10 HLA-A0201+ patients with metastatic RCC and disease progression under cytokine therapy were included. 10 vaccinations with 2.5–40x106 gene-modified irradiated tumor cells which had been produced under GMP conditions were performed s.c. over 22 weeks. Primary endpoints of the study were feasibility, safety and immunological response, secondary endpoint was clinical response. The protocol was approved by the ethics committee, all patients gave informed consent. Results: Gene-modified RCC26 cells produced IL-7 (3.4 ng/106 cells/24h), more than 90% of the cells were CD80+. Vaccination was feasible and safe with no severe toxicity. Local DTH-reactions were observed in 4 patients. Skin biopsies of the vaccination site showed lymphocytic infiltrates dominated by CD4+T cells. In 8 patients vaccination induced HLA- and /or antinuclear antibodies without clinical signs of autoimmunity. Analysis of the T cell response against RCC-associated antigens is under way. No partial or complete responses could be documented. However, 50% of the patients had stable disease with the longest TTP being 69 weeks. Mean TTP in our cohort was 25 weeks (range 4 to 69 weeks). Conclusion: Our results show that vaccination with an allogeneic gene-modified tumor cell line is feasible and safe. Stable disease lasting up to 69 weeks in a substantial proportion of patients suggests immunological activity of the vaccine. Vaccination of patients with a low tumor burden is a promising strategy for the future, i.e. after surgery or treatment with novel multi-kinase inhibitors. No significant financial relationships to disclose.