bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sipuleucel-T (sip-T) induced lytic CD8+ T cell responses to target antigens in men with hormone-sensitive and castration-resistant prostate cancer (CRPC).
Authors: Antonarakis Emmanuel S. , Quinn David I. , Kibel Adam S. , Petrylak Daniel Peter , Vu Tuyen , Campogan Dwayne , Chang Nancy N. , Trager James B. , Sheikh Nadeem Anwar , Drake Charles G. 
Published: November 2019

162 Background: Sip-T is an FDA-approved immunotherapy for patients (pts) with asymptomatic or minimally symptomatic metastatic CRPC. Sip-T is manufactured from autologous peripheral blood mononuclear cells cultured with the immunogen PA2024, a fusion antigen of prostatic acid phosphatase (PAP) conjugated to granulocyte macrophage colony-stimulating factor. After sip-T, antibody and T cell responses to PA2024 and/or PAP correlate with improved survival. To further elucidate the mechanism of sip-T–induced immune responses, we evaluated the proliferative and lytic ability of PA2024- and PAP-specific CD8+ T cells. Methods: Mononuclear blood cells were labeled with the membrane dye carboxyfluorescein succinimidyl ester (CFSE) and cultured with PA2024 or PAP. In vitro proliferative and lytic CD8+ (cytotoxic T lymphocyte [CTL]) T cell responses to these antigens were evaluated by flow cytometry. For proliferation, progressive dilution of CFSE was measured. For CTL activity, the loss of intracellular granzyme B (GzB), indicating exocytosis of this apoptosis-mediating enzyme, was assessed. Samples were from 2 sip-T clinical trials STAND (NCT01431391) and STRIDE (NCT01981122), hormone-sensitive and CRPC pts, respectively. Results: Six wk after sip-T administration, CD8+ PAP- and PA2024-specific responses were observed (n=14 pts assessed). The magnitude of PA2024-specific CD8+ proliferative responses was greater than that for PAP-specific responses. CD8+ T cells from a subset of pts who exhibited PA2024- and/or PAP-specific proliferative responses were assessed for lytic ability. After in vitro antigen stimulation, CTL activity in all evaluated samples (n=14, PA2024; n=13, PAP) was demonstrated by a significant decrease (p<0.05) in intracellular GzB relative to a no-antigen control. Conclusions: Sip-T induced CD8+ CTL proliferation against the target antigens PAP and PA2024. Moreover, antigen-specific CTL activity provides the first direct evidence that sip-T can induce tumor cell lysis. These antigen-specific CD8+ lytic abilities were observed within 6 wk following sip-T, suggesting rapidly generated immune responses. Clinical trial information: NCT01431391; NCT01981122.
Cytokine profile of sipuleucel-T (sip-T) in differentiating reactivation of latent immunity from de novo immune responses.
Authors: Drake Charles G. , Petrylak Daniel Peter , Antonarakis Emmanuel S. , Kibel Adam S. , Chang Nancy N. , Vu Tuyen , Campogan Dwayne , Haynes Heather , Trager James B. , Sheikh Nadeem Anwar , Quinn David I. 
Published: November 2019

163 Background: Sip-T is an FDA-approved immunotherapy for treating patients (pts) with asymptomatic or minimally symptomatic metastatic castration-resistant prostate cancer (mCRPC). It is manufactured from autologous peripheral blood mononuclear cells (PBMCs) cultured with PA2024, a fusion of prostatic acid phosphatase (PAP) and granulocyte macrophage colony-stimulating factor. Survival of sip-T–treated mCRPC pts correlates with immune responses to PA2024 and/or PAP. PA2024- or PAP-specific CD4+ and CD8+ T cell proliferation and cytokine production and release were assessed to better understand sip-T–induced T cell responses. Methods: Pts with biochemical recurrence or mCRPC were from sip-T trials (NCT01431391, NCT01981122). PBMCs collected at baseline through 6 mo post–sip-T were cultured in vitro and stimulated with PA2024 or PAP. CD4+ and CD8+ T cells were assessed (n=19) for proliferation and intracellular IL-2 and IFN-γ. The cytokine profile was confirmed in supernatant with a meso scale discovery assay. P<0.10 was statistically significant. Results: Compared with baseline, PA2024-specific proliferating CD4+ and CD8+ T cells had increased intracellular IL-2 and IFN-γ levels at wk 6 and mo 6, with a similar trend for PAP-specific proliferating T cells (Table 1). Compared with unstimulated controls, a significant >2-fold increase in PA2024-stimulated IL-2 and IFN-γ in supernatant was observed at wk 6 and mo 6 over baseline (p<0.001). PAP-stimulated IL-2 and IFN-γ supernatant levels increased over baseline and were significantly elevated for IFN-γ at wk 6 (p<0.10). Conclusions: Sip-T therapy generated a de novo PA2024-specific T cell response, as indicated by the cytokine release profile. The PAP-stimulated cytokine profile suggests that pre-existing immunity with terminally differentiated T cells are expanded. Thus, sip-T reactivated an anti-PAP response in memory T cells, thereby overcoming immunosuppressive mechanisms in PC. Clinical trial information: NCT01431391; NCT01981122. [Table: see text]
Feasibility study of multiple HLA-class IA restricted peptide vaccines (KRM-19) for metastatic triple negative breast cancer (TNBC).
Authors: Toh Uhi , Saku Shuko , Iwakuma Nobutaka , Okabe Mina , Shichijo Shigeki , Yamada Akira , Itoh Kyogo , Akagi Yoshito 
Published: November 2019

164 Background: Our previous phase II clinical trial has indicated that the therapeutic selected personalized peptide vaccines (PPV) were effective for boosting anticancer immunity and the immune response after PPV was associated with the clinical outcome as a prognostic factor for metastatic breast cancer (mBC). Based on the data from the PPV studies, we have conducted an early phase II study to evaluate the safety and the efficacy of a new regimen using multiple peptide vaccines (KRM-19) for pts with metastatic TNBC. Methods: KRM-19 consisted of 19 mixed peptides which were chosen from the previously reported 31 PPVs according to their anti-tumor immunologic effect and safety profile for mBC pts. All patients had histologically confirmed measurable ER-PgR-Her2- mBC and their human leukocyte antigen (HLA)-A molecules should be each of -A2, A3, A11, A24, A26, A31, or A33. KRM-19 (19mg/ml) was administrated subcutaneously in order with schedule of every week for a total of 6 doses. The concurrent conventional chemo- and/or endocrine therapy was not permitted for the combination, but was available for post-study treatment. Measurement of peptide-specific cytotoxic T lymphocyte (CTL) and IgG responses were conducted before and after vaccination. Levels of IgG reactive to each of the 19 peptides in the pre- and post-vaccination plasma were measured using the LUMINEX system at every 6 vaccinations. Peptide-specific CTL responses were examined by INF-ɤ ELISPOT. Clinical response was evaluated by RECIST criteria. Results: 10 patients with refractory metastatic TNBC received all 6 vaccines and 10/10 patients experienced Grade 1-2 skin reaction at injection site, 5/10 pts had Grade 2-3 liver function disorder, 3/10 and 1/10 pts had Grade 2 bone marrow suppression and nausea, respectively. The clinical responses were assessed in 8 pts included 1 PR case, 4 SD cases, and 3 PD cases. The evaluation of other clinical and immunologic data is underway. Conclusions: Subcutaneous KRM-19 vaccine administration was safe and resulted in a 12.5% objective response and 62.5% clinical benefit rate in TNBC pts, warranting further larger scale study. Clinical trial information: UMIN000014616.
Effect of RELT on regulation of the threshold of T-cell activation during sterile inflammation.
Authors: Kim Seon-Hee , Choi Beom K. , Han Chungyong , Kwon Byoung S. 
Published: November 2019

25 Background: Receptor expressed in lymphoid tissue (RELT) is a type I transmembrane protein, designated TNFRSF19-like, and primarily expressed in lymphoid tissues and immune cells. However, its immunological function has yet to be characterized. Methods: We developed RELT-deficient mice to examine the immunological role of RELT. The RELT–/– mice were exposed to viral and bacterial infection, chemical-induced liver injury, and tumor induction. Results: RELT–/– mice were viable and fertile, and developed normal lymphoid and myeloid cells. RELT transcripts were decreased in T cells and dendritic cells following their activation, and T-cell proliferation was enhanced in the absence of RELT in vitro. Nevertheless, we could not find significant changes in RELT–/– mice infected with virus or bacteria. However, liver injury and inflammation were significantly increased in RELT–/– mice in comparison to RELT+/+ mice after the injection of acetaminophen. Tumor growth rate was also slower in RELT–/– than RELT+/+ mice. Transfer of naïve or activated pmel-1 CD8+ T cells suppressed the growth of B16–F10 tumors and increased host CD8+ tumor-infiltrating cells more efficiently in RELT–/– than in RELT+/+ mice. In particular, naïve pmel-1 CD8+ T cells were present in increased numbers and activated forms in draining lymph nodes of RELT–/– than in RELT+/+ mice, whereas activated pmel-1 CD8+ T cells were not. Conclusions: As RELT–/– mice only showed significant differences in comparison to RELT+/+ mice in pathogen-associated-molecular-pattern-free conditions, these results provide evidence that RELT functions as a negative modulator of T cell responses in a sterile inflammation.
HER2 gene amplification and protein overexpression in uterine clear cell carcinoma and its implications in targeted immunotherapy.
Authors: Zhou Jain , Sukov William R. , Carter Jodi M , Schoolmeester J. Kenneth 
Published: November 2019

29 Background: Uterine clear cell carcinoma (UCCC) is a high-grade endometrial carcinoma. The current treatment is hysterectomy with post-operative chemotherapy and/or radiation. The 5-year disease free survival remains dismal for UCCC with 65% for early-stage and 54% for advanced stage disease. In addition, UCCC may be more resistant to chemotherapy or radiation therapy than the endometrioid subtype. The aim of the current study is to investigate the HER2 gene amplification status in UCCC and its role for targeted therapy in UCCC. Methods: Twenty-nine cases of UCCC were retrieved from surgical pathology archives of Mayo Clinic at Rochester between 2011 and 2015. All cases except one case were hysterectomy specimens. The blocks contain the most characteristic morphology of UCCC were selected and corresponding paraffin sections were subjected to fluorescent in situ hybridization for amplification of HER2 gene (Hercept, Abbott Molecular) and parallel immunohistochemical (IHC) study. Results: A total of 9 (of 29; 31%) UCCCs showed HER2 amplification and 4 (of 29; 14%) were considered equivocal for HER2 amplification by FISH. A total of 3 (10%) tumors showed 3+ HER2 overexpression while 11 (38%) UCCCs showed 2+ HER2 overexpression, 9 (31%) showed 1+ expression with the remaining cases showing no expression of HER2. Importantly, we observed significant intratumoral heterogeneity with regard to HER2 expression. Comparing the results of IHC with HER2 gene status as determined by FISH, 2 (66%) of the 3 cases that showed 3+ HER2 expression also showed amplification for HER2 by FISH, while 1 (33%) was equivocal for HER2 amplification. Of the 11 tumors that showed 2+ HER2 expression, 6 (55%) were amplified by FISH and 1 (9%) was equivocal. Conclusions: This is the largest number of UCCC cases that has been studied on the HER2 amplification and corresponding protein overexpression. Our results indicate that the HER2 overexpression is common in UCCCs and is frequently associated with HER2 amplification. These results also suggest that targeted adjuvant therapy with trastuzumab-based immunotherapy should be evaluated in patients with UCCC showing HER2 protein overexpression or HER2 gene amplification.
Immune characterization of the programmed death receptor pathway in high-risk prostate cancer.
Authors: Alanee Shaheen Riadh , Baas Wesley , Gershburg Svetlana , Dynda Danuta , Delfino Kristin , Robinson Kathy , Yearley Jennifer 
Published: November 2019

40 Background: Programmed Death 1 (PD-1) is a T cell inhibitory receptor critical to a major immunomodulation pathway which has been implicated in tumor evasion of immune response. PD-1 and its ligand PD-L1 have been found to be expressed in many tumor types, and this expression has led to the development of drugs targeting the PD-1 pathway. The goal of this study was to understand the expression of PD-1, and PD-L1 in high grade prostate cancer tissue, and correlate the expression with disease and patients characteristics. Methods: Immunohistochemistry for PD-1 (CD279), PD-L1 (B7-H1), and CD3 was performed on prostatectomy/biopsy tissue samples taken from 25 men with high grade (Gleason 8-10) prostate cancer using anti-PD-L1 clone 22C3 (Merck) and anti-PD-1 clone NAT105 (Cell Marque) . CD3 was used as a specific marker for T cell infiltration. Charts were then retrospectively reviewed for patient and disease characteristics including age at diagnosis, race, Gleason score, prostatic specific antigen (PSA) level at diagnosis, number of positive cores at biopsy, volume of tissue on biopsy and/or prostatectomy involved by cancer, clinical TNM stage, pathologic TNM stage, biochemical recurrence, or metastasis. Statistical analyses were done to correlate these patient and disease characteristics with PD-1, PD-L1, and CD3 expression. Results: A score of 3-5 on the semi-quantitative 0-5 score was deemed “high” expression whereas a score of 0-2 was deemed “low” expression. Of the 25 samples, 2 (8%) scored high for PD-1 expression, 2 (8%) scored high for PD-L1 expression, and 18 (72%) scored high for CD3 expression. With independent t-tests there was found to be no statistically significant correlation between any of the variables we collected and expression of PD-1, PD-L1, or CD3. Conclusions: We found an overall low expression of PD-1 and PD-L1, and a concurrent high expression of CD3+ T cells in high risk prostate cancer tissue. No significant correlations were made between expression of PD-1, PD-L1, or CD3 and patient and disease characteristics. The elevated T cell content in high risk prostate cancer is particularly interesting with the continued emergence of new drugs focused on enhancing efficacy of the immune response in cancer.
Variability in the use of cut-off points with different commercial and lab-developed assays for PD-L1 expression testing in the NSCLC clinical setting in the United States.
Authors: De Richter Pieter 
Published: November 2019

46 Background: Following the recent approval of several companion diagnostic and complementary diagnostic assays for PD-L1 expression testing, as well as the availability of multiple antibodies for use in laboratory testing, pathologists have a range of tools at their disposal for measuring expression levels of this predictive biomarker. While specific cut-off points for positivity are specified by the assay manufacturer and/or recommended based on trial data, clinicians can choose different cut-off points to define positivity. In this study, we explore the variety of test brands and cut-off points that are being used in clinical practice in the US by examining real-world usage data. Methods: Between June and August 2016, a panel of pathologists (n = 27) in the US were asked to report on their practices relating to PD-L1 expression testing for NSCLC patients, through the submission of online de-identified record forms (n = 167 PD-L1-tested samples). Results: Of the 167 samples, 112 (67%) were tested with the Dako 22C3 pharmDx assay, 37 (22%) with the Dako 28-8 pharmDx assay and 16 (10%) with a lab-developed test (LDT). When using 22C3 pharmDx, 72% of samples were tested with the recommended 50% cut-off point, with the remaining 28% using lower cut-off points, ranging from 1% to 25%. For 28-8 pharmDx, both the 1% cut-off point (in 54% of cases) and the 5% cut-off point (in 35% of cases) were frequently used (1% recommended; 5% being investigated in clinical trials e.g. CHEKMATE-026). LDTs were mostly (88%) employed with a cut-off of 1% across antibodies. Conclusions: The use of PD-L1 expression testing to determine eligibility for PD-(L)1 inhibitors in the clinical setting shows significant fragmentation in the US, with multiple different assays being employed. Furthermore, there is a lack of standardization in terms of the cut-off points that are used when testing with each of those assays. This lack of consistency adds a level of complexity to oncologists treatment decisions when choosing whether or not to treat with a PD-(L)1 inhibitor, and which brand to choose.
Differences in immune profiles of metastatic melanoma patients treated with anti-CTLA-4 and anti-PD-1 combined immunotherapy.
Authors: Gide Tuba Nur , Quek Camelia , Madore Jason , Menzies Alexander M. , Dent Rebecca , Carlino Matteo S. , Guminski Alex , Saw Robyn , Thompson John F. , Wilmott James S. , Scolyer Richard A. , Long Georgina V. 
Published: November 2019

51 Background: Immune checkpoint inhibitors targeting the cytotoxic T-lymphocyte antigen 4 (CTLA-4) and programmed cell death 1 (PD-1) receptor have significantly improved the outcomes of metastatic melanoma patients resulting in durable response and longer overall survival. However, while some patients exhibit a complete response to immunotherapy, others have demonstrated little to no response. The aim of this study is to examine the immune cell subsets between the good and poor responders to anti-CTLA-4 and anti-PD-1 combined immunotherapy in order to determine potential mechanisms of response and resistance. Methods: Multiplex immunofluorescent staining was performed on pre-treatment formalin-fixed, paraffin-embedded (FFPE) tissue samples from good and poor responders ( n= 17; n= 11 good responders, n= 6 poor responders) for markers including CD8, PD-1, PD-L1, FOXP3 and SOX10. Spectral image analysis was conducted via the Vectra 3.0 imaging system. Quantitative analysis was carried out using the inForm software. Gene signatures associated with the T-cell signalling pathways were identified following RNA sequencing of FFPE samples. Results: Good responders displayed significantly higher levels of baseline intratumoural PD-1 and PD-L1 expression in comparison to the poor responders ( P= 0.01 and P= 0.01 respectively), highlighting their roles as predictive biomarkers of response. Additionally, the CD8+ T-cell to PD-L1 ratio was significantly higher ( P= 0.001) in the tumours of the poor responders, indicating a distinction in the CD8+ T-cell phenotypes between the good and poor responders. Furthermore, analysis of the RNA sequencing data revealed a panel of immune signatures that have been implicated in prognosis and responsiveness to immunotherapy in melanoma. Conclusions: In summary, these findings provide insight into the possible biomarkers of response and resistance to immunotherapy, thus improving our understanding of the mechanisms driving variations in response.
The prognostic impact of CD3, CD8, FoxP3, and IL17 tumor-infiltrating immune cells in periampullary cancer differs by morphological type and adjuvant chemotherapy.
Authors: Warfvinge Carl Fredrik , Lundgren Sebastian , Elebro Jacob , Heby Margareta , Krzyzanowska Agnieszka , Bjartell Anders , Nodin Bjorn , Eberhard Jakob , Leandersson Karin , Jirstrom Karin 
Published: November 2019

53 Background: The prognostic impact of tumor-infiltrating T lymphocytes has been evaluated in periampullary carcinoma, but previous studies have not taken into consideration the different morphological subtypes of cancers derived from the periampullary region. Herein, we investigated the prognostic impact and distribution of tumor-infiltrating CD3+, CD8+, FoxP3+ T lymphocytes as well as IL-17+cells in periampullary adenocarcinoma, with particular reference to morpohological subtyoe. Methods: Counts of CD3+, CD8+, FoxP3+ T lymphocytes, and IL-17+cells were assessed using immunohistochemistry on tissue microarrays with tumors from a consecutive retrospective cohort of 175 patients with resected periampullary adenocarcinoma; 110 of pancreatobiliary type (PB-type) and 65 of I intestinal type (I-type). Results: Distribution of all evaluated immune cells differed significantly between PB- and I-type tumors. In I-type tumors, the ratios of IL-17+/FoxP3+ and IL-17+/CD3+ were independent factors of poor prognosis. Notably, the unfavorable prognostic impact of IL-17+ was only evident in cases that had not received adjuvant chemotherapy. In PB-type tumors, CD3+was an independent marker of good prognosis. Conclusions: The results from this study highlight that the composition and prognostic importance of various subsets of infiltrating immune cells in periampullary adenocarcinoma differ by morphological subtype, and potentially by adjuvant chemotherapy.
Estimating long-term survival of PD-L1-expressing, previously treated, non-small cell lung cancer patients who received pembrolizumab in KEYNOTE-001 and -010.
Authors: Hellmann Matthew David , Ma Junshui , Garon Edward B. , Hui Rina , Gandhi Leena , Soria Jean-Charles , Anderson Keaven M. , Lubiniecki Gregory M. , Piperdi Bilal , Herbst Roy S. 
Published: November 2019

77 Background: Pembrolizumab showed promising activity in patients with advanced non–small cell lung cancer (NSCLC) in the KEYNOTE-001 study (NCT01295827) and significantly prolonged overall survival (OS) compared with docetaxel in the randomized KEYNOTE-010 study (NCT01905657). Responses to pembrolizumab appear to be remarkably durable, making long-term survival possible in some patients. Typical parametric survival models do not account for the possibility of long-term survival. An alternative, well-established class of statistical models called long-term survival models can be used to directly estimate the percentage of patients achieving long-term survival (>5 years), called “long-term survival rate.” (Berkson J, Gage RP. J Am Stat Assoc. 1952;47:501-515.)(Tsodikov AD et al. J Am Stat Assoc. 2003;98:1063-1078.) Methods: Data from patients with PD-L1–expressing (tumor proportion score ≥1%), previously treated, advanced NSCLC in KEYNOTE-001 and KEYNOTE-010 were used. KEYNOTE-001 data were used to initially estimate the long-term survival rate of pembrolizumab, while KEYNOTE-010 data were used for subsequent independent validation. Point estimates of long-term survival rates with their 95% CIs based on the model described above are reported. Results: Based on the long-term survival model, the estimated long-term survival rate in pembrolizumab-treated population in KEYNOTE-001 (n = 306) is 25.4% (95% CI, 15.2%-33.3%) and the intention-to-treat population who received pembrolizumab in KEYNOTE-010 (n = 690) is 25.3% (95% CI, 8.9%-36.9%). In contrast, the long-term survival rate of docetaxel arm (n = 343) in KEYNOTE-010 is estimated to be 3.2% (95% CI, 0%-17.4%). Conclusions: In 2 independent data sets, it is estimated that 25% of patients with previously treated PD-L1–expressing NSCLC may achieve long-term benefit from pembrolizumab monotherapy. Long-term survival models can provide an early estimate of long-term benefit from pembrolizumab using data with limited follow-up time. Long-term follow-up from these trials will further validate this finding. MDH and JM are co-first authors.