Predictors Of Immune Response Research Articles

Correlation between tumor-infiltrating lymphocyte density and disease-free survival rate in basal cell carcinoma patients at Dr. Soetomo General Hospital, Surabaya

Basal cell carcinoma has a high risk of recurrence and can be associated with other types of cancer linked to ultraviolet radiation, impacting clinical outcomes, increasing morbidity, and shortening Disease-Free Survival (DFS) rates. Tumor-infiltrating lymphocytes (TILs) are considered predictors of immune response that are useful in suppressing tumor growth by infiltrating tumor cells. TILs were used as a predictive factor in the survival rate of basal cell carcinoma, with the result that high density was a good marker of immune system activation around the tumor and was associated with better prognosis. This study employed a retrospective cohort observational analytic research design to correlate the density of TILs with the DFS rate of basal cell carcinoma patients who had undergone wide excision at Dr. Soetomo General Hospital Surabaya from January 2017 to December 2023, with a total sample of 40 patients. Among the 40 samples, a group with strong TILs density was identified from a total of 14 samples, with 9 samples (22.5%) having DFS ≥ 5 years. In the weak TILs density group, there were 26 samples, with 9 samples (22.5%) having DFS for 1 year, 7 samples (17.5%) having DFS for 2 years, and 8 samples (20%) having DFS for 3 years. The analysis of the correlation between TILs density and DFS found that the stronger the TILs density, the longer the DFS time (p value = 0.002, RR 1.683). There was a significant correlation between TILs density and disease-free survival in patients with basal cell carcinoma who had undergone wide excision and/or radiotherapy.

P-2064. COVID-19 Vaccination-infection Status and Immunological Profile: Identification of High-Risk Population for Targeted Booster Immunization In India

Abstract Background The immunological profiles and predictors of immune response in people with hybrid immunity, resulting from a combination of natural infection and vaccination, particularly in regions where non-mRNA vaccines were administered, remain less understood. Methods This was a cross-sectional study done at AIG Hospitals, Hyderabad, India. Consecutive participants were enrolled, and sampled for immunological testing. The study population was divided into six cohorts based on their infection and vaccination status. Immunological assays were performed to measure anti-spike protein and interferon-γ release assay. Hospitalization rates were compared in all groups. Chi-square test was used to compare categorical variables, and the Mann-Whitney U test was used for comparing medians. Post-hoc Dunn test was used for pairwise comparison. Bonferroni adjustment for p-value was performed for multiple pairwise comparisons. Linear regression model was used appropriately to find predictors of antibody titres. Results A total of 1040 patients were enrolled in the study. The population was divided into six cohorts: Unvaccinated, Primary, Booster, Hybrid (Pre-Omicron only), Hybrid (Omicron only), and Hybrid (Both). Antibody titres and IGRA levels were found to be statistically higher in the Hybrid (both infection) group (Table 1 and Figures 1a & 1b). The linear regression model showed factors predicting antibody response (Table 2). Age and hypertension were independent (negative) predictors of antibody levels. During the follow-up (median 30.7 [26.6 – 34.4 months]) of 1008 patients, 770 (76.4%) had a documented COVID infection with Omicron. Hospitalization was evaluated in three groups: Hybrid, vaccination only and unvaccinated. The incidence was significantly lower in the Hybrid group; log-rank test: p < 0.001. (Figure 2) Conclusion The prevalence of hybrid immunity was high in the study population. Individuals above 60 years old and with comorbidities are more likely to have lower antibody titers. Immunized patients were protected against hospitalization. There is a need for prioritized immune-boosting efforts in high-risk individuals. Disclosures All Authors: No reported disclosures

Immune Predictors of Response after Bacillus Calmette–Guérin Treatment in Non-Muscle-Invasive Bladder Cancer

Bacillus Calmette–Guérin (BCG) has been the standard of care for the treatment of high-risk, non-muscle-invasive bladder cancer (NMIBC) for decades, but 49.6% of high-risk and very-high-risk patients will experience progression to muscle-invasive disease in five years. Furthermore, cytology and cystoscopy entail a high burden for both patients and health care systems due to the need for very long periods of follow-up. Subsequent adjuvant treatment using intravesical immunotherapy with BCG has been shown to be effective in reducing tumor recurrence and progression, but it is not free of severe adverse effects that ultimately diminish patients’ quality of life. Because not all patients benefit from BCG treatment, it is of paramount importance to be able to identify responders and non-responders to BCG as soon as possible in order to offer the best available treatment and prevent unnecessary adverse events. The tumor microenvironment (TME), local immune response, and systemic immune response (both adaptive and innate) seem to play an important role in defining responders, although the way they interact remains unclear. A shift towards a proinflammatory immune response in TME is thought to be related to BCG effectiveness. The aim of this review is to collect the most relevant data available regarding BCG’s mechanism of action, its role in modulating innate and adaptive immune responses and the secretion of certain cytokines, and their potential use as immunological markers of response; the aim is also to identify promising lines of investigation.

Comprehensive characterization of pyroptosis phenotypes with distinct tumor immune profiles in gastric cancer to aid immunotherapy.

Pyroptosis is a form of programmed cell death that is essential for immunity. Herein, this study was conducted to uncover the implication of pyroptosis in immunomodulation and tumor microenvironment (TME) in gastric cancer. Prognostic pyroptosis-related genes were extracted to identify different pyroptosis phenotypes and pyroptosis genomic phenotypes via unsupervised clustering analysis in the gastric cancer meta-cohort cohort (GSE15459, GSE62254, GSE84437, GSE26253 and TCGA-STAD). The activation of hallmark gene sets was quantified by GSVA and immune cell infiltration was estimated via ssGSEA and CIBERSORT. Through PCA algorithm, pyroptosis score was conducted. The predictors of immune response (TMB and IPS) and genetic mutations were evaluated. The efficacy of pyroptosis score in predicting immune response was verified in two anti-PD-1 therapy cohorts. Three different pyroptosis phenotypes with different prognosis, biological pathways and tumor immune microenvironment were established among 1275 gastric cancer patients, corresponding to three immune phenotypes: immune-inflamed, immune-desert, and immune-excluded. According to the pyroptosis score, patients were separated into high and low pyroptosis score groups. Low pyroptosis score indicated favorable survival outcomes, enhanced immune responses, and increased mutation frequency. Moreover, low pyroptosis score patients displayed more clinical benefits from anti-PD-1 and prolonged survival time. Our findings uncovered a nonnegligible role of pyroptosis in immunomodulation and TME multiformity and complicacy in gastric cancer. Quantifying the pyroptosis score in individual tumors may tailor more effective immunotherapeutic strategies.

Evaluation of haemolymph phenoloxidase activity from the grub of Zophobas morio as a predictor of immune response.

In insects, enzyme phenoloxidase plays a critical role in cuticular sclerotisation and defensive functions. In the present investigation, haemolymph phenoloxidase activity from the grub of Zophobas morio was attempted to evaluate as a reliable predictor of insect's immunological response. Among the various substrates tested, L-DOPA was chosen as an appropriate substrate due to its high oxidation. The optimum pH and temperature for haemolymph PO activity was found to be 8 and 30°C, respectively. The optimum substrate concentration of L-DOPA was found to be 7.5mM for subsequent PO enzymatic characterisation. Among the various chemical inhibitors and copper chelators, PO activity was significantly reduced in the case of PMSF and thiourea. Preincubation of haemolymph with non-self-molecules showed enhancement of PO activity in the case of LPS from Serratia marcescens. In addition, exogenous proteases like α-chymotrypsin enhanced the PO activity of haemolymph and an increase in PO activity was demonstrated when haemolymph was preincubated with the anionic detergent, SDS and cationic detergent, cetyl pyridium chloride. Alteration of PO activity was observed under agonising conditions of starvation, ligation and microplastics injection at different time intervals. Interestingly, there were no correlation between PO and insect defence under live challenge of microbes. SDS protein profile revealed a significant increase in the 85kDa and 55kDa polypeptides in all the experiments over control after 24h, 48h and 96h. Mass spectrophotometric analysis of the polypeptides revealed their homology to antimicrobial peptides for 55kDa protein and 85kDa protein. A significant increase in 85kDa polypeptide was observed in the haemolymph of the grubs after 72h in the case of starved and microplastics injected groups only. These results demonstrated that PO may not be a reliable benchmark of immunological response in this insect.

Diet‐mediated immune response to parasitoid attacks on a caterpillar with a broad diet breadth

Abstract Bottom‐up (plant) and top‐down (natural enemy) trophic factors can interact to have significant influence on the diet breadth of herbivores. For herbivorous insects that are victim to parasitoid attacks, diet composition can modulate insect immune responses against the parasitoid. However, immune responses are costly and insect herbivores experience a trade‐off between investment in immune defences and other physiological processes. We used a split‐brood laboratory experiment to explore how the diet of the fall webworm (Hyphantria cunea), a species that eats over 400 plant species, affects larval growth and fitness and cellular immune response to attacks from a parasitic wasp. We reared larvae on four different plant diets (apple, alder, chokecherry, cottonwood) and then exposed them to an immune challenge from a parasitoid attack. We found that diet influenced larval development as well as parameters indicative of immune response. Larvae reared on the plant that led to the poorest development also had the fewest granulocytes and the highest odds of containing a parasitoid larva. However, larval growth was not a predictor of immune response. Overall, we show that the bottom‐up effect of diet variability has significant impacts on insect immune response such that larval fitness varies considerably when fed different dietary plant species. Broad diet ranges may offer herbivorous insects the opportunity to exploit a different set of resources depending on the severity of top‐down pressures. Here, we show that this variability in plant quality also has significant impacts on larval immune response.

Pembrolizumab with trastuzumab and chemotherapy (PTC) in HER2-positive metastatic esophagogastric cancer (mEG): Plasma and tumor-based biomarker analysis.

4559 Background: Pembrolizumab can be safely combined with trastuzumab and chemotherapy and has promising activity with median OS 27 months and 91% objective response rate in HER2-positive mEG cancer irrespective of PD-L1 status (NCT02954536; Janjigian ESMO 2019). Tumor biopsies and blood samples were collected in this phase II trial to identify molecular and immune predictors of response and resistance to PTC. Methods: Pre-treatment and post-progression biopsies were analyzed using WES and IHC (HER2, PD-L1). Peripheral blood was collected pre-treatment, every 9 weeks on-treatment and at progression for plasma ctDNA (Guardant 360, Guardant Health, Redwood, CA). Tumor-matched DNA alterations were identified by correlating ctDNA and solid tumor WES results. Landmark PFS analysis was used to compare ctDNA clearance status at 9 weeks post-treatment. Results: Baseline ctDNA was analysed from 31 of 37 patients of whom 84% (26/31) had tumor-matched ctDNA detected at baseline. Patients who cleared ctDNA at 9 weeks (n = 17/23) achieved a longer median PFS than those who did not - mPFS 12.3 months (95% CI 7.44-NA vs 3.9 months (95% CI 2.01-NA) (log-rank p = 0.02). On serial blood monitoring of 16 patients with eventual radiographic progression, ctDNA re-appearance preceded CT detection in 8 (50%) patients. WES was completed in 31 patients with pre-treatment, and 12 patients post-progression, including matched samples from 10 patients. Loss of HER2 over-expression/amplification was noted in 44% (7/16) of post-progression samples by IHC/FISH (2 IHC 0/1, 5 FISH-). In paired post-progression samples on WES, we observed loss of ERBB2 in 2 patients, and new amplifications of CCND1/3, FGF3/4/19, CDK6/12, KRAS, MYC, and MET, as well as mutations in KRAS, PIK3CD and PIK3RA. Plasma analysis at progression demonstrated copy number increases and/or new amplifications in MET, CKD6, PIK3CA, KRAS, FGFR2, EGFR and CCDN1 as well as KRAS, RB1, PTEN, NF1, NOTCH1, BRAF, and FGFR1 mutations. Conclusions: The majority of patients with previously untreated HER2 positive mEG have detectable plasma ctDNA at baseline. The re-appearance of ctDNA during therapy may serve as an early predictor of progression and help identify genetic drivers of acquired resistance. Loss of ERBB2 over-expression/amplification and activating MAPK alterations occur at PTC progression. Evaluation of tumor immune environment by multiplex IHC and additional ctDNA analysis is underway.

Metabolic predictors of individual immune responses to adaptogenic balneotherapy

Background. Earlier four variants of the immune responses to adaptogenic balneotherapy have been identified. All four variants of immune responses are virtually unmistakably (with an accuracy of 97,7%) predicted by a set of 20 predictors including 12 immune blood parameters and one saliva parameter, 4 information parameters, 2 fecal microbiota parameters as well as erythrocyturia. In the next study discriminant analysis revealed that constellation 8 HRV and 5 Endocrine parameters as well as Gender of the patient predicts the nature of the immune response with an accuracy of 90,9%. The purpose of this study is to search for predictors of immune responses among the registered Metabolic parameters. Material and methods. Theobjectofobservationwere 34menand 10 womenaged24-70yearsold,whocametotheTruskavets’ spa for the treatment of chronic pyelonephritis combined withcholecystitis inremission. We determined them theElectrokinetic index, Lipoproteines spectrum, plasma and daily urine levels of Electrolytes and Nitrous metabolites as well as basal and postprandial volume of gall-bladder. Results. Discriminant analysisrevealed 16 parameters as the predictors. Of these, 3 reflect the level of Plasma Electrolytes, 5 - the Urine Electrolytes, another 2 - the levels of plasma and urine Urea. Other predictors were: Very Low Density Lipoprotein Cholesterol plasma level, Body Mass index, fasting gallbladder volume and 30 minutes after cholekinetics, Electrokinetic index, and patient Age. These predictors, taken together, determine the nature of the immune response with an accuracy of 95,5%. Conclusion. The variety of immune responses to adaptogenic balneotherapy is quite strictly conditioned by the initial state of the neuroendocrine-immune complex, microbiota, cholekinetics and metabolism, as well as the age and sex of patients.

Humoral and cellular immune response in mice induced by the classical swine fever virus E2 protein fused to the porcine CD154 antigen

The development of subunit vaccines against classical swine fever is a desirable goal, because it allows discrimination between vaccinated and infected animals. In this study, humoral and cellular immune response elicited in inbred BALB/c mice by immunization with a recombinant classical swine fever virus (CSFV) E2 protein fused to porcine CD154 antigen (E2CD154) was assessed. This model was used as a predictor of immune response in swine. Mice were immunized with E2CD154 emulsified in Montanide ISA50V2 or dissolved in saline on days 1 and 21. Another group received E2His antigen, without CD154, in the same adjuvant. Montanide ISA50V2 or saline served as negative controls for each experimental group. Animals immunized with 12.5 and 2.5 μg/dose of E2CD154 developed the highest titers (>1:2000) of CSFV neutralizing antibodies. Moreover, CSFV specific splenocyte gamma-interferon production, measured after seven and twenty-eight days of immunization, was significantly higher in mice immunized with 12.5 μg of E2CD154. As a conclusion, E2CD154 emulsified in Montanide ISA50 V2 was able to induce a potent humoral and an early cellular immune response in inbred BALB/c mice. Therefore, this immunogen might be an appropriate candidate to elicit immune response in swine, control CSF disease and to eliminate CSFV in swine.

CD4+ levels control the odds of induction of humoral immune responses to tracer doses of therapeutic antibodies.

Rapidly increasing number of therapeutic antibodies are being repurposed to imaging probes for noninvasive diagnosis, as well as monitoring during treatment or disease recurrence. Though antibody-based imaging involves tracer doses (~3 log lower than therapeutic doses), and immune responses are severely reduced in patients with impaired immunity, formation of anti-tracer antibodies (ATA) has been observed hampering further diagnostic monitoring. Here, we explored the potential to develop humoral responses to intravenously administered tracer dose of a monoclonal antibody F(ab΄)2 fragment, and associated with host related immune measures in 49 rhesus macaques categorized into healthy (uninfected controls), SIV-progressors, SIV non-progressors, or total body irradiated (TBI). Antibody fragment administered in tracer amount (~100μg) induced immune responses with significantly lower odds in SIV-progressors or TBI macaques (P<0.005) as compared to healthy animals. Peripheral blood (PB) CD4+ cell counts, but not CD20+ cell levels, were associated with significantly higher risk of developing a humoral response (P<0.001). Doubling the PB CD4+ counts is associated with an odds ratio of developing an immune response of 1.73. Among SIV-infected animals, CD4+ cell count was a stronger predictor of immune response than plasma SIV-RNA levels. Both SIV-progressors and TBI macaques showed higher odds of responses with increasing CD4+ counts, however when compared to healthy or SIV non-progressors with similar CD4+ count, they were still functionally incompetent in generating a response (P<0.01). Moreover, presence of ATA in systemic circulation altered the in vivo biodistribution by increasing hepatic uptake and decreasing plasma radiotracer clearance, with minimal to no binding detected in targeted tissues.

Molecular and immune predictors of response and toxicity to combined CTLA-4 and PD-1 blockade in metastatic melanoma (MM) patients (pts).

9579 Background: Combined treatment with ipilimumab and nivolumab (Ipi/Nivo) achieves clinical responses in &gt; 50% of mm pts. However, responses are not universal and toxicity may be limiting, thus biomarkers of response and toxicity are needed to optimize and personalize this therapy. Methods: Tumor biopsies were collected before (n = 29) and on treatment (n = 7) from mm pts (n = 40) treated with Ipi/Nivo. Whole exome sequencing (WES), gene expression profiling, TCR sequencing, and immunohistochemistry (IHC) were performed to define molecular and immune features of the tumors. Radiographic responses in patients were assessed via RECIST 1.1criteria, and patients were classified as responders (R) deriving clinical benefit (with SD, PR, CR) and non-responders (NR) not deriving clinical benefit (PD). Toxicity was also scored, with patients dichotomized into low toxicity ( &lt; grade 2) versus high toxicity ( &gt; grade 3) re: immune-related (IR) toxicities. Results: In this cohort, the response rate was 80%, with 53% of patients experiencing &gt; grade 3 toxicity. There was no significant difference in baseline mutational load in responders (R) vs non-responders (NR) to Ipi/Nivo, but NR had a higher burden of copy number alterations (CNA; p = 0.013), with frequent alterations detected in PTEN, JAK2, and B2M. There were no significant differences in baseline CD8+ T cell density, expression of immune-related genes, or T cell clonality for R vs NR pts. Ipi/Nivo treatment increased intratumoral T cell clonality, but this did not correlate with response. A more diverse peripheral T cell repertoire at baseline was detected in pts who developed IR toxicity (p &lt; 0.05). Conclusions: This data suggests that responses to Ipi/Nivo in mm may occur in the absence of high mutational load or brisk immune infiltrate at baseline. Putative mechanisms of resistance to Ipi/Nivo include high burden of CNA and alterations in PTEN, JAK2, and B2M. Together these studies identify candidate biomarkers of resistance and toxicity for Ipi/Nivo, though they need to be tested in larger cohorts and across cancer types.

Systemic inflammation before and after antiretroviral therapy initiation as a predictor of immune response among HIV-infected individuals in Manitoba

IntroductionDespite the life-prolonging effects of Highly Active Antiretroviral Therapy (HAART), persons with HIV are still prone to higher rates of non-AIDS related morbidity (such as heart, kidney, and liver disease) than the general public. This is likely due to chronic immune activation and inflammation that persists in HIV-positive persons despite virological suppression.What remains undetermined, however, is whether a link exists between chronic inflammation/immune activation and suboptimal immune recovery on HAART.The hypothesis of the present study is that higher levels of systemic subclinical inflammation and immune activation are linked with suboptimal immune recovery on HAART. MethodsFifteen eligible patients from the Manitoba HIV program were enrolled and followed for up to two years; blood samples were drawn at 4 timepoints each, and concentrations of 21 proinflammatory markers were measured. Patients were grouped according to CD4:CD8 recovery at viral suppression, and the inflammatory profiles of the two groups were compared. Results and conclusionsAPRIL and BAFF are higher in those with poor recovery at the point of viral suppression, but were also higher in this group at the onset of therapy and through the three additional follow-up visits. TNF-R1, CD163, and Osteopontin, were also in higher concentrations at the outset of therapy and beyond. These five molecules could thus see potential use in the future as biomarkers of likely poor immune recovery. Future work should focus on replicating these findings with larger cohorts.

Abstract 767: TCR sequencing can identify and track tumor-specific T cell populations and is a predictive biomarker of response to DC vaccination in glioblastoma patients

Abstract While immunotherapeutic strategies are emerging adjunctive treatments for cancer, sensitive methods of monitoring the immune response after treatment remain to be established. We used a novel next generation sequencing (NGS) approach to determine whether quantitative assessments of tumor infiltrating lymphocyte (TIL) content and the degree of overlap of T cell receptor (TCR) sequences in brain tumors and peripheral blood were predictors of immune response and overall survival in glioblastoma (GBM) patients treated with autologous tumor lysate-pulsed dendritic cell (DC) immunotherapy. A significant correlation was found between a higher estimated TIL content and increased time to progression (TTP) and overall survival (OS). In addition, we were able to assess the proportion of shared TCR sequences between tumor and peripheral blood at time points before and after therapy, and found the level of TCR overlap to correlate with survival outcomes. Higher degrees of overlap, or the development of an increased overlap following immunotherapy, correlated with improved clinical outcome, and may provide insights into the successful, antigen-specific immune response. Citation Format: Shaina Sedighim, Melody Hsu, Tina Wang, Richard G. Everson, Alex Tucker, Joseph P. Antonios, Lin Du, Ryan Emerson, Erik Yusko, Catherine Sanders, Harlan Robins, William Yong, Tom B. Davidson, Gang Li, Linda M. Liau, Robert Prins. TCR sequencing can identify and track tumor-specific T cell populations and is a predictive biomarker of response to DC vaccination in glioblastoma patients. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 767.

Abstract 3207: Influence of tumor mutation burden on response to anti-PD-1 treatment in murine models of melanoma

Abstract Therapeutic antibodies directed against immune checkpoints, such as cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) and programmed cell death-1 (PD-1), have recently shown remarkable clinical benefits, particularly in metastatic melanoma and non-small cell lung cancer (NSCLC), cancers largely caused by chronic exposure to mutagenic agents. Recent studies have focused on identifying genomic and immune predictors of response, but consistent, robust biomarkers have yet to be fully characterized. However, some data suggests a high non-synonymous tumour mutation burden is associated with clinical benefit in melanoma patients who received anti-CTLA-4 therapy. A similar correlation was also found in NSCLC patients treated with anti-PD-1. Using mouse models of BRAFV600E-driven melanoma, we examined the relationship between tumour mutation burden and response to the immune checkpoint inhibitor anti-PD-1. Chronic exposure to ultraviolet radiation (UVR) was used to increase the number of tumour non-synonymous single nucleotide variants (SNVs), and thus mutational load. Responses to PD-1 blockade were compared between tumours having low (∼10 SNVs) and high (&amp;gt;100 SNVs) mutation burden and tumours were subsequently subjected to comprehensive molecular and immune profiling using whole exome sequencing, immunohistochemistry, and flow cytometry. Our initial data suggest there is no significant correlation between tumour mutation burden and response in our model of BRAFV600E-driven melanoma, and we will present a detailed analysis of low- and high-mutation burden tumours in light of their response to PD-1 blockade. Consistent with previous studies, our results highlight that integrated molecular characterization of tumour tissue and associated microenvironment in larger cohorts is needed to identify robust determinants of response to immune checkpoint inhibitors. Citation Format: Elena Galvani, Kate Hogan, Gabriela Gremel, Amaya Viros, Amit K. Mandal, Matthew Smith, Jacqueline Swan, Antonia Banyard, Garry Ashton, Nathalie Dhomen, Richard Marais. Influence of tumor mutation burden on response to anti-PD-1 treatment in murine models of melanoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3207.

IMCT-11NEXT GENERATION T CELL RECEPTOR SEQUENCING CAN IDENTIFY, QUANTIFY, AND TRACK TUMOR-SPECIFIC T CELL POPULATIONS BEFORE AND AFTER DENDRITIC CELL VACCINATION IN GLIOBLASTOMA MULTIFORME PATIENTS

While dendritic cell vaccination is a promising treatment for glioblastoma (GBM), sensitive methods of monitoring the immune response after treatment remain to be established. We sought to determine whether: 1) T cell receptor (TCR) repertoire overlap between peripheral blood and tumor and 2) tumor infiltrating lymphocyte (TIL) content, as assessed by next generation DNA sequencing, are predictors of immune response and survival. Genomic DNA from pre and post treatment tumor, as well as pre and post treatment peripheral blood, of 16 GBM patients enrolled in Phase I and II DC vaccination clinical trials was subjected to next generation sequencing through the TCRVβ region to quantify TIL content and assess the overlap between tumor and blood. Of the 5 patients for whom initial and relapsed tumors were available, two patients with high TCR sequence overlap between tumor and blood prior to treatment demonstrated the longest overall survival (OS) of 71.3 and 28.7 months. Two patients with poor OS (6.3 and 4.5 months) had low blood and tumor TCR overlap both before and after treatment. One multi-focal recurrent patient with intermediate OS (12.6 months) demonstrated an initially low overlap between pre-treatment tumor and blood; however, the TCR overlap increased 200-fold after treatment. Because we did not possess post-treatment, relapsed tumors for many patients, we tested whether the density of pre-treatment TILs was relevant. For the 16 patients from whom we had pre-treatment tumor tissue, the estimated TIL content significantly correlated with time to progression and OS (HR = 0.252, p = 0.0442; HR = 0.277, p = 0.0461), respectively. The estimated TIL content of pre-treatment tumor directly correlates with survival in GBM patients who received autologous tumor lysate-pulsed DC vaccination. Furthermore, the trends of tumor and peripheral blood TCR overlap suggest that this technology can be utilized to track tumor-specific T cell populations without prior knowledge of their antigen specificity.

Needle-free jet injector intradermal delivery of fractional dose inactivated poliovirus vaccine: Association between injection quality and immunogenicity

IntroductionThe World Health Organization recommends that as part of the polio end-game strategy a dose of inactivated poliovirus vaccine (IPV) be introduced by the end of 2015 in all countries currently using only oral poliovirus vaccine (OPV). Administration of fractional dose (1/5 of full dose) IPV (fIPV) by intradermal (ID) injection may reduce costs, but its conventional administration is with Bacillus Calmette-Guerin (BCG) needle and syringe (NS), which is time consuming and technically challenging. We compared injection quality achieved with BCG NS and three needle-free jet injectors and assessed ergonomic features of the injectors. MethodsChildren between 12 and 20 months of age who had previously received OPV were enrolled in the Camaguey, Cuba study. Subjects received a single fIPV dose administered intradermally with BCG NS or one of three needle-free injector devices: Bioject Biojector 2000® (B2000), Bioject ID Pen® (ID Pen), or PharmaJet Tropis® (Tropis). We measured bleb diameter and vaccine loss as indicators of ID injection quality, with desirable injection quality defined as bleb diameter ≥5mm and vaccine loss <10%. We surveyed vaccinators to evaluate ergonomic features of the injectors. We further assessed the injection quality indicators as predictors of immune response, measured by increase in poliovirus neutralizing antibodies in blood between day 0 (pre-IPV) and 21 (post-vaccination). ResultsDelivery by BCG NS and Tropis resulted in the highest proportion of subjects with desirable injection quality; health workers ranked Biojector2000 and Tropis highest for ergonomic features. We observed that vaccine loss and desirable injection quality were associated with an immune response for poliovirus type 2 (P=0.02, P=0.01, respectively). ConclusionsOur study demonstrated the feasibility of fIPV delivery using needle-free injector devices with high acceptability among health workers. We did not observe the indicators of injection quality to be uniformly associated with immune response.

Clinical characteristics and treatment-related biomarkers associated with response to high-dose interleukin-2 in metastatic melanoma and renal cell carcinoma: retrospective analysis of an academic community hospital's experience.

BackgroundImmunotherapy in the treatment of metastatic melanoma and renal cell carcinoma can produce durable therapeutic responses, which may improve survival. We aimed to identify clinical characteristics and biomarkers associated with response to high-dose interleukin-2 therapy (IL-2) in patients with metastatic melanoma and renal cell carcinoma treated at an academic community hospital.Patients/MethodsWe retrospectively analyzed clinical variables and biomarkers of 50 consecutive metastatic melanoma or renal cell carcinoma patients treated at our institution with IL-2 during 2004 – 2012. We evaluated clinical characteristics: metastatic sites of disease, prior therapies, number of IL-2 doses per cycle, response duration, autoimmune phenomena, and peak fever, as well as laboratory biomarkers: baseline LDH, platelet nadir, and baseline and highest absolute lymphocyte count (ALC). Survival outcomes were calculated using Kaplan-Meier curves.ResultsVariables differing between responders (clinical benefit group) and non-responders (no clinical benefit group) in metastatic melanoma included platelet nadir during treatment (p = 0.015), autoimmune phenomena (p = 0.049), and in renal cell carcinoma, platelet nadir (p = 0.026). There were no significant differences between number of doses of IL-2 received per cycle and response in either cancer subtype. Clinical benefit occurred in 25% of patients (9/36) when IL-2 was given as first-line therapy. Median overall survival for the clinical benefit group from the initiation of IL-2 to death or last follow-up was 61 months versus 17 months for the no clinical benefit group (p < 0.001) for metastatic melanoma. In renal cell carcinoma overall survival for clinical benefit patients was 48 months versus 17 months. No treatment-related deaths occurred.ConclusionsHigh-dose IL-2 can be safely administered by an experienced team in a non–intensive care oncology unit. The clinical benefit group developed autoimmune phenomena (melanoma patients), lower platelet nadir, and on average, received the same number of IL-2 doses as the no clinical benefit group, suggesting a response relationship to the patient’s baseline immune status. Further investigation of immune predictors of response may be useful to select appropriate patients for this therapy.

Dynamic changes in biomarkers for immune response to sipuleucel-t (SipT) for metastatic castrate-resistant prostate cancer (mCRPC).

181 Background: SipT prolongs OS in asymptomatic or minimally symptomatic CRPC without regularly inducing PSA declines. Elevated CD54 count, CD54 upregulation (UPREG) and total nucleated cells (TNC), markers of immune activation in the product, have been shown to correlate with longer survival. We prospectively collected clinical characteristics and inflammatory markers as part of an audit to identify possible predictors of immune response in patients receiving SipT. Methods: 92 men with CRPC had blood drawn routinely before and after SipT treatment at USC. Circulating tumor cells (CTC), PSA, prostatic acid phosphatase (PAP), albumin (Alb), hemoglobin (Hb), serum alkaline phosphatase (SAP), LDH, C-reactive protein (CRP) and β2-microglobulin (β2m) were evaluated in a reference lab. SipT product parameters, CD54, UPREG and TNC were measured by Dendreon. These datasets were analyzed using Spearman coefficients for continuous variables and the Kruskal-Wallis test for categorical variables. Results: 92 pts who received SipT were included. Median age was 69 (48-90). 43% had Gleason 8-9. 42% had ECOG Performance Status (PS) 0, 44% PS 1, and 13% PS 2. 26% had received prior chemotherapy and 18% had used opioids. 84% had bone mets, 42% had lymphadenopathy, and 12% had visceral mets. Increases in the PSA, PAP, LDH and Alb was directly correlated with increased CD54 counts, increases in the CTC was directly correlated with increased TNC, whereas decrease in SAP was correlated with increased upregulation of dendritic cells. Conclusions: Our results show that increased tumor markers immediately after treatment are more common in high CD54 product pts. This may represent a “flare” due to immune activation. [Table: see text]

Clinical and biologic factors for immune response to sipuleucel-T (SipT) for metastatic castrate-resistant prostate cancer (mCRPC).

219 Background: SipT prolongs overall survival of men with asymptomatic or minimally symptomatic mCRPC. Recently, it has been demonstrated that SipT immune parameters correlate with survival. We prospectively collected biological and inflammatory markers as part of an audit to identify possible predictors of immune response in patients (pts) receiving SipT. Methods: Circulating tumor cells (CTC), PSA, prostatic acid phosphatase (PAP), albumin (Alb), hemoglobin (Hb), serum alkaline phosphatase (SAP), LDH, C-reactive protein (CRP) and β2-microglobulin (β2m) were evaluated in reference lab before SipT. Product parameters CD54, CD54 upregulation (UPREG) and total nucleated cells (TNC) were measured by Dendreon. These datasets were analyzed using Spearman coefficients for continuous variables and the Kruskal-Wallis test for categorical variables. Results: 92 pts who received SipT were included. Median age was 69 (48-90). 43% had Gleason 8-9. 42% had ECOG Performance Status (PS) 0, 44% PS 1, and 13% PS 2. 26% had received prior chemotherapy and 18% had used opioids. 84% had bone mets, 42% had lymphadenopathy, 12% had visceral mets. Of clinical factors only PS was inversely correlated with UPREG (p=0.014) and TNC (p= 0.023). Biomarkers which significantly correlated with SipT immune parameters are shown in the Table. Age, location of mets, prior chemotherapy or opioid use did not impact the immune response to SipT Conclusions: Better product parameters correlated with good ECOG PS, lower PSA and SAP, and higher Hgb and Alb, suggesting that SipT induces a greater immune response in men with lower disease burden. The correlation of increased CD54 count with lower CRP, a surrogate for IL-6 activity, warrants investigation. [Table: see text]

Psychosocial Predictors of Immune Response Following Bone Marrow Transplantation

This study analyzed the relationship between some psychosocial variables (depression, anxiety, stress, coping strategies, social support, optimism, rationality, and need for harmony) and clinical parameters indicative of immunological response after bone marrow transplantation (BMT; day of engraftment, number of infections and hemoglobin level) while controlling for demographic variables (age, educative level, civil state, and time from cancer diagnosis). Thirty-one post BMT hematological cancer patients were evaluated. Results show that higher educative levels are associated to lower number of infections, while age is associated with a delay in the time of engraftment; coping strategies, specially redefinition of the situation, relaxation, stoicism and passivity, are positively associated with the three clinical indices; depression is positively associated to number of infections during the hospitalization period; and rationality is associated with lower hemoglobin levels. These results suggest that psychosocial variables, especially coping strategies, play an important role in determining the immunological response after BMT.