Markers Of Immune Response Research Articles

Myocardial Iba1, MHC class II, and CD3 are diffusely increased in canine myocarditis: A step toward antemortem myocarditis diagnostics

Canine myocarditis is a rare but serious health concern, potentially causing heart failure and death. Antemortem diagnosis is hampered by the numerous causes, nonspecific course, and dearth of diagnostic criteria. Currently, definitive diagnosis can only be made after death. The current human diagnostic gold standard is endomyocardial biopsy pairing cardiac histopathology with immunohistology to enhance detection of often-multifocal disease. We evaluated immune response markers in the canine heart to establish similar immunohistologic criteria. We hypothesized that myocardial major histocompatibility complex class II (MHCII), cluster of differentiation 3 (CD3), and ionized calcium binding adapter molecule 1 (Iba1), markers increased in human myocarditis, would be increased in canine myocarditis cases. Archived paraffin-embedded myocardial tissue from 22 histopathologically confirmed cases of adult and juvenile myocarditis and 23 controls was analyzed by immunohistochemistry for MHCII, CD3, and Iba1, and the fraction of myocardium with labeling was determined. All 3 markers were significantly increased compared with controls across the entire section: Iba1, 10.1× (P < .0001, Mann-Whitney U test); MHCII, 3.04× (P = .0019); and CD3, 4.4× (P = .0104). To mimic off-target biopsy, samples from 2 mm2 outside of inflammatory foci were analyzed, and these showed significant increases in Iba1 by 3.2× (P = .0036, Mann-Whitney U test) and CD3 by 1.2× (P = .0026). These data show diffusely increased immune response markers with canine myocarditis, with detection potentially independent of tissue sampling. Thus, endomyocardial biopsy and immunohistochemical detection of MHCII, CD3, and Iba1 may permit sensitive antemortem diagnosis of canine myocarditis.

Assessing the role of eosinophil-mediated immune response markers in detecting hookworm infection: A case-control study in Kintampo, Ghana.

Background and AimHuman hookworm disease caused by Ancylostoma duodenale and Necator americanus is a serious public health problem. Hookworm infection activates eosinophil‐mediated tissue inflammatory responses, involving the production of the eosinophil‐specific chemokine (eotaxin), recruitment of eosinophils, secretion of the cationic protein, and production of antiparasite immunoglobulin E (IgE). We investigated eosinophil‐mediated immune response as markers (CCL11, eosinophil cationic protein [ECP], and IgE) for detecting hookworm infection.MethodsThis case‐control study was carried out in hookworm endemic areas within the Kintampo North Municipality.Forty hookworm‐positive subjects and 36 apparently healthy individuals were recruited as cases and controls, respectively. Stool samples were collected for hookworm detection by the Kato–Katz technique and speciation by polymerase chain reaction. Approximately, 5 ml of intravenous blood was used to obtain plasma for the immunological assays.ResultsOf eosinophil‐mediated immune response markers studied, ECP and CCL11 were significantly higher among hookworm patients compared to controls. Increasing CCL11 (β = −0.81, p = 0.015) was associated with a significant decrease hookworm intensity. However, increasing eosinophil count (β = 0.62, p = 0.027) was associated with significant increase in hookworm intensity. In receiver operator characteristics analysis, ECP could significantly detect hookworm infection with a very high area under the curve (AUC) (AUC = 0.97, p < 0.0001). At a cutoff of 39.05, ECP was the best eosinophil‐mediated immune response marker for detecting hookworm infection with a sensitivity of 97.2%, specificity of 87.8%, a positive predictive value of 89.7%, and a negative predictive value of 96.6%.ConclusionECP best predicts eosinophil‐mediated immune response for detecting hookworm infection, while CCL11 and eosinophil count better predict the intensity of hookworm. Moreover, the ECP level is a good indicator of hookworm infection and intensity and may require additional investigations to augment current hookworm diagnostic techniques.

The impact of serum albumin-to-alkaline phosphatase ratio in cervical cancer patients treated with definitive chemoradiotherapy

We retrospectively analysed the prognostic significance of serum albumin, alkaline phosphatase (ALP) and albumin to ALP ratio (AAPR) and other prognostic factors affecting the overall survival (OS) and progression-free survival (PFS) in 200 cervical cancer patients treated with definitive chemoradiotherapy (CRT). The prognostic factors for OS and DFS, in addition to the predictive factors of albumin, ALP and AAPR, were investigated. Older age, lymph node metastasis, non-complete response (CR) to treatment and low serum albumin levels emerged as predictors of poor OS and PFS in multivariate analysis. However, with a cut-off value of 0.51, AAPR was not a significant prognostic factor of survival in multivariable analysis. There were no significant differences in clinicopathological factors between patients with low and high AAPR, except for lymph node metastasis, where lymph node metastasis rate was significantly higher in patients with a low AAPR compared to those with a high AAPR. Patients with CR had a significantly higher serum albumin level and AAPR compared to patients without CR. The pre-treatment serum albumin level was independent predictive for survival; therefore, it could be a suitable biomarker to guide systemic therapy and predict patient outcomes. Impact Statement What is already known on this subject? Two major determinants of tumour progression are nutritional status and inflammation. The albumin-to-alkaline phosphatase ratio (AAPR), which was originally proposed as a marker for nutritional status and immune response, was recently discovered to be a prognostic factor for various cancer types. However, its utility in the treatment of cervical cancer has not been established. What do the results of this study add? Low serum albumin levels were associated with a significantly shorter OS and PFS in cervical cancer patients treated definitively with CRT. AAPR, on the other hand, was not a significant prognostic factor for survival with a cut-off value of 0.51. Regional lymph node metastasis was significantly more common in patients with a low AAPR than in those with a high AAPR. What are the implications of these findings for clinical practice and/or further research? Patients with multiple clinicopathological risk factors and low serum albumin levels had an increased risk of disease recurrence and a poorer prognosis, highlighting the importance of additional adjuvant treatment strategies in these patients. Due to the preliminary nature of our findings, additional research is required to corroborate them.

Neutrophil-lymphocyte ratio and prediction of pathological complete response to neoadjuvant treatment in breast cancer.

e12627 Background: Pathological complete response (pCR) after neoadjuvant therapy in breast cancer is an important prognostic indicator for improved disease free survival. High tumour grade, low ER-status, HER2-positivity predict higher likelihood of a pCR however better predictive biomarkers are needed. The role of the immune system and specifically the presence of tumour infiltrating lymphocytes in increasing response to chemotherapy has been extensively studied across cancer subtypes2. We sought to determine if neutrophil-to-lymphocyte ratio (NLR) as a marker of immune inflammatory response could be informative in predicting likelihood of pCR. Methods: We identified a cohort of patients who had received neoadjuvant chemotherapy between 2012 to 2018. We extracted clinical, pathological and laboratory data for each case. A pretreatment peripheral blood test was used to calculate a neutrophil-to-lymphocyte ratio. We used R-studio to analyse our data. The primary objective was to determine if there was an association between NLR and pCR. Statistical analysis was performed to assess whether NLR ratio correlated with pCR. Results: We identified 107 patients who had received NAC for early stage breast cancer during our study period. The mean NLR in the cohort was 2.95 (range 0.76-11.9). 35 (32.7%) patients achieved a pCR. There was no statistically significant different in NLR in those who achieved pCR (n=35) and those who did not (n=72) (p value= 0.75). Patients with triple negative breast cancer (TNBC) had a higher rate of pCR (45.1%), compared to HER2-positive (42.1%) and ER-positive HER2-negative (25.4%). The NLR was not significant in predicting pCR within the separate histological cohorts. Conclusions: NLR did not predict for pathological complete response in our cohort. Further studies into the predictive value of NLR in this setting with larger cohorts are needed. Investigation into the dynamic changes of NLR both pre, post during treatment may provide further insights as may larger cohorts for analysis in each histological subtype.

Proteomic profiling of serum small extracellular vesicles reveals immune signatures of children with pneumonia.

BackgroundPneumonia is the leading cause of death in young children globally. However, the underlying pathological mechanism of pediatric pneumonia remains unclear. In infection disease contexts, small extracellular vesicles (sEVs) have been shown to be a useful source of markers for pathogenesis and immune response. We hypothesized that functional molecules such as protein harbored by sEVs would provide mechanistic insights into the immune response in children with pneumonia.MethodsWe isolated sEVs from serum collected from children with and without pneumonia, performed proteomic analysis of the sEVs with label-free mass spectrometry, and then conducted functional enrichment analysis of proteomic data.ResultsWe identified fifteen differentially expressed proteins and ten unique proteins in children with pneumonia as compared to healthy children. In the pneumonia group, immune-related processes and pathways were positively enriched as upregulated proteins were involved in neutrophil activation, complement regulation, defense against bacteria, humoral immune response and regulation of immune effector processes However, pathways associated with tissue development and extracellular matrix remodeling were negatively enriched, as downregulated proteins were linked to extracellular matrix structure and cell adhesions.ConclusionsOur findings provided insights into host responses to pathogen infection, which has contributed to understanding the pathogenesis of children with pneumonia. Furthermore, our studies suggested that serum sEVs proteins could be considered a potential source of biomarkers for diagnosing pediatric pneumonia.

Toxic Effects on Oxidative Stress, Neurotoxicity, Stress, and Immune Responses in Juvenile Olive Flounder, Paralichthys olivaceus, Exposed to Waterborne Hexavalent Chromium.

Simple SummaryMetals such as chromium can be exposed at high levels in the marine environment, and exposure to these heavy metals can have a direct effect on marine organisms. High levels of chromium exposure can have a direct impact on organisms in a coastal cage and terrestrial aquaculture. Hexavalent chromium exposure of more than 1.0 and 2.0 mg Cr6+/L induced physiological responses such as antioxidant, neurotransmitter, immune, and stress indicators in Paralichthys olivaceus. Therefore, this study will provide a reference indicator for stable aquaculture production through reference indicators for toxicity due to chromium exposure that may exist in the marine environment.Juvenile Paralichthys olivaceus were exposed to waterborne hexavalent chromium at various concentrations (0, 0.5, 1.0, and 2.0 mg/L) for 10 days. After chromium exposure, the activities of superoxide dismutase and glutathione S-transferase, which are oxidative stress indicators, were significantly increased; however, the glutathione level was significantly reduced. Acetylcholinesterase activity as a neurotoxicity marker was significantly inhibited upon chromium exposure. Other stress indicators, including plasma cortisol and heat shock protein 70, were significantly increased. The immune response markers (lysozyme and immunoglobulin M) were significantly decreased after chromium exposure. These results suggest that exposure to environmental toxicity in the form of waterborne chromium at concentrations higher than 1.0 mg/L causes significant alterations in antioxidant responses, neurotransmitters, stress, and immune responses in juvenile olive flounders. This study will provide a basis for an accurate assessment of the toxic effects of hexavalent chromium on aquatic organisms.

Analysis of possible markers of effective antitumor cellular immune response before starting therapy with immune check-point inhibitors

The aim of the study. to analyse possible markers of an effective antitumor cellular immune response.Material and methods. using the keywords «checkpoint inhibitors, immunotherapy, t-lymphocytes, exhausted t-lymphocytes, anti-tumor immune response», review and original articles (n=34) published from 2005 to 2020 in the Pubmed, Web Of Science, Elsevier databases were selected.Results. the study revealed possible markers reflecting a high activity of an adaptive immune response based on effective recognition of tumor antigens through MHC molecules, a sufficient number of T-lymphocytes and a predominance of T-cytotoxic cells, as well as a low level of expression of inhibitory receptors and small molecules. the presence of single nucleotide polymorphisms in the HLA-I and HLA-II genes encoding MHC-I and MHC-II proteins, respectively, a high level of lymphocytes, among which the most important is the predominance of CD8+ t cells and a low level of T-regulatory cells (T-reg), as well as the presence of single nucleotide polymorphisms in the genes encoding FcγR receptors of T-lymphocytes showed their predictive significance. the diagnostic significance of determining the expression of inhibitory receptors for T-lymphocytes (TIM3, LAG3, TIGIT), especially in combination with the determination of PD-1 expression, was also revealed.Conclusion. the results obtained may be relevant for applying new methods for the assessment of the functional activity of the T-cell immune response before starting therapy with checkpoint inhibitors, as well as for the development of new diagnostic panels, which may be of interest to employees of clinical diagnostic laboratories and research centers.

Association of immune response with overall and disease-free survival in laryngeal squamous cell carcinomas

ObjectivesThis study aimed to examine the relationship between checkpoint receptors (PD-1, PD-L1, PD-L2, CTLA-4) and lymphoid infiltration level (TILs) with prognostic features of patients with laryngeal squamous cell carcinoma (LSCC). MethodsA retrospective study was designed at a tertiary referential university hospital between April 2008 and December 2020. The surgical specimen of the patients who met the eligibility criteria were re-examined histopathological, sociodemographic, clinical, pathological, and follow-up findings of patients were determined. The impact of PD-1, PD-L1, PD-L2, CTLA4, and TILs levels for the presence of cancer recurrence, disease-specific mortality, overall survival (OS), disease-free survival (DFS) was investigated. ResultsForty-five patients with LSCC were included in the study. The mean follow-up period was 48.3 ± 14.3 months (min: 36, max 84). TILs scores were detected significantly lower in patients with distant metastasis and recurrence (p = 0.046 and 0.010). Also, only TILs was a significant risk factor for recurrence and survival among the PD-1, PD-L1, PD-L2, CTLA-4, and TILs (HR = 0.217 CI: 0.070–0.679, p = 0.009 and HR = 0.566, CI: 0,321–980, p = 0.048). Similarly, for the TILs score: > 1 was significant for DFS. (Long-Rank = 0.009). The examined markers and TILs scores were not a significant predictive factor for OS. ConclusionAn increase in TILs density in LSCCs is associated with a better prognosis. However, PD-1, PD-L1, PD-L2, CTLA-4 could not be associated with prognosis. Controlled studies combined with immunotherapy treatment results are needed to reveal their role as a marker and prognostic factor of the anti-tumor immune response.

Low But Recoverable Markers of Humoral Immune Response to BNT162b2 in Elderly LTCF Residents Five to Seven Months After Two-Dose Vaccination.

The immune response is known to wane after vaccination with BNT162b2, but the role of age, morbidity and body composition is not well understood. We conducted a cross-sectional study in long-term care facilities (LTCFs) for the elderly. All study participants had completed two-dose vaccination with BNT162b2 five to 7 months before sample collection. In 298 residents (median age 86 years, range 75–101), anti-SARS-CoV-2 rector binding IgG antibody (anti-RBD-IgG) concentrations were low and inversely correlated with age (mean 51.60 BAU/ml). We compared the results to Health Care Workers (HCW) aged 18–70 years (n = 114, median age: 53 years), who had a higher mean anti-RBD-IgG concentration of 156.99 BAU/ml. Neutralization against the Delta variant was low in both groups (9.5% in LTCF residents and 31.6% in HCWs). The Charlson Comorbidity Index was inversely correlated with anti-RBD-IgG, but not the body mass index (BMI). A control group of 14 LTCF residents with known breakthrough infection had significant higher antibody concentrations (mean 3,199.65 BAU/ml), and 85.7% had detectable neutralization against the Delta variant. Our results demonstrate low but recoverable markers of immunity in LTCF residents five to 7 months after vaccination.

Clinical, FDG-PET and Molecular Markers of Immune Checkpoint Inhibitor Response in Patients with Advanced Merkel Cell Carcinoma

Merkel cell carcinoma (MCC) is a rare, highly aggressive, neuroendocrine cancer of the skin, associated with immunosuppression, Merkel cell polyoma virus (MCPyV) infection and UV-carcinogenesis.Whilst impressive and durable responses to immune checkpoint inhibitors have revolutionised the treatment of advanced MCC.

HPV16 infection decreases vaccine-induced HPV16 antibody avidity: the CVT trial

The HPV vaccine has shown sustained efficacy and consistent stabilization of antibody levels, even after a single dose. We defined the HPV16-VLP antibody avidity patterns over 11 years among women who received one- or three doses of the bivalent HPV vaccine in the Costa Rica HPV Vaccine Trial. Absolute HPV16 avidity was lower in women who received one compared to three doses, although the patterns were similar (increased in years 2 and 3 and remained stable over the remaining 8 years). HPV16 avidity among women who were HPV16-seropositive women at HPV vaccination, a marker of natural immune response to HPV16 infection, was significantly lower than those of HPV16-seronegative women, a difference that was more pronounced among one-dose recipients. No differences in HPV16 avidity were observed by HPV18 serostatus at vaccination, confirming the specificity of the findings. Importantly, point estimates for vaccine efficacy against incident, six-month persistent HPV16 infections was similar between women who were HPV16 seronegative and seropositive at the time of initial HPV vaccination for both one-dose and three-dose participants. It is therefore likely that this lower avidity level is still sufficient to enable antibody-mediated protection. It is encouraging for long-term HPV-vaccine protection that HPV16 antibody avidity was maintained for over a decade, even after a single dose.

Role of triggering receptor expressed on myeloid cells-1 (TREM-1) in COVID-19 and other viral pneumonias: a systematic review and meta-analysis of clinical studies.

BackgroundTriggering receptor expressed on myeloid cells-1 (TREM-1) has emerged as an important inflammatory marker of immune response associated with severity and mortality outcomes in infection diseases, including viral pneumonias.Aim(1) To evaluate the expression of TREM-1 in patients with COVID-19 and other viral pneumonias compared to healthy individuals; and (2) to analyze the levels of these biomarkers according to disease severity.Materials and methodsThis review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline. Searches were performed in PubMed, Scopus, Embase, and Google Scholar. Studies were considered eligible if they were observational studies that provided data on the levels of TREM-1 in humans with viral pneumonia compared to healthy controls. The results of the meta-analysis were expressed as standardized mean difference (SMD) and an effect size of 0.8 was considered a large effect. A subgroup analysis was performed according to the disease severity.ResultsSeven studies were included in this systematic review. Four studies included patients with COVID-19 and three analyzed patients with different viruses. The meta-analysis was performed only with patients with COVID-19, which showed increased levels of soluble form of TREM-1 (sTREM-1) among patients with COVID-19 compared to healthy controls (SMD 1.53; 95% CI 0.53–2.52; p < 0.01). No differences were found between patients with mild-to-moderate COVID-19 and healthy controls, but higher levels of sTREM-1 were shown among patients with severe COVID-19 (SMD 1.83; 95% CI 0.77–2.88; p < 0.01). All three studies including patients with other viral pneumonias showed that TREM-1 levels were significantly elevated in infected patients compared with controls.ConclusionThese findings may provide evidence on the pro-inflammatory role of TREM-1 in these infections, contributing to the inflammatory profile and disease progression.Supplementary InformationThe online version contains supplementary material available at 10.1007/s10787-022-00972-6.

Impact of Chemoembolic Regimen on Immune Cell Recruitment and Immune Checkpoint Marker Expression following Transcatheter Arterial Chemoembolization in a VX2 Rabbit Liver Tumor Model

PurposeTo characterize the effects of commonly used transcatheter arterial chemoembolization (TACE) regimens on the immune response and immune checkpoint marker expression using a VX2 rabbit liver tumor model. Materials and MethodsTwenty-four VX2 liver tumor-bearing New Zealand white rabbits were assigned to 7 groups (n = 3 per group) undergoing locoregional therapy as follows: (a) bicarbonate infusion without embolization, (b) conventional TACE (cTACE) using a water-in-oil emulsion containing doxorubicin mixed 1:2 with Lipiodol, drug-eluting embolic-TACE with either (c) idarubicin-eluting Oncozene microspheres (40 μm) or (d) doxorubicin-eluting Lumi beads (40–90 μm). For each therapy arm (b–d), a tandem set of 3 animals with additional bicarbonate infusion before TACE was added, to evaluate the effect of pH modification on the immune response. Three untreated rabbits served as controls. Tissue was harvested 24 hours after treatment, followed by digital immunohistochemistry quantification (counts/μm2 ± SEM) of tumor-infiltrating cluster of differentiation 3+ T-lymphocytes, human leukocyte antigen DR type antigen-presenting cells (APCs), cytotoxic T-lymphocyte–associated protein-4 (CTLA-4), and programmed cell death protein-1 (PD-1)/PD-1 ligand (PD-L1) pathway axis expression. ResultsLumi-bead TACE induced significantly more intratumoral T-cell and APC infiltration than cTACE and Oncozene-microsphere TACE. Additionally, tumors treated with Lumi-bead TACE expressed significantly higher intratumoral immune checkpoint markers compared with cTACE and Oncozene-microsphere TACE. Neoadjuvant bicarbonate demonstrated the most pronounced effect on cTACE and resulted in a significant increase in intratumoral cluster of differentiation 3+ T-cell infiltration compared with cTACE alone. ConclusionsThis preclinical study revealed significant differences in evoked tumor immunogenicity depending on the choice of chemoembolic regimen for TACE.

Targeting the Gut Microbiota of Vertically HIV-Infected Children to Decrease Inflammation and Immunoactivation: A Pilot Clinical Trial.

Aims: Children with HIV exhibit chronic inflammation and immune dysfunction despite antiretroviral therapy (ART). Strategies targeting persistent inflammation are needed to improve health in people living with HIV. The gut microbiota likely interacts with the immune system, but the clinical implications of modulating the dysbiosis by nutritional supplementation are unclear. Methods: Pilot, double-blind, randomized placebo-controlled trial in which 24 HIV-infected on ART were randomized to supplementation with a daily mixture of symbiotics, omega-3/6 fatty acids and amino acids, or placebo four weeks, in combination with ART. We analyzed inflammatory markers and T-cell activation changes and their correlations with shifts in fecal microbiota. Results: Twenty-four HIV-infected children were recruited and randomized to receive a symbiotic nutritional supplement or placebo. Mean age was 12 ± 3.9 years, 62.5% were female. All were on ART and had HIV RNA < 50/mL. We did not detect changes in inflammatory (IL-6, IL-7, IP-10), microbial translocation (sCD14), mucosal integrity markers (IFABP, zonulin) or the kynurenine to tryptophan ratio, or changes in markers of the adaptive immune response in relation to the intervention. However, we found correlations between several key bacteria and the assessed inflammatory and immunological parameters, supporting a role of the microbiota in immune modulation in children with HIV. Conclusions: In this exploratory study, a four-week nutritional supplementation had no significant effects in terms of decreasing inflammation, microbial translocation, or T-cell activation in HIV-infected children. However, the correlations found support the interaction between gut microbiota and the immune system.

Abstract P4-05-06: High absolute lymphocyte counts are associated with longer overall survival in patients with metastatic breast cancer treated with eribulin as the first-line chemotherpy. Combined analysis of two phase 2 study

Abstract Background: Eribulin demonstrated improving overall survival of HER2-negative metastatic breast cancer in EMBRACE trial. Recently, ad hoc analysis of the trial showed that immune response markers such as absolute lymphocyte count (ALC) was associated with prognosis. However, blood cell count must be influenced by previous chemotherapy because the trial was targeted for late-line treatment.Previously we had conducted two phase 2 trials that estimated efficacy of eribulin as the first-lien chemotherapy for HER2-negative metastatic breast cancer in Japan. Base line ALC and NLR were examined for the participants of these trials to determine whether they were also associated with prognosis in the first-line setting. Patients and Methods: A total of 59 patients were enrolled this study including 35 patients of BIRICHEN trial (UMIN000006086; SpringerPlus 2016;5:164) for only first-line chemotherapy with eribulin and 24 patients who treated as the first-line chemotherapy in OMC-BC03 trial targeted for first and second line chemotherapy (UMIN000009568; Cancer Chemother Pharmacol 2018; 81:923). Pretreatment blood cell counts were collected from case report form and compared with survival data. Cutoff value of ALC was set at 1500/mm3 and that of NLR was set at 3 in accordance with ad hoc analysis of EMBRACE trial. The ethics committees of Osaka Medical College and Osaka City University approved the present study. Results: Median value of ALC was 1690/mm3(Quartile Q1,Q3: 1014, 2012) and that of NLR was 2.17(Q1,Q3: 1.54, 2.99). In comparison with ALC, overall survival (OS) was 132.6 months in the ALC-high group(&amp;gt;=1500;n=33) versus 76.4 months in the ALC-low group (&amp;lt; 1500; n=26). Hazard ratio(HR) was 0.52(95%CI; 0.27-1.01) with border line significancy. Progression free survival(PFS) is 28.0 months in the ALC-high group versus 20.4 months in the ALC-low group. HR was 0.91(95% CI; 0,51-1.60) without statistically significant difference. In NLR, OS was 20.7 months in the NLR-low group(&amp;lt;3;n=45) versus 4.6 months in the NLR-high group. HR was 0.40 (95%CI; 018-0.90) with statistical significance. PFS was 6.2 months in the NLR-low group versus 10.8 months in the NLR-high group (&amp;gt;=3; n=14). HR was 0.57 (95%CI; 0.25-1.30) without statistically significant difference. Conclusions: In the post hoc analysis of the EMBRACE trial, patients who assigned eribulin group with ALC 1500 or higher had better OS than those with ALC less than 1500, but no difference was observed in PFS. In addition, those with NLR less than 3 had better PFS and OS than those with NLR 3 or more. In comparison with the capecitabine group, although low NLR was a good prognostic factor not only in eribulin group but also in capecitabine group, high ALC was suggested to be a particular prognostic factor of eribulin. However, since EMBRACE study was a late line setting, there could be an effect of myelosuppression by pretreatment. Our first-line results did not affect bone marrow function by prior chemotherapy, but similar results were obtained. ALC may be a prognostic factor of eribulin regardless of the treatment line, suggesting that eribulin exerts its effect by acting on the immune microenvironment. Citation Format: Tsutomu Takashima, Kosei Kimura, Hidemi Kawajiri, Shinichiro Kashiwagi, Shinya Tokunaga, Shigehiko Nishimura, Satoru Noda, Hiroyo Oku, Ayana Ikari, Tomo Tominaga, Saki Maezawa, Junna Sakane, Mitsuhiko Iwamoto. High absolute lymphocyte counts are associated with longer overall survival in patients with metastatic breast cancer treated with eribulin as the first-line chemotherpy. Combined analysis of two phase 2 study [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P4-05-06.

Abstract P4-07-11: Prognostic significance of changes in tumor infiltrating lymphocytes and neutrophil-to-lymphocyte ratio after neoadjuvant chemotherapy in early breast cancer patients

Abstract Introduction. There are still 30-40% of patients with early breast cancer (BC) that relapse after neoadjuvant chemotherapy (NAC). New prognostic biomarkers are needed. Current evidence shows that both markers of antitumor local and systemic immune response, such as tumor infiltrating lymphocytes (TIL) and neutrophil-to-lymphocyte ratio (NLR) have prognostic value in early BC. Most works have evaluated baseline levels, but their variation after NAC might be related to treatment resistance and should be explored.The aim of this study was to simultaneously investigate the prognostic significance of the pre-post-NAC variation of TIL and NLR in patients with early BC.Methods. Retrospective, single-center cohort of 121 patients with early BC treated with NAC between 2001 and 2010. Pre- and post-NAC TIL (CD3+) were assessed by tumor tissue microarrays performed in both diagnostic core-needle biopsy and surgical excision specimens, respectively. Pre- and post-NAC NLR were calculated based on blood tests collected within 1 month of cancer diagnosis and at the moment of surgery, respectively. The change on TIL and NLR after NAC was estimated by calculating the absolute difference of post- and pre- values of these parameters (Δpost-pre). We examined the association between the absolute difference of TIL and NLR (both divided into deciles) and survival outcomes by Cox regression. To represent in Kaplan-Meier curves the combined effect of the change of TIL and NLR in OS, we divided patients into 4 groups, depending on the sign of Δpost-pre values of TIL and NLR: 1) no TIL nor NLR increment after NAC (ΔTIL &amp; ΔNRLpost-pre≤0); 2) no TIL but NLR increment (ΔTILpost-pre≤0, ΔNRLpost-pre&amp;gt;0); 3) TIL but no NLR increment (ΔTILpost-pre&amp;gt;0, ΔNRLpost-pre≤0); and 4) TIL and NLR increment following NAC (ΔTIL &amp; ΔNRLpost-pre&amp;gt;0).Results. 47 (40%) out of 121 patients had pre- and post-NAC values of both NLR and TIL. Patients’ baseline characteristics are shown in Table 1. The median follow-up of this group was 11.5 years (IQR: 10.5 - 12.5 years). At the final follow-up date, overall survival (OS) was 87.2 %. In the univariable analysis, the increase in TIL and NLR after NAC among deciles showed a negative prognostic value for OS (TIL: HR 1.54, 95%CI 1.05-2.26, p=0.026; HR 1.32 95%CI 0.98-1.79, p=0.070). In the multivariable analysis, adjusted by cancer staging after NAC, both the increase of TIL and NLR after NAC among deciles add independent predictive value (TIL: HR 1.53, 95%CI 1.00-2.34, p=0.049; RNL: HR 1.42 95%CI 0.95-2.12, p=0.09).The subpopulation of patients with no TIL increment after NAC (Groups 1&amp;2, N=23) showed the best prognosis, with no deaths at 10 years. Among patients with TIL increment (Groups 3&amp;4, N=22), those with no NLR increment (Group 3, N=5) showed an intermediate prognosis (20% deaths at 10 years), whereas patients with concurrent NLR increment (Group 4, N=17) had the lowest OS (29.4% deaths at 10 years). Differences observed between groups were statistically significant (Log Rank p=0.042).Conclusion. The integrated characterization of TIL and NLR variations after NAC identifies different prognostic subgroups in early BC patients. In our cohort, both the increment of TIL and/or NLR after NAC are associated to a worse prognosis. Future validation of these findings in large, multicenter cohorts might allow treatment optimization by means of new strategies such as immunotherapy. Table 1.Patients’ baseline characteristicsN=47Age (median, range)56 (21, 78)Menstrual statusPostmenopausal25 (53,2%)Premenopausal22 (46,8%)Histologic subtypeInvasive ductal carcinoma42 (91,5%)Other subtypes4 (8,5%)Histologic gradeGrade 12 (4,3%)Grade 218 (38,3%)Grade 323 (48,9%)Unknown4 (8,6%)cTcT221 (44,7%)cT324 (51,1%)cT4a-d2 (4,2%)cNcN018 (38,3%)cN114 (29,8%)cN26 (12,8%)cN39 (19,1%)Molecular subtypeHR+ HER2-22 (46,8%)HER2+ HR+8 (17,0%)HER2+ HR-6 (12,8%)Triple negative10 (21,3%)Breast surgeryConservative26 (55,3%)Mastectomy21 (44,7%)Axillary surgerySentinel lymph node biopsy12 (25,5%)Axillary lymph node dissection35 (74,5%)Relapse typeMetastasis7 (14,9%)Local/contralateral1 (0,83%)Deaths6 (12,8%) Citation Format: Esmeralda Garcia-Torralba, Beatriz Álvarez-Abril, Carlos Bravo-Pérez, Esther Navarro Manzano, Pilar de la Morena Barrio, Alejandra Ivars Rubio, Elisa García-Garre, Gema Marín Zafra, Francisco Ayala de la Peña, Elena García-Martínez. Prognostic significance of changes in tumor infiltrating lymphocytes and neutrophil-to-lymphocyte ratio after neoadjuvant chemotherapy in early breast cancer patients [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P4-07-11.

Evaluation of Anti-HBsAg Titres among the High-Risk Population of A Tertiary Care Hospital, Tamil Nadu, India

Health care workers (HCW) are a high-risk population for Hepatitis B infection. Hepatitis B vaccine which is 95% effective confers long-term protection and anti-HBs titre is a marker for protective immune response. Our objective was to assess the status of hepatitis B vaccination and to evaluate the anti-HBsAg titres among health care workers in a tertiary care hospital, Tamil Nadu. It was an observational study conducted among 610 Health care workers in a tertiary care teaching hospital from June to December 2018 after obtaining clearance from IEC. Workers were assessed for their HBV vaccination status and for their anti-HBsAg titre after getting informed consent. The antibody titres were measured using CLIA (chemiluminescent Immunoassay) supplied by Abbott diagnostics. The data was entered and analyzed using a Microsoft Excel sheet. In our study, 80.5% were fully vaccinated, 18.5% of them were defaulters which comprised the HCW with 2 doses and 1 dose of vaccine and 0.9% were not vaccinated. In the fully vaccinated group, 37% showed Anti HBs titres of 10-100 mIU/ml, 59.2% showed the titre of &gt; 100 mIU/ml and 3.6% did not show the protective antibody titre (&lt;10 mIU/ml ). The 3.6% who did not show protective antibody titre were given the booster dose of vaccine. Among those who received a booster dose, 61.1% responded with the titre of 10-100 mIU/ml and 22.2% responded with the titre of &gt; 100 mIU/ ml and 16.6% did not respond even with the booster dose. In the defaulters 86.7% had titres &lt; 10 mIU/ml , 9.7% had titres of 10 mIU/ml and 3.6% had titres &gt; 10 mIU/ml. In the non vaccinated group all had titres &lt; 10 mIU/ml. The present study emphasizes the importance of screening of anti-HBsAg titres to be made mandatory for all the health care workers along with HBV Vaccination.

Multi-OMICs data analysis identifies molecular features correlating with tumor immunity in colon cancer.

There is a current need for new markers with higher sensitivity and specificity to predict immune status and optimize immunotherapy use in colon cancer. We aimed to investigate the multi-OMICs features associated with colon cancer immunity and response to immunotherapy. We evaluated the association of multi-OMICs data from three colon cancer datasets (TCGA, CPTAC2, and Samstein) with antitumor immune signatures (CD8+ T cell infiltration, immune cytolytic activity, and PD-L1 expression). Using the log-rank test and hierarchical clustering, we explored the association of various OMICs features with survival and immune status in colon cancer. Two gene mutations (TERT and ERBB4) correlated with antitumor cytolytic activity found also correlated with improved survival in immunotherapy-treated colon cancers. Moreover, the expression of numerous genes was associated with antitumor immunity, including GBP1, GBP4, GBP5, NKG7, APOL3, IDO1, CCL5, and CXCL9. We clustered colon cancer samples into four immuno-distinct clusters based on the expression levels of 82 genes. We have also identified two proteins (PREX1 and RAD50), ten miRNAs (hsa-miR-140, 146, 150, 155, 342, 59, 342, 511, 592 and 1977), and five oncogenic pathways (CYCLIN, BCAT, CAMP, RB, NRL, EIF4E, and VEGF signaling pathways) significantly correlated with antitumor immune signatures. These molecular features are potential markers of tumor immune status and response to immunotherapy.

Inflamed Tumor Phenotype as Predictor of Long-Term Response to Pembrolizumab in an EGFR-Mutated Non-Small Cell Lung Cancer (NSCLC) Patient with Acquired Resistance to Afatinib: a Case Report and Review of the Literature.

Treatment with immune checkpoint inhibitors (ICIs) that target the programmed cell death 1/programmed cell death ligand-1 (PD-1/PD-L1) axis is usually ineffective in patients with epidermal growth factor receptor (EGFR)-mutated advanced non-small cell lung cancer (NSCLC), either as first-line treatment or in later lines. By contrast, especially for patients with common EGFR mutations (exon 19 deletion/L858R point mutation), an orally bioavailable EGFR tyrosine kinase inhibitor (EGFR-TKI) is the best upfront therapy, being able to provide response rates well above 50% and a median progression-free survival ranging from 11 to 19 months, depending on whether a second-generation (e.g., afatinib) or a third-generation (i.e., osimertinib) EGFR-TKI is used. Unfortunately, treatment options for these patients at the time of acquired resistance are limited. As for afatinib-pretreated patients, those who develop a T790M mutation may benefit from osimertinib, whereas platinum-based chemotherapy is the preferable therapeutic strategy for T790M-negative patients as well as for patients who progress on osimertinib administered as first-line therapy. Here, we describe the case of an exon-19-deleted patient who experienced a complete response to the anti-PD-1 agent pembrolizumab upon the development of T790M-negative acquired resistance to afatinib. Furthermore, we discuss this case in the context of the existing literature, especially focusing on the importance of evaluating multiple markers of immune response post-EGFR-TKI and prior to ICI treatment in order to select the best treatment strategy in this clinical scenario.

TMC5 is Highly Expressed in Human Cancers and Corelates to Prognosis and Immune Cell Infiltration: A Comprehensive Bioinformatics Analysis.

Background: The oncological role of TMC5 in human cancers has only been revealed partially. We performed integrated bioinformatics analysis to provide a thorough and detailed insight of associations between TMC5 and tumorigenesis, cancer progression, and prognosis. Methods: With reference to the accessible online databases, the TMC5 expressions in tumor tissues and corresponding normal tissues, different pathological stages, and various cancer cells were analyzed, while the protein levels of TMC5 in different cancers were also inspected. Meanwhile, the prognostic value of TMC5 expression in multiple cancers as well as in advanced-stage patients was investigated. Furthermore, the mutational data of TMC5 and its correlation with cancer prognosis were assessed. Moreover, the association between the TMC5 level and immune cell infiltration was evaluated. Next, TMC5-related pathway alterations and drug responses were summarized. Finally, the TMC5 based protein network was generated, and relevant enrichment was performed. Results: In our study, the expression level of TMC5 was significantly higher in the tumor tissue than that of the normal tissues in most cancer types. Fluctuations of TMC5 levels were also observed among different pathological stages. In the meantime, the protein level elevated in the tumor tissue in the cancers enrolled. Moreover, the expression of TMC5 was not only prognostic for overall survival (OS) or recurrence free survival (RFS) in various types of cancers but also correlated to OS in patients with more advanced cancers. Additionally, the mutational status of TMC5 is also associated with prognosis in cancer patients. It is worth noting that the TMC5 level was closely related to immune cell infiltrations, especially in ESCA, TGCT, and USC. The TMC5 expression was also identified as an activator for pathways including PI3K/AKT, RAS/MAPK, and TSC/mTOR, proved to be associated with multiple drug responses and assessed to be interactive with the TMEM family. Conclusion: TMC5 might function as a potential marker for cancer survival and immune responses.