Nonspecific Antigen Research Articles

Scalable graphene nanogrid FET biosensors: Fabrication & characterization for sub-femtomolar detection of viral proteins

The development of highly sensitive biosensors has been significantly advanced by graphene nanostructures, particularly nanogrids, due to their remarkable surface area-to-volume ratio, which enhances biomolecule detection at ultra-low concentrations. This study introduces a scalable fabrication technique for graphene nanogrids integrated into field-effect transistor (FET) biosensors, specifically designed for sub-femtomolar detection of viral proteins. Through the optimization of pore size and distribution, the graphene nanogrid FET biosensors demonstrated high signal-to-noise ratios (SNR) and overcame challenges associated with non-specific antigen interference. The surface antigen was detected at 0.25 fM in serum, even in the presence of Hepatitis C. In addition to these advancements, structural analysis techniques, including XPS and TEM, were employed to observe the effects of lattice temperature and core structures on the electronic band gap, which directly influenced electron mobility. Lattice temperature effects were analyzed for their impact on sensitivity, revealing that optimized temperature control significantly improved detection performance, while precise adjustments in pore morphology and material composition reduced device non-linearity and minimized quantification errors. By operating in heterodyne mode (80 kHz–2 MHz) and utilizing a probabilistic neural network (PNN), the sensor mitigated Debye screening effects, achieving a 70 % increase in SNR, detecting 0.20 fM of Hepatitis B viral proteins with 60 % sensitivity and 6 % standard deviation, demonstrating potential for ultra-sensitive biomedical detection with enhanced electron mobility and reduced non-linearity.

Fluorescence immunoassay using triangular carbon dots for detection of the cardiac marker Troponin T in acute myocardial infarction

A cardiac immunosensor was developed to detect heart attacks (myocardial infarctions) at an early stage. This technique, utilizing carboxylic functionalized triangular carbon dots (caf-TCDs) combined with an antibody anti-cardiac Troponin T (anti-cTnT), proved to be a quick and sensitive method for detecting cardiac troponin T (cTnT) in both buffer solutions and biological fluids such as serum. The antibody for cardiac Troponin T (cTnT) was chemically linked to amphiphilic caf-TCDs using a carbodiimide coupling reaction. Exfoliated graphene oxide (GO) sheets effectively bind to anti-cTnT labeled caf-TCDs on their surface, resulting in a non-radiative energy transfer process where the fluorescence is suppressed. This interaction between caf-TCDs and GO follows a nonlinear and increasing Stern-Volmer relationship, indicating a combination of static and dynamic quenching. When antigen (cTnT) is present, the interaction between antibodies (anti-cTnT) and the cTnT molecules hinders the energy transfer process. This interaction also affects the positioning of the nano-couple and causes the detachment of CDs from the GO surface. As a result, the energy transfer process is impeded, leading to the restoration of fluorescence intensity. The sensor exhibits high specificity and low sensitivity to non-specific antigens. It shows a linear reaction to cTnT in the range of 0.05–50 ng mL-1, with a detection limit of 0.046 ng mL-1. Therefore, the anti-cTnT-caf-TCD/GO sensor offers a potential platform for detecting cTnT with increased sensitivity.

Diazo-functionalised immunoelectrochemical sensor for the detection of ochratoxin a in foods

Ochratoxin A (OTA) is a toxic fungal metabolite that is commonly found in cereals and animal feed. It is economically damaging and potentially hazardous to human health. Herein, we propose an electrochemical immunosensor for the rapid detection of OTA using anti-OTA antibodies and diazonium-functionalized, screen-printed electrodes. We attached 4-aminobenzoic acid to an electrode surface, activated the carboxyl groups on the surface with carbodiimide, and attached an antibody to the diazo layer. Subsequently, we used bovine serum protein as a blocker to prevent non-specific antigens from binding to the antibody. We evaluated the performance of the sensor by cyclic voltammetry, electrochemical impedance spectroscopy, and differential pulse voltammetry. The sensor is highly specific and sensitive, has good linear responses in the range 20–200 ng/mL, a limit of detection of 0.5 ng/mL, and good recoveries of 90.5%–100.9% in spiked samples. It can be stored at 4 °C for approximately 2 weeks, and is highly stable, with a current response variation of no more than 4.6%.

Synthesis of carbon nanotubes-chitosan nanocomposite and immunosensor fabrication for myoglobin detection: An acute myocardial infarction biomarker

The use of single-walled carbon nanotubes (SWCNTs) in biomedical applications is limited due to their inability to disperse in aqueous solutions. In this study, dispersed –COOH functionalized CNTs with N-succinylated chitosan (CS), greatly increasing the water solubility of CNTs and forming a uniformly dispersed nanocomposite solution of CNTs@CS. Coupling reagent EDC/NHS was used as a linker with the –COOH groups present on the N-succinylated chitosan which significantly improved the affinity of the CNTs for biomolecules. Myoglobin (Mb) is a promising biomarker for the precise assessment of cardiovascular risk, type 2 diabetes, metabolic syndrome, hypertension and several types of cancer. A high level of Mb can be used to diagnose the mentioned pathogenic diseases. The CNTs@CS-FET demonstrates superior sensing performance for Mb antigen fortified in buffer, with a wide linear range of 1 to 4000 ng/mL. The detection limit of the developed Mb immunosensor was estimated to be 4.2 ng/mL. The novel CNTs@CS-FET immunosensor demonstrates remarkable capability in detecting Mb without being affected by interferences from nonspecific antigens. Mb spiked serum showed a recovery rate of 100.262 to 118.55 % indicating great promise for Mb detection in clinical samples. The experimental results confirmed that the CNTs@CS-FET immunosensor had excellent selectivity, reproducibility and storage stability.

Advancements in Molecular Imaging for the Diagnosis and Management of Hepatocellular Carcinoma

Hepatocellular Carcinoma (HCC) is a growing global health burden with high incidence and mortality rates. Despite advances in surgical techniques and perioperative care, outcomes after surgical treatment have not improved over the past three decades. Molecular imaging is an emerging field that enables researchers to study diseases at the molecular and cellular levels, enabling the detection of elevated serum α-fetoprotein (AFP) and abnormal expressions of various HCC-specific and nonspecific cell surface antigens and intracellular targets. Molecular imaging techniques detect liver lesions at the molecular and cellular level, allowing early detection and accurate staging of HCC. Positron emission tomography (PET) imaging offers greater sensitivity and specificity, while hepatobiliary-specific radiotracers with SPECT imaging provide insights into benign and malignant lesion differentiation. Radiomics and artificial intelligence are vital in deciphering molecular imaging data, with machine learning algorithms boosting diagnostic gains and predicting treatment response. Theranostics, a state-of-the-art application, provides diagnostic and therapeutic leverage following a single imaging agent. By understanding tumor biology in real time, radiopharmaceuticals can be transformed into personalized radiotherapies, enabling clinicians to make science-driven decisions throughout the illness. Future directions include developing novel radiotracers and integrating AI into clinical decision-making. Collaboration between academic researchers, clinicians, and industry colleagues is crucial to converting exciting advances into improved clinical outcomes for HCC patients.

Морфометрический анализ сетчатки глаза крыс при хроническом стрессе в условиях измененной иммунореактивности

Introduction. Chronic stress is an integral part of a modern industrial society. Chronic stress affects both physical and psychological well-being by causing various morphological and functional changes in different organ tissues, including the retina. As a result of an excess of catecholamines and glucocorticoids, retinal neurons become damaged, which leads to visual impairment. There are controversial data on how altered immunoreactivity affects the severity of stress-associated changes in organs and tissues, particularly in the retina. Thus, we aimed to study stress-associated changes in the retina. Materials and methods. We performed experiments on 32 Wistar male rats, which were divided into 4 equal groups, group 1 including intact animals. Group 2 consisted of the intact rats that received one injection of a nonspecific immunogen, i.e., ram erythrocytes (500 million cells per 500 microliters of saline). The rats from group 3 were exposed to light and sound stress for 30 minutes 7 days running. The stressed animals that were injected once with a nonspecific immunogen on day 3 of stress exposure comprised group 4. We assessed changes in the microstructural parameters of the retina using light microscopy and measuring interleukin-6 (IL-6), cortisol, adrenocorticotropic hormone, and corticotropin-releasing hormone in blood serum with enzyme immunoassay. Results. Among the stress-exposed animals, microscopic examination revealed a disruption of the retinal stratification, decreased width of the outer and inner nuclear layers, hypocellularity, “empty” sites in retinal layers, as well as hyperchromically and hydropically altered neurons. In contrast, there were no obvious retinal defects in the stressed animals that received a nonspecific antigen, which was confirmed morphometrically. The immunization was correlated with decreased IL-6 in the blood serum of the stressed rats. The results of blood tests showed a significantly low level of stress-associated hormones, which was not different from that in the intact animals. Conclusion. Nonspecific immunization of laboratory rats under chronic stress leads to preserved retinal cytoarchitectonics and inhibits stress-induced activity of the hypothalamic-pituitary-adrenal system. Keywords: chronic stress, retina, immunization

Developing a diagnostic model for predicting prostate cancer: a retrospective study based on Chinese multicenter clinical data.

The overdiagnosis of prostate cancer (PCa) caused by nonspecific elevation serum prostate-specific antigen (PSA) and the overtreatment of indolent PCa have become a global problem that needs to be solved urgently. We aimed to construct a prediction model and provide a risk stratification system to reduce unnecessary biopsies. In this retrospective study, clinical data of 1807 patients from three Chinese hospitals were used. The final model was built using stepwise logistic regression analysis. The apparent performance of the model was assessed by receiver operating characteristic curves, calibration plots, and decision curve analysis. Finally, a risk stratification system of clinically significant prostate cancer (csPCa) was created, and diagnosis-free survival analyses were performed. Following multivariable screening and evaluation of the diagnostic performances, a final diagnostic model comprised of the PSA density and Prostate Imaging-Reporting and Data System (PI-RADS) score was established. Model validation in the development cohort and two external cohorts showed excellent discrimination and calibration. Finally, we created a risk stratification system using risk thresholds of 0.05 and 0.60 as the cut-off values. The follow-up results indicated that the diagnosis-free survival rate for csPCa at 12 months and 24 months postoperatively was 99.7% and 99.4%, respectively, for patients with a risk threshold below 0.05 after the initial negative prostate biopsy, which was significantly better than patients with higher risk. Our diagnostic model and risk stratification system can achieve a personalized risk calculation of csPCa. It provides a standardized tool for Chinese patients and physicians when considering the necessity of prostate biopsy.

AlGaN/GaN HEMT Based Biosensor for Detection of the HER2 Antigen Spiked in Human Serum

This work reports the development of a gallium nitride-based high electronic mobility transistor (GaN HEMT)-based biosensor to detect human epidermal growth factor receptor-2 (HER2) antigen in spiked human serum. The platform devices have been fabricated with a source to drain distance, gate length, and unit gate width of 25, 3, and <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$100 ~\mu \text{m}$ </tex-math></inline-formula> , respectively. The cysteine methyl ester (CME)-based chemistry has been utilized to immobilize the antiHER2 antibody on the sensing region of the sensor. The formation of the CME layer was confirmed through Raman spectroscopy, which shows a peak around a wavelength of 262 cm <sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">−1</sup> . The antibody immobilization on the gold surface has been confirmed through enzyme-linked immunosorbent assay (ELISA). The sensor has been electrically characterized before and after each step of the functionalization process. The sensor shows a significant change in the drain current of 0.95 and 1.7 mA at a <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">${V}_{{\mathrm {ds}}} $ </tex-math></inline-formula> of +5 V for 50- and 200-ng/ml concentration of HER2 in spiked human serum. A minor change in the drain current of the sensor, when tested with a nonspecific antigen, suggested the high specificity of the sensor for HER2. The sensor has been tested for a broad range concentration of HER2 antigen from 0.7 pg/ml to 200 ng/ml.

Development of a rapid diagnostic test for the detection of antibodies or antigens to Coronavirus (COVID-19)

The global health crisis caused by COVID-19 has overwhelmed both healthcare settings and economies globally. While mass population testing has improved drastically, recent reviews of existing methods have highlighted various shortcomings with these methods.&amp;nbsp;The aim of this project was to investigate whether the LAA could be modified and utilised as a rapid detection test which either matched or exceeded the existing sensitivity and specificity values.&amp;nbsp; &amp;nbsp;The latex agglutination assay (LAA) investigated whether the COVID-19 spike protein could be detected in samples. COVID-19-specific&amp;nbsp; IgM and IgG were used in conjunction with a series of non-specific antigens. Control or AG-containing samples were mixed with AB-microsphere complexes on glass microscope slides.&amp;nbsp;Manual visualisation identified various levels of agglutination. Light microscopy and spectrophotometry at 405 nm determined that the LAA could detect at least 2.3 ng of spike protein.&amp;nbsp;&amp;nbsp;The particle-counting tool of ImageJ was utilised to obtain a data set which was subjected to statistical analysis which indicated that there was a significant difference between control samples and live tests, P = 0.000102 for the spike protein assay and P = 0.254 for the non-specific assay respectively.&amp;nbsp;The results obtained fell in line with a similar study conducted by Buffin et al. in 2018.&amp;nbsp;The analytical methods used in this project twinned with data obtained in previous studies supports the significant difference between control values and live test values. The LAA is easier, quicker to use (results in ≤ 30 minutes) and cheaper, with potentially better sensitivity to existing methods. This could benefit high- and low-income countries alike upon further research and optimisation.&amp;nbsp;

Research progress of anti-γ-aminobutyric acid B receptor encephalitis and a case report of paraneoplastic associated encephalitis and treatment analysis.

Encephalitis is one of the common diseases in neurology. Early diagnosis and appropriate treatments are essential. Autoimmune encephalitis (AE) generally refers to a type of encephalitis mediated by autoimmune mechanisms. It is gradually considered to be an important cause of reversible encephalitis caused by noninfectious factors. It can occur in children, adolescents, and adults, and is clinically characterized by multifocal or diffuse brain damage such as personality changes, seizures, and cognitive impairment, with an overall good effect of immunotherapy. According to the clinical features of the patients, blood and cerebrospinal fluid tests, neuroelectrophysiology, cranial imaging, treatment and prognosis, AEs can be broadly divided into specific antigen (antibody)-related AEs and nonspecific antigen (or antibody) -related AEs. With the development of AEs research, more and more anti-neuron antibodies have been found, which provides an important reference for the diagnosis and treatment of AEs. Understanding the knowledge about AEs is important to discover new diseases and deepen the understanding of the immunopathological mechanisms of existing central nervous system diseases. Anti-γ-aminobutyric acid B (GABA-B) receptor encephalitis is a type of AE, but this disease is rare in AE, often develop to the clinical manifestations of marginal encephalitis, accompanied by obvious seizures or status epilepticus, Some patients had tumors, mainly small-cell carcinoma, prompt diagnosis, early immunotherapy and, if necessary, tumor treatment resulted in complete or partial neurological improvement in most patients.

CAR-T Regulatory (CAR-Treg) Cells: Engineering and Applications.

Regulatory T cells are critical for maintaining immune tolerance. Recent studies have confirmed their therapeutic suppressive potential to modulate immune responses in organ transplant and autoimmune diseases. However, the unknown and nonspecific antigen recognition of polyclonal Tregs has impaired their therapeutic potency in initial clinical findings. To address this limitation, antigen specificity can be conferred to Tregs by engineering the expression of transgenic T-cell receptor (TCR) or chimeric antigen receptor (CAR). In contrast to TCR Tregs, CAR Tregs are major histocompatibility complex (MHC) independent and less dependent on interleukin-2 (IL-2). Furthermore, CAR Tregs maintain Treg phenotype and function, home to the target tissue and show enhanced suppressive efficacy compared to polyclonal Tregs. Additional development of engineered CAR Tregs is needed to increase Tregs’ suppressive function and stability, prevent CAR Treg exhaustion, and assess their safety profile. Further understanding of Tregs therapeutic potential will be necessary before moving to broader clinical applications. Here, we summarize recent studies utilizing CAR Tregs in modulating immune responses in autoimmune diseases, transplantation, and gene therapy and future clinical applications.

Electrocardiographic and echocardiographic findings in children with dengue infection.

ABSTRACTBackground:The mechanism of myocardial damage in dengue could be the release of inflammatory mediators or the direct action of the dengue virus on myocytes leading to myocarditis. The release of inflammatory mediators is more in a severe form of the disease that correlates to the higher incidence of cardiac manifestations in patients with severe dengue.Aim:To determine the electrocardiographic and two-dimensional (2D)-echocardiographic findings in children with dengue infection and to find a correlation with disease severity.Materials and Methods:A total of 150 children between 1 month and 12 years of age seropositive for dengue Non-specific antigen 1 (NS1) Enzyme linked immunosorbent assay (ELISA) or dengue Immunoglobulin M (IgM) ELISA were studied in the Department of Pediatric Medicine of a tertiary care government hospital. The patients were undertaken for 12 lead electrocardiograms (ECGs) and echocardiograms.Results:Out of the 150 dengue seropositive cases, 61 cases were of mild dengue, 67 cases were of dengue with warning signs (DWSS), and 22 cases were severe dengue cases. Abnormal ECG was found in 78 cases (52%) in a total of 150 cases in terms of rate, prolonged PR interval (interval between atrial depolarization and ventricular activation), ST (ventricular repolarisation)-segment depression, and low-voltage complexes. Abnormal 2D- echocardiography (ECHO) was found in 70 (46.6%) out of 150 in terms of the ejection fraction (EF) <55%, Early diastole/atrial contraction (E/A) ratio <1, and the presence of pericardial effusion.Conclusion:Nearly 50% of the patients have abnormal ECG and ECHO findings, more so in the severe dengue group. There is a statistically significant association of the EF between mild dengue, DWWS with severe dengue (P =0.001).

Risk assessment of latent tuberculosis infection through a multiplexed cytokine biosensor assay and machine learning feature selection

Accurate detection and risk stratification of latent tuberculosis infection (LTBI) remains a major clinical and public health problem. We hypothesize that multiparameter strategies that probe immune responses to Mycobacterium tuberculosis can provide new diagnostic insights into not only the status of LTBI infection, but also the risk of reactivation. After the initial proof-of-concept study, we developed a 13-plex immunoassay panel to profile cytokine release from peripheral blood mononuclear cells stimulated separately with Mtb-relevant and non-specific antigens to identify putative biomarker signatures. We sequentially enrolled 65 subjects with various risk of TB exposure, including 32 subjects with diagnosis of LTBI. Random Forest feature selection and statistical data reduction methods were applied to determine cytokine levels across different normalized stimulation conditions. Receiver Operator Characteristic (ROC) analysis for full and reduced feature sets revealed differences in biomarkers signatures for LTBI status and reactivation risk designations. The reduced set for increased risk included IP-10, IL-2, IFN-γ, TNF-α, IL-15, IL-17, CCL3, and CCL8 under varying normalized stimulation conditions. ROC curves determined predictive accuracies of > 80% for both LTBI diagnosis and increased risk designations. Our study findings suggest that a multiparameter diagnostic approach to detect normalized cytokine biomarker signatures might improve risk stratification in LTBI.

Comparative characteristics of the state of the immune system in patients with coronary artery disease with stable angina pectoris and acute coronary syndrome

The aim – to assess the relationship between the state of the immune system and the development of acute coronary syndrome in patients with IHD.Materials and methods. The first group consisted of 64 patients with ST-segment elevation acute coronary syndrome, mean age 54 (49–64) years; the second group – 223 patients with coronary artery disease with stable exertional angina, FC II–III, mean age 56 (49–63) years; the third group – 47 patients with acute coronary syndrome without ST segment elevation, mean age 61 (52–65) years. The material for the immunological study was peripheral venous blood. To determine the parameters of cellular and humoral innate and adaptive immunity in blood serum and supernatants of mononuclear cells, enzyme immunoassay was used.Results and discussion. In patients with coronary artery disease with acute coronary syndrome with ST segment elevation compared with patients with coronary artery disease with stable angina pectoris, the levels of indicators of the immune status in the blood were: CRP – 9.3 (5.3–12.0) versus 4.8 (2.4–8.1) mg/L (p=0.0001), sICAM – 785 (690–830) versus 565 (406–744) ng/ml (p=0.0001), IL-10 in blood mononuclear cells – 48 (1–228) versus 194 (21–758) pg/ml (p=0.0007), circulating immune complexes – 90 (70–108) versus 76 (54–105) od. (p=0.045), lymphocytes with apoptosis (CD95) – 16 (9–27) versus 11 (8–17) % (p=0.029), spontaneous oxygen-dependent metabolism of monocytes – 16 (12–21) versus 13 (9–17) (p=0.001). The levels of indicators of the immune system in the blood in patients with coronary artery disease with acute coronary syndrome with ST segment elevation compared with patients with coronary artery disease with acute coronary syndrome without ST segment elevation were: T-helpers – 37 (32–41) versus 42 (37–48) % (p=0.0006) (R=–0.33; p=0.0005), reaction of lymphocyte blast transformation to nonspecific antigen – 38 (32–47) versus 50 (42–61) % (p=0.0004) (R=–0.37; p=0.0003).Conclusions. The development of acute coronary syndrome is directly combined with increased activity of the immune system, as evidenced by the high production of proinflammatory CRP, IL-8, sICAM with a low level of anti-inflammatory IL-10, a pronounced humoral adaptive immune response (in terms of antibodies to the myocardium and vascular tissues, CD40, circulating immune complexes) and active functional state of monocytes (according to cNCT test, functional reserve, phagocytosis) in patients with coronary artery disease with acute coronary syndrome, regardless of the position of the ST segment in comparison with patients with stable coronary artery disease. Elevated levels of antibodies to the myocardium in patients with stable coronary heart disease indicate moderate myocardial damage due to temporary ischemia in angina attacks, even with a stable course of the disease. In patients with acute coronary syndrome, high levels of antibodies to the myocardium indicate myocardial damage due to increased ischemia in plaque destabilization much earlier than the clinical manifestations of acute coronary syndrome. In acute coronary syndrome with ST-segment elevation, compared with ACS patients without ST-segment elevation, activation of neutrophils and suppression of the activity of adaptive T-cell immunity is noted (by the level of T-helpers, sCD40L, blast transformation of lymphocytes, γ-interferon in mononuclear cells, apoptosis of lymphocytes).

Induction of Therapeutic Protection in an HPV16-Associated Mouse Tumor Model Through Targeting the Human Papillomavirus-16 E5 Protein to Dendritic Cells

HPV E5 is an oncoprotein mainly expressed in premalignant lesions, which makes it an important target for a vaccine to prevent or cure cervical cancer (CC). In this study, we evaluated whether E5 targeted to DEC-205, present in dendritic cells (DCs), could induce a therapeutic protection against HPV16-induced tumor cells in a mouse model. The HPV-16 E5 (16E5) protein was cross-linked to a monoclonal antibody (mAb) specific to mouse DEC-205 (anti-DEC-205:16E5) or to an isotype control mAb (isotype:16E5). Rotavirus VP6 was cross-linked to the mouse anti-DEC-205 mAb (anti-DEC-205:VP6) as a non-specific antigen control. BALB/c mice were inoculated subcutaneously (s.c.) with the 16E5-expressing BMK-16/myc tumor cells, and 7 and 14 days later the mice were immunized s.c. with the conjugates, free 16E5 or PBS in the presence of adjuvant. Tumor growth was monitored to evaluate protection. A strong protective immune response against the tumor cells was induced when the mice were inoculated with the anti-DEC-205:16E5 conjugate, since 70% of the mice controlled the tumor growth and survived, whereas the remaining 30% developed tumors and died by day 72. In contrast, 100% of the mice in the control groups died by day 30. The anti-DEC-205:16E5 conjugate was found to induce 16E5-specific memory T cells, with a Th1/Th17 profile. Both CD4+ and CD8+ T cells contributed to the observed protection. Finally, treating mice that had developed tumors with an anti-PD-1 mAb, delayed the tumor growth for more than 20 days. These results show that targeting 16E5 to DEC-205, alone or combined with an immune checkpoint blockade, could be a promising protocol for the treatment of the early stages of HPV-associated cancer.

Boosting immune response with GM-CSF optimizes primary cryotherapy outcomes in the treatment of prostate cancer: a prospective randomized clinical trial.

We explored the association of prostate cryotherapy and immunomodulation with granulocyte-macrophage colony-stimulating factor (GMCSF) in the generation of detectable tumor-specific T- and B-cell responses in men with prostate cancer. A randomized pilot study of patients assigned to either cryotherapy alone (Control group) or in combination with GMCSF (Treatment group). The impact of therapy on the development of T- and B-cell responses against tumor-related antigens was studied using enzyme-linked immune absorbent spot (ELISpot) and protein microarray panels (Sematrix) assays, respectively. Fold changes in response to treatment were calculated by normalization of post-treatment ELISpot values against the mean pre-cryoablation response. Student t tests between treatment and control groups at 4 weeks and 12 weeks across all the antigens were performed. A total of 20 patients were randomized to either control or treatment arm. At 4 weeks after cryotherapy, the treatment group demonstrated an average fold change in cancer antigen-related antibodies of 2.8% above their mean baseline values, whereas controls averaged an 18% change below mean baseline (p < 0.05). At 12 weeks, antibody response in treatment group increased to 25% above baseline, while the average of control group patients remained 9% below baseline (p < 0.05). Patients in treatment group displayed, on average, higher ELISPOT readings for the 4- and 12-week times points (527 vs 481 for PSA and 748 vs 562 for PAP). GMCSF appeared to broadly elevate antibodies against prostate-specific and nonspecific antigens. Prostate antigen-specific T-cell responses were more enhanced over non-prostate-specific responses, preferentially in the treatment group. Our findings suggest a possible therapeutic effect of adjuvant immunotherapy in association with cryotherapy for the treatment of prostate cancer.

Image-Guided Intratumoral Delivery of Immunotherapeutics in Gastrointestinal Malignancies

AbstractIntratumoral (IT) administration of immunotherapy is a promising treatment strategy under clinical development for gastrointestinal malignancies. Due to its targeted nature, IT immunotherapies can generate regional proinflammatory microenvironments that result in the focal recruitment of tumor-specific immune cells. Precision targeting of tumors via IT immunotherapy injection theoretically produces a more robust immune response to the treated tumor itself and to distant metastatic tumors that share tumor-specific antigens with those of the treated tumor, while also minimizing the priming of the adaptive immune system to nonspecific antigens. Diverse arrays of IT immunotherapeutic agents including but not limited to lyophilized bacteria, viral vectors, cellular-based agents, molecules, and peptides, both as monotherapies and in combination with systemic immunotherapies, are in various stages of preclinical and clinical development. In this review, we summarize the current state of the art for IT immunotherapy and highlight potential future directions and their relevance to image-guided interventionalists.

Pathology of extranodal marginal zone lymphoma at different anatomic sites

: Extranodal marginal zone lymphomas arise within acquired lymphoid tissue at a variety of extranodal sites. They have characteristic architectural and morphological features that recapitulate those seen in constitutive lymphoid tissue in extranodal locations that is best exemplified by the mucosa (gut) associated lymphoid tissue (MALT) present as Peyer’s patches in the terminal ileum. The morphology of these MALT lymphomas is constant across the various extranodal sites in which they are encountered although there is some variation between cases in the detailed cellular morphology with variable proportions of cells with centrocyte-like, monocytoid and plasmacytoid/plasmacytic appearances. Architecturally there is inter-case variation in the degree and type of follicular colonisation. The appearance of lymphoepithelial lesions varies between sites. There is variable association with light chain deposition disease and nodular amyloidosis which appear to be more frequently associated with a pulmonary origin. The immunophenotype is uniform across sites with a rather non-specific B-cell antigen expression with aberrant CD43 expression in a proportion of cases. There is infrequent expression of CD5 and this appears more frequent in lymphomas arising in the ocular/ocular adnexal region.

PULMONARY MUCORMYCOSIS: A RARE CAUSE OF HYDROPNEUMOTHORAX

SESSION TITLE: Medical Student/Resident Chest Infections Posters SESSION TYPE: Med Student/Res Case Rep Postr PRESENTED ON: October 18-21, 2020 INTRODUCTION: Mucormycosis is a rare but often fatal fungal infection. These fungi are ubiquitous in nature and most commonly cause rhino-orbital-cerebral infections. Patients who are immunocompromised are at higher risk (1). CASE PRESENTATION: A 76 year-old never-smoker with a medical history significant for type 2 diabetes (HbA1c 5.8%) and Coombs negative hemolytic anemia was admitted with 3 months of progressive dyspnea. She had an associated nonproductive cough for 10 days. She denied fevers, night sweats, chest pain or hemoptysis. Two months prior to presentation, she started prednisone 60 mg daily for hemolytic anemia. Prednisone was slowly tapered to 30 mg daily at the time of presentation. She received one dose of Rituximab due to persistent anemia. Evaluation 2 weeks prior to presentation revealed a right upper lobe, 5 cm cavitary lesion on chest CT. On arrival she was tachycardic (heart rate 110) and tachypneic (respiratory rate 32) without hypoxemia. Labs were notable for a hemoglobin of 9.5 g/dl, WBC of 13.6, and creatinine of 1.85 mg/dl. Chest x-ray showed a loculated right hydropneumothorax. Chest CT showed the right hydropneumothorax with progression of the right upper lobe cavitary lesion. A 14 French pigtail catheter was placed at bedside via ultrasound guidance. Pleural studies demonstrated exudative fluid with negative cytology and cultures. She underwent CT-guided biopsy of the cavitary lesion, which revealed inflammation, necrosis, and numerous broad-ribbon-like, pauci-septate hyphae favoring mucormycosis. IV liposomal amphotericin and posaconazole were initiated. Thoracic surgery declined operative intervention due to her age and comorbidities. She developed respiratory and renal failure and passed away 1 month after presentation. DISCUSSION: This patient developed pulmonary mucormycosis in the setting of glucocorticoid and Rituximab therapy. The diagnosis of mucormycosis can be difficult due to nonspecific symptoms (1) and lack of serologic tests. Commonly used fungal antigen tests such as 1,3-beta-D-glucan are traditionally negative (2). Chest CT findings are nonspecific with the classic halo or reversed halo sign present in only a minority of cases (1). A definitive diagnosis is made with histopathology (1). Successful treatment requires early diagnosis, reversal of predisposing factors, surgical debridement, and prompt initiation of antifungal therapy (3). Liposomal amphotericin is first-line pharmacologic therapy (1); however, antifungal therapy without surgical debridement is unlikely to be successful. CONCLUSIONS: Our patient presented with an unusual case of pulmonary mucormycosis; a high suspicion and histopathology are required to make the diagnosis. CT findings are non-specific and common fungal antigen tests can be negative. Early diagnosis and prompt initiation of antifungal therapy with surgical debridement improves the chance for survival. Reference #1: Farmakiotis D, Kontoyiannis DP. Mucormycoses. Infect Dis Clin North Am 2016; 30:143. Reference #2: Ostrosky-Zeichner L, Alexander BD, Kett DH, et al. Multicenter clinical evaluation of the (1-->3) beta-D-glucan assay as an aid to diagnosis of fungal infections in humans. Clin Infect Dis 2005; 41:654. Reference #3: Spellberg B, Walsh TJ, Kontoyiannis DP, et al. Recent advances in the management of mucormycosis: from bench to bedside. Clin Infect Dis 2009; 48:1743. DISCLOSURES: No relevant relationships by Meghan Hill, source=Web Response author/editior relationship with UpToDate Please note: $5001 - $20000 Added 04/01/2020 by David Midthun, source=Web Response, value=Royalty No relevant relationships by Kelly Pennington, source=Web Response

Fasciola hepatica Extracellular Vesicles isolated from excretory-secretory products using a gravity flow method modulate dendritic cell phenotype and activity.

Parasite-released extracellular vesicles (EVs) deliver signals to the host immune system that are critical to maintaining the long-term relationship between parasite and host. In the present study, total EVs (FhEVs) released in vitro by adults of the helminth parasite Fasciola hepatica were isolated using a recently described gravity flow method that protects their structural integrity. The FhEVs molecular cargo was defined using proteomic analysis and their surface topology characterised by glycan microarrays. The proteomic analysis identified 618 proteins, 121 of which contained putative N-linked glycosylation sites while 132 proteins contained putative O-linked glycosylation sites. Glycan arrays revealed surface-exposed glycans with a high affinity for mannose-binding lectins indicating the predominance of oligo mannose-rich glycoproteins, as well as other glycans with a high affinity for complex-type N-glycans. When added to bone-marrow derived dendritic cells isolated FhEV induced a novel phenotype that was categorised by the secretion of low levels of TNF, enhanced expression of cell surface markers (CD80, CD86, CD40, OX40L, and SIGNR1) and elevation of intracellular markers (SOCS1 and SOCS3). When FhEV-stimulated BMDCs were introduced into OT-II mice by adoptive transfer, IL-2 secretion from skin draining lymph nodes and spleen cells was inhibited in response to both specific and non-specific antigen stimulation. Immunisation of mice with a suspension of FhEV did not elicit significant immune responses; however, in the presence of alum, FhEVs induced a mixed Th1/Th2 immune response with high antigen specific antibody titres. Thus, we have demonstrated that FhEVs induce a unique phentotype in DC capable of suppressing IL-2 secretion from T-cells. Our studies add to the growing immuno-proteomic database that will be an important source for the discovery of future parasite vaccines and immunotherapeutic biologicals.