Field Of Immunology Research Articles

Problems of creating antibody phage libraries and their solutions

Phage display has become an efficient, reliable and popular molecular technique for generating libraries encompassing millions or even billions of clones of divergent peptides or proteins. The method is based on the correspondence between phage genotype and phenotype, which ensures the presentation of recombinant proteins of known amino acid composition on the surface of phage particles. The use of affinity selection allows one to choose variants with affinity for different targets from phage libraries. The implementation of the antibody phage display technique has revolutionized the field of clinical immunology, both for developing tools to diagnose infectious diseases and for producing therapeutic agents. It has also become the basis for efficient and relatively inexpensive methods for studying protein–protein interactions, receptor binding sites, as well as epitope and mimotope identification. The antibody phage display technique involves a number of steps, and the final result depends on their successful implementation. The diversity, whether natural or obtained by combinatorial chemistry, is the basis of any library. The choice of molecular techniques is critical to ensure that this diversity is maintained during the phage library preparation step and during the transformation of E. coli cells. After a helper phage is added to the suspension of transformed E. coli cells, a bacteriophage library is formed, which is a working tool for performing the affinity selection procedure and searching for individual molecules. Despite the apparent simplicity of generating phage antibody libraries, a number of subtleties need to be taken into account. First, there are the features of phage vector preparation. Currently, a large number of phagemid vectors have been developed, and their selection is also of great importance. The key step is preparing competent E. coli cells and the technology of their transformation. The choice of a helper phage and the method used to generate it is also important. This article discusses the key challenges faced by researchers in constructing phage antibody libraries.

Immune Checkpoint Inhibitors in Geriatric Oncology.

This manuscript will update prior reviews of immune checkpoint inhibitors (ICIs) in light of basic science, translational, and clinical discoveries in the field of cancer immunology and aging. ICIs have led to significant advancements in the treatment of cancer. Landmark trials of ICIs have cited the efficacy and toxicity experienced by older patients, but most trials are not specifically designed to address outcomes in older patients. Underlying mechanisms of aging, like cellular senescence, affect the immune system and may ultimately alter the host's response to ICIs. Validated tools are currently used to identify older adults who may be at greater risk of developing complications from their cancer treatment. We review changes in the aging immune system that may alter responses to ICIs, report outcomes and toxicities in older adults from recent ICI clinical trials, and discuss clinical tools specific to older patients with cancer.

The complement system and its involvement in inhibition of Batrachochytriym dendrobatidis, a lethal fungal pathogen of amphibians

The field of ecological immunology, or ecoimmunology, has provided valuable insights on the immune responses of diverse host organisms threatened by infectious diseases in many different environments. One infectious disease that has been particularly notable for its impacts on host populations is amphibian chytridiomycosis, which has been linked with amphibian declines around the world. Amphibian immune responses to the pathogen that causes chytridiomycosis (Batrachochytriym dendrobatidis) are not well understood but thought to involve innate immune factors, including the complement system. In this study, we tested the ability of complement proteins to inhibit B. dendrobatidis in in vitro challenge assays. We found that complement proteins from amphibian plasma that were not heat inactivated reduced the viability and growth of B. dendrobatidis. The inhibitory efficacy was similar to effects on Pseudomonas fluorescens, a bacterium that is known to be inhibited by complement protein activation. These findings suggest inhibition of B. dendrobatidis that is consistent with the involvement of the complement system. In addition, we provide methods for standardizing pathogen killing assays, and set a foundation for further investigations on the amphibian complement system and other immune responses to amphibian chytridiomycosis.

Alzheimer's Disease Immunotherapy: Current Strategies and Future Prospects.

Alzheimer's disease (AD) is an extremely complex and heterogeneous pathology influenced by many factors contributing to its onset and progression, including aging, amyloid-beta (Aβ) plaques, tau fibril accumulation, inflammation, etc. Despite promising advances in drug development, there is no cure for AD. Although there have been substantial advancements in understanding the pathogenesis of AD, there have been over 200 unsuccessful clinical trials in the past decade. In recent years, immunotherapies have been at the forefront of these efforts. Immunotherapy alludes to the immunological field that strives to identify disease treatments via the enhancement, suppression, or induction of immune responses. Interestingly, immunotherapy in AD is a relatively new approach for non-infectious disease. At present, antibody therapy (passive immunotherapy) that targets anti-Aβ aimed to prevent the fibrillization of Aβ peptides and disrupt pre-existing fibrils is a predominant AD immunotherapy due to the continuous failure of active immunotherapy for AD. The most rational and safe strategies will be those targeting the toxic molecule without triggering an abnormal immune response, offering therapeutic advantages, thus making clinical trial design more efficient. This review offers a concise overview of immunotherapeutic strategies, including active and passive immunotherapy for AD. Our review encompasses approved methods and those presently under investigation in clinical trials, while elucidating the recent challenges, complications, successes, and potential treatments. Thus, immunotherapies targeting Aβ throughout the disease progression using a mutant oligomer-Aβ stimulated dendritic cell vaccine may offer a promising therapy in AD.

Comparison of ELISA with automated ECLIA for IL-6 determination in COVID-19 patients: An Italian real-life experience

ObjectivesCoronavirus disease 2019 (COVID-19) has a wide spectrum of clinical severity. A cytokine storm is associated with COVID-19 severity. Of these, IL-6 is significantly associated with higher mortality and is also a marker for predicting disease prognosis. IL-6 may act as a target for therapeutics and, a blockade of IL-6 function by Tocilizumab has been described as a treatment of the inflammatory process COVID-19-related. The aim of this study is to describe our experience comparing two different methods, in detail Human IL-6 Instant ELISA and the Elecsys IL-6 based on ECLIA, for the IL-6 assessment. Desing and methodsIL-6 levels from serum samples of 104 COVID-19 patients, admitted to the AOU Careggi (Hospital in Florence -Italy), were assessed by using the two above-mentioned methods and the results were analysed through Passing-Bablok regression fit and Bland-Altman plot. ResultsThe regression exhibited a linear relation between the methods with a regression equation (y = - 0.13 + 0.63 x; 95 % C.I. intercept = − 0.13 to 4.55; 95 % C.I. slope = 1.03 to 1.26 with R2 = 0.89, p > 0.05), showing a positive slope. The agreement of the two methods reported a bias of −25.0 pg/mL. Thus, the two methods correlate, but do not agree in terms of numeric results. ConclusionsThe two assays showed good comparability. However, because of the extremely wide linear range of the ECLIA, its throughput and its capacity for immune profiling, it represents an interesting emerging technology in the immunology field.

Evaluation of Antibacterial, Antioxidant and Immunomodulatory Potential of Mitragyna parvifolia Leaves-Assisted Fe2O3 Nanoparticles

This work includes the use of leaves of Mitragyna parvifolia for green synthesis of Fe2O3 nanoparticles (Fe2O3 NPs), which offers a sustainable eco-friendly approach. Characterization of synthesized Fe2O3 NPs was done by UV-visible, FTIR spectroscopy, XRD and FESEM studies. FTIR exhibited the existence of Fe-O, O–H and COO–Fe stretching owing to the occurrence of the carboxylic group. FESEM analysis revealed that the average size of Fe2O3 NPs was 92 nm, which varies in their shape. The study also includes the determination of antibacterial, in vitro antioxidant and immunomodulatory efficacy of synthesized Fe2O3 NPs. Results suggested that Fe2O3 NPs exhibit potent antibacterial efficacy against B. subtilis and S. aureus, gram-positive as well as E. coli and P. aureginosa, gram-negative bacterial strains used in this study. The study exhibited significant (p < 0.01) in vitro antioxidant and immunomodulatory potential of synthesized Fe2O3 NPs against concanavalin-A activated Wistar albino rat’s splenocytes. The green synthesized Fe2O3 NPs using M. parvifolia leaves may offer a novel approach in the medicinal and immunological field to overcome oxidative stress as well as enhance immune responses for potential health benefits of human beings.

Side Effects and Solutions of CAR-T Treatment

Cancer poses a significant threat to human health. It is a kind of complex and severe diseases characterized by the expanding of abnormal cells in the whole body due to uncontrolled growth. If left untreated, cancer with metastasis can invade nearby tissues, spread to other parts of the body, and impair the normal functioning of organs and systems. There are some treatment methods for it, such as surgery, chemical therapy and radiation. The field of immunology has made tremendous strides in understanding how the immune system works and how it can be harnessed to fight disease. Immunotherapy has become a rising solution in recent years. CAR-T therapy belongs to immunotherapy using genetically modified T cells with receptors of tumor antigens to specifically eliminate cancer cells, which is a promising approach for cancer patients. However, there are still some issues that remain to be resolved to make it wider using in treatment of cancer patients. For example, CAR-T therapy occurs severe side effects sometimes, even induced patients die, although it has been approved application in clinic since 2017, however, limited studies investigated its adverse reactions. Therefore, in the present, the author provides a comprehensive overview of the side effects of CAR-T therapy from reduction of therapeutic effects of antigen escape, to induction patients die of severe cytokine storms, then discusses corresponding treatment measures, which purpose is to help guide treatment for patients, make clinical staff focus on its side effects and provide some ideas for research and development in the future.

Abstract 118: Development of a novel murine model, NCG-M/hIL15, for enhanced post-HSC transplantation immunoreconstruction

Abstract Advances in the field of immunology and hematopoietic stem cell (HSC) transplantation have prompted the development of murine models that faithfully recapitulate human immune responses. We present the NCG-M/hIL15 model, an innovative advancement in the realm of preclinical research, addressing two key challenges associated with existing models: the limited capacity for human natural killer (NK) cell regeneration in NCG-M and the suboptimal reconstitution of T cells, myeloid lineage cells in NCG-hIL15 mice. Compared to the conventional NCG-M model, the NCG-M/hIL15 model demonstrates a significant improvement in its ability to regenerate human NK cells post-HSC transplantation. This advancement offers a valuable tool for studying NK cell biology, immune surveillance, and antitumor responses. Furthermore, when compared to the NCG-hIL15 model, the NCG-M/hIL15 model outperforms in the reconstitution of human T cells and myeloid lineage cells, contributing to a more comprehensive simulation of human immune responses. The overall survival of mice in this model is notably stable, further enhancing its utility for long-term immunological studies. The NCG-M/hIL15 model not only advances our understanding of human immune responses but also has significant implications for the development of novel immunotherapies and the preclinical evaluation of HSC transplantation strategies. This novel murine model offers a more accurate representation of the human immune system, with the potential to expedite the development of innovative therapeutic interventions. We present this model as a valuable addition to the preclinical research toolkit, promising a deeper understanding of immunology and a brighter future for clinical applications. Citation Format: Huiyi Wang, Jun Xing, Jialu Fan, Yuan Fan, Jing Zhao, Xiang Gao, Cunxiang Ju. Development of a novel murine model, NCG-M/hIL15, for enhanced post-HSC transplantation immunoreconstruction [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 118.

Biopterin metabolism and nitric oxide recoupling in cancer.

Tetrahydrobiopterin is a cofactor necessary for the activity of several enzymes, the most studied of which is nitric oxide synthase. The role of this cofactor-enzyme relationship in vascular biology is well established. Recently, tetrahydrobiopterin metabolism has received increasing attention in the field of cancer immunology and immunotherapy due to its involvement in the cytotoxic T cell response. Past research has demonstrated that when the availability of BH4 is low, as it is in chronic inflammatory conditions and tumors, electron transfer in the active site of nitric oxide synthase becomes uncoupled from the oxidation of arginine. This results in the production of radical species that are capable of a direct attack on tetrahydrobiopterin, further depleting its local availability. This feedforward loop may act like a molecular switch, reinforcing low tetrahydrobiopterin levels leading to altered NO signaling, restrained immune effector activity, and perpetual vascular inflammation within the tumor microenvironment. In this review, we discuss the evidence for this underappreciated mechanism in different aspects of tumor progression and therapeutic responses. Furthermore, we discuss the preclinical evidence supporting a clinical role for tetrahydrobiopterin supplementation to enhance immunotherapy and radiotherapy for solid tumors and the potential safety concerns.

Improved characterization and translation of NK cells for canine immunotherapy.

The field of cancer immunology has seen a meteoric rise in interest and application due to the discovery of immunotherapies that target immune cells, often leading to dramatic anti-tumor effects. However, successful cellular immunotherapy for solid tumors remains a challenge, and the application of immunotherapy to dogs with naturally occurring cancers has emerged as a high yield large animal model to bridge the bench-to-bedside challenges of immunotherapies, including those based on natural killer (NK) cells. Here, we review recent developments in the characterization and understanding of canine NK cells, a critical springboard for future translational NK immunotherapy research. The characterization of canine NK cells is exceptionally pertinent given the ongoing challenges in defining them and contextualizing their similarities and differences compared to human and murine NK cells compounded by the limited availability of validated canine specific reagents. Additionally, we summarize the current landscape of the clinical and translational literature employing strategies to capitalize on endogenous and exogenous NK cell immunotherapy in canine cancer patients. The insights regarding efficacy and immune correlates from these trials provide a solid foundation to design and test novel combinational therapies to enhance NK cell activity with the added benefit of motivating comparative work to translate these findings to human cancers with extensive similarities to their canine counterparts. The compilation of knowledge from basic canine NK phenotype and function to applications in first-in-dog clinical trials will support the canine cancer model and enhance translational work to improve cancer outcomes for both dogs and humans.

Vaccine Immunogenicity versus Gastrointestinal Microbiome Status: Implications for Poultry Production

At the turn of the 21st century, the importance of maintaining a balanced microbiome was brought to the forefront of the microbiology, immunology, and physiology research fields. Exploring the complex interactions between vaccine administration, mucosal microbiome, oral tolerance, and enteric inflammation in health and disease is challenging since environmental factors (such as diet and sanitation) have major influences on gut microbiota composition. High enteric pathogen load has been shown to contribute to dampened cell-mediated and humoral immune responses to vaccines in human case studies, either through elevated enteric inflammation or increased tolerance to environmental microbes. Although antibiotic and probiotic interventions have been evaluated in human health as well as research animal models, effective measures to mediate vaccine hyporesponsiveness are still ill-defined. Research in this field is becoming increasingly important for managing flock health in commercial poultry production, especially as antibiotic-free production is more prevalent and vaccination programs remain extensive during the first weeks of a bird’s life. By understanding the cellular interactions between commensal microbiota, vaccine antigens, and the host immune system (particularly in avian models), advancements in bacterial and viral vaccine immunogenicity could lead to improved flock health in meat-type and egg-type chickens in the future.

Complements from the Male Reproductive Tract: A Scoping Review

The organs of the male reproductive tract, including the testis, epididymis, prostate, seminal vesicles, and semen, must provide an immunoregulatory environment conducive to germ cell viability and successful fertilization. Many immune components, such as immune cells, have been investigated regarding reproductive immunology and function; however, the investigation of the role of complement in this system has only more recently been gaining traction in research. This review focuses on complement in the male reproductive tract, with the goal of compiling information currently known about complement components detected in male reproductive organs and identifying areas in need of further research. Considering the recent and upcoming research about the noncanonical functions of complement, this information is relevant and applicable in the fields of reproductive immunology, fertility, and immune regulation.

HLA class I peptide polymorphisms contribute to class II DQβ0603:DQα0103 antibody specificity

Antibodies reactive to human leukocyte antigens (HLA) represent a barrier for patients awaiting transplantation. Based on reactivity patterns in single-antigen bead (SAB) assays, various epitope matching algorithms have been proposed to improve transplant outcomes. However, some antibody reactivities cannot be explained by amino acid motifs, leading to uncertainty about their clinical relevance. Antibodies against the HLA class II molecule, DQβ0603:DQα0103, present in some candidates, represent one such example. Here, we show that peptides derived from amino acids 119-148 of the HLA class I heavy chain are bound to DQβ0603:DQα0103 proteins and contribute to antibody reactivity through an HLA-DM-dependent process. Moreover, antibody reactivity is impacted by the specific amino acid sequence presented. In summary, we demonstrate that polymorphic HLA class I peptides, bound to HLA class II proteins, can directly or indirectly be part of the antibody binding epitope. Our findings have potential important implications for the field of transplant immunology and for our understanding of adaptive immunity.

Life-threatening infections in human newborns: Reconciling age-specific vulnerability and interindividual variability

In humans, the interindividual variability of clinical outcome following exposure to a microorganism is immense, ranging from silent infection to life-threatening disease. Age-specific immune responses partially account for the high incidence of infection during the first 28 days of life and the related high mortality at population level. However, the occurrence of life-threatening disease in individual newborns remains unexplained. By contrast, inborn errors of immunity and their immune phenocopies are increasingly being discovered in children and adults with life-threatening viral, bacterial, mycobacterial and fungal infections. There is a need for convergence between the fields of neonatal immunology, with its in-depth population-wide characterization of newborn-specific immune responses, and clinical immunology, with its investigations of infections in patients at the cellular and molecular levels, to facilitate identification of the mechanisms of susceptibility to infection in individual newborns and the design of novel preventive and therapeutic strategies.

Abstract PR005: Identification of DNA Damage Repair genes controlling the immune landscape of breast tumors via spatial functional genomics

Abstract Immune checkpoint blockade (ICB) has revolutionized the field of cancer immunology, yet only 17-20% of patients who receive this treatment derive any clinical benefit. There is an unmet need to either identify the subset of patients who would respond to ICB or identify targets for combination therapies. By assessing frequently mutated pathways in these patients, we can identify potential biomarkers and candidates for combinatorial therapy. One such pathway is the DNA Damage Response (DDR) pathway. Faults in DDR pathways can lead to genome instability, higher tumor mutational burden (TMB), and accumulation of neoantigens in tumors. Consequently, tumors with one or more deficiencies in DDR (DDRd) have been shown to respond better to therapy and are associated with increased immune cell infiltration. However, previous work from our lab has shown that different gene alterations, even in the same DDR pathway, can polarize immune outcomes differently. To address this issue, we employed Perturb-map, a first-of-its-kind functional genomics approach, which utilizes protein barcodes (pro-Codes) to enable spatial resolution of CRISPR screens. We generated a Pro-Code/CRISPR library targeting frequently mutated DDR genes and introduced the library into 4T1 breast cancer cells, a syngeneic model of triple negative breast cancer. We introduced the cells into immunocompetent mice and performed multiplex imaging on metastatic lesions in the lung to spatially resolve each gene knockout tumor. We observed that loss of Rbbp8 and Polq led to larger tumors, whereas loss of Pms2 led to smaller tumors. Strikingly, Rbbp8 knockout tumors were also less infiltrated by CD4 T cells compared to control tumors. This system was also used to find therapeutic targets by performing synthetic lethality screens using mass cytometry. We transduced the library in isogenic Brca2 deficient 4T1 cells along with their wildtype (WT) counterparts and looked for enrichment or loss of pro-Codes across the different cell lines. These screens consistently identified known synthetic lethality partners like Parp1 in the Brca2 deficient cell lines and identified additional targets like Mre-11a, Bard1 and Rbbp8, which are being profiled further. Taken together, we have developed a scalable system to study the effect of DDR mutations on tumor cell intrinsic and extrinsic properties, which can not only help characterize immune outcomes but also identify potential therapeutic partners. This system proves to be a useful tool that can be leveraged to profile a number of different genes across multiple cancer types. Citation Format: Prerna Suri, Juhana Habib, Gurkan Mollaoglu, Luisanna Pia, Alessia Baccarini, Brian Brown, Robert Samstein. Identification of DNA Damage Repair genes controlling the immune landscape of breast tumors via spatial functional genomics [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: DNA Damage Repair: From Basic Science to Future Clinical Application; 2024 Jan 9-11; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2024;84(1 Suppl):Abstract nr PR005.

Necroptosis in Pneumonia: Therapeutic Strategies and Future Perspectives.

Pneumonia remains a major global health challenge, necessitating the development of effective therapeutic approaches. Recently, necroptosis, a regulated form of cell death, has garnered attention in the fields of pharmacology and immunology for its role in the pathogenesis of pneumonia. Characterized by cell death and inflammatory responses, necroptosis is a key mechanism contributing to tissue damage and immune dysregulation in various diseases, including pneumonia. This review comprehensively analyzes the role of necroptosis in pneumonia and explores potential pharmacological interventions targeting this cell death pathway. Moreover, we highlight the intricate interplay between necroptosis and immune responses in pneumonia, revealing a bidirectional relationship between necrotic cell death and inflammatory signaling. Importantly, we assess current therapeutic strategies modulating necroptosis, encompassing synthetic inhibitors, natural products, and other drugs targeting key components of the programmed necrosis pathway. The article also discusses challenges and future directions in targeting programmed necrosis for pneumonia treatment, proposing novel therapeutic strategies that combine antibiotics with necroptosis inhibitors. This review underscores the importance of understanding necroptosis in pneumonia and highlights the potential of pharmacological interventions to mitigate tissue damage and restore immune homeostasis in this devastating respiratory infection.

Nanoparticle technology for mRNA: Delivery strategy, clinical application and developmental landscape.

The mRNA vaccine, a groundbreaking advancement in the field of immunology, has garnered international recognition by being awarded the prestigious Nobel Prize, which has emerged as a promising prophylactic and therapeutic modality for various diseases, especially in cancer, rare disease, and infectious disease such as COVID-19, wherein successful mRNA treatment can be achieved by improving the stability of mRNA and introducing a safe and effective delivery system. Nanotechnology-based delivery systems, such as lipid nanoparticles, lipoplexes, polyplexes, lipid-polymer hybrid nanoparticles and others, have attracted great interest and have been explored for mRNA delivery. Nanoscale platforms can protect mRNA from extracellular degradation while promoting endosome escape after endocytosis, hence improving the efficacy. This review provides an overview of diverse nanoplatforms utilized for mRNA delivery in preclinical and clinical stages, including formulation, preparation process, transfection efficiency, and administration route. Furthermore, the market situation and prospects of mRNA vaccines are discussed here.

Dissecting the Immune SystemthroughGene Regulation.

The immune system plays a dual role in human health, functioningboth as a protector against pathogensand, at times, as a contributor to disease. This featureemphasizes the importanceto uncover the underlying causes of its malfunctions,necessitatingan in-depth analysisin both pathological and physiological conditionsto betterunderstand the immune system and immune disorders. Recent advancesin scientifictechnology have enabledextensive investigationsinto gene regulation, a crucialmechanismgoverningcellular functionality. Studying gene regulatory mechanisms within the immune system is a promising avenue for enhancingour understanding of immune cells and the immune systemas a whole. The gene regulatory mechanisms, revealed through various methodologies, and their implications in the field of immunology are discussed in this chapter.

Estimating nitric oxide (NO) from MDSCs by Griess method.

The functional importance of nitric oxide (NO) in the fields of immunology concerning its antimicrobial, anti-tumoral, anti-inflammatory, and immunosuppressive effects have made it inevitable to study its secretion from various cells. Nitrogen oxide synthase (NOS) is the enzyme responsible for synthesizing NO and its three isoforms function in a cell-dependent manner. NO is oxidized rapidly to Reactive nitrogen oxide species (RNOS) through which the roles of NO are being carried out. One of the major immune cells secreting NO is myeloid-derived suppressor cells (MDSCs). The function of these MDSCs in the suppression of T-cell proliferation as well as T-cell differentiation is found to be dependent on NO secretion. Apart from T-cell suppressive activity, NO is also known to interfere with natural killer (NK) cell functions. A convenient method to estimate NO secretion is by using Griess reagent named after Johann Peter Griess. In this method, NO reacts with the reagents to form a colored azo dye detectable using a microplate reader at a wavelength of 548nm. In this chapter, we summarized the detailed method of estimating NO from MDSCs by the Griess method.

Immunology, toxicology, and immunotoxicology: An overview

Immunology, toxicology, and immunotoxicology are three different fields of science. Immunology is the branch of science that deals with the studies of the immune system. On the other hand, toxicology is the branch of science which deals with chemicals, toxic substances, or polluted environments and their ill effect on living creatures as well as the environment. Immunotoxicology deals with the both fields of immunology and toxicology. Immunotoxicology is an active area of toxicology, but this is still a relatively small area. Over the past 30 years, the main focus of immunotoxicology has been the aspects of immunotoxicity, from a mechanistic or regulatory process.