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We present a method for conjugating antigens to gold nanoparticles (GNPs) during their synthesis via gas plasma, eliminating the need for chemical linkers and significantly speeding up the process (taking only 15 min). This fast, linker-free method produces biocompatible and stable GNPs, with potential for immunotherapy applications, such as antigen and antibody conjugation and drug delivery. We demonstrate the conjugation of the antigen Nestin (NES), a tumor marker, to GNPs using two approaches. The first method involves synthesizing GNPs with citrate, followed by NES conjugation via plasma. The second method synthesizes and conjugates GNPs to NES simultaneously using plasma treatment. Conjugation was confirmed by Enzyme-Linked Immunosorbent Assay , Zeta-sizer , UV-vis spectroscopy , and Transmission Electron Microscopy . In addition, the toxicity of the prepared samples was investigated in vitro using peripheral blood mononuclear cells (PBMCs) and flow cytometry , which proved the nontoxicity of the samples.

Despite great advancements in the discovery of genetic variants underlying inborn errors of immunity, functional assessment of the immunological profile of patients in routine clinical practice remains challenging. The lymphocyte proliferation assay using 3 H-thymidine incorporation has been the gold standard for decades for functional evaluation of T cells in the clinical laboratory, however, recently developed flow cytometry methods allowing for single cell analysis provide non-radioactive alternatives. Understanding the technical and analytical challenges of test development, validation and maintenance is essential for correct interpretation and test utilization, to assure appropriate and timely patient care. This review will discuss the technological aspects, validation and clinical utilization of in vitro lymphocyte proliferation assays performed in the clinical immunology laboratory, providing a diagnostic readout for T lymphocyte function, an essential hallmark of a functional cellular immune response, and allowing for the detection of impaired responses such as in patients with functional T cell defects., (Copyright © 2025. Published by Elsevier B.V.)

Since their inception, therapeutic or prophylactic vaccines have emerged as promising candidates for the prevention or treatment of infections and various diseases, including cancer and autoimmune disorders. In recent times, gold nanoparticles (GNPs) have acquired active roles in the field of vaccine development due to their intrinsic capacity to adjust and enhance the immune response. Due to their characteristics, GNPs can exert optimal effects as both delivery vehicles and adjuvants. Despite their significant importance in vaccinology, numerous obstacles need to be overcome before GNPs can be used in the formulations of vaccines in clinical settings. The current review summarizes the latest and successful use of gold nanoparticles as a viable method for developing a new generation of vaccines., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Bovine adenoviruses (BAdVs) are major contributors to the bovine respiratory disease complex (BRDC). A systematic review and meta-analysis were carried out to explore the epidemiology of BAdV across diverse cattle populations using different detection methods. The study showed a higher BAdV prevalence of 0.66 in general cattle populations using antibody detection, compared with 0.28 in cattle showing clinical signs. The study identified significant prevalence differences between BAdV-3 (0.87) and BAdV-7 (0.21) in general cattle populations. However, in clinical cattle, BAdV-3 and BAdV-7 showed similar prevalence at 0.27 and 0.32, respectively. Moreover, a high herd-based BAdV seroprevalence of 0.82 was observed. When nucleic acid detection methods were used in general cattle populations, a lower BAdV (0.05) prevalence was observed, in contrast to the higher prevalence (0.32) in cattle exhibiting clinical signs. In contrast, using antigen detection in cattle with clinical signs of disease showed a prevalence of 0.06, compared to 0.32 with nucleic acid methods, indicating detection method-specific sensitivity and specificity. The study also highlighted the role of BAdV in BRDC, particularly BAdV-3 and BAdV-7. Existing empirical evidence on BAdV epidemiology and pathobiology is scarce and requires further investigation; however, the current findings offer insights into the epidemiology of BAdV and its role in the BRDC, which could potentially inform and enhance disease control strategies., Competing Interests: Conflict of Interest The authors declare no conflict of interest., (Copyright © 2025 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Aims: The screening and diagnosis of dengue virus infection play a crucial role in controlling the epidemic of dengue fever, highlighting the urgent need for a highly sensitive, simple, and rapid laboratory testing method. This study aims to assess the clinical performance of MAGLUMI Denv NS1 in detecting dengue virus NS1 antigen., Methods: A retrospective study was conducted to assess the sensitivity and specificity of MAGLUMI Denv NS1 using residual samples. Dengue-confirmed and excluded samples, validated by qPCR, were subjected to testing with MAGLUMI Denv NS1 in accordance with the manufacturer's instructions. The linear range, endogenous interference, and cross-reactivity of MAGLUMI Denv NS1 were verified, and a consistency analysis with commercial comparator products was carried out., Results: The diagnostic specificity of MAGLUMI Denv NS1 is 98.41% (62/63), and the sensitivity is 98.32% (117/119). It effectively detects various serotypes of dengue virus, with no observed endogenous interference or cross-reactivity. Additionally, the consistency of NS1, IgM, and IgG tests on the MAGLUMI platform compared to commercial comparator reagents reaches 85.71%, 99.25%, and 98.97%, respectively., Conclusions: The MAGLUMI Denv NS1 represents a highly sensitive laboratory testing method capable of enhancing the diagnostic accuracy and efficiency of dengue virus infection detection.

Accurate rapid diagnostic tests (RDTs) are needed to diagnose lymphatic filariasis (LF) in global elimination programmes. We evaluated the performance of the new STANDARD Q Filariasis Antigen Test (QFAT) against the Bioline Filariasis Test Strip (FTS) for detecting W. bancrofti antigen (Ag) in laboratory conditions, using serum (n = 195) and plasma (n = 189) from LF-endemic areas (Samoa, American Samoa and Myanmar) and Australian negative controls (n = 46). The prior Ag status of endemic samples (54.9% Ag-positive) was determined by rapid test (ICT or FTS) or Og4C3 ELISA. The proportion of samples testing positive at 10 min was similar for QFAT (44.8%) and FTS (41.3%). Concordance between tests was 93.5% (kappa 0.87, n = 417) at 10 min, and it increased to 98.8% (kappa 0.98) at 24 h. The sensitivities of QFAT and FTS at 10 min compared to the prior results were 92% (95% CI 88.0-96.0) and 86% (95% CI 80.0-90.0), respectively, and they increased to 97% and 99% at 24 h. Specificity was 98% for QFAT and 99% for FTS at 10 min. Both tests showed evidence of cross-reaction with Dirofilaria repens and Onchocerca lupi but not with Acanthochilonema reconditum or Cercopithifilaria bainae. Under laboratory conditions, QFAT is a suitable alternative RDT to FTS.

Artificially designed, functional nanostructured surfaces, called metasurfaces, are an emerging platform for biosensing. Two major types of metasurface biosensors have been reported: one is based on resonant-wavelength shift and the other is specialized for fluorescence (FL) detection. The all-dielectric metasurfaces that composed of periodic arrays of silicon nanocolumns have a series of optical magnetic-mode resonances, some of which were found to significantly enhance capability for FL detection of diverse target biomolecules, ranging from nucleic acid to antigens and antibodies. Here, we mainly address the recent advances in productive metasurface FL biosensors, provide an overview of the pivotal results, and discuss the future prospects, including artificial-intelligence-driven big data analysis for the next-generation healthcare services., Competing Interests: The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2025 Iwanaga.)

To battle seasonal outbreaks of influenza B virus infection, which continue to pose a major threat to world health, new and improved vaccines are urgently needed. In this article, we discuss the current state of next-generation influenza B vaccine development, including both advancements and challenges. This review covers the shortcomings of existing influenza vaccines and stresses the need for more-effective and broadly protective vaccines and more-easily scalable manufacturing processes. New possibilities for vaccine development have emerged due to recent technical developments such as virus-like particle (VLP) platforms, recombinant DNA technologies, and reverse genetics. By using these methods, vaccines can be developed that elicit stronger and longer-lasting immune responses against various strains of influenza B virus. Vaccines may be more effective and immunogenic when adjuvants and new delivery mechanisms are used. Progress has been made in the development of influenza B vaccine mRNA vaccines, nanoparticle-based vaccines, and vector-based vaccines. However, there are still several obstacles to overcome before next-generation influenza B vaccines can be widely used, including the challenge of antigenic drift, the extinction of the B/Yamagata lineage, and difficulties in strain selection. There are also other challenges related to public acceptance, vaccine distribution, manufacturing complexity, and regulations. To overcome these challenges, scientists, politicians, and pharmaceutical firms must work together to expedite the development and licensing of vaccines and the establishment of immunization programs. The need for constant monitoring and quick adaptation of vaccines to match the currently circulating strains is further highlighted by the appearance of novel influenza B virus variants. To be ready for future pandemics and influenza B outbreaks, we need better vaccines and better monitoring systems., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Background: Ringworm is highly prevalent in our setting and is frequently observed in our routine clinical practice. Diagnostic confirmation depends on techniques that are not always accessible (PCR), with highly variable sensitivity depending on the observer (direct microscopy) or delayed results (culture, histopathology). Recently, an immunochromatography-based rapid test (Diafactory®) for the antigenic detection of dermatophytes has been developed. This diagnostic tool can help diagnose ringworm, allowing early initiation of treatment and fewer consultation visits., Objective: To determine the sensitivity and specificity of the rapid antigen detection test compared to conventional culture., Material and Methods: For a full year, 333 nail samples were collected from patients with suspected onychomycosis. The rapid test and the conventional culture were simultaneously performed on each sample. Those with a positive antigenic test result began treatment early. The remaining patients had appointments for serial cultures and subsequent medical consultation to evaluate the results., Results: Compared to conventional culture, the sensitivity and specificity rates of the rapid antigen detection test are 97.2% and 80.7%, respectively., Conclusion: The effectiveness of the rapid antigen detection test is similar to that of conventional culture for the detection of dermatophytes in nail samples. It is a quick and simple diagnostic technique that reduces the number of patient visits to the hospital, and allows early treatment start., (Copyright © 2024 AEDV. Publicado por Elsevier España, S.L.U. All rights reserved.)

We conducted a systematic review and meta-analysis of studies and reports comparing the performance of antigen rapid diagnostic tests (Ag RDT) for diagnosing Ebola disease (EVD). We searched PubMed, EMBASE, and Web of Science for diagnostic studies published between 1976 and 2023, evaluating them with QUADAS-2. Using a bivariate random-effects model, we estimated the pooled sensitivity and specificity of Ag RDTs. Of 64 eligible full studies and reports, 16 met the inclusion criteria. Pooled sensitivity and specificity were 82.1% (95%CI: 75.2 - 88.0) and 97.0% (95%CI: 95.1-98.2), respectively. We conducted subgroup analysis on 4 Ag RDTs, 3 RT-PCR tests, and 4 sample types, showing varied performance. The high specificity and positive predictive value of Ag RDTs support their use to "rule-in" patients with EVD. However, high-sensitivity RDTs suitable for field settings and capable of detecting multiple ebolavirus species are needed., Competing Interests: Declaration of competing interest That there is no conflict of interest for this paper, (Copyright © 2024. Published by Elsevier Inc.)

The culture-attenuated alcelaphine herpesvirus 1 (AlHV-1) C500 strain can be grown to high titre and has been used successfully as a candidate vaccine for wildebeest-associated malignant catarrhal fever (MCF). This vaccine virus was also used to develop an indirect ELISA to allow monitoring of virus-specific antibodies in vaccinated cattle. However the extraction method was expensive and time-consuming, and the resulting test was not suitable for use in sheep. Here we describe an improved antigen extraction method that also broadens the application of the assay, allowing its application to sheep samples. The updated assay was tested using control samples from cattle and sheep, and showed a high level of accuracy in both species. This novel assay should prove to be a useful tool in MCF diagnosis and in evaluation of MCF vaccine responses., Competing Interests: Declaration of Competing Interest I have nothing to declare, (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Sepsis is a life-threatening immune response to infection in the body, eventually resulting in fatal organ failure. Current methods utilize blood cultures and quick-Sequential-Organ-Failure-Assessment (qSOFA), but there is a need for more accurate and time-sensitive diagnostic methods to improve survival rates. We present a 3D-printed microfluidic chip that bioconjugates antibodies CD69, CD64, and CD25 to channel surfaces to capture sepsis cells in blood samples and validate it with clinical samples (n = 125 septic, n = 10 healthy). Other variables were taken such as healthy volunteer blood samples and patient demographics to validate and confirm our device's diagnostic ability. Statistical differences were found between healthy volunteer and sepsis patient antigen cell counts (CD69 p-value < 0.001, CD64 p-value < 0.004, CD25 p-value < 0.0009), and were confirmed using principal component analysis. Demographics such as length of stay, age, culture results, and need for surgery also factored into sepsis detection on a smaller scale than the antigen cell counts. The receiver operating characteristic (ROC) analysis showed an area under the curve (AUC) of 0.989, 0.988, and 0.992 for CD69, CD64, and CD25, respectively, and a combined biomarker panel of 0.997. Overall, the device performed within a shorter time frame of 4 h compared to standard blood culture tests and was validated for use in detecting sepsis in patients., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: D Pappas reports financial support was provided by National Institutes of Health. D Pappas reports a relationship with NaMi Diagnostics that includes: non-financial support. D Pappas has patent #10,761,093 licensed to NaMi Diagnostics. Dr. Pappas’s patent on this technology is licensed through NaMi Diagnostics, who have no competing interest in this work. Dr. Pappas has no financial position in NaMi Diagnostics. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024 Elsevier B.V. All rights reserved.)

A growing body of research indicates the promising potential of vaccines in both preventing and treating atherosclerosis (AS). To gain a comprehensive understanding of the current research landscape and emerging trends in this field, this study conducted a bibliometric analysis of publications on AS vaccines using the Web of Science Core Collection (WoSCC) database, based on the "bibliometric" R package and VOSviewer software. From 1991 to 2024, a total of 462 publications were identified in the WoSCC. The United States appeared as the leading contributor in terms of both total publications and citations. The Vaccine journal exhibited the highest publications output. Nilsson J from the Lund University in Sweden was the author with the most published articles, total citations and Hirsch index (H-index). Keywords analysis and thematic maps analysis revealed the passive immunotherapy (AS protective antibodies vaccines) was a hot mature theme, the active immunotherapy (AS antigens vaccines) was an emerging and booming theme, while the efficacy and safety of AS vaccines was a niche and well-developed theme. These findings offered valuable insights into the AS vaccination and provided guidance for future research in this domain.

Histoplasmosis poses a significant risk to HIV patients, particularly in regions with limited access to antiretroviral therapy. Antigen detection assays are crucial in these settings for timely diagnosis and treatment, which can reduce mortality. While commercial antigen detection kits have performed well in diagnosing histoplasmosis, their effectiveness in monitoring treatment remains unclear. This study aimed to evaluate the correlation between urine antigen levels and clinical response using the clarus Histoplasma Galactomannan (GM) enzyme immunoassays (EIA) kit. The study followed 27 HIV patients diagnosed with histoplasmosis over 24 weeks, measuring urinary Histoplasma antigen (Ag) levels and clinical outcomes. Patients received amphotericin B as induction therapy, followed by maintenance with itraconazole. Results showed a significant decrease in Ag levels over time, with clinical scores improving in correlation with the decline in Ag levels. Four patients exhibited atypical Ag patterns due to immune reconstitution inflammatory syndrome or issues with itraconazole bioavailability. Despite these challenges, all patients showed improvement by week 24. The findings suggest that the clarus Histoplasma GM EIA kit could be a valuable tool for monitoring and evaluating the response to antifungal therapy in histoplasmosis patients., (© The Author(s) 2024. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.)

Bovine tuberculosis (bTB), a global zoonotic disease, causes negative effects on human and animal health. PhoP protein is a key regulator of pathogenic phenotypes in members of the Mycobacterium tuberculosis complex, which includes the causative agent of bTB. Despite extensive research on this protein focused in deciphering its regulatory role, little was explored about it as a diagnostic antigen. In humans, a novel role of anti-PhoP antibodies as a possible marker for the diagnosis of TB was demonstrated. However, this issue was not addressed in bovines. In this study, antigenic properties of the PhoP protein were evaluated in naturally Mycobacterium bovis (M. bovis) infected bovines. A high homology of PhoP (≥ 75 %) was observed in environmental mycobacterial species and other genera such as Salmonella and Pasteurella. Using the IFN-gamma release assay (IGRA), we detected cell-mediated immune response against PhoP in cattle from infected herds (25 %; IC 95 % 3.2-65.1), although it was significantly lower than that evoked by the reference antigens, ESAT-6/CFP-10/Rv3615c (75 %; IC95 % 34.9-96.8), and the purified protein derivative (87.5 %; IC 95 % 47.4-99.7) (p < 0.05)). Animals from a bTB free area showed no response against PhoP when analyzed by IGRA. Although, the humoral response detected 62.5 % (CI95% 24.5-91.5) of naturally infected animals, there was 100 % cross-reactivity among TB-free cattle. These results suggest that the PhoP protein is not a promising candidate for bTB diagnosis, due to it had relatively low levels of test sensitivity in the IGRA test, and very low specificity in a humoral antibody western blot assay., Competing Interests: Declaration of competing interest The authors have declared no conflict of interest., (Copyright © 2024. Published by Elsevier Ltd.)

Introduction: Cryptococcal meningitis is a major cause of death in HIV/AIDS patients due to the existence of Cryptococcus neoformans in the central nervous system. Our objective was to evaluate the prevalence of Cryptococcus antigenuria in a population of HIV-infected patients in Libreville, Gabon. Patients and Methods : This study was conducted from April to October 2021 at the Infectious Diseases ward of the Centre Hospitalier Universitaire de Libreville. Hospitalized patients with HIV were included. The detection of cryptococcal antigen (CrAg) in urine was performed using the Pastorex Crypto Plus Kit. Results : Out of the 255 PLHIV, 142 benefited from the CrAg detection. The prevalence of urine CrAg was 24.6% (n = 35). The majority of CrAg+ patients (82.8%; n = 29) were under 55 years old. Almost three-quarters of them (n = 25; 71.4%) had CD4 counts < 200, and 80.0% (n = 28) were at WHO clinical stages III and IV. All patients with neck stiffness at admission had a CrAg positive test. Conclusion : This study showed a non-negligible prevalence of Cryptococcal urinary antigen in HIV-infected patients with neurological symptoms. These data underline the importance of CrAg screening in routine care for better management of PLHIV.

Background: The development of effective and safe vaccines and their timely delivery to the public play a crucial role in preventing and managing infectious diseases. Many vaccines have been produced and distributed globally to prevent COVID-19 infection. However, establishing effective vaccine development platforms and evaluating their safety and immunogenicity remains critical to increasing health security, especially in developing countries. Objectives: Therefore, we developed a local subunit vaccine candidate, RENOVAC, and reported its toxicity and immunogenicity profile in animal models. Methods: First, the synthetic gene-coding tandem RBD linked with the GS linker was cloned into the expression vector and expressed in CHO cells. The protein was then purified and filter sterilized, and 10 µg/dose and 25 µg/dose formulations were finally examined for the 14-day repeated dose toxicity followed by the immunogenic profile in preclinical studies. Results: When administered to Sprague Dawley rats by intramuscular route, the vaccine was well tolerated up to and including the dose of 25 µg/animal, and no toxicologically adverse changes were noted. The observed change in weight of the thymus and spleen might be related to the immunological response to the vaccine. The dimer RBD vaccine demonstrated the ability to generate high levels of specific immunoglobulins (IGs) and neutralization antibodies (NAbs). Finally, changes in the amounts of specific T cells and cytokines after vaccination suggested that the vaccine mainly triggers an immune response by activating CD4+ Th2-cells, which then activate B-cells to provide humoral immunity. Conclusions: The study suggests that, based on its reliable immunogenicity and acceptable safety, the vaccine can be further directed for clinical trials.

Background: Colorectal cancer (CRC) is the third most prevalent cancer worldwide, with numerous risk factors contributing to its development. Recent research has illuminated the significant role of the gut microbiota in CRC pathogenesis, identifying various microbial antigens as potential targets for vaccine development., Aim: This review aimed at exploring the potential sources of microbial antigens that could be harnessed to create effective CRC vaccines and understand the role of microbiome-CRC interactions in carcinogenesis., Methods: A comprehensive search of original research and review articles on the pathological links between key microbial candidates, particularly those more prevalent in CRC tissues, was conducted. This involved extensive use of the PubMed and Medline databases, as well as the Google Scholar search engine, utilizing pertinent keywords. A total of one hundred and forty-three relevant articles in English, mostly published between 2018 and 2024, were selected., Results: Numerous microbes, particularly bacteria and viruses, are significantly overrepresented in CRC tissues and have been shown to promote tumorigenesis by inducing inflammation and modulating the immune system. This makes them promising candidates for antigens in the development of CRC vaccines., Conclusion: The selection of microbial antigens focuses on their capacity to trigger a strong immune response and their link to tumor presence and progression. Identifying and validating these antigens through preclinical testing is essential in developing a CRC vaccine., Competing Interests: The authors declare that there are no financial or non-financial competing interests., (© 2024 Ibeanu et al.)

Background/objectives: Since 2022, outbreaks of monkeypox have raised widespread concern and have been declared a public health emergency of international concern by the World Health Organization. There is an urgent need to develop a safe and effective vaccine against the monkeypox virus (MPXV). Recombinant protein vaccines play a significant role in the prevention of infectious diseases due to their high safety and efficacy., Methods: We used the A29, E8, M1, A35, and B6 proteins of MPXV as candidate antigens to generate a panel of multi-component MPXV vaccine candidates, which were administered subcutaneously to immunize mice., Results: The results showed that the vaccine candidates Mix-AEM, Mix-AEMA, Mix-AEMB, and Mix-AEMAB effectively elicited strong neutralizing antibody responses and demonstrated significant protection against vaccinia virus (VACV) infection in a murine model. The vaccine candidate Mix-AEM induced significantly higher levels of neutralizing antibodies, cellular immunity capacity, and virus clearance compared to the vaccine candidate Mix-AE (lacking M1). Single-component immunization showed that M1 induced higher levels of neutralizing antibodies than A29 and E8. These results indicated that M1 is a critical and essential antigen in the MPXV vaccine. The number of cells secreting IFN-γ was significantly increased in the Mix-AEMA and Mix-AEMAB groups compared to the A35-deficient vaccine candidates, demonstrating the important role of A35 in inducing IFN-γ secreting. In addition, the neutralizing antibodies induced by these multi-component vaccine candidates were maintained at high levels six months after the third immunization., Conclusions: In summary, this study lays the groundwork for combining antigens to develop multi-component subunit vaccines.

Invasive aspergillosis (IA) is a life-threatening infection. Early and specific diagnosis is pivotal to ensure adequate therapy. Antigen testing from blood is a widespread and convenient diagnostic approach. Various tests for the detection of Aspergillus antigen as well as for the panfungal antigen β-1,3-D-glucan (BDG) are available, for which comprehensive comparisons are still lacking. Blood samples of 82 proven/probable (11/71) IA patients and 52 controls were tested using two enzyme-linked immunosorbent assays (ELISAs) (Bio-Rad and Euroimmun), one chemiluminescent immunoassay (CLIA) (Vircell), one BDG assay (Fujifilm Wako), and two point of care (PoC) assays (Immy sōna and OLM). PoC assays were evaluated visually and used automated read out systems. Of the 82 IA patients, 37 had received solid organ transplantation (SOT) and 25 hematopoietic stem cell transplant (HSCT). Sensitivities and specificities for the eight test systems ranged from 27% to 71% and from 64% to 100%. Estimating a 10% prevalence of IA, test performance would have resulted in positive and negative predictive values of 14%-100% and 91%-95%. Areas under the curve (AUCs) for all tests except GM were below 0.7. When the cut-off values for quantitative tests were normalized to a specificity close to 95%, sensitivities ranged from 14% to 40%. The use of automated read out systems for the PoC assays had a significant impact. Combining different tests did not result in better test strategies. Sensitivity of Aspergillus antigen testing from single serum samples is low. Due to specificity issues, the majority of tests is not suited for screening purposes. The different assays can meet different needs in different diagnostic settings., Competing Interests: J.W. received financial support (research grant) from Pfizer outside of this study, technical and financial support by Fujifilm Wako Chemicals Europe and Euroimmun Medizinische Labordiagnostika for past projects outside this study, and speaker fees from Pfizer, Wako, und Gilead. J.P. serves as President of the Austrian Society of Medical Mycology, received speaker fees from Gilead, Pfizer, Swedish Orphan BioVitrum, and Associates of Cape Cod, and holds stock in AbbVie Inc. and Novo Nordisk, all unrelated to the submitted work. M.H. received research funding from Gilead, Astellas, MSD, IMMY, Mundipharma, Scynexis, F2G, and Pfizer, all outside of the submitted work. J.H. received speaker fees from Pfizer, Gilead, Associates of Cape Cod, BD, and Biomerieux, research funding from Pfizer and Gilead, and technical support for projects outside this study from Associates of Cape Cod, Vircell, Virion\Serion, IMMY, and OLM. K.D. received technical and financial support by Fujifilm Wako Chemicals Europe and by Euroimmun Medizinische Labordiagnostika for past projects outside this study and technical support by Vircell, OLM, and IMMY for past projects outside this study. T.S., I.K., and S.S. have no conflicts of interest.

Background: Immune correlates of protection are ideal tools to predict treatment or vaccine efficacy. However, the accuracy of the immune correlate and the capability to robustly predict the outcome of a vaccine candidate are determined by the performance of the in vitro immunoassay used. Several Theileria parva sporozoite seroneutralization assays have previously been used to assess antibody functional activities; however, a common limitation has been the need for fresh material, target cells and sporozoites, and operator-to-operator bias. An improved assay represents a positive step toward overcoming challenges associated with variability and it might provide a more reliable means of establishing an immune correlate with protection after sub-unit vaccine administration., Methods: Herein, we describe key improvements, among them, (1) the use of frozen parasites and target cells to avoid batch-to-batch variations and (2) the development of a new assay read-out based on the detection of infected cells through flow cytometry, instead of the use of Giemsa staining and microscopic evaluation, in order to improve the reproducibility of the results., Results: The improved seroneutralization assay is not only able to detect the individual neutralizing capacity of antibodies; it also detects the additive effect of antibody combinations., Conclusions: This effect is described for the first time in Theileria parva and is of great interest for new antigen discovery and/or the epitope discovery of already known antigens like p67, opening a new avenue for the identification of ECF immune correlates of protection and the in vitro down-selection of new Theileria parva vaccine candidates, thereby contributing to reducing the use of animals in challenge experiments.

Cryptococcus neoformans is an opportunistic fungal pathogen that is a continuous global health concern, especially for immunocompromised populations. The World Health Organization recognized C. neoformans as one of four critical fungal pathogens, thus emphasizing the need for increased research efforts and clinical resource expansion. Currently, there are no fungal vaccines available for clinical use. Exciting new findings in cryptococcal vaccine development have identified whole cell-based and subunit-based vaccinations to help mitigate health risks and make commercialization attainable. Importantly, recent work has focused on how different cryptococcal cell-wall antigens modified in these vaccine candidates allow us to manipulate their immunogenicity to produce a desired long-term protective anti-fungal immune response. In this review, we discuss the different cryptococcal cell immunogens, namely the polysaccharide capsule, glucans, chitin/chitosan, mannoproteins, and extracellular vesicles, and their role in novel cryptococcal vaccination approaches. Additionally, we examine the immunological mechanisms responsible for protection in these vaccine candidates and the similar host response-stimulation pathways induced through different immunogen exposure.

Glycopeptides derived from the glycoprotein mucin-1 (MUC1) have shown potential as tumor-associated antigens for cancer vaccine development. However, their low immunogenicity and non-selective conjugation to carriers present significant challenges for the clinical efficacy of MUC1-based vaccines. Here, we introduce a novel vaccine candidate based on a structure-guided design of an artificial antigen derived from MUC1 glycopeptide. This engineered antigen contains two non-natural amino acids and has an α-S-glycosidic bond, where sulfur replaces the conventional oxygen atom linking the peptide backbone to the sugar N-acetylgalactosamine. The glycopeptide is then specifically conjugated to the immunogenic protein carrier CRM 197 (Cross-Reactive Material 197), a protein approved for human use. Conjugation involves selective reduction and re-bridging of a disulfide in CRM 197 , allowing the attachment of a single copy of MUC1. This strategy results in a chemically defined vaccine while maintaining both the structural integrity and immunogenicity of the protein carrier. The vaccine elicits a robust Th1-like immune response in mice and generates antibodies capable of recognizing human cancer cells expressing tumor-associated MUC1. When tested in mouse models of colon adenocarcinoma and pancreatic cancer, the vaccine is effective both as a prophylactic and therapeutic use, significantly delaying tumor growth. In therapeutic applications, improved outcomes were observed when the vaccine was combined with an anti-programmed cell death protein 1 (anti-PD-1) checkpoint inhibitor. Our strategy reduces batch-to-batch variability and enhances both immunogenicity and therapeutic potential. This site-specific approach disputes a prevailing dogma where glycoconjugate vaccines require multivalent display of antigens., (© 2024 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.)

Protein phosphorylation is a crucial process in various cellular functions, and its irregularities have been implicated in several diseases, including cancer. Antibodies are commonly employed to detect protein phosphorylation in research. However, unlike the extensive studies on recognition mechanisms of the phosphate group by proteins such as kinases and phosphatases, only a few studies have explored antibody mechanisms. In this study, we produced and characterized two rabbit monoclonal antibodies that recognize a monophosphorylated Akt peptide. Through crystallography, thermodynamic mutational analyses, and molecular dynamics simulations, we investigated the unique recognition mechanism that enables higher binding affinity and selectivity of the antibodies compared to other generic proteins with lower binding affinity to phosphorylated epitopes. Our results demonstrate that molecular dynamics simulations provide novel insights into the dynamic aspects of molecular recognition of posttranslational modifications by proteins beyond static crystal structures, highlighting how specific atomic level interactions drive the exceptional affinity and selectivity of antibodies., Competing Interests: Conflict of interest The authors used the patented technology Wiz-Amp (US 9,890,414), invented by S. C. J. O. and T. M. for antibody acquisition., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

In this study, heavy chain genes of IgD and IgT were sequenced and characterized their gene expression in rock bream (Oplegnathus fasciatus). Rock bream (RB)-IgD cDNA is 3319 bp in length and encodes a leader region, variable domains, a μ1 domain, and seven constant domains (CH1-CH7). A membrane-bound (mIgT) and secretory form (sIgT) of RB-IgT cDNAs are 1902 bp and 1689 bp in length, respectively, and encode a leader region, variable domains, four constant domains (CH1-CH4) and C-terminus. Their predicted 3D-structure and phylogenetic relation were similar to those of other teleost. In healthy fish, RB-IgD and mIgT gene expressions were higher in major lymphoid organs and blood, while RB-sIgT gene was more highly expressed in midgut. IgT expressing cells were detected in melano-macrophage centers (MMC) of head kidney in immunohistochemistry analysis. Under immune stimulation in vitro, RB-IgD and IgT gene expressions were upregulated in head kidney and spleen cells by bovine serum albumin or a rock bream iridovirus (RBIV) vaccine. In vivo, their expressions were significantly upregulated in head kidney, blood, and gill upon vaccination. Especially, RB-mIgT gene expression in head kidney and blood was upregulated at day 3 after vaccination while upregulated at earlier time point of day 1 by challenge with RBIV. This may suggest that memory cells might be produced during the primary response by vaccination and rapidly proliferated by secondary immune response by viral infection. RB-sIgT gene expression was highly upregulated in peripheral blood in vaccinated fish after viral infection, indicating that IgT plays an important role in systemic immune response as well as mucosal immune system. Our findings provide information on the role of RB-IgT in adaptive immunity during vaccination and viral infection in the vaccinated fish., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Reverse vaccinology is a novel vaccine development technology that uses genome and proteome analyses through bioinformatics to select antigenic epitopes capable of eliciting an immunological and protective response through a quick and cheap methodology. However, data on its use in animal health are scant and further research is advocated. Therefore, this systematic review aimed to evaluate the evidence of the efficiency of reverse vaccinology in the search for antigens against bovine parasites, as well as its perspectives and limitations. One hundred seventy-four studies were found, of which 95 were selected for full reading following the PRISMA guidelines and considering all databases. After the last evaluation and reading of the references, only 19 studies were included and evaluated for methodological quality and biases. The studies applied reverse vaccinology to bacteria, protozoa, and ectoparasites that affect cattle, emphasizing on the tick species Rhipicephalus microplus and the protozoa of the genus Babesia that use it as a vector. Most studies evaluated the acquisition of an immune response through ELISA, WB and IFAT analyses to measure predominantly IgG. In addition, many studies did not examine the complete proteome of the parasites and are carried out only in silico, in vitro, or even with unrelated animals, the reason why they were excluded from our systematic review. Due to lack of studies that met the eligibility criteria, in this systematic review we also included studies carried out with different groups and species of parasites, providing a broad overview of the application of this technique in cattle farming. Conversely, this also resulted in variable methodologies, which makes comparison among studies difficult. Despite that, the application of reverse vaccinology in cattle farming has shown promising results in the development of immunological and protective responses in cattle. However, research methodologies need to be improved to reduce biases and obtain reliable results, in addition to clarity of data and methodologies to enable reproducibility., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2024. Published by Elsevier B.V.)

Immunotherapy is one of the rising stars in the field of anticancer regiments. Aimed at reinvigorating immune cytotoxicity, this platform is capable of bulking up memory subsets by which protection against tumors is served. The most commonly applied immunotherapy is immune checkpoint inhibitor (ICIs) which received FDA approval for non-small lung cancer (NSLC) in 2014. The response toward ICI is closely related to the antigen processing machinery (APM) within which antigens are processed prior to loading onto the human leukocyte antigen (HLA) to induce cascade mechanisms for immune clearance. APM allows immune cell infiltration thus strengthens immunogenicity. Impaired components of the APM are frequently found in tumors because tumor progression requires tumor cells to acquire immune recognition evasion. Alterations in tumors' APM result in downregulation of HLA molecules and transformation of antigenic peptide repertoire presented to the T lymphocytes. Interactions of processed antigens (peptide)-HLA complex are critical for successful T cell priming and differentiation into cytotoxic effector cells. The interaction underlies not only ICI-related mechanism but also anticancer immunity in general where T cell subset can induce antitumor recognition only if a proper peptide-HLA complex is present. This feature, unfortunately, is missing in tumors. This Review highlights presentation of tumor-specific antigens to T cells in HLA-restricted manner which leads to their eradication. This is a pivotal point but in most cases is overlooked which might add some volume to the off-target and less functional of anticancer immunotherapy.

Haemonchus contortus (Barber pole worm) is one of the dominant helminth parasitic infections in small ruminants which is economically important and causes severe losses in the livestock industry, particularly in tropical and subtropical regions. This parasite resides in the abomasum and is responsible for severe blood loss, leading to anemia, emaciation, hypoproteinemia, weight loss, and potentially death. The economic impact of H. contortus on the livestock industry necessitates effective control measures, including early diagnosis and the development of effective vaccines. H. contortus secretes a variety of excretory and secretory proteins (ESPs), which are glycoproteins that play a crucial role in modulating the host's immune response. These ESPs are not only vital for understanding the immunological interactions between the parasite and the host but also serve as potential diagnostic tools and vaccine candidates. Similar ESPs have been identified in other parasitic species such as Cooperia spp, Ostertagia ostertagia, Teladorsagia circumcincta, Ascaris sum, Schistosoma japonicum, and Echinococcus multilocularis, underscoring their importance in both detection and vaccine development. In addition, there is a lack of highly potential specific proteins which having immunogenic properties that can be used for the accurate, early diagnosis serologically and serve as a potential candidate for the vaccine development against H. contortus. Recent research highlights that T H -9 stimulated proteins from H. contortus are emerging as promising candidates for vaccine development due to their immunomodulatory effects. These proteins have been shown to induce a T H -9 immune response, characterized by increased production of interleukin-9 (IL-9), which is critical for enhancing protective immunity against helminth infections. It is suggested to investigate T H -9 stimulated protein as potential candidates for vaccine development and diagnostic antigen., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Antibodies in oncology are being equipped with toxic cargoes and effector functions that can kill cells at very low concentrations. A key challenge is that most targets on cancer cells are also present on at least some healthy cells. Shared targets can result in off-tumor binding and compromise the safety and potential of therapeutic candidates. In this review, we survey strategies that can help direct biologics to cancer sites more selectively. These strategies are becoming increasingly feasible thanks to advances in molecular design and engineering. The objective is to create therapeutics that exploit changes in cancer and leverage the human body infrastructure, enabling therapeutics that discriminate not just self from non-self but diseased from healthy tissue., Competing Interests: Declaration of interests None declared by authors., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Exposure to allogenic red blood cells (RBCs), either through pregnancy or transfusion, can result in alloimmunization, which can lead to severe hemolytic transfusion reactions and pregnancy complications. Passively administered antibodies can be used to prevent alloimmunization, where steric hindrance of allogeneic epitopes has been postulated as one mechanism whereby antibody engagement may prevent RBC alloimmunization. However, the dynamics of antibody engagement on the RBC surface has remained difficult to study. To examine this, we leveraged the HOD (HEL, OVA and Duffy) model system and Fcγ receptor knockout recipients to define the dynamics of antibody engagement of the Duffy antigen in the absence of RBC clearance or antigen modulation. Using this approach, the on-rate of antibody engagement of HOD RBCs was very similar in vivo and in vitro , with high levels of antibody binding observed within minutes of HOD RBC exposure. In contrast, the off-rate of HOD RBC bound antibody was relatively slow, with appreciable dissociation not being observed for an hour. However, the dynamics of antibody interactions with HOD changed significantly when antibody decorated HOD RBCs were exposed to free antibody. Despite the presence of prebound antibody, free antibody rapidly associated with HOD RBCs, with the rate of free antibody association observed being faster in vivo than in vitro . Importantly, antibody association and dissociation occurred in the absence of any appreciable changes in RBC clearance, antigen modulation or complement deposition, suggesting that differences in antibody levels observed reflected actual differences in the dynamics of antibody binding. These results suggest that while antibodies appear to be relatively static on the cell surface once bound, antibody engagement can be quite dynamic, especially in the face of free antibody in solution. These results not only have implications in the mechanisms of antibody-mediated immunosuppression, but also the potential use of other antibody-based approaches designed to prevent hemolytic transfusion reactions or target antigens in vivo in general., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Jajosky, Ayona, Mener, Stowell and Arthur.)

Background: Epstein-Barr virus (EBV) is associated with nasopharyngeal carcinoma (NPC), but the existence of NPC protective antibody against EBV-associated antigens remains inconclusive., Methods: NPC cases and matched controls were identified from prospective cohorts comprising 75,481 participants in southern China. ELISA and conditional logistic regression were applied to assess effects of gp42-IgG on NPC. The expression of HLA-II, the gp42 receptor, in nasopharyngeal atypical dysplasia and its impact on EBV infecting epithelial cells were evaluated., Findings: gp42-IgG titers were significantly lower in NPC cases compared to controls across various follow-up years before NPC diagnosis (P<0.05). Individuals in the highest quartile of gp42-IgG titers had a 71% NPC risk reduction comparing to those in the lowest quartile (odds ratios [OR]Q4vsQ1=0.29, 95% confidence intervals [CIs]=0·15 to 0·55, P<0.001). Each unit antibody titer increase was associated with 34% lower risk of NPC (OR=0.66, 95% CI=0.54 to 0.81, Ptrend <0.001). Their protective effect was observed in cases diagnosed ≥5 years, 1-5 years and <1 year after blood collection (P<0.05). HLA-II expression was detected in 13 of 27 nasopharyngeal atypical dysplasia and its overexpression substantially promoted epithelial-cell-origin EBV infection., Conclusion: Elevated EBV gp42-IgG titers can reduce NPC risk, indicating gp42 as a potential EBV prophylactic vaccine design target., Trial Registration: NCT00941538, NCT02501980, ChiCTR2000028776, ChiCTR2100041628., Funding: Noncommunicable Chronic Diseases-National Science and Technology Major Project, National Natural Science Foundation of China, Local Innovative and Research Teams Project of Guangdong Pearl River Talents Program, Central Financial Transfer Payment Projects of the Chinese Government, Cancer Research Grant of Zhongshan City.

Antibody-mediated immunity is crucial in the vertebrate immune system. Nanobodies, also known as VHH or single-domain antibodies (sdAbs), are emerging as promising alternatives to full-length antibodies due to their compact size, precise target selectivity, and stability. However, the limited availability of nanobodies (Nbs) for numerous antigens (Ags) presents a significant obstacle to their widespread application. Understanding the interactions between Nbs and Ags is essential for enhancing their binding affinities and specificities. Experimental identification of these interactions is often costly and time-intensive. To address this issue, we introduce NABP-BERT, a deep-learning model based on the BERT architecture, designed to predict NANOBODY®-Ag binding solely from sequence information. Furthermore, we have developed a general pretrained model with transfer capabilities suitable for protein-related tasks, including protein-protein interaction tasks. NABP-BERT focuses on the surrounding amino acid contexts and outperforms existing methods, achieving an AUROC of 0.986 and an AUPR of 0.985., (© The Author(s) 2024. Published by Oxford University Press.)

Follow-up of previously healthy patients surviving cryptococcal meningitis found that cryptococcal antigen could be detected for >1 year in serum from 38 of 44 (86%) patients and in cerebrospinal fluid (CSF) from 20 of 31 patients (67%), far beyond the time of culture conversion. The speed of titer decline, measured as the number of days for a 2-fold drop in titer to occur, was slower in serum than in CSF. The speed of decline of antigen titers was much slower in serum and CSF for patients infected with Cryptococcus gattii than Cryptococcus neoformans. The speed of decline in CSF and serum titers was also much slower in patients who had received a ventriculoperitoneal shunt for increased intracranial pressure. The variable and extraordinarily slow rate of clearance in our patients did not appear to reflect differences in disease control but rather differences in species and shunting for increased intracranial pressure., Competing Interests: Potential conflicts of interest. Both authors: No reported conflicts. Both authors have submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest., (Published by Oxford University Press on behalf of Infectious Diseases Society of America 2024.)

This article recapitulates the discoveries and anti-tumoural therapeutical proposals by Francesco Sanfelice, who in 1931 published an essay entitled The Treatment of Cancer and Sarcoma with Cancrocidin (paraneoforming Blastomycetes). Sanfelice's discoveries are contextualised with subsequent scientific discoveries, especially with those by L. Scott McDaniel and G. Cozad, who evaluated the functionality of murine peritoneal macrophages previously sensitised precisely with Blastomyces dermatitidis antigen extracts. Finally, recent research on the topic of intratumoural microbiota is mentioned showing how Sanfelice's ideas, albeit partly outdated, can still inspire current biomolecular research., (© 2024. The Author(s).)

Hepatocellular carcinoma (HCC) ranks among the most prevalent cancers worldwide, highlighting the urgent need for improved diagnostic and therapeutic methodologies. The standard treatment regimen generally involves surgical intervention followed by systemic therapies; however, the median survival rates for patients remain unsatisfactory. Chimeric antigen receptor (CAR) T-cell therapy has emerged as a pivotal advancement in cancer treatment. Both clinical and preclinical studies emphasize the notable efficacy of CAR T cells in targeting HCC. Various molecules, such as GPC3, c-Met, and NKG2D, show significant promise as potential immunotherapeutic targets in liver cancer. Despite this, employing CAR T cells to treat solid tumors like HCC poses considerable challenges within the discipline. Numerous innovations have significant potential to enhance the efficacy of CAR T-cell therapy for HCC, including improvements in T cell trafficking, strategies to counteract the immunosuppressive tumor microenvironment, and enhanced safety protocols. Ongoing efforts to discover therapeutic targets for CAR T cells highlight the need for the development of more practical manufacturing strategies for CAR-modified cells. This review synthesizes recent findings and clinical advancements in the use of CAR T-cell therapies for HCC treatment. We elucidate the therapeutic benefits of CAR T cells in HCC and identify the primary barriers to their broader application. Our analysis aims to provide a comprehensive overview of the current status and future prospects of CAR T-cell immunotherapy for HCC., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Zhou, Wei, Xu, Wu, Dou, Li, Zhang and Zhang.)

Although therapeutic nanovaccines have made a mark in cancer immunotherapy, the shortcomings such as poor homing ability of lymph nodes (LNs), low antigen presentation efficiency and low antitumor efficacy have hindered their clinical transformation. Accordingly, we prepared advanced nanovaccines (CMB and CMC) by integrating carbon dots (CDs) with tumor-associated antigens (B16F10 and CT26). These nanovaccines could forwardly target tumors harbouring LNs, induce strong immunogenicity for activating cytotoxic T cells (CTLs), thereby readily eliminating tumor cells and suppressing primary/distal tumor growth. This work provides a promising therapeutic vaccination strategy to enhance cancer immunotherapy., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Objectives: This study aimed to determine the true frequency of weak ABO subgroups and to investigate the serological characteristics of various subgroup alleles in the Chinese population., Methods: A total of 2,945,643 blood samples were collected from January 2009 to December 2017. After initial screening and re-examination using automatic blood group analyzers, all ABO-discrepant samples were confirmed by standard serological analysis and molecular detection by DNA sequencing. The true frequency of weak ABO subgroups was determined by the number of ABO subgroup donors and the missed detection rate. The ABO antigen expressions corresponding to subgroup alleles were analyzed by the agglutination intensity of red blood cells., Results: The detection rate of ABO subgroups was 0.031 % (927/2,945,643). Considering the missed detection rate (27.81 %), the true frequency of ABO subgroups in the Chinese population was 0.044 %. The three most common genetic variations among blood donors in Shanghai were BA.04, BW.12 and BA.02. BW.03 showed the weakest B antigen expression (6.00 ± 1.97) and B var- 1 the strongest (9.20 ± 1.10)., Conclusion: Many ABO subgroups were missed. BA.04 was found to be the most common subgroup allele in the Chinese population. Different ABO subgroup alleles exhibit different ABO antigen expression patterns., Competing Interests: Conflicts of interest We declare that we have no conflicts of interest relevant to the content of this paper., (Copyright © 2024. Published by Elsevier España, S.L.U.)

Objectives: Despite surgical resection, chemoradiation, and targeted therapy, brain tumors remain a leading cause of cancer-related death in children. Immunotherapy has shown some promise and is actively being investigated for treating childhood brain tumors. However, a critical step in advancing immunotherapy for these patients is to uncover targets that can be effectively translated into therapeutic interventions., Methods: In this study, our team performed a transcriptomic analysis across pediatric brain tumor types to identify potential targets for immunotherapy. Additionally, we assessed components that may impact patient response to immunotherapy, including the expression of genes essential for antigen processing and presentation, inhibitory ligands and receptors, interferon signature, and overall predicted T cell infiltration., Results: We observed distinct expression patterns across tumor types. These included elevated expression of antigen genes and antigen processing machinery in some tumor types while other tumors had elevated inhibitory checkpoint receptors, known to be associated with response to checkpoint inhibitor immunotherapy., Conclusion: These findings suggest that pediatric brain tumors exhibit distinct potential for specific immunotherapies. We believe our findings can guide investigators in their assessment of appropriate immunotherapy classes and targets in pediatric brain tumors.

It is a fact that sperm possess antigenic properties. Substantial scientific research suggests that specific antibodies that attach to sperm antigens can induce infertility in both humans and other species. Antisperm antibodies (ASA) represent a significant etiology of infertility in humans, leading to immunoinfertility. The association between ASA and infertility is multifaceted. The observation of sperm agglutination, although not conclusive for the diagnosis of immunological infertility, may suggest the presence of ASA. Nevertheless, ASA may also manifest in the lack of any sperm agglutination. Managing ASA from an andrological perspective depends on the underlying cause and the specific approaches healthcare professionals adopt. The precise etiology of male infertility resulting from ASA remains unclear. Current research has examined the impact of ASA and its prevalence among infertile males to understand the relationship between ASA and changes in semen parameters. However, the findings have been inconclusive. Numerous techniques have been documented for the management of immunoinfertility. This review examines the importance of ASA in the context of infertility, encompassing the postulated mechanisms underlying the development of ASA, the various assays employed for detecting them, and the available treatments., (© 2024. The Author(s), under exclusive licence to Society for Reproductive Investigation.)

Avian influenza viruses (AIV) are capable of infecting a considerable proportion of the world's population each year, leading to severe epidemics with high rates of morbidity and mortality. The methods now used to diagnose influenza virus A include the Western blot test (WB), hemagglutination inhibition (HI), and enzyme-linked immunosorbent assays (ELISAs). But because of their labor-intensiveness, lengthy procedures, need for costly equipment, and inexperienced staff, these approaches are considered inappropriate. The present review elucidates the recent advancements in the field of avian influenza detection through the utilization of nanomaterials-based immunosensors between 2014 and 2024. The classification of detection techniques has been taken into account to provide a comprehensive overview of the literature. The review encompasses a detailed illustration of the commonly employed detection mechanisms in immunosensors, namely, colorimetry, fluorescence assay, surface plasmon resonance (SPR), surface-enhanced Raman spectroscopy (SERS), electrochemical detection, quartz crystal microbalance (QCM) piezoelectric, and field-effect transistor (FET). Furthermore, the challenges and future prospects for the immunosensors have been deliberated upon. The present review aims to enhance the understanding of immunosensors-based sensing platforms for virus detection and to stimulate the development of novel immunosensors by providing novel ideas and inspirations. Therefore, the aim of this paper is to provide an updated information about biosensors, as a recent detection technique of influenza with its details regarding the various types of biosensors, which can be used for this review., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Oz virus (OZV) belongs to the Orthomyxoviridae family which includes viruses with a negative-sense, single-stranded, and segmented RNA genome. OZV is a zoonotic pathogen, particularly since the virus can cause deadly illness when injected intracerebrally into nursing mice. OZV is an emerging pathogen with the potential to spark a pandemic as there is no preventive and licensed treatment against this virus. The goal of this study was to develop a novel multi-epitope vaccination against OZV proteins utilizing immunoinformatics and immunological simulation analysis. This work evaluated immunological epitopes (B cells, MHC-I, and MHC-II) to identify highly antigenic OZV target proteins. Shortlisted epitopes were joined together by using appropriate linkers and adjuvants to design multi-epitope vaccine constructs (MEVC). The vaccine models were designed, improved, validated, and the globular regions and post-translational modifications (PTMs) were also evaluated in the vaccine's structure. Molecular docking analysis with the Toll-like receptor (TLR4) showed strong interactions and appropriate binding energies. Molecular dynamics (MD) simulation confirmed stable interactions between the vaccines and TLR4. Bioinformatics tools helped optimize codons, resulting in successful cloning into appropriate host vectors. This study showed that the developed vaccines are stable and non-allergenic in the human body and successfully stimulated immunological responses against OZV. Finally, a mechanism of action for the designed vaccine construct was also proposed. Further experimental validations of the designed vaccine construct will pave the way to create a potentially effective vaccine against this emerging pathogen., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.)

Background: Dormancy survival regulator (DosR) and resuscitation-promoting factor (Rpf) antigens of Mycobacterium tuberculosis are activated during dormant phase of tuberculosis (TB). This study evaluates the differential immunogenicity potentials of DosR and Rpf antigens in individuals with latent tuberculosis infection (LTBI) and active TB patients., Methods: After a literature search in electronic databases, studies were selected by following precise eligibility criteria. Outcomes were synthesized systematically, and meta-analyses were performed to estimate standardized mean differences (SMDs) in interferon-gamma (IFNγ) levels, and IFNγ positive immune cells between individuals with LTBI and active TB patients., Results: Twenty-six studies (1278 individuals with LTBI and 1189 active TB patients) were included. DosR antigens Rv0569 (Standardized mean difference; SMD 2.44 [95%CI: 1.21, 3.66]; p < 0.0001), Rv1733c (SMD 0.60 [95%CI: 0.14, 1.07]; p = 0.011), Rv1735c (SMD 1.16 [95%CI: 0.44, 1.88]; p = 0.002), Rv1737c (SMD 1.26 [95%CI: 0.59, 1.92]; p < 0.0001), Rv2029c (SMD 0.89 [95%CI: 0.35, 1.42]; p = 0.002), RV2626c (SMD 1.24 [95%CI: 0.45, 2.02); p = 0.002), and Rv2628 (SMD 0.65 [95%CI: 0.38, 0.91]; p < 0.0001) and Rpf antigens Rv0867c (SMD 1.33 [95%CI: 0.48, 2.18]; p = 0.002), Rv1009 (SMD 0.65 [95%CI: 0.05, 1.25]; p = 0.034), and Rv2450c (SMD 1.54 [95%CI: 0.92, 2.16]; p < 0.0001) elicited higher IFNγ levels in individuals with LTBI in comparison with active TB patients. IFNγ-positive immunoresponsive cells were significantly higher in individuals with LTBI than in active TB patients for antigens Rv1733c (SMD 1.02 [95%CI: 0.15, 1.88]; p = 0.021), Rv2029c (SMD 0.57 [95%CI: 0.05, 1.09]; p = 0.031), and Rv2628 [SMD 0.38 [95%CI: 0.15, 0.61]; p = 0.001)., Conclusion: DosR antigens Rv0569, Rv1733c, Rv1735c, Rv1737c, RV2626c, Rv2628, and Rv2029c, and Rpf antigens Rv0867c, Rv1009, and Rv2450c are found to elicit immune responses differently in individuals with LTBI and active TB patients., (© 2024. The Author(s).)

Tuberculosis (TB) is one of the leading causes of death from infectious diseases, killing approximately 1.3 million people worldwide in 2022 alone. The current vaccine for TB contains a live attenuated bacterium, Mycobacterium bovis BCG (Bacille Calmette-Guérin). The BCG vaccine is highly effective in preventing severe forms of childhood TB but does not protect against latent infection or disease in older age groups. A new or improved BCG vaccine for prevention of pulmonary TB is urgently needed. In this study, we infected murine bone marrow derived dendritic cells from C57BL/6 mice with M. bovis BCG followed by elution and identification of BCG-derived MHC class I and class II-bound peptides using tandem mass spectrometry. We identified 1436 MHC-bound peptides of which 94 were derived from BCG. Fifty-five peptides were derived from MHC class I molecules and 39 from class II molecules. We tested the 94 peptides for their immunogenicity using IFN- γ ELISPOT assay with splenocytes purified from BCG immunized mice and 10 showed positive responses. Seven peptides were derived from MHC II and three from MHC class I. In particular, MHC class II binding peptides derived from the mycobacterial surface lipoprotein Mpt83 were highly antigenic. Further evaluations of these immunogenic BCG peptides may identify proteins useful as new TB vaccine candidates., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Introduction: ER aminopeptidase 1 (ERAP1) is a polymorphic intracellular aminopeptidase with key roles in antigen presentation and adaptive immune responses. ERAP1 allotype 10 is highly protective toward developing some forms of autoimmunity and displays unusual functional properties, including very low activity versus some substrates., Methods: To understand the molecular mechanisms that underlie the biology of allotype 10, we studied its enzymatic and biophysical properties focusing on its unique polymorphisms V349M and Q725R., Results: Compared to ancestral allotype 1, allotype 10 is much less effective in trimming small substrates but presents allosteric kinetics that ameliorate activity differences at high substrate concentrations. Furthermore, it is inhibited by a transition-state analogue via a non-competitive mechanism and is much less responsive to an allosteric small-molecule modulator. It also presents opposite enthalpy, entropy, and heat capacity of activation compared to allotype 1, and its catalytic rate is highly dependent on viscosity. Polymorphisms V349M and Q725R significantly contribute to the lower enzymatic activity of allotype 10 for small substrates, especially at high substrate concentrations, influence the cooperation between the regulatory and active sites, and regulate viscosity dependence, likely by limiting product release., Conclusions: Overall, our results suggest that allotype 10 is not just an inactive variant of ERAP1 but rather carries distinct enzymatic properties that largely stem from changes at positions 349 and 725. These changes affect kinetic and thermodynamic parameters that likely control rate-limiting steps in the catalytic cycle, resulting in an enzyme optimized for sparing small substrates and contributing to the homeostasis of antigenic epitopes in the ER., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Georgaki, Mpakali, Trakada, Papakyriakou and Stratikos.)

BACKGROUNDMost humans have been infected with cytomegalovirus (CMV) by midlife without clinical signs of disease. However, in settings in which the immune system is undeveloped or compromised, the virus is not adequately controlled and consequently presents a major infectious cause of both congenital disease during pregnancy as well as opportunistic infection in children and adults. With clear evidence that risk to the fetus varies with gestational age at the time of primary maternal infection, further research on humoral responses to primary CMV infection during pregnancy is needed.METHODSHere, systems serology tools were applied to characterize antibody responses to CMV infection in pregnant and nonpregnant women experiencing either primary or chronic infection.RESULTSWhereas strikingly different antibody profiles were observed depending on infection status, limited differences were associated with pregnancy status. Beyond known differences in IgM responses used clinically for identification of primary infection, distinctions observed in IgA and FcγR-binding antibodies and among antigen specificities accurately predicted infection status. Machine learning was used to define the transition from primary to chronic states and predict time since infection with high accuracy. Humoral responses diverged over time in an antigen-specific manner, with IgG3 responses toward tegument decreasing over time as typical of viral infections, while those directed to pentamer and glycoprotein B were lower during acute and greatest during chronic infection.CONCLUSIONIn sum, this work provides insights into the antibody response associated with CMV infection status in the context of pregnancy, revealing aspects of humoral immunity that have the potential to improve CMV diagnostics.FUNDINGCYMAF consortium and NIH NIAID.

Background: Recent articles have explored the effect of worms on cancer cells. This review focused on various cell cultures employed to understand which cells are more commonly and less utilized., Methods: The present review analyzed studies published between 2013 and 2023 to obtain information about different cell cultures used in cancer studies involving helminths. Databases such as PubMed, Google Scholar, HINARI, and the Cochrane Library were searched., Results: This search yielded 130 records, but 97 papers were excluded because they were either irrelevant to the research topic (n = 72) or contradicted the research idea (n = 25).The remaining twenty-one articles focused on different types of worms, such as Echinococcus granulosus, Clonorchis sinensis, Opisthorchis felineus, Opisthorchis viverrini, Trichinella spiralis, Toxocara canis, and Heligmosomoides polygyrus., Conclusion: Due to the presence of numerous antigens, parasites at different growth stages can impact various cells through unknown mechanisms. Given the high diversity of antigens and their effects, artificial intelligence can assist in predicting initial outcomes for future studies., (© 2024. The Author(s).)

Immunoassays based on the specific antigen-antibody interactions are efficient tools to detect various compounds and estimate their content. Usually, these assays are implemented in water-saline media with composition close to physiological conditions. However, many substances are insoluble or cannot be molecularly dispersed in such media, which objectively creates problems when interacting in aquatic environments. Thus, obtaining immunoreactants and implementing immunoassays of these substances need special methodological solutions. Hydrophobicity of antigens as well as their limited ability to functionalization and conjugation are often overlooked when developing immunoassays for these compounds. The main key finding is the possibility to influence the behavior of hydrophobic compounds for immunoassays, which requires specific approaches summarized in the review. Using the examples of two groups of compounds-surfactants (alkyl- and bisphenols) and fullerenes, we systematized the existing knowledge and experience in the development of immunoassays. This review addresses the challenges of immunodetection of poorly soluble substances and proposes solutions such as the use of hydrotropes, other solubilization techniques, and alternative receptors (aptamers and molecularly imprinted polymers).

Klebsiella pneumoniae (K. pneumoniae) is an opportunistic pathogen and the major cause of healthcare-associated infections, which are increasingly complicated by the prevalence of highly invasive and hyper-virulent K. pneumoniae strains, necessitating the development of alternative strategies for combatting infections caused by this bacterium. In this study, we successfully constructed a fusion antigen called KP-Ag1, comprising three antigens (GlnH, FimA, and KPN&#95;00466) that were previously identified through reverse vaccinology. Immunization with KP-Ag1 formulated with Al(OH) 3 adjuvant elicited robust humoral and cellular immune response in mice, and conferred protective immunity in a murine model of K. pneumoniae lung infection. Further analysis of serum IgG subtypes from mice immunized with KP-Ag1 revealed a predominant IgG1 response, indicating that KP-Ag1 predominantly induces a Th2-biased immune response. Additionally, opsonophagocytic killing assay suggested that humoral immune responses play a pivotal role in mediating protection conferred by KP-Ag1. Moreover, KP-Ag1 was found to promote the activation and maturation of BMDCs in vitro, which is essential for subsequent efficient antigen presentation. More importantly, vaccination with KP-Ag1 demonstrated cross-protective efficacy against clinical isolates of K. pneumoniae varying in serotypes, antibiotic resistance, and virulence profiles. Therefore, KP-Ag1 holds promise as a candidate for K. pneumoniae vaccine development., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)