The aim of this study was to evaluate the diagnostic potential of human papillomavirus type 18 (HPV18) E6/E7-specific T cell receptor (TCR)-like antibodies for the early detection of HPV-associated cervical cancer. An integrated in silico and in vitro approach was employed, in which three fully human TCR-like antibodies (H11\Ab, E5\Ab, and G8\Ab) were analysed by molecular docking to assess antibody-peptide-major histocompatibility complex (P-MHC) class I HLA-A24:02 interactions, followed by experimental validation using SiHa and C33A cervical cancer cell lines. Docking analyses demonstrated stable, high-affinity interactions between the antibodies and their corresponding peptide-MHC complexes, consistent with experimental observations. In vitro assays confirmed strong binding specificity and functional activity in HPV18-positive cells, with no detectable reactivity in HPV-negative controls. These findings demonstrate that TCR-like antibodies targeting HPV18 E6/E7 peptide-MHC complexes enable highly specific detection of HPV-associated cervical cancer and support their potential application as early diagnostic biomarkers.

Mitochondrial dysfunction is a central pathological feature of a wide range of inherited and acquired disorders and is characterized by impaired oxidative phosphorylation, disrupted cellular energy metabolism, and excessive oxidative stress. Although advances in molecular diagnostics have improved disease recognition, effective disease-modifying therapies remain limited, and clinical outcomes are often suboptimal, highlighting the need for novel therapeutic strategies. Mesenchymal stromal cells (MSCs) and their extracellular vesicles (MSC-EVs) have emerged as promising candidates for targeting mitochondrial dysfunction due to their regenerative, immunomodulatory, and metabolic regulatory properties. In this review, we provide a comprehensive overview of recent in vitro and in vivo studies investigating the capacity of MSCs and MSC-EVs to restore mitochondrial function by enhancing mitochondrial respiration, improving cellular bioenergetics, and reducing oxidative stress across diverse disease models. We further discuss the underlying mechanisms involved, including mitochondrial transfer, delivery of functional mitochondrial components, and modulation of the cellular microenvironment. Finally, we highlight the key advantages, translational potential, and remaining challenges associated with MSC- and MSC-EV-based therapies for mitochondrial dysfunction.

This review aims to evaluate the potential of endometriosis models, especially patient-derived iPSC models, to gain deeper insights into the disease, thereby advancing our understanding and treatment of endometriosis. This comprehensive narrative review utilized a structured search of the PubMed, Scopus, and Web of Science databases, primarily covering literature published between January 2000 and May 2025. An expansive search strategy was employed to capture the full breadth of the field using keywords such as "endometriosis," "induced pluripotent stem cells (iPSCs)," "patient-derived organoids," "disease modeling," and "epigenetics" without restrictive filtering, ensuring the integration of both foundational theories and emerging biotechnological advances. In total, over 170 peer-reviewed publications were analyzed, ranging from landmark genomic meta-analyses that have identified significant risk loci to state-of-the-art 3D-culture systems for modeling patient-specific endometrial disease. By synthesizing these diverse sources, the review bridges the gap between traditional anatomical classifications and modern molecular modeling to evaluate the potential of iPSC platforms for personalized medicine and therapeutic discovery. Endometriosis is a multifactorial gynecological condition that affects 176 million women worldwide and can significantly impair quality of life. It occurs when endometrium-like tissue grows outside the uterus, responsive to ovarian hormones, causing inflammation, pain, and discomfort, and leading to fibrotic tissue. World Health Organization estimates indicate that 6-10% of women suffer from this disorder, which can cause infertility and increase the risk of developing various types of cancer and autoimmune disorders. The use of patient-derived iPSC models serves to gain deeper insights into the disease by mimicking the endometrial tissue or lesions observed in affected individuals, thereby advancing our understanding and treatment of endometriosis.

Efficient bacterial production of recombinant antibody fragments is critical for structural characterization and subsequent functional studies. In this study, a systematically optimized workflow was developed for the expression, extraction, and purification of soluble T-cell receptor (TCR)-like antibody fragments in Escherichia coli (E. coli). Two expression hosts, the K-12–derived HB2151 strain and the B-lineage BL21(DE3) pLysS strain, were comparatively evaluated. Although both strains supported recombinant expression, BL21(DE3) pLysS produced higher levels of soluble protein. However, conventional osmotic shock extraction resulted in limited protein recovery. To improve extraction efficiency, a combined lysis approach incorporating freeze–thaw cycles, lysozyme treatment, and sonication was applied, increasing total protein recovery in cell lysates to 149.2–178.4 mg/L of culture. Purification was carried out using Protein A affinity chromatography, and the purification conditions were further optimized to enhance protein recovery. Optimal binding was achieved using 20 mM sodium phosphate buffer (pH 6.8), while elution with 100 mM citric acid (pH 2.9) improved protein yield while maintaining protein stability during purification. Under these conditions, purified soluble TCR-like antibody fragments were obtained at final yields of 6.16–7.08 mg/L of culture. The purified proteins migrated at the expected molecular weight ( 15 kDa) on SDS–PAGE and were specifically detected by dot blot analysis. Overall, this study provides a reproducible and scalable workflow for improving the expression, extraction, and purification of recombinant soluble TCR-like antibody fragments in E. coli.

Tumor-infiltrating lymphocytes (TILs) are recognized as a key component of anticancer immunity and serve as an important prognostic factor in cancer progression. In this review, the latest updates and perspectives on the diverse populations of TILs and their roles in cancer immunity are discussed. The presence and balance between anti-tumorigenic and pro-tumorigenic immune cells in the tumor microenvironment (TME) largely determine tumor progression and fate. Thus, the properties of TILs were reviewed to provide a better insight into the roles of these immune cells within the TME. Additionally, the different factors influencing immune cell infiltration in solid tumors are also described to suggest novel immunotherapeutic approaches for improved TIL infiltration. These recent approaches are then summarised as recommendations to improve infiltration and potentially achieve better clinical outcomes. Overall, this review highlights the critical role of TILs, factors governing immune cell homing and infiltration, and strategic approaches to improve TIL infiltration.

Cervical cancer remains one of the most prevalent malignancies among women worldwide, with persistent infection by high-risk human papillomavirus (HPV), particularly types 16 and 18, as the principal etiological factor. The viral oncoproteins E6 and E7 are well-established drivers of cellular transformation and the maintenance of malignant phenotypes. This study aimed to generate T-cell receptor-like antibodies (TCR-like antibodies) capable of recognizing HPV16-derived E6 and E7 peptides presented by the major histocompatibility complex class I molecule HLA-A2402, with the goal of developing a diagnostic platform for early detection of HPV16-associated disease. Peptide-major histocompatibility complex (P-MHC) antigenic complexes were constructed and characterized using molecular docking, followed by experimental validation. A fully human domain antibody phage display library was screened against these complexes through iterative biopanning, supported by polyclonal and monoclonal enzyme-linked immunosorbent assays (ELISA). Structural evaluation and binding affinity analyses were performed using molecular docking, Western blotting, TCR-like antibody ELISA, and cell-based ELISA. Two novel TCR-like antibodies, designated D2\Ab and A12\Ab, were identified and demonstrated strong and specific binding to HPV16 E6 and E7 P-MHC complexes, confirming high affinity and antigen specificity. Functional validation across multiple assays demonstrated selective recognition of target complexes without reactivity toward irrelevant controls. In conclusion, this study demonstrates the feasibility of generating fully human TCR-like antibodies for HPV-associated malignancies and highlights their potential application in early detection and targeted diagnostic strategies for cervical cancer.

The emergence and re-emergence of infectious diseases have caused great concern for global organizations due to their destructive impacts on health, the economy, and human survivability, following the fast-evolving and adaptive characteristics of pathogens. This concern highlights the urgent need for the development of point-of-care diagnostic tools for rapid and reliable screening of pathogens, which can be applied widely without geographic barriers. An electrochemical impedance spectroscopy-based immunosensor is considered an effective alternative to conventional methods for decentralized diagnostics, owing to its cost efficiency, simplicity, sensitivity, and label-free potential. Incorporating nanotechnology into an impedimetric immunosensor can enhance reliability, but it requires careful consideration of electrode fabrication and surface modification. Thus, this review provides insights into the integration of notable nanomaterials, gold nanoparticles, highlighting their important characteristics such as high surface areas, conductivity, and biocompatibility in enhancing the overall performance of the EIS-based immunosensor. This review further discussed the recent application of this integrated system in diagnosing infectious diseases, including viral, bacterial, and parasitic infections. Despite different AuNP modification strategies, studies reported levels of detection ranging from 5.77 × 10⁻7 to 3.54 × 103 ng mL⁻1 and 15 CFU mL⁻1, high sensitivity, and short detection times (15 min to 60 min), crucial for POC tool development. This review also discussed the current limitations and future directions for establishing a robust, clinically validated diagnostic platform. Overall, the potential of integrating nanotechnology into immunosensors is emphasized to advance biotechnology and translate it into healthcare settings. Summary of the integration of AuNP-modified electrode surface into EIS-based immunosensor and its current application in detecting different infectious diseases in humans carried by viruses, bacteria, and parasites. The performance of nano-modified immunosensors was evaluated, which is crucial for advancing the development of a POC diagnostic platform, in accordance with the ‘ASSURED’ criteria introduced by the WHO.

Immune profiling has become a transformative tool in oncology, offering comprehensive information on tumor immune interactions and facilitating precision medicine. Recent advances such as mass cytometry (CyTOF), single-cell RNA sequencing (scRNA-seq), spatial transcriptomics, and liquid biopsy have greatly enhanced our ability to characterize immune heterogeneity and predict treatment responses. These innovations support the identification of new biomarkers, therapeutic targets, and resistance mechanisms, refining patient stratification and clinical results. Additionally, artificial intelligence (AI) driven models are now being employed to integrate multi-omics datasets and create predictive insights, thereby linking the gap between research and clinical decision-making. This review studies the evolution of immune profiling technologies, their integration into real-world oncology practice, and the associated technical and analytical challenges, including sample variability, data harmonization, and multi-omics integration. Although challenges such as cost, throughput, and standardization persist, the merging of advanced technologies, bioinformatics, and clinical frameworks promises to reshape cancer diagnosis, therapy selection, and disease monitoring through personalized and data-driven strategies.

Tuberculosis (TB) remains as a significant global health threat to date, with latent TB infection (LTBI) serving as a major reservoir for future active disease cases. A practical approach to an effective control and eradication of TB hence, requires an explicit identification of infected patient whom are at high risk of progressing from latent to active TB, particularly in those recently infected individuals. Current diagnostic tools however, including Tuberculin Skin Test and Interferon-Gamma Release Assays, are still lacking for their ability to critically distinguish between recent and remote infections, leading to insufficiency in optimizing targeted preventive treatment strategies. This review examines the limitations of current diagnostic tools and explores novel biomarkers to enhance distinction within the infection timeline in LTBI diagnostics. Advancement in immune profiling, dormancy antigen, along with molecular and transcriptomic approaches holds great promise to develop a diagnostic tools with better accuracy to differentiate recent from remote infections, thereby optimizing targeted interventions to improve TB control strategies. These underscores the need for further research into these emerging diagnostic tools to facilitate an effective public health strategies and contribute to the united efforts in End TB Strategy.

Autoimmune diseases are driven by dysregulated cytokine networks, where excessive cytokines such as TNF-α, IL-6, IL-17, and IL-23 promote chronic inflammation and tissue damage. While monoclonal antibodies effectively neutralise these cytokines, they face limitations including high production costs, glycosylation requirements, limited tissue penetration, and immunogenicity. Nanobodies; small, single-domain antibodies derived from camelids, offer a promising alternative due to their high stability, solubility, microbial expression, and low immunogenicity. This review examines the structural and functional advantages of nanobodies in cytokine neutralisation, with targets including TNF-α, IL-6, IL-17 A, chemokines (e.g., CXCL10), VEGF, and intracellular proteins like STAT3. Advances in artificial intelligence (AI) and machine learning (ML) for nanobody discovery, from de novo library design to affinity optimization, are highlighted. We also discuss Chemistry, Manufacturing, and Controls (CMC), regulatory pathways (FDA/EMA), and innovative delivery strategies such as oral and nanoparticle systems. Furthermore, the advantages and limitations of nanobody-based cytokine neutralisation are critically evaluated. Lastly, we review the current status of clinical trials and future directions to enhance efficacy and safety of nanobody-based therapies in autoimmune diseases.

The human leukocyte antigen (HLA, also known as the major histocompatibility complex or MHC) system, is responsible for immune monitoring of the intracellular proteome of all nucleated cells. The presentation of antigen peptides separates malignant or infected cells from their healthy counterparts and forms aberrant cells tagged as the foundation for identification. Therefore, peptide-MHC molecules can give potential diagnostic targets for cancer or infection. TCR-like antibodies recognize specific peptides that bind to MHC molecules, allowing them to target Such inaccessible cytoplasmic or nuclear tumors or virus-associated antigens. It binds to MHC, presenting peptides found on the surface of target cells. These antibodies have shown promising clinical applications in diagnosing and imaging cancer and infected cells. This review presents the current situation of TCR-like antibodies and its prospects for application in the field of intracellular antigen diagnostics. It also lists the potential application targets of TCR, like antibodies in various disease diagnoses, providing valuable information for developing diagnostic reagents and selecting targets in the future.

Cancer treatment faces ongoing challenges owing to the diverse cancer types and drawbacks of traditional therapies, prompting the search for less invasive alternatives. Photodynamic therapy (PDT) is a non-intrusive approach that uses photosensitisers (PS) to accumulate in target cancer cells and induce cell death upon light activation. This review critically evaluates the applications of two progressive PS, aminolevulinic acid (ALA) and indocyanine green (ICG), in PDT. It synthesizes findings from extensive studies and clinical trials to assess the current landscape of PDT and its potential in combination therapies. Additionally, this review explores the feasibility and rationale for integrating PDT with alternative therapies, such as sonodynamic therapy (SDT), radiodynamic therapy (RDT), photopheresis, photothermal therapy (PTT), and LED-phototherapy, to enhance treatment efficacy. ALA and ICG demonstrate significant potential for PDT owing to their selective accumulation in cancer cells and favorable safety profiles. Clinical trials have shown promising results, indicating PDT’s efficacy of PDT for various cancer types. Moreover, the combination of alternative therapies underscores PDT’s versatility and capacity for synergistic effects in cancer treatment. ALA and ICG are promising PS candidates that offer targeted cell death with minimal toxicity during PDT. The integration of PDT with alternative therapies presents a compelling avenue for enhancing the treatment outcomes and therapeutic potential in oncology. Further research and clinical validation are warranted to refine the full potential of PDT and its synergistic applications in cancer therapy.

Aging involves a series of complex physiological changes that progressively impair cellular function. While chronological aging is inevitable, biological aging is influenced by modifiable factors such as oxidative stress, telomere shortening, chronic inflammation, and mitochondrial dysfunction. Recent research highlights the potential of medicinal plants in managing age-related conditions due to their rich phytochemical content. These bioactive compounds can promote cellular repair, scavenge reactive oxygen species (ROS), enhance telomerase activity, and support tissue regeneration. Polyalthia longifolia var. angustifolia (Thw.), a member of the Annonaceae family traditionally used for rejuvenation, has demonstrated significant anti-aging properties in both yeast and animal models. In vitro and in vivo studies, in particular, provide valuable insights into the anti-aging activity of this plant and its potential applications. This review explores the aging process, outlines the pharmacological profile of P. longifolia, and highlights its key anti-aging constituents, particularly flavonoids, tannins, phenolics, and carbohydrates, which are recognized for their well-documented antioxidant, anti-inflammatory, and cellular protective properties. Moreover, P. longifolia has shown promising effects against various age-associated disorders, including diabetes, hypertension, liver and kidney dysfunctions, inflammation, and oxidative damage. These benefits are largely attributed to its ability to modulate inflammatory pathways, minimize oxidative stress, and regulate abnormal cell proliferation, thereby supporting healthy aging. With its diverse pharmacological properties and abundant bioactive compounds, P. longifolia emerges as a promising natural agent in the field of anti-aging research. Its ethnobotanical significance, phytochemical richness, and therapeutic applications suggest strong potential for development into a sustainable, plant-based strategy to mitigate aging and related health issues.

Aptamer-based immunotherapy can be a new hope for treating solid tumors with personalized and specific approaches toward cancer therapies. Aptamers are small synthetic single-stranded nucleic acids that may bring in a paradigm shift in treating solid tumors. These are highly selective drugs applied in cellular immunotherapy, cytokine modulation, and immune checkpoint suppression. This review provides an overview of the recent advances in aptamer-based technologies with specific key clinical trials involving AON-D21 and AM003. Aptamers are potently active in immune regulation and tumor targeting. However, aptamer stability and bioavailability are seriously compromised by the issues relating to renal clearance and rapid degradation through nucleases. The latter are reviewed here along with novel improvements, some of which involve chemical modifications that greatly enhance stability and prolong the circulation time; exemplary such modifications are PEGylation, cholesterol conjugation, and the synthesis of circular nucleic acids. The regulatory aspect is also crucial. For example, in addition to specific strategies to prevent drug-drug interactions (DDIs) in cancer remediation medications, this paper underscores the need of risk assessment, particularly because of immunogenicity and organ failure. The use of aptamers is expanded by the development of SOMAmers, X-aptamers, and bioinformatics. To make aptamer-based drugs a major part of cancer treatment, future research should concentrate more on resolving existing issues and expanding their beneficial uses.

Cervical cancer is the fourth most prevalent cancer among female patients globally, largely due to persistent infections with high‑risk human papillomavirus (HPV). Viral oncoproteins E6 and E7, produced by HPV, serve a role in driving cellular transformation and maintaining the malignant phenotype. T cell receptor (TCR)‑like antibodies serve as a potential diagnostic tool to capture the oncogenic peptide that is present in MHC. As these antibodies serve as innate antigen detectors, orchestrating immune responses against both cell surface and intracellular proteins. In the present study, a human domain antibody (DAB) phage library was screened by evaluating synthesized HPV18 (E6 and E7) peptide‑major histocompatibility complexes (p‑MHC‑A24) to identify target‑specific TCR‑like antibodies. The present study successfully identified three TCR‑like DABs that specifically target HPV18 (E6 and E7) p‑MHC‑A24 complexes. Characterization of the amino acid sequences in the complementarity‑determining regions 1, 2 and 3 was performed using VBASE2 and the international ImMunoGeneTics information system®/vquest databases. Evaluation of soluble TCR‑like antibodies confirmed strong and selective affinity for the targets through western blotting and ELISA. The present study aimed to clarify the specificity of TCR‑like antibodies against specific targets and demonstrated that TCR‑like antibodies may serve as early diagnostic and immunotherapeutic tools for HPV‑associated cervical cancer.

Exosomes, containing molecular constituents of their cell of origin, including proteins and nucleic acids, were first discovered in immature red blood cells in 1983. Excellent intercell communication can be achieved by shuttling these various molecules between cells. Stem cell-derived exosomes (SC-Exos) contain paracrine-soluble factors that play important roles in tissue development, homeostasis, and regeneration. This paracrine activity of SC-Exos has been found to be a predominant mechanism by which stem cell-based therapies mediate their effects on degenerative, autoimmune and/or inflammatory diseases. Compared to other types of stem cells, human embryonic stem cells (hESCs), human induced pluripotent stem cells (hiPSCs), human mesenchymal stem cells (hMSCs) are the most popular because of their efficient immunomodulatory effects. The advantages and disadvantages of using exosomes isolated from the stem cell trio for therapeutic applications are further discussed in this review.

The promising field of regenerative medicine is thrilling as it can repair and restore organs for various debilitating diseases. Mesenchymal stem cells are one of the main components in regenerative medicine that work through the release of secretomes. By adopting the use of the secretome in cell-free-based therapy, we may be able to address the challenges faced in cell-based therapy. As one of the components of cell-free-based therapy, secretome has the advantage of a better safety and efficacy profile than mesenchymal stem cells. However, secretome has its challenges that need to be addressed, such as its bioprocessing methods that may impact the secretome content and its mechanisms of action in clinical settings. Effective and standardization of bioprocessing protocols are important to ensure the supply and sustainability of secretomes for clinical applications. This may eventually impact its commercialization and marketability. In this review, the bioprocessing methods and their impacts on the secretome profile and treatment are discussed. This improves understanding of its fundamental aspects leading to potential clinical applications.

Lentiviruses are a highly robust gene delivery system capable of in vitro and in vivo gene transfer into multiple cell types. Recent fourth-generation lentiviral systems have been designed for enhanced safety, however, the increased recombination events required to produce infective lentiviral particles may reduce production efficiency. A set protocol for all types of target cells is not recommended and optimization of conditions for gene transfer into different target cells is required. In this study, we aim to evaluate the efficiency and reproducibility of lentiviral production using a fourth-generation lentiviral packaging system and identify optimal parameters for successful transduction in two different cell models, adherent and suspension cells. Lentiviral production, effect of viral volume, sustained gene expression and transduction adjuvants on adherent and suspension gene- cell models were evaluated. Transfection and transduction efficiency of lentiviruses was evaluated by fluorescence microscopy and flow cytometry. This study demonstrates that production of green fluorescent protein (GFP)- lentiviruses using the fourth-generation lentiviral packaging is consistent and reproducible. Optimal transduction of adherent cell types is achieved at lower multiplicity of infection (MOI) compared to suspension cells and produces GFP-expressing cells with higher intensity. Expression of GFP is sustained in all cell types over multiple passages. Polycation DEAE-dextran was determined to improve transduction in suspension cells, however, provides similar transduction efficiency as polybrene in adherent cells. In conclusion, fourth generation lentiviral system reproducibly generates high titre lentiviruses capable of infecting multiple cell types, however transduction protocols for different cell types require further optimization.