Insect cell lines are a cornerstone of recombinant protein production, providing a versatile platform for biopharmaceutical and research applications. In the early 20th century, scientists first attempted to culture insect cells in vitro, developing continuous cell lines to produce the first insect cell-derived recombinant protein, IFN-β. Initial successes, along with advancements in the use of insect cells for recombinant protein manufacturing, primarily relied on baculovirus expression vector systems (BEVSs), which enable heterologous gene expression in infected cells. Today, growing attention is focused on baculovirus-free systems based on the transfection of insect cells with plasmid DNA. This approach simplifies the final product purification process and facilitates the development of stable monoclonal cell lines that produce recombinant proteins or protein complexes, particularly virus-like particles (VLPs). Thanks to advancements in genetic engineering and the application of adaptive laboratory evolution (ALE) methods, significant strides have been made in overcoming many limitations associated with insect cell BEVSs, ultimately enhancing the reliability, yield, and quality of the biomanufacturing process. Our manuscript discusses the history of developing insect cell lines, presents various recombinant protein production systems utilizing these cells, and summarizes modifications aimed at improving insect cell lines for recombinant protein biomanufacturing. Finally, we explore their implications in pharmaceutical production, particularly on Nuvaxovid^®/Covovax, which is the latest approved vaccine developed using insect cell BEVSs for protection against SARS-CoV-2.

Galleria mellonellacationic protein 8 (GmCP8) is a hemolymph protein previously identified as an opsonin and an inhibitor of fungal proteases. In this work, we showed its bactericidal activity towardPseudomonas entomophila,Pseudomonas aeruginosa,Bacillus thuringiensis,Staphylococcus aureus, andEscherichia coliand against yeast-like fungiCandida albicans. The activity againstE. coliwas correlated with bacterial membrane permeabilization. In turn, in the case ofP. entomophila,B. thuringiensis,andC. albicans, the atomic force microscopy analysis of the microbial surface showed changes in the topography of cells and changes in their nanomechanical properties. GmCP8 also showed the inhibitory activity toward the serine protease trypsin and the metalloproteinase thermolysin. The expression of the gene encoding the GmCP8 protein did not increase either in the gut or in the fat body ofG. mellonellaafter oral infection withP. entomophila. Similarly, the amount of GmCP8 in the hemolymph ofG. mellonelladid not change in immune-challenged insects. However, when GmCP8 was injected into theG. mellonellahemocel, a change in the survival curve was observed in the infected larvae. Our results shed new light on the function of GmCP8 protein in insect immunity, indicating its role in humoral defence mechanisms.

Chemosensory protein 16 was identified in the hemolymph of Galleria mellonella as a protein with an amount increasing after oral infection with 10^3 CFU of Pseudomonas entomophila, and decreasing after infection with a higher dose (10^5 CFU) of bacteria. The expression of the CSP16 gene occurred in the fat body and in the gut and correlated with changes in the protein level in the hemolymph. The CSP16 protein inhibited P. entomophila growth in the concentration range from 0.15 to 6 nM. Additionally, the CSP16 protein showed bactericidal activity against P. entomophila, Bacillus thuringiensis, and Escherichia coli in the range of 2-18 μM, but only in the presence of protease inhibitors, otherwise it was degraded by extracellular proteases secreted by P. entomophila. We demonstrated that the bactericidal activity of CSP16 was related to its ability to perforate bacterial cellular membranes in a dose-dependent manner. The antimicrobial properties of this protein were also confirmed with the use of Atomic Force Microscopy, which showed significant changes in the topology of different bacterial cell surfaces. Finally, when CSP16 was injected in vivo into G. mellonella larvae one hour after infection with P. entomophila, more survivors were observed at particular time-points. Taking into account its immune properties and putative ability to bind bacteria-derived compounds, the possible function of CSP16 in the host-pathogen interaction is discussed.

Oncolytic viruses (OVs) are gaining traction as advanced tools in cancer therapy. They are distinguished by their ability to destroy malignant cells while sparing normal tissue specifically. In addition to their direct tumor-lysing properties, an essential benefit of oncolytic virus therapy is its capacity to activate both the innate and adaptive immune systems. To enhance these therapeutic actions, many OVs have been genetically engineered to encode immune-modulating factors that reestablish or strengthen antitumor immune responses. Recent studies show that combining OVs with other forms of immunotherapy—such as immune checkpoint inhibitors, CAR-T cells, specific T-cell receptor therapies, or autologous tumor-infiltrating lymphocytes—offers significant advances in cancer treatment. This article reviews how OVs work, discusses strategies to enhance their immunogenicity further, and presents the latest rational combinations of oncolytic viruses with other immunotherapies based on current preclinical and clinical research.

Fungal exposure and sensitization are linked to asthma severity, but their clinical significance varies by host, exposure context, and disease phenotype. This review integrates epidemiological, immunological, and clinical evidence on fungal-associated asthma, distinguishing environmental exposure, allergic sensitization, and persistent airway colonization. We examine how fungal allergens, proteases, and cell-wall components engage pattern-recognition receptors, disrupt epithelial barriers, and activate overlapping type 2 and type 17 inflammatory pathways. Studies of the airway and intestinal mycobiome offer additional perspectives on host–fungal interactions, although most human findings remain associative and do not establish causality or clinical utility. Severe asthma with fungal sensitization (SAFS) and allergic bronchopulmonary aspergillosis (ABPA) are distinct clinical contexts that require different diagnostic and therapeutic approaches. Antifungal treatment has an established role in ABPA, whereas evidence in SAFS is inconsistent; biologic selection should be guided by clinical phenotype and established eligibility criteria rather than fungal sensitization alone. We also synthesize our published biochemical, epithelial-cell, and murine studies of seven plant-associated fungal and oomycete taxa from eastern Poland. These findings support further investigation of their allergenic and pro-inflammatory potential but do not establish clinical allergenicity or predict treatment response in humans. Priorities include standardized exposure assessment, validated diagnostic reagents, and longitudinal studies linking fungal measurements to clinical outcomes. Expanded allergen panels and mycobiome-based approaches require prospective validation before routine clinical implementation.