Human Beta-defensin Research Articles

In-silico immunoinformatic vaccine design for Treponema denticola ergothionase.

Treponema denticola, a well-studied oral spirochete, adheres, invades, and damages periodontal tissues - gram-negative, anaerobic Treponema denticola. In previous research, sub-gingival spirochetes have correlated positively with dental plaque score, pocket, and clinical attachment level measurements. Hence, the study aims to design an immunoinformatic vaccine using a reverse vaccinology approach against Treponema denticola ergothionase. Protein Data Bank provided the FASTA amino acid sequence of Treponema denticola. Antigenicity, toxicity, and stability of discovered T-cell epitopes were evaluated to develop 6S7Q B and A multiepitope vaccination design. The Vaccine's dual major histocompatibility complex (MHC I and II) binding epitopes were also predicted. The designed Vaccine's identified epitope sequence and secondary structure were then predicted and validated. Protein-protein interactions involving ergothionase and human beta-defensins were investigated using molecular docking. The designed Vaccine had high antigenicity, toxicity, and stability. The Vaccine's three-dimensional structure demonstrated a significant association with beta-defensin. Its low binding energy score of -827.6 kcal/mol indicates that the immune system will respond favorably to the antigen. In this research, we employed immunoinformatic techniques to create a reverse vaccination effort to develop an in-silico vaccine.

Synergistic role of Mycobacterium indicus pranii and human beta Defensin-2 as adjunctive therapy against Mycobacterium tuberculosis

Host-directed therapies (HDT) via modulation of specific host responses like inflammation can limit mycobacterial infection. HDTs could be included in current TB therapy as an adjunct to increase bacterial clearance and limit tissue damage to control spread. Individually, Mycobacterium indicus pranii (MIP) and human beta defensin-2 (hBD-2) are promising therapies for tuberculosis (TB). They can directly target the TB bacilli and enhance cell-mediated immune responses, which is limiting with conventional drugs. Therefore, our study investigated the combined application of MIP and hBD-2 to evaluate their efficacy in clearing infections caused by Mycobacterium smegmatis (M.smeg) and Mycobacterium tuberculosis (M.tb) (both avirulent; H37Ra and virulent strain; H37Rv) in THP-1 cells and human monocyte-derived macrophages (MDMs). A strong pro-inflammatory response was observed against the combination of MIP and hBD-2 which also correlated with a significant reduction in the bacterial load. This combination further showed protection against M.tb by enhancing pyroptosis in the infected cells. The study suggests the combined use of these potent immunomodulators, which could be employed as an effective mode of therapy as adjuvants against mycobacterial infections after validation in a suitable animal model.

Correlation and Clinical Significance of HBD-2 and CXCL-1/2 Levels at Skin Lesions with Psoriasis Vulgaris Severity

ABSTRACT Objective This study was performed to explore the clinical significance of the expression of human beta-defensin 2 (HBD-2) and chemokine ligand 1/2 (CXCL-1/2) in psoriasis vulgaris. Methods This study retrospectively included the study group (n = 160) and control group (n = 100) for analysis. The levels of inflammatory indicators, blood biochemical indicators, and immune indicators using ELISA. The psoriasis area and severity index (PASI) was used to evaluate disease severity. Levels of HBD-2, CXCL-1, CXCL-2 and CCL20 were determined by RT-PCR. The correlations of HBD-2, CXCL-1 and CXCL-2 levels with CCL20 and PASI scores were analyzed. The diagnostic value of HBD-2, CXCL-1 and CXCL-2 in psoriasis vulgaris was analyzed by ROC curve. Results HBD-2, CXCL-1 and CXCL-2 were highly expressed in the lesions of psoriasis vulgaris patients, and were positively correlated with CCL20 and PASI score. HBD-2, CXCL-1 and CXCL-2 alone or in combination had high diagnostic value for psoriasis vulgaris and severe psoriasis, and the combined diagnostic value of the three was higher than that of a single indicator. Conclusion HBD-2, CXCL-1, and CXCL-2 levels are closely related to the severity of psoriasis vulgaris and can effectively diagnose the occurrence and progression of psoriasis vulgaris.

Targeting Polyprotein to Design Potential Multiepitope Vaccine against Omsk Hemorrhagic Fever Virus (OHFV) by Evaluating Allergenicity, Antigenicity, and Toxicity Using Immunoinformatic Approaches.

Omsk Hemorrhagic Fever Virus (OHFV) is an RNA virus with a single-stranded, positive-sense genome. It is classified under the Flaviviridae family. The genome of this virus is 98% similar to the Alkhurma hemorrhagic fever virus (AHFV), which belongs to the same family. Cases of the virus have been reported in various regions of Saudi Arabia. Both OHFV and AHFV have similarities in pathogenic polyprotein targets. No effective and licensed vaccines are available to manage OHFV infections. Therefore, an effective and safe vaccine is required that can activate protective immunity against OHFV. The current study aimed to design a multiepitope subunit vaccine against the OHFV utilizing several immunoinformatic tools. The polyprotein of OHFV was selected and potent antigenic, non-allergenic, and nontoxic cytotoxic T-lymphocyte (CTL), helper T-lymphocyte (HTL), and linear B-lymphocyte (LBL) epitopes were chosen. After screening, eight (8) CTL, five (5) HTL, and six (6) B cell epitopes were joined with each other using different linkers. Adjuvant human beta defensin-2 was also linked to the epitopes to increase vaccine antigenic and immunogenic efficiency. The designed vaccine was docked with Toll-like receptor 4 (TLR4) as it activates and induces primary and secondary immune responses against OHFV. Codon optimization was carried out, which resulted in a CAI value of 0.99 and 53.4% GC contents. In addition, the construct was blindly docked to the TLR4 immune receptor and subjected to conformational dynamics simulation analysis to interpret the intricate affinity and comprehend the time-dependent behavior. Moreover, it was predicted that immune responses to the developed vaccine construct reported formation of strong humoral and cellular immune cells. Therefore, the proposed vaccine may be considered in experimental assays to combat OHFV infections. Laboratory experiments for the above predictions are essential in order to evaluate the effectiveness, safety, and protective properties of the subject in question.

Enhanced Antimicrobial Peptide Response Following Bacillus Calmette-Guerin Vaccination in Elderly Individuals.

Antimicrobial peptides are an important component of host defense against Mycobacterium tuberculosis. However, the ability of BCG to induce AMPs as part of its mechanism of action has not been investigated in detail. We investigated the impact of Bacillus Calmette-Guerin (BCG) vaccination on circulating plasma levels and TB-antigen stimulated plasma levels of AMPs in a healthy elderly population. We assessed the association of AMPs, including Human Beta Defensin 2 (HBD-2), Human Neutrophil Peptide 1-3 (HNP1-3), Granulysin, and Cathelicidin (LL37), in circulating plasma and TB-antigen stimulated plasma (using IGRA supernatants) at baseline (pre-vaccination) and at Month 1 and Month 6 post vaccination. Post BCG vaccination, both circulating plasma levels and TB-antigen stimulated plasma levels of AMPs significantly increased at Month 1 and Month 6 compared to pre-vaccination levels in the elderly population. However, the association of AMP levels with latent TB (LTB) status did not exhibit statistical significance. Our findings indicate that BCG vaccination is linked to heightened circulating levels of AMPs in the elderly population, which are also TB-antigen-specific. This suggests a potential mechanism underlying the immune effects of BCG in enhancing host defense against TB.

Human Beta Defensin-2 mRNA and Proteasome Subunit β Type 8 mRNA Analysis, Useful in Differentiating Skin Biopsies from Atopic Dermatitis and Psoriasis Vulgaris Patients.

(1): Atopic dermatitis and psoriasis vulgaris are chronic, inflammatory diseases. Clinical presentation usually leads to a proper diagnosis, but sometimes neither clinical examination nor histopathological evaluation can be conclusive. Therefore, we aimed to build up a novel diagnostic tool and check it for accuracy. The main objective of our work was to differentiate between healthy skin (C), atopic dermatitis (AD) and psoriasis vulgaris (PV) biopsies on the base of involucrin (IVL) and human β-defensin-2 (hBD-2) concentrations and their mRNA, as well as mRNA for TPP2 and PSMB8. (2): ELISA for IVL and hBD-2 proteins and Real-time PCR for the relative expression of mRNA for: IVL (IVL mRNA), hBD-2 (hBD-2 mRNA), PSMB8 (PSMB8 mRNA) and TPP2 (TPP2 mRNA), isolated from skin biopsies taken from AD and PV patients and healthy volunteers were performed. (3): hBD-2 mRNA and PSMB8 mRNA correlated with some parameters of clinical assessment of inflammatory disease severity. hBD-2 mRNA expression, exclusively, was sufficient to distinguish inflammatory skin biopsies from the healthy ones. (4): hBD-2 mRNA and PSMB8 mRNA analysis were the most valuable parameters in differentiating AD and PV biopsies.

Acute salivary antimicrobial peptide secretion response to different exercise intensities and durations.

Antimicrobial peptides, key players of innate mucosal immunity in the oral cavity, exert antibacterial and bacteriolytic effects. This study aimed to clarify the effects of acute exercise at different intensities and durations on salivary antimicrobial peptide levels. In a randomized crossover trial, 14 young healthy untrained men performed intensity trials (cycling at 35%, 55%, and 75% of maximal oxygen uptake [VO2max] for 30 min) and duration trials (cycling at 55% VO2max for 30, 60, and 90 min). Saliva samples were collected at baseline and 0 and 60 min after exercise. In intensity trials, the change in salivary Lactoferrin levels from baseline to 0 min after 30-min exercise was greater at 75% VO2max exercise intensity compared to that at 35% VO2max. Furthermore, the change in salivary human beta defensin-2 (HBD-2) levels was greater at 75% VO2max compared to that at 35% and 55% VO2max. Salivary Lysozyme levels increased after exercise, independent of exercise intensity. However, salivary LL-37 levels did not change after exercise at any intensity. Additionally, in duration trials, the change in salivary levels of LL-37 and HBD-2 from baseline to 0 min after exercise at 55% VO2max was greater after 60 min and 90 min of exercise compared to that after 30 min of exercise. However, salivary Lactoferrin and Lysozyme levels increased after exercise, independent of exercise duration. Our findings suggest that secretory responses to acute exercise with exercise intensity and duration differ among salivary antimicrobial peptides.

Systemic Factors Effecting Human Beta-Defensins in Oral Cavity.

Human beta-defensins are host defense peptides with broad antimicrobial and inflammatory functions. In the oral cavity, these peptides are produced mainly by the keratinocytes of the epithelium; however, fibroblasts, monocytes, and macrophages also contribute to oral human beta-defensin expressions. The resident and immune cells of the oral cavity come into contact with various microbe-associated molecular patterns continuously and simultaneously. The overall antimicrobial cellular response is highly influenced by local and environmental factors. Recent studies have produced evidence showing that not only systemic chronic diseases but also systemic factors like hyperglycemia, pregnancy, the long-term use of certain vitamins, and aging can modulate oral cellular antimicrobial responses against microbial challenges. Therefore, the aim of this narrative review is to discuss the role of systemic factors on oral human beta-defensin expressions.

Exploring the nuclear proteins, viral capsid protein, and early antigen protein using immunoinformatic and molecular modeling approaches to design a vaccine candidate against Epstein Barr virus

The available Epstein Barr virus vaccine has tirelessly harnessed the gp350 glycoprotein as its target epitope, but the result has not been preventive. Right here, we designed a global multi-epitope vaccine for EBV; with special attention to making sure all strains and preventive antigens are covered. Using a robust computational vaccine design approach, our proposed vaccine is armed with 6–16 mers linear B-cell epitopes, 4–9 mer CTL epitopes, and 8–15 mer HTL epitopes which are verified to induce interleukin 4, 10 & IFN-gamma. We employed deep computational mining coupled with expert intelligence in designing the vaccine, using human Beta defensin-3—which has been reported to induce the same TLRs as EBV—as the adjuvant. The tendency of the vaccine to cause autoimmune disorder is quenched by the assurance that the construct contains no EBNA-1 homolog. The protein vaccine construct exhibited excellent physicochemical attributes such as Aliphatic index 59.55 and GRAVY − 0.710; and a ProsaWeb Z score of − 3.04. Further computational analysis revealed the vaccine docked favorably with EBV indicted TLR 1, 2, 4 & 9 with satisfactory interaction patterns. With global coverage of 85.75% and the stable molecular dynamics result obtained for the best two interactions, we are optimistic that our nontoxic, non-allergenic multi-epitope vaccine will help to ameliorate the EBV-associated diseases—which include various malignancies, tumors, and cancers—preventively.

An integrated comparative genomics, subtractive proteomics and immunoinformatics framework for the rational design of a Pan-Salmonella multi-epitope vaccine.

Salmonella infections pose a significant global public health concern due to the substantial expenses associated with monitoring, preventing, and treating the infection. In this study, we explored the core proteome of Salmonella to design a multi-epitope vaccine through Subtractive Proteomics and immunoinformatics approaches. A total of 2395 core proteins were curated from 30 different isolates of Salmonella (strain NZ CP014051 was taken as reference). Utilizing the subtractive proteomics approach on the Salmonella core proteome, Curlin major subunit A (CsgA) was selected as the vaccine candidate. csgA is a conserved gene that is related to biofilm formation. Immunodominant B and T cell epitopes from CsgA were predicted using numerous immunoinformatics tools. T lymphocyte epitopes had adequate population coverage and their corresponding MHC alleles showed significant binding scores after peptide-protein based molecular docking. Afterward, a multi-epitope vaccine was constructed with peptide linkers and Human Beta Defensin-2 (as an adjuvant). The vaccine could be highly antigenic, non-toxic, non-allergic, and have suitable physicochemical properties. Additionally, Molecular Dynamics Simulation and Immune Simulation demonstrated that the vaccine can bind with Toll Like Receptor 4 and elicit a robust immune response. Using in vitro, in vivo, and clinical trials, our findings could yield a Pan-Salmonella vaccine that might provide protection against various Salmonella species.

In-silico formulation of a next-generation polyvalent vaccine against multiple strains of monkeypox virus and other related poxviruses.

Mpox (formerly known as monkeypox) virus and some related poxviruses including smallpox virus pose a significant threat to public health, and effective prevention and treatment strategies are needed. This study utilized a reverse vaccinology approach to retrieve conserved epitopes for monkeypox virus and construct a vaccine that could provide cross-protection against related viruses with similar antigenic properties. The selected virulent proteins of monkeypox virus, MPXVgp165, and Virion core protein P4a, were subjected to epitope mapping for vaccine construction. Two vaccines were constructed using selected T cell epitopes and B cell epitopes with PADRE and human beta-defensins adjuvants conjugated in the vaccine sequence. Both constructs were found to be highly antigenic, non-allergenic, nontoxic, and soluble, suggesting their potential to generate an adequate immune response and be safe for humans. Vaccine construct 1 was selected for molecular dynamic simulation studies. The simulation studies revealed that the TLR8-vaccine complex was more stable than the TLR3-vaccine complex. The lower RMSD and RMSF values of the TLR8 bound vaccine compared to the TLR3 bound vaccine suggested better stability and consistency of hydrogen bonds. The Rg values of the vaccine chain bound to TLR8 indicated overall stability, whereas the vaccine chain bound to TLR3 showed deviations throughout the simulation. These results suggest that the constructed vaccine could be a potential preventive measure against monkeypox and related viruses however, further experimental validation is required to confirm these findings.

The Significance of Measuring Human Beta Defensin-2 in Patients with Diabetic Foot Ulcer

Background: Approximately one out of every four diabetic patients will acquire a diabetic foot ulcer (DFU) in their lifetime. Human beta-defensin (HBD) promotes wound healing. Objective: To find the correlation between HBD-2 and ulcer grade, diabetic foot infection, and the type of bacterial isolates recovered from bacteriological culture. Methods: We included forty-nine patients with DFU and obtained blood samples and wound swabs from each participant between October 2023 and December 2023. We measure HBA1c using the ARCHITECT c4000 system, and HBD-2 using the ELISA technique. The classification of DFU was done based on Wagner’s method. Swabs from foot ulcers are used for isolation and preliminary identification of bacteria based on standard guidelines. The VITEK® 2 system confirmed the diagnosis. Results: The patients' mean age was 57.31 years, and the male/female ratio was 1.57. Grade 3 was the most common type (57.1%). We observed the highest significant level of HBD-2 in grade one, non-infected DFU patients, and ulcers infected with gram-positive bacteria. Patients infected with Staphylococcus aureus showed the highest HBD-2 level according to the type of isolate, while patients infected with Proteus mirabilis showed the lowest level. Conclusions: HBD-2 levels might reflect the impaired or dysregulated immune response in patients with type 2 diabetes mellitus (T2DM) and have a negative impact on wound healing. The type of bacteria influenced this level, with Staphylococcus aureus infections reporting the highest level.

Effectiveness of 650 nm red laser photobiomodulation therapy to accelerate wound healing post tooth extraction

After tooth extraction, there can be consequences involving injury to the tissue surrounding the extracted tooth, which may lead to severe problems such as inflammation and infection. The wound healing process comprises inflammation, proliferation, and remodeling phases. Photobiomodulation is a therapy form that utilizes the interaction of a light source with tissue. This interaction can activate an increase in Adenosine Triphosphate (ATP), which subsequently triggers a chain reaction leading to the creation of new blood vessels and an increase in the number of fibroblasts. This study used a red laser light source with a power of 3.32 ± 0.01 mW, delivering a dose of 3.5 J to patients for extraction indications. The parameters observed included Interleukin 1_ (IL-1_), Prostaglandin E2 (PGE2), Human Beta defensin 2 (HBD2), and Gingival Index (GI). The results of testing saliva samples using the enzyme-linked immunosorbent test (ELISA) for the parameters IL-1_, PGE2, and HBD2 show a significant influence between the control and therapy groups. Meanwhile, GI revealed a significant influence of therapy on the wound-healing process. Using the Mann-Whitney U test, on day 1, the p-value was found to be 0.32, indicating no significant deference between the control and therapy groups. However, on the third day after the therapy was administered, the p-value was obtained as 0.01, signifying a significant deference between the control and therapy groups. On day 5, a p-value of 0.034 was obtained, signifying a significant deference between the control and therapy groups. Based on the research results, it can be observed that there is a decrease in the values of IL-1_, PGE2, HBD2, and GI. This indicates that local immune cells, including resident macrophages, are activated by pro-inflammatory mediators released in response to injury, and they play an essential role in accelerating wound healing.

Status of salivary human beta defensin-2 in oral potentially malignant disorders and oral cancer: Quest for a novel non-invasive biomarker

Background: Oral cancer ranks sixth among all the types of cancers globally and contributes to significant mortality and morbidity. Inflammation is known to play an important role in tumorigenesis. Human Beta Defensins are a type of AMP that play a role as chemo attractive, antimicrobial, and antitumor agents and also act as immunomodulators. They have also been demonstrated in cancer cell lines. Beta defensins act as tumor suppressor genes by manipulating the tumor microenvironment. The existing literature on human beta defensin-2 activity is scarce. There exists no literature on the comparison of the level of salivary human beta defensin-2 between subjects with oral potentially malignant disorders and oral cancer. Saliva contains constituents that reflect the physiologic state of the body. This can be utilized for rapid and atraumatic diagnosis of diseases owing to its non-invasive nature of collection. Aims and Objectives: The aim of the present study was to assess the level of human beta defensin-2 in the saliva of subjects with oral potentially malignant disorders and oral cancer and compare them with levels in healthy subjects. Materials and methods: The study sample included 75 subjects who were divided into three groups consisting of healthy subjects, subjects with oral potentially premalignant disorders and subjects with oral cancer. Results: The mean salivary Human beta defensin-2 level in subjects with oral cancer was significantly higher than in healthy controls and subjects with oral potentially malignant disorders. The level was highest in the oral cancer group and least in the control group. This difference among the 3 groups was statistically significant. In the group with premalignant disorders, the variation in the level of salivary human beta defensin-2 according to the type of lesion was not statistically significant. Conclusion: This study highlights the diagnostic role of hBD-2 in saliva. The presence of Human beta defensin-2 in the saliva of healthy controls points to its role in the maintenance of mucosal integrity. Elevation in the level of hBD-2 in oral potentially malignant disorders and a further increase in oral cancer indicate the potential use of hBD-2 as a biomarker in early diagnosis of oral cancer. Use of saliva as the diagnostic fluid aids in establishing a non-invasive and atraumatic means of diagnosis.

Change in Tissue Microbiome and Related Human Beta Defensin Levels Induced by Antibiotic Use in Bladder Carcinoma.

It is now generally accepted that the success of antitumor therapy can be impaired by concurrent antibiotic therapy, the presence of certain bacteria, and elevated defensin levels around the tumor tissue. The aim of our current investigation was to identify the underlying changes in microbiome and defensin levels in the tumor tissue induced by different antibiotics, as well as the duration of this modification. The microbiome of the tumor tissues was significantly different from that of healthy volunteers. Comparing only the tumor samples, no significant difference was confirmed between the untreated group and the group treated with antibiotics more than 3 months earlier. However, antibiotic treatment within 3 months of analysis resulted in a significantly modified microbiome composition. Irrespective of whether Fosfomycin, Fluoroquinolone or Beta-lactam treatment was used, the abundance of Bacteroides decreased, and Staphylococcus abundance increased. Large amounts of the genus Acinetobacter were observed in the Fluoroquinolone-treated group. Regardless of the antibiotic treatment, hBD1 expression of the tumor cells consistently doubled. The increase in hBD2 and hBD3 expression was the highest in the Beta-lactam treated group. Apparently, antibiotic treatment within 3 months of sample analysis induced microbiome changes and defensin expression levels, depending on the identity of the applied antibiotic.

The Papain-like Protease Domain of Severe Acute Respiratory Syndrome Coronavirus 2 Conjugated with Human Beta-Defensin 2 and Co1 Induces Mucosal and Systemic Immune Responses against the Virus.

Most of the licensed vaccines against SARS-CoV-2 target spike proteins to induce viral neutralizing antibodies. However, currently prevalent SARS-CoV-2 variants contain many mutations, especially in their spike proteins. The development of vaccine antigens with conserved sequences that cross-react with variants of SARS-CoV-2 is needed to effectively defend against SARS-CoV-2 infection. Given that viral infection is initiated in the respiratory mucosa, strengthening the mucosal immune response would provide effective protection. We constructed a mucosal vaccine antigen using the papain-like protease (PLpro) domain of non-structural protein 3 of SARS-CoV-2. To potentiate the mucosal immune response, PLpro was combined with human beta-defensin 2, an antimicrobial peptide with mucosal immune adjuvant activity, and Co1, an M-cell-targeting ligand. Intranasal administration of the recombinant PLpro antigen conjugate into C57BL/6 and hACE2 knock-in (KI) mice induced antigen-specific T-cell and antibody responses with complement-dependent cytotoxic activity. Viral challenge experiments using the Wuhan and Delta strains of SARS-CoV-2 provided further evidence that immunized hACE2 KI mice were protected against viral challenge infections. Our study shows that PLpro is a useful candidate vaccine antigen against SARS-CoV-2 infection and that the inclusion of human beta-defensin 2 and Co1 in the recombinant construct may enhance the efficacy of the vaccine.

The Pseudomonas aeruginosa 1244 pilin glycan increases susceptibility to human beta defensin 2 but enhances twitching motility.

SHANIA DAVIS & JOSEPH HORZEMPA, Department of Biomedical Sciences, West LibertyUniversity, West Liberty, WV USA. The Pseudomonas aeruginosa 1244 pilin glycan increasessusceptibility to human beta defensin 2 but enhances twitching motility. Pseudomonas aeruginosa is an ESKAPE (highly drug resistant) pathogen that is commonlyacquired during hospital stays. This bacterium produces type IV pili which are adhesins andmotor appendages that mediate a surface motility referred to as “twitching.” These pili arepolymers of protein subunits referred to as pilin. The pilin subunit of P. aeruginosa 1244 isglycosylated. Previous studies have shown that the pilin glycan affects the efficiency oftwitching motility under certain conditions and the surface polarity of these fibers. Becausemodulation of the bacterial surface charge has been shown to mediate defensin resistance inother organisms, we tested whether pilin glycosylation affected sensitivity to human betadefensin 2 (HBD-2). Interestingly, the isogenic mutant strain lacking the pilin glycan (1244G7)showed increased resistance to HBD-2 compared to wild type bacteria and those completelylacking a type IV pilus (1244.47). This increased resistance to HBD-2 could explain data from arecent study in which various P. aeruginosa clinical strains were isolated that had pilinglycosylation defects. We also confirmed and extended previous findings that indicated that the1244 pilin glycan enhances twitching motility and we showed that this is especially true onpositively charged surfaces (poly L-lysine coated plastic). However, the positively chargedsurface was also capable of enhancing twitching of strains producing non-glycosylated pili or piliwith an incomplete glycan (1244.2.1).

Multiepitope subunit vaccine against Colorado tick fever virus by using reverse vaccinology approach

Despite being a medically significant tick-borne virus, little research has been done on the Colorado tick fever virus. This virus is a member of the Spinareoviridae family and is mainly spread in the western parts of the United States and southwest Canada. It is classified as a Coltivirus. The main vector of the virus, Dermacentor andersoni, a wood tick found in the Rocky Mountains, determines patterns of viral spread. Furthermore, no effective antiviral treatment or vaccine has yet been created to guard against viral infections. An alternate method of preventing viral infections is vaccination. To stop a potential pandemic in the future, a thermodynamically stable and economical vaccination should be discovered. The goal of this study was to develop a potential vaccine against the Colorado tick fever virus. We seek to identify epitopes within two critical proteins, the Colorado tick fever virus's structural protein VP9 and the RNA-directed RNA polymerase VP1, to predict the precise locations that B and T cells will recognize by using a variety of reverse vaccinology and Computational techniques. Through modelling, the epitopes were deliberately engineered to connect with their corresponding HLA molecules, serving as the foundation for a multi-epitope subunit vaccination. This meticulously crafted vaccination candidate was created by joining B and T cell epitopes that had been extensively examined and linked with the appropriate linkers. Numerous physiological characteristics, including molecular weight, instability index, and aliphatic index, were assessed for the vaccine model, it was 3D modelled to dock with TLR-4. Previous research suggested that adding human beta-defensin as an adjuvant to the vaccine sequence at the N-terminus might improve immunogenicity. Additionally, the C-IMMSIM server ran immunological simulations to confirm the final vaccine design. The immunological simulation analysis anticipated a natural immune response. E. coli was used as the host for in-silico cloning, and the 0.96 CAI value obtained indicates that the vaccine construct will express itself to its fullest extent in the E. coli host. The binding mode of vaccine construct has been investigated via molecular docking approach against TLR4 human receptor protein. Vaccine created in this study shown efficacious and immunogenic response during immune simulation. Furthermore, this was more accurately validated through molecular dynamic simulations where the receptor molecule strongly bonds to the active residues of the epitope during 200 ns simulation time intervals. No certain changes were inferred among both receptor molecules and epitope during this time interval and remains static. Thus, these investigations result in determining the effectiveness in treating infections caused by the Colorado tick fever virus, which will allow experimentalists for its further validation to tackle this disease.

Abstract 3105: Computational integration with phage library to optimize clones of DEFB4A antibodies for cancer diagnostics and therapeutics

Abstract DEFB4A, a human beta-defensin, functions as a potential cancer biomarker, with elevated expression observed in various cancers, making it a candidate for diagnostic and prognostic applications. Additionally, its role in immune modulation suggests that DEFB4A may hold promise as a target for novel cancer treatments, particularly in the context of immunotherapy. This study aimed to develop a stable and high-affinity DEFB4A antibody for clinical and pre-clinical use. To achieve this, a combination of high-diversity phage libraries and computation-assisted analysis was employed to generate single-chain variable fragments (ScFv) clones specifically targeting DEFB4A. This research presents a robust approach for the generation of DEFB4A-targeting single-chain variable fragments (ScFv) from a phage library characterized by a high diversity of 109 unique clones and high titer of 1013 cfu/mL. Computational optimization was then applied to enhance the affinity, stability, and expression of these candidates through rational design and virtual mutagenesis programs. The binding activity of the optimized mutants was validated using biophysical methods such as surface plasmon resonance (SPR) and thermal stability assays. The process showed a remarkable 100-fold increase in affinity and stability. Additionally, the selected ScFv clone was further employed in immunohistochemistry (IHC) to detect DEFB4A in breast, prostate, and gastric cancers, showing higher expression levels compared to initial clones. This study highlights the synergy of high-diversity phage libraries and computational methods in crafting DEFB4A antibodies with enhanced affinity, stability, and expression. The findings advance the field of antibody engineering, emphasizing the crucial integration of experimental and computational approaches. This work lays the groundwork for the development of high-performance biologics with diagnostic and prognostic applications across diverse cancer types. Furthermore, it demonstrates the possibility of integrating computational approaches in antibody development, offering the promise of expediting the process in the future. Citation Format: Yichen Guo, Jina Yom, Andy (Xi) Han, Zhaoying Guo, Eden Zewdu, Rachel Gonzalez, Tianli Qu, Bailey Gilmore, Xuan Liu, Wei Fu, Xiaomin Hu. Computational integration with phage library to optimize clones of DEFB4A antibodies for cancer diagnostics and therapeutics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3105.

Serum levels of antimicrobial peptides (Cathelicidins and Beta Defensins-1) in patients with periodontitis

Background: periodontitis is a multifactorial oral inflammatory disease characterized by the gradual loss of bone and eventual tooth loss. It starts with microbes and is then influenced by the environment. A diverse family of host defense major compounds known as antimicrobial peptides react quickly to combat microbial invasion and challenge. These little cationic peptides are crucial for the development of innate immunity. The goal of this study was to evaluate the blood levels of healthy individuals and patients with periodontitis for cathelicidins and human beta-defensin-1. In this case-control study, 35 healthy volunteers (matched exactly by age and sex to the patients) and 50 periodontitis patients (aged 20 to 59) participated. In this investigation, periodontal parameters such as plaque index, gingival index bleeding on probing, probing pocket depth, and clinical attachment loss were employed. The levels of cathelicidins and human beta-defensin-1 in patients and controls were estimated using ELISA after blood samples from all individuals were taken. The current findings showed that the mean levels of cathelicidin and human beta defensin-1 were significantly higher (P˂0.01) in the patient group compared to the control group, and that there was no significant correlation with all clinical periodontal parameters. These findings support the notion that antimicrobial peptides play a crucial role in periodontitis' inflammatory process.