Biology subjects

Kolla, H. B.

Publications and source records attributed to Kolla, H. B..

10 recordsLinked to original sources

bioRxiv2026

Phylogenetic Analysis of cpn60 UT region reveal oligonucleotide regions for the detection of prominent Enterobacteriaceae pathogens

The Enterobacteriaceae pathogens like E. coli, Shigella, Klebsiella and Salmonella cause a wide range of gastrointestinal and other mucosal infections when the contaminated food and water are consumed. Conventional microbiological methods are time-consuming and laborious process to detect these pathogens. Molecular assays are fast and sensitive to detect the pathogens. But a potential target for the detection is very essential to develop a molecular assay for the rapid detection. In this study, we explored the cpn60 UT region as a potential target for the detection of significant Enterobacteriaceae members like E. coli, Shigella sp, Salmonella, Klebsiella etc. We did the phylogenetic analysis of cpn60 UT region among the Enterobacteriaceae pathogenic groups. This analysis has revealed the presence of 13 clusters and two oligonucleotide regions with highest nucleotide conservation. Targeting these regions we have designed a set of oligonucleotide primer pairs and developed a SYBR green real time PCR assay to detect the Enterobacteriaceae pathogenic members. The degenerate oligonucleotide primers designed amplified a 132 bp UT region of cpn60 gene present in Enterobacteriaceae pathogens. The sensitivity of the assay was initially investigated by 10-fold dilution series of overnight broth culture of E. coli ATCC 10536 and had a detection limit of 101 CFU/mL. The specificity was assessed by comparing the amplification plot of a set of Enterobacteriaceae pathogens and non-Enterobacteriaceae members like Staphylococcus aureus (S. aureus) and Listeria monocytogenes (L. monocytogenes) and found to detect all the Enterobacteriaceae cultures included in the study whereas amplification was not seen in S. aureus and L. monocytogenes.

microbiology↗

bioRxiv2026

Co-administration of Staphylococcus aures Fibronectin Binding Protein A N1-3 region and Lipoteichoic acid antigen mixture enhance antibody response in BALB/c mice

Staphylococcus aureus (S. aureus) is a major human pathogen that causes a wide range of superficial skin infections to the life-threatening systemic diseases. Emergence and increasing prevalence of anti-biotic resistance S. aureus strains have accelerated the need for effective vaccines. As S. aureus pathogenesis involve its surface antigens to facilitate the colonization and invasion into host, targeting these surface antigens represent a promising strategy for S. aureus vaccine development. In this study, we evaluated the efficacy of two surface antigens of S. aureus -fibronectin-binding protein A (FnBPA) and lipoteichoic acid (LTA) as vaccine candidates. We have generated a recombinant fusion protein r-FR by fusing the N-terminal region of FnBPA and rhizavidin and expressed in Escherichia coli. The recombinant protein was mixed with the biotinylated LTA to generate r-FR+LTA antigen mixture, emulsified with Freunds adjuvant as r-FR+LTA antigen mixture or r-FR alone and immunized to BALB/c mice. Mice immunized with r-FR+LTA mixture displayed a strong antibody response, strong bacterial growth inhibition in vitro and increased survival following lethal S. aureus challenge compared with mice immunized with r-FR alone. These findings suggest that incorporation of the S. aureus surface antigens like FnBPA with LTA provides improved protective efficacy and highlights the potential of a multi-antigen vaccine strategy for preventing S. aureus infections.

microbiology↗

bioRxiv2025

Active Site Heterogeneity Drives Differential Sialic Acid Metabolism in Enterobacteriaceae Pathogens

Enterobacteriaceae pathogens like E. coli, Shigella and Salmonella are mainly associated with the food poisoning and various mucosa associated infections. These pathogens invade the host and rely on the host derived nutrients for their survival. One such nutrient is sialic acid (N-acetylneuraminic acid is the common one) which is present in the mucosal linings throughout the body. Ability of these pathogens to breakdown and utilize sialic acid for their growth is a crucial factor for their survival and pathogenesis. However, the heterogeneity among these enteric pathogens remains unstudied. In this study, we explored the difference in the sialic acid utilization pattern among these pathogens and found that the interactions between the N-acetylneuraminic acid (Neu-5-Ac) and the NanA, an enzyme that catalyze the breakdown of sialic acid play a key role for this heterogeneity. Additionally, we found that the Neu-5-Ac interacted differently with the NanA from each strain because of the variations in the active sites/contact sites which made the strains to utilize sialic acid different. Finally, our results provide an insight into the current understanding of sialiobiology research by highlighting the active site/contact sites differences drived the pathogens ability to adapt to diverse host environments, potentially altering their pathogenicity and resistance to host immune responses. This study provides new insights into the evolution of Enterobacteriaceae and suggest potential targets for therapeutic intervention aimed at disrupting sialic acid utilization.

microbiology↗

bioRxiv2025

Targeted αGalCer/CD1d Therapy Engages Invariant Natural Killer T Cells to Facilitate Cardiac Healing in Chronic Doxorubicin-Induced Cardiomyopathy

Invariant natural killer T (iNKT) cells are a unique subset of innate immune cells activated by T cell receptor (TCR) signaling through CD1d molecules presenting lipid antigens. By localizing to specific tissues, iNKT cells play critical roles in maintaining homeostasis and regulating pathophysiology, making them attractive therapeutic targets for inflammatory diseases. However, their frequency in the heart and potential roles in mitigating inflammation-induced cardiac damage remain poorly understood. In this study, we examined the distribution of cardiac iNKT subsets and their response to TCR-directed alpha Galactosyl ceramide/cluster of differentiation 1d (GalCer/CD1d) complexes in a mouse model of Doxorubicin (Dox)-induced cardiotoxicity. In healthy mice, the heart harbored distinct iNKT subsets, dominated by pro-inflammatory iNKT1 cells, followed by iNKT2 subsets. Treatment with GalCer/CD1d complexes preferentially expanded the iNKT1 subset. In Dox-induced cardiotoxicity, this treatment unexpectedly increased both IFN-{gamma}, a hallmark cytokine of iNKT1 cells, and IL-10, a key anti-inflammatory mediator typically associated with regulatory iNKT subsets. Despite the expansion of iNKT1 cells and elevated IFN-{gamma} levels, GalCer/CD1d treatment did not exacerbate cardiac damage and showed modest clinical improvements compared to controls. These findings highlight the complexity of iNKT-mediated immune regulation in the heart and emphasize the need for modified TCR-directed immunotherapeutics to skew immune responses toward a regulatory phenotype. Such approaches could hold promise for treating inflammatory cardiac diseases, including Dox-induced myocarditis, myocardial infarction, and autoimmune myocarditis.

immunology↗

bioRxiv2024

In vivo Protective Efficacy of Emodin in Swiss Albino Mice Induced with Dalton Ascitic Lymphoma

Lymphoma is a tumor that affects lymphoid tissues in the body. Treating lymphoma has become challenging because of the complexity of disease pathology, drug resistance mechanisms and side effects of existing chemo and radiation therapies. Treating cancers/tumors with plant based natural compounds is gaining interest recently because of their less toxicity profiles and efficiency in controlling the disease severity. Emodin is one such compound with such anti-cancer/tumor properties. It has immunosuppressive and anti-cancer properties through multiple ways. In this study, we have studied the therapeutic effect of emodin molecule in the DAL induced lymphoma, a well-established murine model to study and test the anti-lymphoma drugs. Our data has shown an outstanding therapeutic effect of emodin in controlling the lymphoma readouts in DAL induce Swiss Albino mice. These effects were studied in comparison with a standard drug molecule called methotrexate. Furthermore, the in-silico analysis has shown that emodin as a potential drug candidate for lymphoma based on the Lipinskis rule of 5.

cancer biology↗

bioRxiv2024

Advancing Tolerogenic Immunotherapy: A Multi-Epitope Vaccine Design Targeting the CYP2D6 Autoantigen in Autoimmune Hepatitis Through Immuno-Informatics

Juvenile autoimmune hepatitis (JAIH) is a rare autoimmune disorder affecting children, characterized by the immune systems misguided attack on liver cells, primarily targeting the CYP2D6 autoantigen. This repeated attack leads to hepatic inflammation, fibrosis, and eventual liver failure. Current therapeutic strategies predominantly rely on immunosuppressive agents or whole B cell depletion antibodies, which render patients susceptible to infections and cancers. Hence, there is an urgent need for antigen-specific therapies to mitigate the severity of autoimmune hepatitis. Tolerogenic antigens represent a promising avenue in immunotherapy, capable of dampening autoimmunity. Here, we present a novel computationally designed multi-epitope tolerogenic vaccine tailored to target CYP2D6, aimed at inducing tolerogenic dendritic cells (DCs) and halting autoimmune progression in JAIH patients. To validate our approach, we have developed a similar vaccine for testing in mouse models of JAIH. The selected tolerogenic epitopes exhibit antigenicity without allergenicity or toxicity, and specifically induce IL-10 production (restricted to CD4+ T cell epitopes). In our vaccine design, tolerogenic poly-epitopes are linked with Toll-like receptor (TLR)-4-agonist, the 50S ribosomal unit, and IL-10, effectively programming DCs towards a tolerogenic state. Molecular docking and dynamic simulations have confirmed strong binding affinities and stable complexes between the vaccine structures, TLR4 and IL-10 receptor alpha (IL-10RA), indicating their potential for in vivo DC interaction and programming. Consequently, this innovative vaccine approach demands further exploration through wet lab experiments to assess its tolerogenicity, safety, and efficacy, thereby laying the groundwork for potential application in clinical settings.

immunology↗

bioRxiv2024

Unleashing the Immune Arsenal: Development of Broad-spectrum Multiepitope Bluetongue Vaccine Targeting Conserved T Cell Epitopes of Structural Proteins

Bluetongue (BT) is a severe arboviral disease affecting sheep, cows, and other wild ruminants, caused by the Bluetongue virus (BTV). The virus has evolved into over 32 serotypes, rendering existing vaccines less effective. While the structural proteins of this virus represent promising targets for vaccine development, they unfortunately exhibit high amino acid polymorphism and are laden with numerous inhibitory epitopes. Structural proteins such as VP1 and VP7 are highly conserved and may contain epitopes capable of triggering cross-reactive cell-mediated immunity (CMI). In this study, we identified highly conserved MHC-I and -II-restricted T cell epitopes within VP1, VP5, and VP7 BTV proteins and developed an effective in silico-immuno-informatics-based broad-spectrum BT multiepitope vaccine for bovine and laboratory mouse systems. The conserved epitopes utilized in the vaccines are highly antigenic, non-allergenic, non-toxic, and capable of inducing IFN-{gamma} (only CD4+ T cell epitopes). Both mouse and bovine vaccines were tethered with Toll-like receptor (TLR)-4-agonist adjuvants, beta-defensin 2, and the 50s ribosomal unit to stimulate innate immunity for CMI development. Protein-protein docking analysis revealed strong binding affinities, while extensive 100-nanosecond molecular dynamics simulations indicated stable complexes between the vaccine structures and TLR4. Vaccination simulation studies demonstrated their ability to trigger proinflammatory responses. Therefore, these novel vaccine designs necessitate further exploration through wet lab experiments to evaluate their immunogenicity, safety, and effectiveness for practical deployment in livestock.

immunology↗

bioRxiv2024

In Vitro Anti-cancer potential of Aloe emodin against Oral Squamous Cell Carcinoma

Oral squamous cell carcinoma is caused due to smoking, chewing of tobacco, high exposure of UV radiation and infectious agents such as human papilloma virus (HPV). Epigenetic dysregulation and uncontrolled cell growth are main factors that drives towards tumor development. Risk factors such as smoking, tobacco and viral agents up regulates its activity and leads to the cancer. Although chemotherapy, surgery and radiation were preferred for treating cancer but patients, feel extreme psychological stress and depression. Natural products are been investigated for their numerous diseases and in that they can be treated against cancer without side effects. Aloe emodin is one such natural molecule available, which possess various pharmacological activities for human beneficial. In this current study, we investigated the anti-cancer activity of aloe emodin in vitro. The KB 3-1 cell line were treated with different concentrations of Aloe emodin and its anti-cancer potential was determined through DNA fragmentation and cell viability assays.

cancer biology↗

bioRxiv2023

Immuno-informatics Study Identifies Conserved T Cell Epitopes in Non-structural Proteins of Bluetongue Virus Serotypes: Formulation of Computationally Optimized Next-Generation Broad-spectrum Multiepitope Vaccine

Bluetongue (BT) is a significant arboviral disease affecting sheep, cattle, goats, and wild ruminants, posing serious economic challenges to livestock industry. Control efforts have been hampered by the existence of over 32 distinct BT virus (BTV) serotypes and the absence of broad-spectrum vaccines. Some key non-structural proteins of BTV, including NS1, NS2, and NS3, exhibit notable amino acid sequence conservation. Our findings reveal that mouse MHC class I (MHC-I) CD8+ T cell epitopes are highly conserved in NS1 and NS3, while MHC-II epitopes are prevalent in all the three non-structural NS 1-3 proteins. Similarly, both class I and II Bovine Leukocyte antigen-restricted CD8+ and CD4+ T cell epitopes are conserved within NS1, NS2, and NS3 proteins. To construct in silico broad-spectrum vaccine, we subsequently screened these conserved epitopes based on antigenicity, allergenicity, toxicity, and solubility. Modeling and Refinement of the 3D structure models of vaccine constructs were achieved using protein modeling web servers. Our analysis revealed promising epitopes that exhibit strong binding affinities with low energies against two TLR receptors (TLR3 and TLR4). To ensure atomic-level stability, we evaluated the docking complexes of epitopes and receptors through all-atom molecular dynamics simulations (MDS). Encouragingly, our 100 nanoseconds MDS showed stable complexes with minimal RMSF values. Our study offers valuable insights into these conserved T cell epitopes as promising candidates for a broad-spectrum BT vaccine. We therefore encourage for their evaluation in animal models and natural hosts to assess their immunogenicity, safety, and efficacy for field use in the livestock.

bioinformatics↗

bioRxiv2020

Nutri rich cattle feed (NuCa feed) supplement with improved fiber content free of anti-nutritional factor Phytic acid

Most of the commercial ruminant feeds comprise Corn meal, Groundnut cake, Maize etc., which contain anti-nutritional factors called Phytates. They have high binding affinity to cationic minerals. This results in decrease in activity of the cattle by making minerals unavailable for absorption in the intestine. Many of the feed industries add Phytase enzyme that degrades the Phytic acid. Addition of Phytase enzyme is very complex method and increases the cost of the feed. If Phytase is not added, Phytic acid enters the ecosystem through the dung and due to microbial degradation, the Phytic acid gets converted into Phosphates. Phosphate abundance in water bodies leads to Eutrophication, which is a serious ecological issue. And also, in many parts of the country, post-harvest Paddy straw is burnt unused. This intentional stubble burning causes air pollution. We combined paddy straw and Azolla along with jaggery to prepare animal feed supplement. The feed pellets were evaluated for their nutritional composition and efficacy. The efficacy of the developed feed was evaluated by feeding it to milch cows and compared with regular feed for a period of 15 days. The Azolla-Hay feed improved the milk yield and also quality of the milk in comparison to the control group. This study revealed that Azolla in combination with paddy straw powder can be low-cost feed alternative especially during the lean period where the availability of feed is scarce.

plant biology↗