Gayathri Ravichandran

A mini review on immune role of chemokines and its receptors in snakehead murrel Channa striatus

ABSTRACT Chemokines are ubiquitous cytokine molecules involved in migration of cells during inflammation and normal physiological processes. Though the study on chemokines in mammalian species like humans have been extensively studied, characterization of chemokines in teleost fishes is still in the early stage. The present review provides an overview of chemokines and its receptors in a teleost fish, Channa striatus. C. striatus is an air breathing freshwater carnivore, which has enormous economic importance. This species is affected by an oomycete fungus, Aphanomyces invadans and a Gram negative bacteria Aeromonas hydrophila is known to cause secondary infection. These pathogens impose immune changes in the host organism, which in turn mounts several immune responses. Of these, the role of cytokines in the immune response is immense, due to their involvement in several activities of inflammation such as cell trafficking to the site of inflammation and antigen presentation. Given that importance, chemokines in fishes do have significant role in the immunological and other physiological functions of the organism, hence there is a need to understand the characteristics, activities and performace of these small molecules in details. Graphical abstract Figure. No caption available. HighlightsInformation of chemokines in teleost is still in the early stage.This review provides an overview of chemokines and its receptors in a teleost, Channa striatus.Pathogen in the teleost mounts an immune response involving cytokines.Chemokines in fishes have significant role in the immunological functions.Chemokine receptors have a comprehensive perception on the chemokine immune complex.

Pellino-1 derived cationic antimicrobial prawn peptide: Bactericidal activity, toxicity and mode of action

The antimicrobial peptides (AMPs) are multifunctional molecules which represent significant roles in the innate immune system. These molecules have been well known for decades because of their role as natural antibiotics in both invertebrates and vertebrates. The development of multiple drug resistance against conventional antibiotics brought a greater focus on AMPs in recent years. The cationic peptides, in particular, proven as host defense peptides and are considered as effectors of innate immunity. Among the various innate immune molecules, functions of pellino-1 (Peli-1) have been recently studied for its remarkable role in specific immune functions. In our study, we have identified Peli-1 from the cDNA library of freshwater prawn Macrobrachium rosenbergii (Mr) and analyzed its features using various in-silico methods. Real time PCR analysis showed an induced expression of MrPeli-1 during white spot syndrome virus (WSSV), bacteria (Vibrio harveyi) and lipopolysaccharide (LPS) from Escherichia coli challenge. Also, a cationic AMP named MrDN was derived from MrPeli-1 protein sequence and its activity was confirmed against various pathogenic bacteria. The mode of action of MrDN was determined to be its membrane permeabilization ability against Bacillus cereus ATCC 2106 as well as its DNA binding ability. Further, scanning electron microscopic (SEM) images showed the membrane disruption and leakage of cellular components of B. cereus cells induced by MrDN. The toxicity of MrDN against normal cells (HEK293 cells) was demonstrated by MTT and hemolysis assays. Overall, the results demonstrated the innate immune function of MrPeli-1 with a potential cationic AMP in prawn.

Gene expression and in silico analysis of snakehead murrel interleukin 8 and antimicrobial activity of C-terminal derived peptide WS12

Chemokines have been known for their wide range of functions including chemoattractant property in humans and other vertebrate organisms. They act as a bridge between innate and adaptive immune system. In the present study, we have identified a CXC chemokine from the cDNA library of C. striatus; on the basis of orthology study, it was found highly identical to interleukin 8 (IL8). The bioinformatics analysis of the chemokine revealed the presence of a typical γ-core domain and a CXC motif at the N-terminal region of the molecule. Based on the amphipathic nature at the C terminal helical region of CstIL8 and their antimicrobial propensity observed during bioinformatics analysis, a short peptide namely WS12 comprising 12 amino acid residues was predicted and synthesized to determine its antimicrobial activity. The peptide WS12 was active against Bacillus cereus, a Gram positive bacterium. Scanning electron microscopy (SEM) results showed bleb-like formation on the surface of the bacteria after the treatment of WS12. Additionally, WS12 did not exhibit any cytotoxic activity against the fish leukocytes. Further, the gene expression studies also revealed that CstIL8 was expressed significantly in liver of Channa striatus (Cst) at basal level. The immune challenge studies with pathogens and immune-stimulants revealed an increase in the mRNA levels at different time points post-challenge. Hence, it is possible to conclude that WS12 was a potent antimicrobial agent and it was significantly expressed during the pathogen stress.

A Cumulative Strategy to Predict and Characterize Antimicrobial Peptides (AMPs) from Protein Database

Antimicrobial peptides (AMPs) are indivisible part of the innate immune system in invertebrates. AMPs have been proven to have crucial role with a wide range of biological activities, mainly with immunomodulatory and broad spectrum of antimicrobial activity against various pathogens. The unique and salient features of the AMPs show its exceptional nature of therapeutic activity and serves as an alternative agent for conventional antibiotics. The search for potential AMPs persist, as the emergence of multiple drug resistant bacterial strains has been spreading in higher number. Here, the putative antimicrobial peptide sequences were identified from 19,915 sequences of prawn transcriptome and analyzed with various in silico tools such as EXPASY, AMPA, and helical wheel projection and so on. The characteristic antimicrobial properties have been determined for 660 putative AMPs with above mentioned tools. We have demonstrated an efficient bioinformatics approach to derive and analyze the AMPs from the transcriptome data of Macrobrachium rosenbergii. Even though, 660 peptide regions were identified among those five peptide sequences were demonstrated comprehensively with each characteristic property contributes the antimicrobial activity. In this study, we have proposed a rapid and successful protocol that would help to predict AMP in sequential procedure using various in silico methods. Also, we have shown a distinctive method to shortlist the AMPs based on their various physico-chemical properties. Until now, no sequential protocol has been developed to identify and characterize the AMPs from protein database.

Bactericidal and fungistatic activity of peptide derived from GH18 domain of prawn chitinase 3 and its immunological functions during biological stress

Chitinases play a vital role during the pathogenic invasion and immunosuppression in various organisms including invertebrates and vertebrates. In this study, we have investigated the participation of MrChit-3 (Macrobrachium rosenbergii Chitinase-3) during host-pathogenic interaction in freshwater prawn, M. rosenbergii. Quantitative real-time PCR analysis showed that the expression of MrChit-3 was up-regulated during bacterial, viral and laminarin challenge. Moreover, to understand the antimicrobial role of the GH18 domain, a putative membrane-targeting antimicrobial peptide (MrVG) was identified from the GH18 domain region of the protein and it was chemically synthesized. Physico-chemical features of the GH18 derived antimicrobial peptide (AMP) was assessed by various in silico tools and the antimicrobial property of the peptide was confirmed from in vitro studies. The membrane targeting mechanism of the peptide was determined by flow cytometry (FACS) and scanning electron microscope (SEM) analysis. Interestingly, the peptide was able to inhibit the growth of a chitinolytic fungal pathogen, Aspergillus niger, which was isolated from the shells of M. rosenbergii. The toxicity studies such as hemolysis activity on human blood erythrocytes and cell viability assay with primary kidney cells, HEK293 of MrVG revealed that the peptide was not involved in inducing any toxicity.