In Vitro, In Vivo and In Silico Assessment of the Antimicrobial and Immunomodulatory Effects of a Water Buffalo Cathelicidin (WBCATH) in Experimental Pulmonary Tuberculosis

Tuberculosis (TB) is considered the oldest pandemic in human history. The emergence of multidrug-resistant (MDR) strains is currently considered a serious global health problem. As components of the innate immune response, antimicrobial peptides (AMPs) such as cathelicidins have been proposed to have efficacious antimicrobial activity against Mycobacterium tuberculosis (Mtb). In this work, we assessed a cathelicidin from water buffalo, Bubalus bubalis, (WBCATH), determining in vitro its antitubercular activity (MIC), cytotoxicity and the peptide effect on bacillary loads and cytokines production in infected alveolar macrophages. Our results showed that WBCATH has microbicidal activity against drug-sensitive and MDR Mtb, induces structural mycobacterial damage demonstrated by electron microscopy, improves Mtb killing and induces the production of protective cytokines by murine macrophages. Furthermore, in vivo WBCATH showed decreased bacterial loads in a model of progressive pulmonary TB in BALB/c mice infected with drug-sensitive or MDR mycobacteria. In addition, a synergistic therapeutic effect was observed when first-line antibiotics were administered with WBCATH. These results were supported by computational modeling of the potential effects of WBCATH on the cellular membrane of Mtb. Thus, this water buffalo-derived cathelicidin could be a promising adjuvant therapy for current anti-TB drugs by enhancing a protective immune response and potentially reducing antibiotic treatment duration.

[1]  G. Sotgiu,et al.  Novel treatments in multidrug-resistant tuberculosis. , 2021, Current opinion in pharmacology.

[2]  J. Ivanyi,et al.  Editorial: Advances in Immunotherapeutic Approaches to Tuberculosis , 2021, Frontiers in Immunology.

[3]  R. Hernández-Pando,et al.  Experimental Pulmonary Tuberculosis in the Absence of Detectable Brain Infection Induces Neuroinflammation and Behavioural Abnormalities in Male BALB/c Mice , 2020, International journal of molecular sciences.

[4]  Munehiro Asally,et al.  The Microbiologist's Guide to Membrane Potential Dynamics. , 2020, Trends in microbiology.

[5]  Wuyuan Lu.  Antimicrobial peptides. , 2019, Seminars in cell & developmental biology.

[6]  A. Hollmann,et al.  Antimicrobial Peptides: Interaction With Model and Biological Membranes and Synergism With Chemical Antibiotics , 2018, Front. Chem..

[7]  Philippe Glaziou,et al.  Global Epidemiology of Tuberculosis , 2018, Seminars in Respiratory and Critical Care Medicine.

[8]  R. Hernández-Pando,et al.  The use of immunotherapy for the treatment of tuberculosis , 2018, Expert review of respiratory medicine.

[9]  F. Torres-Juárez,et al.  Antimicrobial peptides for the treatment of pulmonary tuberculosis, allies or foes? , 2018, Current pharmaceutical design.

[10]  F. Niyonsaba,et al.  Tissue-specific Regulation of Innate Immune Responses by Human Cathelicidin LL-37. , 2018, Current pharmaceutical design.

[11]  K. V. Chanu,et al.  Antimicrobial peptides of buffalo and their role in host defenses , 2018, Veterinary world.

[12]  C. Delerue-Matos,et al.  Synergistic and antibiofilm properties of ocellatin peptides against multidrug-resistant Pseudomonas aeruginosa. , 2018, Future microbiology.

[13]  A. Jordaan,et al.  Interactions of the antimicrobial peptide nisin Z with conventional antibiotics and the use of nanostructured lipid carriers to enhance antimicrobial activity. , 2017, International journal of pharmaceutics.

[14]  J. Sánchez-Céspedes,et al.  Perspectives for clinical use of engineered human host defense antimicrobial peptides , 2017, FEMS microbiology reviews.

[15]  K. Poluri,et al.  Diversity, Antimicrobial Action and Structure-Activity Relationship of Buffalo Cathelicidins , 2015, PloS one.

[16]  J. Andrew McCammon,et al.  Gaussian Accelerated Molecular Dynamics: Unconstrained Enhanced Sampling and Free Energy Calculation , 2015, Journal of chemical theory and computation.

[17]  K. Mdluli,et al.  The tuberculosis drug discovery and development pipeline and emerging drug targets. , 2015, Cold Spring Harbor perspectives in medicine.

[18]  Y. Tor,et al.  Antibiotics and Bacterial Resistance in the 21st Century , 2014, Perspectives in medicinal chemistry.

[19]  Estela Isabel Bini and Rogelio Hernandez-Pando.  New Chemotherapy and Immunotherapy for Tuberculosis , 2014 .

[20]  V. P. Panicker,et al.  Identification of three novel myeloid cathelicidin cDNAs and their predicted peptides in buffalo (Bubalus bubalis). , 2013, Indian journal of biochemistry & biophysics.

[21]  R. Raqib,et al.  Oral intake of phenylbutyrate with or without vitamin D3 upregulates the cathelicidin LL-37 in human macrophages: a dose finding study for treatment of tuberculosis , 2013, BMC Pulmonary Medicine.

[22]  Kevin W Eliceiri,et al.  NIH Image to ImageJ: 25 years of image analysis , 2012, Nature Methods.

[23]  B. Mishra,et al.  Cathelicidin is involved in the intracellular killing of mycobacteria in macrophages , 2011, Cellular microbiology.

[24]  C. Serrano,et al.  Kinetics and cellular sources of cathelicidin during the course of experimental latent tuberculous infection and progressive pulmonary tuberculosis , 2010, Clinical and experimental immunology.

[25]  R. Hancock,et al.  Human Cathelicidin Peptide LL-37 Modulates the Effects of IFN-γ on APCs1 , 2009, The Journal of Immunology.

[26]  Aaron Weinberg,et al.  The roles of antimicrobial peptides in innate host defense. , 2009, Current pharmaceutical design.

[27]  R. Gallo,et al.  AMPed up immunity: how antimicrobial peptides have multiple roles in immune defense. , 2009, Trends in immunology.

[28]  Taehoon Kim,et al.  CHARMM‐GUI: A web‐based graphical user interface for CHARMM , 2008, J. Comput. Chem..

[29]  Thomas D. Schmittgen,et al.  Analyzing real-time PCR data by the comparative CT method , 2008, Nature Protocols.

[30]  R. Modlin,et al.  Cutting Edge: Vitamin D-Mediated Human Antimicrobial Activity against Mycobacterium tuberculosis Is Dependent on the Induction of Cathelicidin1 , 2007, The Journal of Immunology.

[31]  F. Blecha,et al.  Canine cathelicidin (K9CATH): gene cloning, expression, and biochemical activity of a novel pro-myeloid antimicrobial peptide. , 2007, Developmental and comparative immunology.

[32]  V. Tsutsumi,et al.  beta-Defensin gene expression during the course of experimental tuberculosis infection. , 2006, The Journal of infectious diseases.

[33]  D. Ercolini,et al.  Genetic Resistance to Brucella abortus in the Water Buffalo (Bubalus bubalis) , 2006, Infection and Immunity.

[34]  S. Said-Fernández,et al.  Bactericidal activity of organic extracts from Flourensia cernua DC against strains of Mycobacterium tuberculosis. , 2006, Archives of medical research.

[35]  J. Ellner,et al.  Human β-Defensin 2 Is Expressed and Associated with Mycobacterium tuberculosis during Infection of Human Alveolar Epithelial Cells , 2005, Infection and Immunity.

[36]  M. Zanetti,et al.  The cathelicidins--structure, function and evolution. , 2005, Current protein & peptide science.

[37]  I. Nagaoka,et al.  Epithelial cell-derived antibacterial peptides human beta-defensins and cathelicidin: multifunctional activities on mast cells. , 2003, Current drug targets. Inflammation and allergy.

[38]  Dong-Kuk Lee,et al.  Mechanism of lipid bilayer disruption by the human antimicrobial peptide, LL-37. , 2003, Biochemistry.

[39]  James M. Wilson,et al.  Cathelicidins - a family of multifunctional antimicrobial peptides , 2003, Cellular and Molecular Life Sciences CMLS.

[40]  M. Tollin,et al.  Phylogeny, processing and expression of the rat Cathelicidin rCRAMP: a model for innate antimicrobial peptides , 2003, Cellular and Molecular Life Sciences CMLS.

[41]  R. Gallo,et al.  Cathelicidins, essential gene-encoded mammalian antibiotics , 2002, Journal of Molecular Medicine.

[42]  J. Ventura,et al.  Ceramide promotes the death of human cervical tumor cells in the absence of biochemical and morphological markers of apoptosis. , 2002, Biochemical and biophysical research communications.

[43]  Kenneth Huttner,et al.  Processing site and gene structure for the murine antimicrobial peptide CRAMP , 2001, Peptides.

[44]  A. Waring,et al.  RL-37, an Alpha-Helical Antimicrobial Peptide of the Rhesus Monkey , 2001, Antimicrobial Agents and Chemotherapy.

[45]  C. Vogelmeier,et al.  Rhesus Monkey (Macaca mulatta) Mucosal Antimicrobial Peptides Are Close Homologues of Human Molecules , 2001, Clinical Diagnostic Laboratory Immunology.

[46]  R. Gennaro,et al.  Structural and Functional Analysis of Horse Cathelicidin Peptides , 2001, Antimicrobial Agents and Chemotherapy.

[47]  H. Jörnvall,et al.  The human antimicrobial and chemotactic peptides LL-37 and alpha-defensins are expressed by specific lymphocyte and monocyte populations. , 2000, Blood.

[48]  R. Gennaro,et al.  Structural features and biological activities of the cathelicidin-derived antimicrobial peptides. , 2000, Biopolymers.

[49]  R. Lehrer,et al.  Purification and Properties of Proline-Rich Antimicrobial Peptides from Sheep and Goat Leukocytes , 1999, Infection and Immunity.

[50]  C. Subbalakshmi,et al.  Mechanism of antimicrobial action of indolicidin. , 1998, FEMS microbiology letters.

[51]  M. Zanetti,et al.  Structural organization of the bovine cathelicidin gene family and identification of a novel member 1 , 1997, FEBS letters.

[52]  A. Sampieri,et al.  Correlation between the kinetics of Th1, Th2 cells and pathology in a murine model of experimental pulmonary tuberculosis. , 1996, Immunology.

[53]  C. E. Ooi,et al.  Antibacterial 15-kDa protein isoforms (p15s) are members of a novel family of leukocyte proteins. , 1993, The Journal of biological chemistry.