Show simple item record

dc.contributor.advisorKennedy, Hendrick
dc.contributor.authorJimenez-Martinez, R
dc.contributor.authorIkonomopoulou, M
dc.contributor.authorHaigh, O
dc.contributor.authorLutzky, V
dc.contributor.authorLeal-Rojas, I
dc.contributor.authorRyan, R
dc.contributor.authorRatnatunga, C
dc.contributor.authorWong, Y
dc.contributor.authorLopez, A
dc.contributor.authorFry, B
dc.contributor.authorHerzig, V
dc.contributor.authorKing, G
dc.contributor.authorMiles, JJ
dc.date.accessioned2018-10-31T01:25:17Z
dc.date.available2018-10-31T01:25:17Z
dc.date.issued2016
dc.identifier.issn0014-2980
dc.identifier.doi10.25904/1912/2838
dc.identifier.urihttp://hdl.handle.net/10072/380987
dc.description.abstractThe millions of bioactive compounds found within animal venoms are a veritable war chest of novel immunotherapeutics. Discoveries have resulted in six venom derived drugs currently approved by the FDA for diseases such as blood disorders and diabetes. Venom-derived compounds are known to be highly potent and highly selective for ion channels as well as other key biological pathways. To date only an infinitesimally small fraction of venom-derived compounds have been investigated for medical applications. The aim of this project was the systematic scan of a vast venom biorepository (over 300 different venoms; including 217 different spider venoms, 92 snake venoms, 33 cone snail venoms and 2 jellyfish venoms, and 21 venom peptides) to characterize novel venom-derived peptides with immunomodulatory function. Using resting and stimulated human leucocytes, we performed high throughput, multiplex scanning of venom activity using Cytometric Bead Array readout (CBA). During my doctorate, this experimental platform was optimized and used for venom and the venom fraction screening. The CBA scanning allowed the detection and quantification of nine different human cytokines/chemokines. In Chapter 1, I provide an overview of what is currently known around immune modulation using venoms. Here, I detail previously described immunomodulation from snake, scorpion, bee and sea anemone venoms/toxins and provide an outline of what is known in the field. In Chapter 2, I detailed the breadth of materials and methods used in this doctorate including immunology and proteomics methodologies. In Chapter 3, I present the mass CBA scanning across spider, snake, jellyfish, and cone snail on primary human leucocytes. In this Chapter, I present cytokine/chemokine mastergraphs of IL-1β, IL-6, IL-8, IL-10, TNF-α, IFN-γ and MIP-1β release from primary human leucocytes after the addition of venom. In this Chapter, I also present mastergraphs of cytokine release from PMA/Ionomycin-stimulated primary human leucocytes after the addition of venom. These experiments identified T cell “accelerants” that enhanced cytokine production such as the spider P57 Badumna Insignis, P332 Pamphobeteus wuschi and the snake Boiga Dendrophila. In Chapter 3, I also present mastergraphs of cytokine/chemokine release from LPS-stimulated primary human leucocytes after the addition of venom. These experiments identified myeloid cell “accelerants” with enhanced cytokine production such as Pseudonaja affinis and Psudonaja Nuchalis. In this Chapter, I used principal component analysis (PCA) to identify venoms with similar cytokine/chemokine release patterns. I found that in general the PCA clustered venoms that produced pro-inflammatory cytokines IL-1β, IL-6, IL-8, MIP-1β and the anti-inflammatory cytokine IL-10 in one group and venoms that produced T cell related cytokines such as IL-2, IL-4, TNF-α and IFN-γ in a different group. I next performed Pearson’s correlation between cytokine/chemokine release and known LD50 data from venom. This was to investigate whether there was a correlation between release of a particular cytokine and lethality of the venom. I observed a strong positive correlation between IL-10 and intraperitoneal administered LD50 in the presence of Phorbol 12-Myristate 13-Acetate (PMA) and Ionomycin (r=0.9325) (P ≤ 0.05). In Chapter 4, I identified the top five venoms that enhanced the immune response mediated by cytokines (P204, P175, P332, P57 and P168) by testing venoms across multiple genetically unrelated donors. I attempted to map immunologically active peptides in the most bioactive venoms using Reversed Phase-High Performance Liquid Chromatography (RP-HPLC), CBA, Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), transcriptomics and Edman degradation. The sequences of three original peptides derived from spider venom were identified. In Chapter 5, I investigated the ability of venoms and venom-derived peptides to augment the human immune response in vitro. Here, I used monocyte and dendritic cell stimulation, mixed lymphocyte reactions and dextran uptake to quantify antigen presenting cell activation profiles, T cell stimulation and antigen processing. I found that the venom peptides 16 and 19 increased the CD83 expression marker and the peptide P332 increased dendritic cell antigen uptake. In Chapter 6, I provide an overall summary of my findings and discuss how they fit in the wider literature. In this Chapter I also discuss possible future directions of my findings to progress research to the preclinical phase. In summary, I anticipate this research will help catalyse development of new classes of venom-derived peptide based immune stimulatory compounds to help augment vaccine effectiveness and enhance the effects of current and future immunotherapeutic strategies for cancer, infectious disease and autoimmunity.
dc.languageEnglish
dc.language.isoen
dc.publisherGriffith University
dc.publisher.placeBrisbane
dc.relation.ispartofconferencenameInternational Congress of Immunology (ICI)
dc.relation.ispartofconferencetitleEUROPEAN JOURNAL OF IMMUNOLOGY
dc.relation.ispartofdatefrom2016-08-21
dc.relation.ispartofdateto2016-08-26
dc.relation.ispartoflocationMelbourne, AUSTRALIA
dc.relation.ispartofpagefrom396
dc.relation.ispartofpageto397
dc.relation.ispartofvolume46
dc.subject.fieldofresearchImmunology
dc.subject.fieldofresearchcode1107
dc.subject.keywordsVenom-derived molecules
dc.subject.keywordsHuman immune function
dc.subject.keywordsPeptides
dc.subject.keywordsImmunomodulatory function
dc.subject.keywordsCytometric Bead Array readout
dc.titleInvestigating venom-derived molecules that augment human immune function
dc.typeGriffith thesis
gro.facultyScience, Environment, Engineering and Technology
gro.rights.copyrightThe author owns the copyright in this thesis, unless stated otherwise.
gro.hasfulltextFull Text
dc.contributor.otheradvisorMiles, John
dc.contributor.otheradvisorBurrows, Scott
gro.thesis.degreelevelThesis (PhD Doctorate)
gro.thesis.degreeprogramDoctor of Philosophy (PhD)
gro.departmentSchool of Environment and Sc
gro.griffith.authorJimenez Martinez, Rocio


Files in this item

This item appears in the following Collection(s)

Show simple item record