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You searched for +publisher:"University of Otago" +contributor:("McLellan, Alexander Donald"). Showing records 1 – 3 of 3 total matches.

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University of Otago

1. Bouwer, Anthea Lynne. Role of NK cells in DC-based immunotherapy of melanoma .

Degree: 2011, University of Otago

Natural killer (NK) cells were first identified by their ability to kill tumour or virally infected cells without prior sensitization. In spite of this, the actual role of NK cells in tumour immunotherapy remains controversial. This study therefore set out to investigate the potential of Streptococcus salivarius K12, a gram-positive bacterium that has a history of commercial application as a probiotic in New Zealand, for use as a NK cell adjuvant, applying the therapy using B16.OVA melanoma as a model. To confirm that S. salivarius K12 was able to induce efficient activation of NK cells, I first screened a number of gram-positive and gram-negative bacteria for their ability to induce IFNγ release from NK cells. Using ELISA and fluorescence activated cell sorting (FACS) I found that gram-positive bacteria stimulated a rapid release (<24 hours) of IFNγ from dendritic cell: NK cell co-cultures. Cytotoxicity assays showed that despite optimal activation of NK cells by S. salivarius K12, their cytotoxic activity was not enhanced above that of the naive NK cells. Dissecting NK cells into two subsets based on their CD27 expression using FACS, it was discovered that S. salivarius K12-NK activation was predominantly exerted on the mature CD27high NK cell subset and was dependent on membrane-contact with DC and IL-12/IL-18 expression. NK cell activation was found to be independent of Ly49A expression, a marker that can be used in C57BL/6 mice to discriminate between 'unlicensed' NK cells, and those that had been 'licensed' through interaction with self MHC during development. Therefore having a setting where the addition of S. salivarius K12 activates NK cells, I investigated whether these NK cells were recruited to the draining lymph nodes where they could potentially influence the adaptive immune response. A range of adjuvant-activated and S. salivarius K12-activated DC were injected subcutaneously into the flanks of mice and tested their ability to recruit NK cells to the draining lymph node. The adjuvants differed markedly in their ability to recruit NK cells with S. salivarius K12 being the most effective. To determine if activated NK cells would be of benefit in tumour immunotherapy, I investigated the ability of bacterially activated DCs to elicit anti-tumour responses in a B16.OVA melanoma model. Utilizing a therapeutic tumour model where treatment was started three days following tumour inoculation, I found a significant delay of tumour growth in mice that were immunized with ovalbumin-pulsed DC that had been treated for 4 hours with S. salivarius K12 as opposed to other adjuvants tested. I also determined that in vivo depletion of NK cells completely abolished the benefit of DC immunotherapy. A therapeutic tumour experiment where DC were primed in the presence or absence of tumour antigen showed that while NK cells were critical for the antigen-dependent anti-tumour response they did not appear to exert an effector function. To investigate the role of NK cells in priming the anti-tumour response I next utilized a… Advisors/Committee Members: McLellan, Alexander Donald (advisor).

Subjects/Keywords: NK cells; immunotherapy

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Bouwer, A. L. (2011). Role of NK cells in DC-based immunotherapy of melanoma . (Doctoral Dissertation). University of Otago. Retrieved from http://hdl.handle.net/10523/1619

Chicago Manual of Style (16th Edition):

Bouwer, Anthea Lynne. “Role of NK cells in DC-based immunotherapy of melanoma .” 2011. Doctoral Dissertation, University of Otago. Accessed April 15, 2021. http://hdl.handle.net/10523/1619.

MLA Handbook (7th Edition):

Bouwer, Anthea Lynne. “Role of NK cells in DC-based immunotherapy of melanoma .” 2011. Web. 15 Apr 2021.

Vancouver:

Bouwer AL. Role of NK cells in DC-based immunotherapy of melanoma . [Internet] [Doctoral dissertation]. University of Otago; 2011. [cited 2021 Apr 15]. Available from: http://hdl.handle.net/10523/1619.

Council of Science Editors:

Bouwer AL. Role of NK cells in DC-based immunotherapy of melanoma . [Doctoral Dissertation]. University of Otago; 2011. Available from: http://hdl.handle.net/10523/1619


University of Otago

2. Czepluch, Wenzel. Factors mediating successful oral vaccination with lipid-encapsulated Mycobacterium bovis BCG .

Degree: 2012, University of Otago

During the course of this thesis factors mediating successful oral vaccination with lipidencapsulated Mycobacterium bovis BCG were examined. Mice were fed 2x10E07 CFU BCG encapsulated into a lipid matrix to prevent destruction by the gastrointestinal tract and to allow passage through the gut epithelia. In order to trace the vaccine following oral vaccination, mice were sacrificed at various time points ranging from 6 hours to 8 weeks post vaccination, and macerated lymphatic and non-lymphatic organs plated on solid agar. Initially, BCG was distributed widely in lymphatic and non-lymphatic organs, however, BCG was cleared quickly from most organs and formed small populations of less than 500 CFU/mouse in the mesenteric and cervical lymph nodes, as well as the Peyer’s patches 8 weeks post vaccination. Immuno-histochemistry and confocal microscopy, showed that BCG was absent from the follicles, but instead resided in the T cell containing intra-follicular areas. Very rarely BCG was associated with small CD11b+ cells that did not resemble typical macrophages and lacked peroxidase activity. Instead the majority of BCG could be found forming extracellular groups of 1-4 rods. This was confirmed using cell sorting of leukocytes isolated from alimentary tract lymphatics of orally vaccinated mice and only showed a minority of BCG to be associated with CD11c+ cells. Therefore, BCG is absent from typical antigen presenting cells, but instead might reside in CD11c+CD11b+ myeloid DC. Additionally, Ziehl-Neelsen staining revealed groups of intracellular coccoid forms of BCG. These were located toward the subcapsular space of draining lymph nodes where they were associated with sub-capsular macrophages. Interestingly, cocci proved to be non-platable using solid agar but instead required resuscitation in liquid media and therefore might resemble a form of dormancy. The presence of extracellular rods and intracellular cocci in on-professional antigen presenting cells might highlight the importance of secreted factors as an antigen source promoting successful activation of the immune system. Following oral vaccination, IFN-γ producing cells almost exclusively resided in the spleen. In order to characterize these cells, splenocytes of orally vaccinated mice were isolated 6 weeks post vaccination on the basis of surface marker expression using fluorescence activated cell sorting (FACS). Antigen-specific release of IFN-γ was monitored using ELISA and ELISpot assays and IFN-γ producing T cells were characterized as T effector memory cells expressing CD44, but not CD62L and lacked the expression of mucosal homing markers such as CD103 or α4β7. In addition, Lincoplex assays revealed the production of IL-17 by splenocytes. These did not express CD4+ but rather the γδ T cell eceptor. Together these results show that antigen reservoirs of BCG present in the draining lymphatics contain small numbers of typical filamentous BCG. A larger population of coccoid forms leaves open the possibility that coccoid forms are a major source of antigen… Advisors/Committee Members: McLellan, Alexander Donald (advisor).

Subjects/Keywords: BCG vaccination; T cells DC

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Czepluch, W. (2012). Factors mediating successful oral vaccination with lipid-encapsulated Mycobacterium bovis BCG . (Doctoral Dissertation). University of Otago. Retrieved from http://hdl.handle.net/10523/2086

Chicago Manual of Style (16th Edition):

Czepluch, Wenzel. “Factors mediating successful oral vaccination with lipid-encapsulated Mycobacterium bovis BCG .” 2012. Doctoral Dissertation, University of Otago. Accessed April 15, 2021. http://hdl.handle.net/10523/2086.

MLA Handbook (7th Edition):

Czepluch, Wenzel. “Factors mediating successful oral vaccination with lipid-encapsulated Mycobacterium bovis BCG .” 2012. Web. 15 Apr 2021.

Vancouver:

Czepluch W. Factors mediating successful oral vaccination with lipid-encapsulated Mycobacterium bovis BCG . [Internet] [Doctoral dissertation]. University of Otago; 2012. [cited 2021 Apr 15]. Available from: http://hdl.handle.net/10523/2086.

Council of Science Editors:

Czepluch W. Factors mediating successful oral vaccination with lipid-encapsulated Mycobacterium bovis BCG . [Doctoral Dissertation]. University of Otago; 2012. Available from: http://hdl.handle.net/10523/2086

3. Coutinho, Frazer Paul. An investigation of CD147 expression in apoptotic vesicles .

Degree: 2014, University of Otago

Our laboratory has recently found that glycoprotein CD147 (Basigin, EMMPRIN) is highly enriched on the surface of apoptotic vesicles (Apo-V) isolated from EL4 and B16 tumour cells. The CD147 protein can appear at a range of sizes due to the presence of multiple isoforms and glycosylation processes. CD147 has been described as an anti-apoptotic molecule and is involved in the invasion and metastasis of metastatic cancers. Apoptotic vesicles (Apo-V) have been shown to bind the CD169 sialic acid-receptor expressed by macrophages in the sub-capsular sinus of lymph nodes and the marginal zone of the spleen. It has been suggested that this interaction of vesicles with the CD169 receptor may have the potential to suppress immune responses. In this investigation, the expression and role of CD147 in Apo-V was observed. After many failed attempts to knockdown CD147 expression in EL4 cells using small hairpin RNA (shRNA) techniques, an alternate pathway was undertaken using B16 melanoma cells. The observed knockdown in cellular fractions of B16 was also replicated in corresponding Apo-V fractions. No significant changes in Apo-V production in the CD147 shRNA knock-down B16 cell line were observed, neither were there major changes in Coomassie blue bands as analysed by polyacrylamide gel electrophoresis. This suggested that CD147 knockdown did not affect apoptotic vesicle production quality. The binding of Apo-V to spleen and lymph node sections was not affected by the knockdown of CD147, suggesting that the CD147 glycoprotein is not the major ligand for CD169 in lymphatic tissues. To further investigate the potential of CD147 to bind CD169, a CD169-Fc-FLAG protein was produced. However, no specific binding of CD169-Fc-FLAG to CD147 was detected. Since CD147 has been implicated in tumour metastasis, the metastatic potential of CD147 knockdown B16 cell lines was next tested to confirm the knockdown. This resulted in functional changes in the cell line behaviour. However, no conclusive evidence for an effect of CD147 knockdown on metastasis was noted. Advisors/Committee Members: McLellan, Alexander Donald (advisor).

Subjects/Keywords: Apo-V; apoptotic; vesicles; CD147; CD169; cancer; melanoma; B16; EL4; tumour; metastasis; transfection; knockdown; GFP; FLAG; spleen; lymph; node; immunology; immunity; glycoproteins

…x28;AVF; 100 ng/ml; provided to us by Dr. Lyn Wise; University of Otago, Dunedin, New… 

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Coutinho, F. P. (2014). An investigation of CD147 expression in apoptotic vesicles . (Masters Thesis). University of Otago. Retrieved from http://hdl.handle.net/10523/4607

Chicago Manual of Style (16th Edition):

Coutinho, Frazer Paul. “An investigation of CD147 expression in apoptotic vesicles .” 2014. Masters Thesis, University of Otago. Accessed April 15, 2021. http://hdl.handle.net/10523/4607.

MLA Handbook (7th Edition):

Coutinho, Frazer Paul. “An investigation of CD147 expression in apoptotic vesicles .” 2014. Web. 15 Apr 2021.

Vancouver:

Coutinho FP. An investigation of CD147 expression in apoptotic vesicles . [Internet] [Masters thesis]. University of Otago; 2014. [cited 2021 Apr 15]. Available from: http://hdl.handle.net/10523/4607.

Council of Science Editors:

Coutinho FP. An investigation of CD147 expression in apoptotic vesicles . [Masters Thesis]. University of Otago; 2014. Available from: http://hdl.handle.net/10523/4607

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