Immunogenicity of Leishmania Derived Antigens in Peripheral Blood Mononuclear Cells Using Blood Obtained from Previously Treated Visceral Leishmaniasis Patients Of Southwest Ethiopia

Mihiretie, Gebreselassie Demeke (2015) Immunogenicity of Leishmania Derived Antigens in Peripheral Blood Mononuclear Cells Using Blood Obtained from Previously Treated Visceral Leishmaniasis Patients Of Southwest Ethiopia. Masters thesis, Addis Ababa University.

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Abstract

Background:-Visceral leishmaniasis is a public health problem and it is fatal if untreated. Disease burden is associated with poverty, lack of effective, affordable, and minimally toxic treatments. An effective vaccine is needed to combat this disease. Method: - Peripheral blood mononuclear cell-based assays were set-up in blood samples obtained from 5 healthy controls and 18 drug-cured VL patients (DC-VL), who are putatively immune to re-infection. For PBMCs stimulation nine Leishmania derived antigens were selected, and together with whole promastigote lysate antigen and Phytohaemagglutinin (PHA), and assessment of immunogenicity was carried out by cytokine assays using ELISA kit. The assay aimed at measuring IFN-gamma, TNF-alpha, IL-10 and IL-5 in culture supernatants of PBMCs stimulated in vitro. The association of cytokine production with immunogenicity difference of antigens was determined by using non-parametric tests (Mann Whitney & Tukey’s Multiple Comparison tests) found in Graph Pad Prism version 6. Result: - Drug-cured visceral leishmaniasis (VL) patients develop protection mediated by Th1-type cellular responses against new infections. IFN-gamma, TNF-alpha and IL-10 production were higher in drug-cured groups, but neither drug-cured nor healthy controls produced IL-5. The data suggest the mounting of Th1 type responses upon cure from visceral leishmaniasis, while small amounts of IL-10 were also measurable. NS, S, PNS and NSL were more immunogenic than other antigens as determined by levels of IFN-gamma, TNF-alpha and IL-10. Conclusion:-This study indicates that T cell recall ability of humans cured from VL by chemotherapy was conspicuous and related to Th1 type immune response. Further, the data confirm that high levels of IFN-gamma and TNF-alpha in the presence of low levels of interleukin-10 (IL-10) could be proxy indicators of protective immunity in drug-cured visceral leishmaniasis patients. Among tested antigens NS, NSL, PNS and S were more immunogenic and elicited high levels of IFN-gamma and TNF-alpha in PBMCs of drug-cured patients, with supernatant levels ranging from16 to1826 mg/ml.

Item Type: Thesis (Masters)
Uncontrolled Keywords: Antigen immunogenicity, visceral leishmaniasis
Subjects: Q Science > Q Science (General)
Q Science > QR Microbiology > QR180 Immunology
Divisions: Africana
Depositing User: Vincent Mpoza
Date Deposited: 13 Jul 2018 09:14
Last Modified: 13 Jul 2018 09:14
URI: http://thesisbank.jhia.ac.ke/id/eprint/7403

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