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1-Post challenging effects of new formulation of leishmania major antigen in Balb/c mice.
Latifynia A. *1, Gharagozlou M.J²
1: Dep. of Immunology, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
²Department of Pathobiology, Veterinary Medicine, University of Tehran, Tehran, Islamic Republic of Iran,
*1: Corresponding author: Latifynia Afshineh.
E.mail addresses: alatifynia@sina.tums.ac.ir afltn52@yahoo.com
Keywords: Leishmania major antigen, challenge,spleen
Abstract:
Background/Purpose: Cutaneous leishmaniasis (CL) is a zoonotic disease transmitted between rodents and canines, mainly induced by phelebotomus sand flies. In southern of Iran, the incidence of this protozoan disease has doubled over the last decade. Human leishmaniasis is distributed worldwide, but mainly in the topics and subtropics, with a prevalence of 12 milion cases and an approximated incidence of 0.5 milion cases of VL and 1.5 milion cases of cutaneous leishmaniasis (CL) . The aim of this study was comparing the protective effects of a candidate cocktail vaccine encoding various leishmania major antigens in highly susceptible (Balb/c) mice after challenge with live leishmania. In this regards we selected two previous study’s successfull doses (100 & 200 μg/o.1ml), three injection groups : Leishmania plus BCG (LB), Leishmania plus Teucrium Polium as a new adjuvant [LT], leishmania plus BCG and Teucrium Polium (LBT),and one type susceptible mice(Balb/c) which measured expansion of white pulp size after challenge with live leishmania. Methods: A new formulation antigen evaluated in susceptible mice (Balb/c).Leishmania major promastigotes which cultured and harvested at different growth stagesThese harvested organisms modified and combined of five different methods to produce cocktail crud Antigen antigen. It was tested for sterility and protein levelsmeasured by Lowry method. This crud antigen injected intradermally to Balb/c mice. We have one injection both two same booster doses with one week interval. After 20 days after third leishmania injection challenge or re-exposure was performed with live Leishmania. The protective response was evaluated by observation of inducing lesion, and progress of it or another critical signal, at least survival of injected mice for every week, over 70 days . After this all mice were euthanized with diethyl ether, their spleens removed and prepared Histological sections of them and stained with hematoxylin & eosin .After that expantion of spleeny white pulp (SWP) were studied microscopically. Results: Our results show control group white pulps compared with others, have different structure and size. The SWP size increases is dependent on the injection group .There was a remarkable expansion of lymphoid follicles in the treated groups in Balb/c mice. Conclusion: This new formulation antigen was able to stimulate and expand the lymphoid constituents of spleen tissue after challenge with live leishmania. The SWP is where immune responses and antibodies are produced .Therefore, the effect of antigen preparations on secondary immune responses, adaptive immunity, and antibody production is important in determining the susceptibility of mice to cutaneous leishmaniasis and the induction of immunity lead to protectivity encounter to challenge with live Leishmania major.
2-The effect of new leishmania vaccine against Th1, Th2 as well as increasing of spleen white pulp size in Bulb /c mice after repeated exposure
Short Title: ŮŽ
A comparison study between Th1, Th2 response and spleen's white pulp size against new leishmania vaccine.
Latifynia A. *1, , Hajaran H²., , Mir Amin Mohammady³, Gheflati Z¹., N. Khansari1..
1 Dept.of Immunulogy, Faculty of medicine, Tehran University of Medical Sciences, Tehran, Islamic Republic of Iran .
²Department of Pathobiology,Veterinary Medicine,Tehran University, Tehran, Islamic Republic of Iran
³Leprosy and Dermal disease Center,Tehran University of Medical Sciences, Tehran, Islamic Republic of Iran,
.
*1: Corresponding author
e.mail addresses : alatifynia@sina.tums.ac.ir
afltn52@yahoo.com
Keywords: Leishmania major,T helper 1,T helper 2,spleen,Teukrium Butirium, BCG,
Abstact:
Introduction: Human leishmaniasis is distributed worldwide, but mainly in the tropics and subtropics, with a prevalence of 12 million cases and an approximately incidence of 0.5 million cases of VL and 1.5 million cases of cutaneous leishmaniasis (CL). Leishmania parasites are vector-born protozoan pathogens found in tropical and subtropical regions of both the old and new world. The disease in human can be divided into cutaneous, visceral, and mucosal syndromes. The aim of this study was more experiments over our previous new formulation modify leishmania major antigen that had satisfactory results, before.
Material and Method: In this study we have made preliminarily new vaccine with the same methodology again and selected two injection doses (100 & 200 μg/0.1ml), three injection groups: Leishmania plus BCG (LB), Leishmania plus new adjuvant (Teucrium Polium)[LT], leishmania plus BCG and Teucrium Polium (LBT),and one susceptible mice(Balb/c) and measure two type cytokines:Th1(IFN- γ, IL-12)and Th2(IL-4,IL-10) and expansion of white pulp size after challenge with live leishmania.
Results: results show that both doses over LBT group have highest IL-12, lowest IL-10 and highest increasing in spleen’s white pulp size, whether; other groups have lower IL-12, higher IL-10 and lowest increased size in spleen’s white pulp in susceptible mice. Correlation to IL-12 and Il-10, IFN- γ and IL-4 against to spleen’s white pulp expansion is significant.
Conclusion: Our study show that in Balb/c mice, best injection group that produced highest IL-12 and lowest IL-10 which have significant differences, is LBT group, and adjuvants BCG both Teucrium which have synergic effect with together. And also, higher spleen’s white pulp size increasing was seen in LBT group.
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