IJFANS International Journal of Food and Nutritional Sciences

ISSN PRINT 2319 1775 Online 2320-7876

LIPOSOMAL ANTIGEN DELIVERY SYSTEM: ISOLATION, PREPARATION, CHARACTERIZATION, IN-VITRO STUDIES, HUMORAL IMMUNITY (HI) AND CELL MEDIATE IMMUNITY (CMI) ASSESSMENT (TH-1/TH-2 IMMUNE RESPONSE INDUCED BY IMMUNOMODULATORY LIPOSOMAL ANTIGEN OF B. MALAYI)

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Neeraj Jain, Neelam Jain, Jagat Pal Yadav, Shubhrat Maheshwari, Rajesh Nath, Pratiksha Jayaswal

Abstract

Objective: The present study was aimed on developing and characterizing liposomal delivery system loaded with antigen of filaria parasite for better sustain release, immunomodulation effect of isolated antigen. Methods: Liposomes were prepared by reverse phase evaporation (REV) method with slight modification using molar ratio of Soya PC:PE:Cholesterol in different molar concentration. Results: In the present study percent of residual antigen remain in liposomes by assuming the initial content to be 100%, in Soya PC:PE:CH liposomes only 14-15% antigen was lost at temperature 25±1ºC and 5-6% antigen was lost on storage at 4±1ºC. Antigen integrity was evaluated by performing the SDS -PAGE of the liposome formulations (Optimized CL3 formulation stored at 4±1ºC) after 30 Days. Antigens were found to be intact in the formulation stored at 4±1ºC after 30 days. The levels of F6 specific IgG1, IgG2a and IgG2b antibodies were found to be elevated in immunized animals over non-immunized controls. Analysis of IgG-subclasses revealed that all the subclasses at (1:25 dilution) increased several folds over the controls with IgG1 showing the greatest increase (25.0-fold) followed by IgG2b (3.0fold). Antibodies titers showed the many fold increment of titers on liposomised antigen groups (Gr.I; without booster dose and Gr.IV; with booster dose).IgG showed about 2.2 fold increment in Gr. IV than control group (Gr.V). IgG1 after booster dose showed about 25-fold increment followed by IgG2b than IgG2a. NO release from peritoneal macrophages of the animals (Gr.I, II, III, IV and V) was increased by exposure to LPS or no exposure to any stimulants in-vitro as compared to cells of non-immunized animals (Gr.V). In summary, F6 was able to induce greater NO production. The TNF-α release in cells of F6 immunized animals was elevated in response to F6, LPS or no stimulation in-vitro over non-immunized ones. The IFN-γ release in cells of F6 immunized animals was elevated in response to F6 or without any stimulation in-vitro in comparison to non-immunized ones. Up-regulation in Th-I responses and down-regulation in Th-II responses show that the immunological cytokines were in function and cause triggers to body immunity to destroy the parasite, the cytokines production checked at mRNA transcription level using RT-PCR.

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