Nunuk Dyah Retno Lastuti

Humoral and cellular immune response induced by antigenic protein of Sarcoptes scabiei var. caprae

Aim: Scabies is one of the most important diseases in goats and caused by a complex hypersensitivity process that involves both humoral and cell-mediated immune responses. This phenomenon shows that the variety of Sarcoptes scabiei has different characteristics of specific antigenic protein or different immune-dominant. This research aims to detect the humoral and cellular immune response of rabbits which were immunized with the protein of S. scabiei var. caprae. Materials and Methods: This research was done as follows, identification and collection of Sarcoptes scabiei var caprae from goat infected with scabies, separation of protein antigen from S. scabiei mites with ultrasonic sonicator, measurement of protein content with spectrophotometry, rabbit injection with 500 μg dose of antigen protein which was repeated 5 times (5x booster) every 2 weeks. Measurement of IgG titer using indirect ELISA, whereas to detect the expression of cellular immune response (TLR-9, CD4, and CD8) using Direct Immunofluorescence assay. Results: Based on the statistical analysis, it showed that there was a significant enhancement (p<0.05) for optical density value or antibody titer and cellular immune response was shown by TLR-9, CD4, and CD8 expression in rabbit T lymphocytes which appear yellow to green fluorescent color using fluorescence microscope. The amount of fluorescence T lymphocytes showed a significant difference (p<0.05) between the control and various boosters. Conclusion: Antigenic protein of S. scabiei var. caprae contains ligands, which are involved in the pathogen-associated molecular pattern that has an ability to induce humoral and cellular immune response in rabbit. Specifically, that TLR-9 is not only involved in innate immunity but also in adaptive immunity and can be used as alternative adjuvant development research.

Antigenic Protein of Leucocytozoon caulleryi schizont Inducing Cellular Immune Response: TLR-2 and CD4 as Marker

Leucocytozoonosis is caused by Leucocytozoon caulleryi and is responsible for death in chickens by bleeding. Leucocytozoonosis is an endemic disease in Indonesia and incidences have been reported in several regions in East and Central Java. The financial losses caused by this disease include growth disorders in chick, decreased egg production, higher mortality rate and also a higher production cost. This research aims to detect TLR-2 and CD4 expression as a cellular immune response in rabbits immunised by the protein of L.caulleryi schizont. It is needed as a preliminary research study for molecular vaccine development which is considerably effective when it comes to preventing leucocytozoonosis occurance in Indonesia. This research study was performed in several stages i.e. the isolation of L.caulleryi schizont from a chicken liver infected with leucocytozoonosis to be based on the clinical signs observed, microscopic examination, and the pathological changes in the other chicken organs. The purification of the soluble protein of L.caulleryi schizont including the immunisation of the rabbits. Each of the experimental rabbits was injected with 500 μg of L.caulleryi schizont protein and added adjuvant complete with a ratio of 1:1. Every two weeks the injection was performed with the same protein with a dosage of 500 μg each and an added adjuvant that was incomplete (the booster was performed 5 times in 2 weeks). The examination of the cellular immune response of CD4 and TLR-2 expression in the rabbits’ T cells using an immunocytochemistry method visualised by fluorescein isothiocyanate. The examination of the results was done by immunocytochemistry showing TLR-2 and CD4 expression as yellow to green fluorescent colour, mainly in the 5 booster where the activation of the CD4 coreceptor and TLR-2 occurred. The conclusion shows that the antigenic protein of L.caulleryi schizont has the ability to induce a cellular immune response through the co-receptors CD4 and TLR-2 in the rabbits’ T cells as the preliminary research in the sub-unit vaccine development for leucocytozoonosis in chickens. KEY WORD: antigenic protein, Leucocytozoon caulleryi, schizont, TLR-2, CD4.