I am trying to perform a co-infection experiment on activated macrophage and I want to know discrimination of Immune regulations. What kind of experimental assaying outlines can resolve my research questions?
Ex: Primary infection with parasites, secondary infection with Bacteria. Could you add me a note of explanation? I appreciate a favorable answer to gain experience and knowledge.
Cell type: Defensing cell line (THP1 -Macrophage)
Infecting bugs: Primary infection with -Parasite
Secondary Infection with Bacteria
You can use a luciferase assay . The principle of the assay is based on the fact that luciferase is bifunctional enzyme activity as it catalyses reactions in two different pathways: a bioluminescence pathway and a CoA-ligase pathway. This will result in light is emission when luciferase acts on the appropriate luciferin substrate . Photon emission can be detected by light sensitive apparatus such as a luminometer or modified optical microscopes. This allows observation of biological processes. I Suggest you read, An In vitro Co-infection Model to Study Plasmodium falciparum-HIV-1 Interactions in Human Primary Monocyte-derived Immune Cells,Guadalupe Andreani, Dominic Gagnon, Robert Lodge, Michel J. Tremblay, Dave Richard
J Vis Exp. 2012, as they used similar techniques.
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