Encyclopedia of Experiments: Immunology
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Transcript
Place cervical uterine mucosa in a petri dish. Separate the ectocervix and endocervix mucosa from the submucosa. Slice the mucosal layer into smaller explants and transfer them into a conical tube.
Add the human immunodeficiency virus type 1, or HIV-1 into the explant tube. The HIV-1 virus particles approach the exposed mucosal layer and bind to the CD4 receptor and the CXCR4 co-receptor on the host cell.
Upon binding, the viral envelope fuses with the host cell membrane, releasing a viral capsid containing viral RNA and reverse transcriptase enzyme into the cytoplasm. Inside the cytoplasm, the reverse transcriptase converts the viral RNA into complementary DNA, or cDNA.
The cDNA moves into the host cell nucleus, where integrase incorporates viral DNA into the host genome. The viral DNA directs the synthesis of new viral RNA and proteins which assemble into new virus particles. These particles bud off from the host cell, acquiring a lipid envelope.
Rinse the explants with a suitable buffer to remove any unbound virus. Finally, place the explants on a gelatin sponge in a multiwell plate with a suitable medium and incubate.
The air-liquid interface maximizes oxygen exposure and nutrient accessibility through the sponge capillaries, enhancing model viability for downstream analysis.
