The gene encoding HAVCR1/TIM in humans has been shown to be associated with susceptibility to asthma and allergy. Infection with HAV has shown novel results indicating a great reduction in the risk of developing asthma and allergy as an outcome of its interaction with HAVCR1/TIM. HAVCR1/TIM is thought to be associated with regulation of T- cell differentiation and development of atopy. However, normal functions of HAVCR1/TIM in absence of HAV infection are not fully understood.
Researchers used an expression cloning strategy. They transfected dog cells with a human lymph node cDNA library which gives the ability of transfected dog cells to produce human immunoglobulins. Binding of transfected dog cells to HAVCR1/TIM Fc fusion protein is studied. Soluble form of HAVCR1/TIM containing the HAVCR1/TIM Ig variable-like (IgV) region fused to the Fc fragment of human IgG1 antibody [HAVCR1/TIM(IgV)-Fc].
Materials and Methods:
Cells and virus. Perro6D cells derived from canine osteogenic sarcome D-17 cells transfected with EBNA-1 cDNA are sistant to the antibiotic G418 and have an increased transfection efficiency for episomal plasmids containing and Epstein-Barr virus P1 origin of replication are used to show possib...
... middle of paper ...
...sis carried on by researchers to determine if IgA by itself is enough to bind to HAVCR1/TIM. They coated plates with goat anti- human IgG Fc, which will bind to HAVCR1/TIM(IgV)-Fc since the Fc portion in this fusion protein is derived from human IgG. Afterwards, they introduced human IgA to the well plates and detected binding of IgA to HAVCR1/TIM(IgV)-Fc. Results showed that IgA itself is able to bind HAVCR1/TIM without requiring other cell proteins and components. This confirmed that IgA is indeed a natural ligand of this receptor.
Since IgA and HAV are ligands of the same receptor, researchers thought that IgA binding to HAVCR1/TIM would inhibit binding of HAV to the receptor. To answer this controversy, they treated GL37 cells with 1, 5, or 10 μg of IgA for 30 min and then infected the cells with HAV at multiplicity of infection (MOI) of 1 to 5 TCID50 per cell.
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