Construction and characterization of an adenoviral vector which expresses interleukin 6 to study its role in cortical inflammation

EMILIA HABERFELD; CARINA C FERRARI; BERENICE ANABEL SILVA; FARIAS, MARIA I.; FERNANDO J PITOSSI; MARIA CELESTE LEAL

Buenos Aires

Congreso; SAN 2022 Meeting; 2022

Sociedad Argentina de Investigacion en Neurociencias

Multiple sclerosis (MS) is a chronic demyelinating disease that causes neurological disabilities in young adults. Although it has been described as an autoimmune disorder, its aetiology is still unknown. By overexpressing interleukin 1 ß in the cerebral cortex, our group has developed a rat MS model that presents the characteristic inflammatory and demyelinated cortical lesions of the progressive forms of the disease. Interleukin 6 (IL-6) was found among the cytokines whose expression was induced in these lesions. Therefore, the aim of this work is to study the effects of IL-6 on the cortex. In order to obtain a long-term overexpression of IL-6 in the cortex of rats, we constructed a non-replicating adenoviral vector which encodes IL-6 and GFP genes with an IRES in between, downstream a strong constitutive promoter. To produce a viral stock, we used the HEK293A complementing cell line, achieving a standard titer. In addition, we obtained a viral stock of an adenoviral vector which expresses ß-galactosidase and GFP to use as a control. With the purpose of analysing the efficiency of transduction of the vectors and the expression levels of the genes of interest, we transduced the non-complementing cell line HeLa. Finally, we injected stereotaxically different doses of the vectors into the prefrontal cortex of adult rats, observing inflammatory lesions after 7 days. These results suggest a fundamental role of IL-6 in the development of inflammatory lesions in the cortex.