Construction of AAV-rat-IL4 and Evaluation of its Modulating Effect on Aβ (1-42)-Induced Proinflammatory Cytokines in Primary Microglia and the B92 Cell Line by Quantitative PCR Assay

Marzieh Jamalidoust; Mehrdad Ravanshad; Mandana Namayandeh; Maryam Zare; Sadaf Asaei; Mazyar Ziyaeyan

doi:10.5812/jjm.30444

Construction of AAV-rat-IL4 and Evaluation of its Modulating Effect on Aβ (1-42)-Induced Proinflammatory Cytokines in Primary Microglia and the B92 Cell Line by Quantitative PCR Assay

Jundishapur J Microbiol. 2016 Mar 5;9(3):e30444. doi: 10.5812/jjm.30444. eCollection 2016 Mar.

Authors

Marzieh Jamalidoust¹, Mehrdad Ravanshad², Mandana Namayandeh³, Maryam Zare³, Sadaf Asaei³, Mazyar Ziyaeyan³

Affiliations

¹ Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IR Iran; Alborzi Clinical Microbiology Research Center, Nemazi Hospital, Shiraz University of Medical Sciences, Shiraz, IR Iran.
² Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IR Iran.
³ Alborzi Clinical Microbiology Research Center, Nemazi Hospital, Shiraz University of Medical Sciences, Shiraz, IR Iran.

Abstract

Background: Interleukin-4 (IL-4), as the most prominent anti-inflammatory cytokine, plays an important role in modulating microglial activation and inflammatory responses in Alzheimer's disease (AD), a chronic inflammatory disorder.

Objectives: The current study aimed to develop a new recombinant Adeno-associated viral (rAAV) vector that delivers IL-4 and then assess the counterbalancing effect of the new construct along with recombinant IL-4 (rIL-4) protein in in-vitro models of AD.

Materials and methods: The rAAV-IL4 was originally prepared and then employed along with rIL-4 protein to counter Amyloid β (1-42)-induced proinflammatory cytokines in a primary microglia cell culture and the B92 rat microglia continuous cell line, using relative Real-Time PCR assay.

Results: Aβ (1-42) stimulated the production of the proinflammatory cytokines IL6, IL1β, TNFα, and IL18 in both the primary microglia cell culture and the B92 cell line. Both the rAAV-IL4 construct and the rIL-4 protein were found to inhibit production of the most important Aβ (1-42)-induced proinflammatory cytokine mRNAs in the two types of cells with different patterns.

Conclusions: It seems that the new construct can serve as an appropriate option in the modulation of Aβ-induced proinflammatory cytokine gene expression and microglia activation in patients affected by AD.

Keywords: Adeno-Associated Viral Vector; Alzheimer Disease; Amyloid β (1-42); B-92 Cell Line; IL-4; Primary Microglia Cell; Proinflammatory Cytokines.