Fusion to the Lysosome Targeting Signal of the Invariant Chain Alters the Processing and Enhances the Immunogenicity of HIV-1 Reverse Transcriptase

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Abstract

Intracellular processing of the antigen encoded by a DNA vaccine is one of the key steps in generating an immune response. Immunization with DNA constructs targeted to the endosomal-lysosomal compartments and to the MHC class II pathway can elicit a strong immune response. Herein, the weakly immunogenic reverse transcriptase of HIV-1 was fused to the minimal lysosomal targeting motif of the human MHC class II invariant chain. The motif fused to the N-terminus shifted the enzyme intracellular localization and accelerated its degradation. Degradation of the chimeric protein occurred predominantly in the lysosomal compartment. BALB/c mice immunized with the plasmid encoding the chimeric protein demonstrated an enhanced immune response, in the form of an increased antigen-specific production of Th1 cytokines, INF-γ and IL-2, by mouse splenocytes. Moreover, the majority of the splenocytes secreted both cytokines; i.e., were polyfunctional. These findings suggest that retargeting of the antigen to the lysosomes enhances the immune response to DNA vaccine candidates with low intrinsic immunogenicity.

About the authors

E. S. Starodubova

Engelhardt Institute of Molecular Biology Russian Academy of Sciences; D.I. Ivanovsky Institute of Virology, Ministry of Health of the Russian Federation

Author for correspondence.
Email: estarodubova@yandex.ru
Russian Federation

М. G. Isaguliants

D.I. Ivanovsky Institute of Virology, Ministry of Health of the Russian Federation; Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet

Email: estarodubova@yandex.ru
Russian Federation

Y. V. Kuzmenko

Engelhardt Institute of Molecular Biology Russian Academy of Sciences

Email: estarodubova@yandex.ru
Russian Federation

A. A. Latanova

Engelhardt Institute of Molecular Biology Russian Academy of Sciences

Email: estarodubova@yandex.ru
Russian Federation

О. А. Krotova

Engelhardt Institute of Molecular Biology Russian Academy of Sciences; D.I. Ivanovsky Institute of Virology, Ministry of Health of the Russian Federation

Email: estarodubova@yandex.ru
Russian Federation

V. L. Karpov

Engelhardt Institute of Molecular Biology Russian Academy of Sciences

Email: estarodubova@yandex.ru
Russian Federation

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Copyright (c) 2014 Starodubova E.S., Isaguliants М.G., Kuzmenko Y.V., Latanova A.A., Krotova О.А., Karpov V.L.

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