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| Author(s): |
Maria Clara Martins Ferreira
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | São Paulo. |
| Institution: | Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI) |
| Defense date: | 2021-03-03 |
| Examining board members: |
Bruna Cunha de Alencar Bargieri;
Simone Gonçalves da Fonseca;
Daniela Santoro Rosa;
Luis Lamberti Pinto da Silva
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| Advisor: | Bruna Cunha de Alencar Bargieri |
| Abstract | |
HIV-1 has CD4 lymphocytes as its main target cells. However, myeloid cells can also become infected with HIV-1 through interaction with CD4, CCR5 and CXCR4 receptors. Macrophages can survive infected for long periods and present compartments containing viruses (VCCs), where HIV-1 accumulates and remains infectious. For these reasons, macrophages may be potential viral reservoirs. On the other hand, dendritic cells (DCs) rarely become infected with HIV-1, but are able to capture viruses and keep them in compartments where they are not degraded. DCs are also able to perform viral transfer to CD4 T cells through the trans-infection process. The capture of HIV-1 by mature DCs during trans-infection is mediated by a surface protein, Siglec1, which binds to ganglioside sialic acid in the viral envelope. Siglec1 is also expressed by macrophages, and its importance in these cells has been linked to the formation of VCCs. However, the molecular mechanisms of internalization and signaling involving the lectin Siglec1 are still not well understood. It is not yet known if there are proteins that help Siglec1 in these processes. Therefore, in this work, we aimed to identify proteins with potential for interaction with Siglec1, in DCs and macrophages, after incubation with HIV-1. For that, co-immunoprecipitation experiments were performed from lysate of DCs and macrophages derived from human monocytes after incubation with HIV-1 virus-like particles. Co-immunoprecipitated proteins obtained by mass spectrometry were identified and after in silico analysis of the data, it was possible to demonstrate that the cell cytoskeleton and its associated proteins appear to play an important role in helping Siglec1 in the capture and internalization of HIV-1. After mass spectrometry analysis, some of the proteins identified were validated by Western Blotting and fluorescence microscopy. Understanding the molecular mechanisms involved in the interaction of Siglec1 with HIV-1 may pave the way for the development of new therapeutic strategies focusing on the dispersion of the virus by DCs and reservoirs in macrophages. (AU) | |
| FAPESP's process: | 19/01733-8 - Siglec1 interactome after HIV-1 binding |
| Grantee: | Maria Clara Martins Ferreira |
| Support Opportunities: | Scholarships in Brazil - Master |