Trypanosoma cruzi invades non-professional phagocytic cells by subverting their membrane repair process, which is dependent on membrane injury and cell signaling, intracellular calcium increase, and lysosome recruitment. Cells lacking lysosome-associated membrane proteins 1 and 2 (LAMP1 and LAMP2) are less permissive to parasite invasion but more prone to parasite intracellular multiplication. Several passages through a different intracellular environment can significantly change T. cruzi’s gene expression profile. Here, we evaluated whether one single passage through LAMP-deficient (KO) or wild-type (WT) fibroblasts, thus different intracellular environments, could influence T. cruzi Y strain trypomastigotes’ ability to invade L6 myoblasts and WT fibroblasts host cells. Parasites released from LAMP2 KO cells (TcY-L2−/−) showed higher invasion, calcium signaling, and membrane injury rates, for the assays in L6 myoblasts, when compared to those released from WT (TcY-WT) or LAMP1/2 KO cells (TcY-L1/2−/−). On the other hand, TcY-L1/2−/− showed higher invasion, calcium signaling, and cell membrane injury rates, for the assays in WT fibroblasts, compared to TcY-WT and TcY-L1/2−/−. Albeit TcY-WT presented an intermediary invasion and calcium signaling rates, compared to the others, in WT fibroblasts, they induced lower levels of injury, which reinforces that signals mediated by surface membrane protein interactions also have a significant contribution to trigger host cell calcium signals. These results clearly show that parasites released from WT or LAMP KO cells are distinct from each other. Additionally, these parasites’ ability to invade the cell may be distinct depending on which cell type they interact with. Since these alterations most likely would reflect differences among parasite surface molecules, we also evaluated their proteome. We identified few protein complexes, membrane, and secreted proteins regulated in our dataset. Among those are some members of MASP, mucins, trans-sialidases, and gp63 proteins family, which are known to play an important role during parasite infection and could correlate to TcY-WT, TcY-L1/2−/−, and TcY-L2−/− biological behavior.
Bibliografisk noteFunding Information:
Below are the funding resources for the development of this research: Conselho Nacional de Desenvolvimento Cientıfí co e Tecnológico (CNPq) (MCTIC/CNPq No 28/2018—Universal 429635/2018-4) Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG—APQ-02974-17), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES / PROEX). None of the fundings received allow for publication fees, the resources being destined only to the acquisition of reagents for the development of the methodological assays.
We are especially grateful to Dr. Paul Saftig and Dr. Renato Mortara for providing the LAMP1/2 and LAMP2 knockout cells that have been used in this work, and the Laboratório de Equipamentos Multiusuários for the use of the Synergy2 and Jamil Silvano de Oliveira from Laboratório de Biologia Estrutural for technical assistance with the ultracentrifuge—Department of Biochemistry and Immunology, ICB/UFMG. We are grateful to the Laboratório de Citometria from the Centro de Laboratórios Multiusuários (CELAM-ICB) and to Daniela Silva dos Reis for technical assistance. We are also grateful to the funding agencies: Conselho Nacional de Desenvolvimento Cientıfí co e Tecnológico— CNPq (MCTIC/CNPq No. 28/2018—Universal 429635/2018-4), Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG—APQ-02974-17) and Coordenacão̧ de Aperfeicoamento̧ de Pessoal de Nıv́ el Superior (CAPES fellowship).
We are especially grateful to Dr. Paul Saftig and Dr. Renato Mortara for providing the LAMP1/2 and LAMP2 knockout cells that have been used in this work, and the Laborat?rio de Equipamentos Multiusu?rios for the use of the Synergy2 and Jamil Silvano de Oliveira from Laborat?rio de Biologia Estrutural for technical assistance with the ultracentrifuge?Department of Biochemistry and Immunology, ICB/UFMG. We are grateful to the Laborat?rio de Citometria from the Centro de Laborat?rios Multiusu?rios (CELAM-ICB) and to Daniela Silva dos Reis for technical assistance. We are also grateful to the funding agencies: Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico?CNPq (MCTIC/CNPq No. 28/2018?Universal 429635/2018-4), Funda??o de Amparo ? Pesquisa do Estado de Minas Gerais (FAPEMIG?APQ-02974-17) and Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES fellowship).
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