Quantification of Live Bacterial Sensing for Chemotaxis and Phagocytosis and of Macropinocytosis
Initial immunological defense mechanisms to pathogen invasion rely on innate pathways of chemotaxis and phagocytosis, original to ancient phagocytes. Although chemotaxis has been well-studied in mammalian and model systems using purified chemoattractants in defined conditions, directed movement towa...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-03-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/article/10.3389/fcimb.2018.00062/full |
| _version_ | 1818548247316135936 |
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| author | Netra P. Meena Alan R. Kimmel |
| author_facet | Netra P. Meena Alan R. Kimmel |
| author_sort | Netra P. Meena |
| collection | DOAJ |
| description | Initial immunological defense mechanisms to pathogen invasion rely on innate pathways of chemotaxis and phagocytosis, original to ancient phagocytes. Although chemotaxis has been well-studied in mammalian and model systems using purified chemoattractants in defined conditions, directed movement toward live bacteria has been more difficult to assess. Dictyostelium discoideum is a professional phagocyte that chemotaxes toward bacteria during growth-phase in a process to locate nutrient sources. Using Dictyostelium as a model, we have developed a system that is able to quantify chemotaxis to very high sensitivity. Here, Dictyostelium can detect various chemoattractants at concentrations <1 nM. Given this exceedingly sensitive signal response, Dictyostelium will migrate directionally toward live gram positive and gram negative bacteria, in a highly quantifiable manner, and dependent upon bacterially-secreted chemoattractants. Additionally, we have developed a real-time, quantitative assay for phagocytosis of live gram positive and gram negative bacteria. To extend the analyses of endocytic functions, we further modified the system to quantify cellular uptake via macropinocytosis of smaller (<100 kDa) molecules. These various approaches provide novel means to dissect potential for identification of novel chemoattractants and mechanistic factors that are essential for chemotaxis, phagocytosis, and/or macropinocytosis and for more detailed understanding in host-pathogen interactive defenses. |
| first_indexed | 2024-12-12T08:17:25Z |
| format | Article |
| id | doaj.art-b9a164aaa6c04d938652a142506358ab |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-12-12T08:17:25Z |
| publishDate | 2018-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-b9a164aaa6c04d938652a142506358ab2022-12-22T00:31:32ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882018-03-01810.3389/fcimb.2018.00062323050Quantification of Live Bacterial Sensing for Chemotaxis and Phagocytosis and of MacropinocytosisNetra P. MeenaAlan R. KimmelInitial immunological defense mechanisms to pathogen invasion rely on innate pathways of chemotaxis and phagocytosis, original to ancient phagocytes. Although chemotaxis has been well-studied in mammalian and model systems using purified chemoattractants in defined conditions, directed movement toward live bacteria has been more difficult to assess. Dictyostelium discoideum is a professional phagocyte that chemotaxes toward bacteria during growth-phase in a process to locate nutrient sources. Using Dictyostelium as a model, we have developed a system that is able to quantify chemotaxis to very high sensitivity. Here, Dictyostelium can detect various chemoattractants at concentrations <1 nM. Given this exceedingly sensitive signal response, Dictyostelium will migrate directionally toward live gram positive and gram negative bacteria, in a highly quantifiable manner, and dependent upon bacterially-secreted chemoattractants. Additionally, we have developed a real-time, quantitative assay for phagocytosis of live gram positive and gram negative bacteria. To extend the analyses of endocytic functions, we further modified the system to quantify cellular uptake via macropinocytosis of smaller (<100 kDa) molecules. These various approaches provide novel means to dissect potential for identification of novel chemoattractants and mechanistic factors that are essential for chemotaxis, phagocytosis, and/or macropinocytosis and for more detailed understanding in host-pathogen interactive defenses.http://journal.frontiersin.org/article/10.3389/fcimb.2018.00062/fullcell migrationBacillusPseudomonasE. colipterincAMP |
| spellingShingle | Netra P. Meena Alan R. Kimmel Quantification of Live Bacterial Sensing for Chemotaxis and Phagocytosis and of Macropinocytosis Frontiers in Cellular and Infection Microbiology cell migration Bacillus Pseudomonas E. coli pterin cAMP |
| title | Quantification of Live Bacterial Sensing for Chemotaxis and Phagocytosis and of Macropinocytosis |
| title_full | Quantification of Live Bacterial Sensing for Chemotaxis and Phagocytosis and of Macropinocytosis |
| title_fullStr | Quantification of Live Bacterial Sensing for Chemotaxis and Phagocytosis and of Macropinocytosis |
| title_full_unstemmed | Quantification of Live Bacterial Sensing for Chemotaxis and Phagocytosis and of Macropinocytosis |
| title_short | Quantification of Live Bacterial Sensing for Chemotaxis and Phagocytosis and of Macropinocytosis |
| title_sort | quantification of live bacterial sensing for chemotaxis and phagocytosis and of macropinocytosis |
| topic | cell migration Bacillus Pseudomonas E. coli pterin cAMP |
| url | http://journal.frontiersin.org/article/10.3389/fcimb.2018.00062/full |
| work_keys_str_mv | AT netrapmeena quantificationoflivebacterialsensingforchemotaxisandphagocytosisandofmacropinocytosis AT alanrkimmel quantificationoflivebacterialsensingforchemotaxisandphagocytosisandofmacropinocytosis |