The unique tropism of Mycobacterium leprae to the nasal epithelial cells can be explained by the mammalian cell entry protein 1A
| Autor: | Naoya Sato, Andini Febriyanda, Hiroaki Takimoto, Irfan Idris, Vienza Beby Aftitah, Viesta Beby Fadlitha, Takao Fujimura, Haslindah Dahlan, Fuki Yamamoto |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Bacterial Diseases Physiology Protein Expression Respiratory System Colony Count Microbial Cultured tumor cells medicine.disease_cause Biochemistry Epithelium law.invention HeLa 0302 clinical medicine law Animal Cells Medicine and Health Sciences Mycobacterium leprae Latex beads biology Chemistry lcsh:Public aspects of medicine Mycobacterium Leprae Microspheres Recombinant Proteins Body Fluids Actinobacteria medicine.anatomical_structure Infectious Diseases Recombinant DNA Cell lines Antibody Cellular Types Anatomy Biological cultures Research Article Neglected Tropical Diseases DNA Bacterial lcsh:Arctic medicine. Tropical medicine lcsh:RC955-962 030231 tropical medicine Genetic Vectors Cell Line Mycobacterium tuberculosis 03 medical and health sciences Bacterial Proteins Leprosy medicine Escherichia coli Gene Expression and Vector Techniques Humans HeLa cells Molecular Biology Techniques Molecular Biology Tropism Nasal Septum Molecular Biology Assays and Analysis Techniques Bacteria Public Health Environmental and Occupational Health Organisms Biology and Life Sciences Proteins Epithelial Cells lcsh:RA1-1270 Cell Biology biology.organism_classification Cell cultures Tropical Diseases Molecular biology Research and analysis methods Nasal Mucosa 030104 developmental biology Biological Tissue Cell culture biology.protein Mycobacterium Tuberculosis |
| Zdroj: | PLoS Neglected Tropical Diseases, Vol 13, Iss 3, p e0006704 (2019) PLoS Neglected Tropical Diseases |
| DOI: | 10.1101/373324 |
| Popis: | Leprosy is a chronic infection where the skin and peripheral nervous system is invaded by Mycobacterium leprae. The infection mechanism remains unknown in part because culture methods have not been established yet for M. leprae. Mce1A protein (442 aa) is coded by mce1A (1326 bp) of M. leprae. The Mce1A homolog in Mycobacterium tuberculosis is known to be associated with M. tuberculosis epithelial cell entry, and survival and multiplication within macrophages. Studies using recombinant proteins have indicated that Mce1A of M. leprae is also associated with epithelial cell entry. This study is aimed at identifying particular sequences within Mce1A associated with M. leprae epithelial cell entry. Recombinant proteins having N-terminus and C-terminus truncations of the Mce1A region of M. leprae were created in Escherichia coli. Entry activity of latex beads, coated with these truncated proteins (r-lep37 kDa and r-lep27 kDa), into HeLa cells was observed by electron microscopy. The entry activity was preserved even when 315 bp (105 aa) and 922 bp (308 aa) was truncated from the N-terminus and C-terminus, respectively. This 316–921 bp region was divided into three sub-regions: 316–531 bp (InvX), 532–753 bp (InvY), and 754–921 bp (InvZ). Each sub-region was cloned into an AIDA vector and expressed on the surface of E. coli. Entry of these E. coli into monolayer-cultured HeLa and RPMI2650 cells was observed by electron microscopy. Only E. coli harboring the InvX sub-region exhibited cell entry. InvX was further divided into 4 domains, InvXa—InvXd, containing sequences 1–24 aa, 25–46 aa, 47–57 aa, and 58–72 aa, respectively. Recombinant E. coli, expressing each of InvXa—InvXd on the surface, were treated with antibodies against these domains, then added to monolayer cultured RPMI cells. The effectiveness of these antibodies in preventing cell entry was studied by colony counting. Entry activity was suppressed by antibodies against InvXa, InvXb, and InvXd. This suggests that these three InvX domains of Mce1A are important for M. leprae invasion into nasal epithelial cells. Author summary Mce1A protein is a cell surface protein encoded by the mce1A region of mce1 locus of M. tuberculosis and M. leprae, and is involved in the bacteria’s invasion into epithelial cells. Using cloned sub domains of mce1A and peptides synthesized for these sub domains, cell entry studies and binding studies were performed. The present study revealed that the active sequence of M. leprae involved in the invasion into nasal mucosa epithelial cells is present in the 316–531 bp region of mce1A. The Mce1A protein is a good candidate as a contributor to invasion of M. leprae and M. tuberculosis into epithelial cells. The comparative data between Mce1A of M. leprae and M. tuberculosis was relied on to further elucidate the role of specific regions within Mce1A. The most important region of Mce1A protein involved in the invasion of M. tuberculosis into human epithelial cells is called the InvIII region, which is located between amino acids at position 130 to 152. The InvIII region of M. tuberculosis corresponds to InvXb of M. leprae. The sequences of these regions are identical between amino acids at positions 1 to position 22 as counted from the N terminus, except that amino acids at positions 1 to 3, 5, 8, 9, 13 are different between M. leprae and M. tuberculosis. |
| Databáze: | OpenAIRE |
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