Chapter title |
CD4+ T Cells Mediate Aspergillosis Vaccine Protection
|
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Chapter number | 19 |
Book title |
Vaccines for Invasive Fungal Infections
|
Published in |
Methods in molecular biology, June 2017
|
DOI | 10.1007/978-1-4939-7104-6_19 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7103-9, 978-1-4939-7104-6
|
Authors |
Diaz-Arevalo, Diana, Kalkum, Markus, Diana Diaz-Arevalo, Markus Kalkum |
Editors |
Markus Kalkum, Margarita Semis |
Abstract |
Adaptive effector CD4(+) T cells play essential roles in the defense against fungal infections, especially against invasive aspergillosis (IA). Such protective CD4(+) T cells can be generated through immunization with specialized antifungal vaccines, as has been demonstrated for pulmonary Aspergillus fumigatus infections in mouse experiments. Adaptive transfer of fungal antigen-specific CD4(+) T cells conferred protection onto non-immunized naive mice, an experimental approach that could potentially become a future treatment option for immunosuppressed IA patients, focusing on the ultimate goal to improve their otherwise dim chances for survival. Here, we describe the different techniques to analyze CD4(+) T cell immune responses after immunization with a recombinant fungal protein. We present three major methods that are used to analyze the role of CD4(+) T cells in protection against A. fumigatus challenge. They include (1) transplantation of CD4(+) T cells from vaccinated mice into immunosuppressed naive mice, observing increasing protection of the cell recipients, (2) depletion of CD4(+) T cells from vaccinated mice, which abolishes vaccine protection, and (3) T cell proliferation studies following stimulation with overlapping synthetic peptides or an intact protein vaccine. The latter can be used to validate immunization status and to identify protective T cell epitopes in vaccine antigens. In the methods detailed here, we used versions of the well-studied Asp f3 protein expressed in a bacterial host, either as the intact full length protein or its N-terminally truncated version, comprised of residues 15-168. However, these methods are generally applicable and can well be adapted to study other protein-based subunit vaccines. |
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