T-Cell Epitope Mapping in Mycobacterium tuberculosis Using PepMixes Created by Micro-Scale SPOT™− Synthesis

Frieder, Marisa; Lewinsohn, David

doi:10.1007/978-1-59745-450-6_27

Cited by 7 publications

(4 citation statements)

References 27 publications

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“…As a result, just 12 vaccine epitopes representing each of these 12 MHC‐I supertypes would lead to almost complete population coverage. To date, however, only CTL epitopes restricted by a limited number of HLA molecules have been identified 19 …”

Section: Introductionmentioning

confidence: 99%

Identification of MHC class II restricted T-cell-mediated reactivity against MHC class I binding Mycobacterium tuberculosis peptides

et al. 2011

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Summary Major histocompatibility complex (MHC) class I restricted cytotoxic T lymphocytes (CTL) are known to play an important role in the control of Mycobacterium tuberculosis infection so identification of CTL epitopes from M. tuberculosis is of importance for the development of effective peptide‐based vaccines. In the present work, bioinformatics technology was employed to predict binding motifs of 9mer peptides derived from M. tuberculosis for the 12 HLA‐I supertypes. Subsequently, the predicted peptides were synthesized and assayed for binding to HLA‐I molecules in a biochemically based system. The antigenicity of a total of 157 peptides with measured affinity for HLA‐I molecules of KD ≤ 500 nm were evaluated using peripheral blood T cells from strongly purified protein derivative reactive healthy donors. Of the 157 peptides, eight peptides (5%) were found to induce T‐cell responses. As judged from blocking with HLA class I and II subtype antibodies in the ELISPOT assay culture, none of the eight antigenic peptides induced HLA class I restricted CD8+ T‐cell responses. Instead all responses were blocked by pan‐HLA class II and anti‐HLA‐DR antibodies. In addition, CD4+ T‐cell depletion before the 10 days of expansion, resulted in total loss of reactivity in the ELISPOT culture for most peptide specificities. FACS analyses with intracellular interferon‐γ staining of T cells expanded in the presence of M. tuberculosis peptides confirmed that the responsive cells were indeed CD4+. In conclusion, T‐cell immunity against HLA‐I binding 9mer M. tuberculosis‐derived peptides might in many cases turn out to be mediated by CD4+ T cells and restricted by HLA‐II molecules. The use of 9mer peptides recognized by both CD8+ and CD4+ T cells might be of importance for the development of future M. tuberculosis peptide‐based vaccines.

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Section: Introductionmentioning

confidence: 99%

Identification of MHC class II restricted T-cell-mediated reactivity against MHC class I binding Mycobacterium tuberculosis peptides

et al. 2011

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“…For example, whole genome analysis can be performed in order to identify candidate vaccine components, or alternatively, bioinformatics tools that select proteins according to their secretion characteristics can be used to narrow the number of proteins evaluated in the next step [14][15][16]. Frieder et al used pepmixes created by MicroScale SPOT TM to map T cell epitopes in 389 proteins of Mycobacterium tuberculosis [17]. These proteins are sorted by their known or likely known function such as PPE/PE, cell wall, cell processes, virulence, detoxification, and adaptation.…”

Section: Selection Of Antigens From Mycobacterium Tuberculosismentioning

confidence: 99%

Identification of CD8+ T Cell Epitopes Against Mycobacterium tuberculosis

Chen¹,

Gao²,

Qi³

2012

Understanding Tuberculosis - Analyzing the Origin of Mycobacterium Tuberculosis Pathogenicity

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“…Although possible, the mapping of the discontinuous binding sites using PEPscan may sometimes turn more difficult due to the lower affinities of the individual fragments with the corresponding partner [ 26 ]. One of the first applications of PEPscan was the identification of epitopes recognized by monoclonal antibodies [ 27 ] as well as the recognition of T and B cells epitopes [ 5 , 28 , 29 , 30 ]. The epitopes recognized were mapped using a series of 15 mer peptides synthesized using the SPOT method and with a shift of five amino acids between two peptides.…”

Section: Introductionmentioning

confidence: 99%

Pepscan Approach for the Identification of Protein–Protein Interfaces: Lessons from Experiment

2021

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PEPscan is an old approach that has recently gained renewed interest for the identification of interfering peptides (IPs), i.e., peptides able to interfere with protein–protein interactions (PPIs). Its principle is to slice a protein sequence as a series of short overlapping peptides that are synthesized on a peptide array and tested for their ability to bind a partner, with positive spots corresponding to candidate IPs. PEPscan has been applied with a rather large success in various contexts, but the structural determinants underlying this success remain obscure. Here, we analyze the results of 14 PEPscan experiments, and confront the in vitro results with the available structural information. PEPscan identifies candidate IPs in limited numbers that in all cases correspond to solvent-accessible regions of the structures, their location at the protein–protein interface remaining to be further demonstrated. A strong point of PEPscan seems to be its ability to identify specific IPs. IPs identified from the same protein differ depending on the target PPI, and correspond to patches not frequently involved in the interactions seen in the 3D structures available. Overall, PEPscan seems to provide a cheap and rapid manner to identify candidate IPs, that also comes with room for improvement.

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T-Cell Epitope Mapping in Mycobacterium tuberculosis Using PepMixes Created by Micro-Scale SPOT™− Synthesis

Cited by 7 publications

References 27 publications

Identification of MHC class II restricted T-cell-mediated reactivity against MHC class I binding Mycobacterium tuberculosis peptides

Identification of MHC class II restricted T-cell-mediated reactivity against MHC class I binding Mycobacterium tuberculosis peptides

Identification of CD8+ T Cell Epitopes Against Mycobacterium tuberculosis

Pepscan Approach for the Identification of Protein–Protein Interfaces: Lessons from Experiment

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