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HLA class I-restricted MYD88 L265P-derived peptides as specific targets for lymphoma immunotherapy

View Article: PubMed Central - PubMed

ABSTRACT

Genome sequencing has uncovered an array of recurring somatic mutations in different non-Hodgkin lymphoma (NHL) subtypes. If affecting protein-coding regions, such mutations may yield mutation-derived peptides that may be presented by HLA class I proteins and recognized by cytotoxic T cells. A recurring somatic and oncogenic driver mutation of the Toll-like receptor adaptor protein MYD88, Leu265Pro (L265P) was identified in up to 90% of different NHL subtype patients. We therefore screened the potential of MYD88L265P-derived peptides to elicit cytotoxic T cell responses as tumor-specific neoantigens. Based on in silico predictions, we identified potential MYD88L265P-containing HLA ligands for several HLA class I restrictions. A set of HLA class I MYD88L265P-derived ligands elicited specific cytotoxic T cell responses for HLA-B*07 and -B*15. These data highlight the potential of MYD88L265P mutation-specific peptide-based immunotherapy as a novel personalized treatment approach for patients with MYD88L265P+ NHLs that may complement pharmacological approaches targeting oncogenic MyD88 L265P signaling.

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Spontaneous memory T cell responses are detectable in a leukemia patient. The presence of memory T cell responses in leukemia and lymphoma patients was analyzed using 12-d recall IFNγ ELISPOT assays. (A) In a single (out of 22 tested) MYD88L265P+ NHL patients (CLL-05-R) IFNγ secretion was observed after stimulation with the MYD88L265P-derived peptides P5B*15/B*40 (HQKRPIPI) and P1C*03 (RPIPIKYKAM). (B) Representative example of a MYD88L265P+ patient (CLL-03-R) where no IFNγ secretion was observed after stimulation with the MYD88L265P-derived peptides P5B*15 (HQKRPIPI) and P1C*03 (RPIPIKYKAM). An EBV epitope mix containing the frequently recognized peptides BRLF1 109–117 YVLDHLIVV (HLA-A*02) and EBNA3 247–255 RPPIFIRRL (HLA-B*07) served as positive control. Benign-tissue derived peptide DDX5 YLLPAIVHI (HLA-A*02) served as negative control. The dotted line indicates the 3-fold number of spot forming unit of the negative control. Error bars indicate ± SEM of two independent replicates. Abbreviations: SFU, spot forming unit; neg., negative; pos., positive; SEM, standard error of the mean.
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f0002: Spontaneous memory T cell responses are detectable in a leukemia patient. The presence of memory T cell responses in leukemia and lymphoma patients was analyzed using 12-d recall IFNγ ELISPOT assays. (A) In a single (out of 22 tested) MYD88L265P+ NHL patients (CLL-05-R) IFNγ secretion was observed after stimulation with the MYD88L265P-derived peptides P5B*15/B*40 (HQKRPIPI) and P1C*03 (RPIPIKYKAM). (B) Representative example of a MYD88L265P+ patient (CLL-03-R) where no IFNγ secretion was observed after stimulation with the MYD88L265P-derived peptides P5B*15 (HQKRPIPI) and P1C*03 (RPIPIKYKAM). An EBV epitope mix containing the frequently recognized peptides BRLF1 109–117 YVLDHLIVV (HLA-A*02) and EBNA3 247–255 RPPIFIRRL (HLA-B*07) served as positive control. Benign-tissue derived peptide DDX5 YLLPAIVHI (HLA-A*02) served as negative control. The dotted line indicates the 3-fold number of spot forming unit of the negative control. Error bars indicate ± SEM of two independent replicates. Abbreviations: SFU, spot forming unit; neg., negative; pos., positive; SEM, standard error of the mean.

Mentions: Functional characterization of the predicted candidate HLA class I MYD88L265P-derived NHL-specific ligands was performed by 12-d recall IFNγ ELISPOT assays using PBMCs obtained from MYD88L265P-mutated and MYD88WT patients (Table S1). In one (out of 22 tested) MYD88L265P-mutated NHL patients, memory T cell responses targeting two different MYD88L265P-derived HLA class I ligands were detected by IFNγ ELISPOT (Fig. 2). Importantly, one of the peptides (P5) is a predicted ligand for both, HLA-B*15 and -B*40, which are both expressed by the patient. Therefore, we could not resolve which peptide:HLA combination is responsible for the observed IFNγ secretion. Importantly, no IFNγ secretion was observed for any tested ligand in MYD88WT patients. The frequency of memory T cell responses in 1/22 MYD88L265P-mutated NHL patients appears low, but it is important to be aware of the HLA allotype-specific frequencies. The peptide P1, which is among others analyzed in further immunogenicity experiments, leads to a detectable memory T cell response in 1/3 (33%) tested HLA-matched MYD88L265P-mutated NHL patients. For the other positive peptide P5 memory T cell responses could be detected in 1/4 (25%) HLA-B*15 patients and in 1/3 (33%) HLA-B*40 patients. Furthermore, only two-digit HLA typings were available for the NHL patients. Therefore, it is possible that a negative response is due to a different four-digit HLA restriction of the tested peptide and the HLA type of the patient. Table S3 summarizes all tested MYD88L265P-derived HLA class I ligands.Figure 2.


HLA class I-restricted MYD88 L265P-derived peptides as specific targets for lymphoma immunotherapy
Spontaneous memory T cell responses are detectable in a leukemia patient. The presence of memory T cell responses in leukemia and lymphoma patients was analyzed using 12-d recall IFNγ ELISPOT assays. (A) In a single (out of 22 tested) MYD88L265P+ NHL patients (CLL-05-R) IFNγ secretion was observed after stimulation with the MYD88L265P-derived peptides P5B*15/B*40 (HQKRPIPI) and P1C*03 (RPIPIKYKAM). (B) Representative example of a MYD88L265P+ patient (CLL-03-R) where no IFNγ secretion was observed after stimulation with the MYD88L265P-derived peptides P5B*15 (HQKRPIPI) and P1C*03 (RPIPIKYKAM). An EBV epitope mix containing the frequently recognized peptides BRLF1 109–117 YVLDHLIVV (HLA-A*02) and EBNA3 247–255 RPPIFIRRL (HLA-B*07) served as positive control. Benign-tissue derived peptide DDX5 YLLPAIVHI (HLA-A*02) served as negative control. The dotted line indicates the 3-fold number of spot forming unit of the negative control. Error bars indicate ± SEM of two independent replicates. Abbreviations: SFU, spot forming unit; neg., negative; pos., positive; SEM, standard error of the mean.
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f0002: Spontaneous memory T cell responses are detectable in a leukemia patient. The presence of memory T cell responses in leukemia and lymphoma patients was analyzed using 12-d recall IFNγ ELISPOT assays. (A) In a single (out of 22 tested) MYD88L265P+ NHL patients (CLL-05-R) IFNγ secretion was observed after stimulation with the MYD88L265P-derived peptides P5B*15/B*40 (HQKRPIPI) and P1C*03 (RPIPIKYKAM). (B) Representative example of a MYD88L265P+ patient (CLL-03-R) where no IFNγ secretion was observed after stimulation with the MYD88L265P-derived peptides P5B*15 (HQKRPIPI) and P1C*03 (RPIPIKYKAM). An EBV epitope mix containing the frequently recognized peptides BRLF1 109–117 YVLDHLIVV (HLA-A*02) and EBNA3 247–255 RPPIFIRRL (HLA-B*07) served as positive control. Benign-tissue derived peptide DDX5 YLLPAIVHI (HLA-A*02) served as negative control. The dotted line indicates the 3-fold number of spot forming unit of the negative control. Error bars indicate ± SEM of two independent replicates. Abbreviations: SFU, spot forming unit; neg., negative; pos., positive; SEM, standard error of the mean.
Mentions: Functional characterization of the predicted candidate HLA class I MYD88L265P-derived NHL-specific ligands was performed by 12-d recall IFNγ ELISPOT assays using PBMCs obtained from MYD88L265P-mutated and MYD88WT patients (Table S1). In one (out of 22 tested) MYD88L265P-mutated NHL patients, memory T cell responses targeting two different MYD88L265P-derived HLA class I ligands were detected by IFNγ ELISPOT (Fig. 2). Importantly, one of the peptides (P5) is a predicted ligand for both, HLA-B*15 and -B*40, which are both expressed by the patient. Therefore, we could not resolve which peptide:HLA combination is responsible for the observed IFNγ secretion. Importantly, no IFNγ secretion was observed for any tested ligand in MYD88WT patients. The frequency of memory T cell responses in 1/22 MYD88L265P-mutated NHL patients appears low, but it is important to be aware of the HLA allotype-specific frequencies. The peptide P1, which is among others analyzed in further immunogenicity experiments, leads to a detectable memory T cell response in 1/3 (33%) tested HLA-matched MYD88L265P-mutated NHL patients. For the other positive peptide P5 memory T cell responses could be detected in 1/4 (25%) HLA-B*15 patients and in 1/3 (33%) HLA-B*40 patients. Furthermore, only two-digit HLA typings were available for the NHL patients. Therefore, it is possible that a negative response is due to a different four-digit HLA restriction of the tested peptide and the HLA type of the patient. Table S3 summarizes all tested MYD88L265P-derived HLA class I ligands.Figure 2.

View Article: PubMed Central - PubMed

ABSTRACT

Genome sequencing has uncovered an array of recurring somatic mutations in different non-Hodgkin lymphoma (NHL) subtypes. If affecting protein-coding regions, such mutations may yield mutation-derived peptides that may be presented by HLA class I proteins and recognized by cytotoxic T cells. A recurring somatic and oncogenic driver mutation of the Toll-like receptor adaptor protein MYD88, Leu265Pro (L265P) was identified in up to 90% of different NHL subtype patients. We therefore screened the potential of MYD88L265P-derived peptides to elicit cytotoxic T cell responses as tumor-specific neoantigens. Based on in silico predictions, we identified potential MYD88L265P-containing HLA ligands for several HLA class I restrictions. A set of HLA class I MYD88L265P-derived ligands elicited specific cytotoxic T cell responses for HLA-B*07 and -B*15. These data highlight the potential of MYD88L265P mutation-specific peptide-based immunotherapy as a novel personalized treatment approach for patients with MYD88L265P+ NHLs that may complement pharmacological approaches targeting oncogenic MyD88 L265P signaling.

No MeSH data available.


Related in: MedlinePlus