Diverse HLA-I Peptide Repertoires of the APC Lines MUTZ3-Derived Immature and Mature Dendritic Cells and THP1-Derived Macrophages

Nyambura, Lydon Wainaina; Jarmalavicius, Saulius; Baleeiro, Renato B.; Walden, Peter

doi:10.4049/jimmunol.1600762

Cited by 9 publications

(11 citation statements)

References 44 publications

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“…2.3 x 10 9 THP1MΦi cells were lysed, and affinity chromatography and LC-MS/MS was used to isolate the HLA class I molecules and analyze the HLA-bound peptides. A total of 86 non-redundant HLA class I self-ligands were identified from 82 source proteins of THP1MФi ( S1 Table ) compared to 347 non-redundant HLA-I self-ligands derived from 282 source proteins identified for 2.8 x 10 9 THP1MΦ cells at the same time and reported earlier [ 26 ]. Only 17 HLA-I self-peptide sequences and 18 source proteins were found to be shared between THP1MФ and THP1MФi.…”

Section: Resultsmentioning

confidence: 83%

“…Thus, in both THP1MФ and THP1MФi the HLA I-bound peptides were dominated by nonapeptides, though the percentage in THP1MФi was slightly higher by 4%. Nonapeptides are the optimum lengths of HLA I-bound peptides [ 25 , 26 , 35 – 37 ]. The slight increase of nonapeptides in THP1MФi compared to THP1MФ could suggest a shift in antigen processing towards the more optimum peptide lengths for MHC I binding.…”

Section: Resultsmentioning

confidence: 99%

“…The heterogeneity and individuality in the HLA I self-peptides and source proteins identified in THP1MФ and THP1MФi depicts differences in protein expression, processing and presentation, as was in other cells and tumor samples [ 25 , 26 , 35 , 36 ]. The nonapeptides are the optimum lengths for MHC class I binding [ 26 ] and though the infection of THP1MФ by LD did not change the nonapeptides dominance in the identified HLA I-bound peptides, profound differences in antigen processing and presentation were evident, firstly, in the HLA restriction of identified peptides. For THP1MΦ the percentage of the peptides identified for the different HLA-restrictions ranked in the order HLA-A*02:01 > HLA-B*15:11 > HLA-C*03:03 while in THP1MФi they were in the order HLA-B*15:11 > HLA-C*03:03 > HLA-A*02:01.…”

Section: Discussionmentioning

confidence: 99%

“…Isolation of MHC I molecules was carried out as previously described [ 25 , 26 ]. In brief, 2.3 x 10 9 shock frozen cells were lyzed in 0.3% CHAPS, 0.2% NP-40, 145 mM NaCl, 1 mM EDTA, 1mM Pefabloc, 20 mM Tris-HCl buffer at pH 7.4 and ultracentrifuged for 1h at 100,000 x g. HLA I molecules were purified from the supernatants using monoclonal antibody of irrelevant specificity for preclearing and HLA class I-specific mAb W6/32, respectively, coupled to CNBr-activated sepharose (Amersham Biosciences, Uppsala, Sweden).…”

Section: Methodsmentioning

confidence: 99%

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Impact of Leishmania donovani infection on the HLA I self peptide repertoire of human macrophages

Nyambura¹,

Jarmalavicius²,

Walden³

2018

PLoS ONE

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Macrophages are specialized antigen-presenting cells that process and present self-antigens for induction of tolerance, and foreign antigens to initiate T cell-mediated immunity. Despite this, Leishmania donovani (LD) are able to parasitize the macrophages and persist. The impact of this parasitizing and persistence on antigen processing and presentation by macrophages remains poorly defined. To gain insight into this, we analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS) and compared the HLA-I self-peptidomes, proteasome compositions, HLA expression and activation states of non-infected and LD-infected THP1-derived macrophages. We found that, though both HLA-I peptidomes were dominated by nonapeptides, they were heterogeneous and individualized, with differences in HLA binding affinities and anchor residues. Non-infected and LD-infected THP1-derived macrophages were able to sample peptides from source proteins of almost all subcellular locations and involved in various cellular functions, but in different proportions. In the infected macrophages, there was increased sampling of plasma membrane and extracellular proteins, and those involved in immune responses, cell communication/signal transduction and metabolism/energy pathways, and decreased sampling of nuclear and cytoplasmic proteins and those involved in protein metabolism, RNA binding and cell growth and/or maintenance. Though the activation state of infected macrophages was unchanged, their proteasome composition was altered.

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Section: Resultsmentioning

confidence: 83%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Impact of Leishmania donovani infection on the HLA I self peptide repertoire of human macrophages

Nyambura¹,

Jarmalavicius²,

Walden³

2018

PLoS ONE

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“…In a prior study 30 , we used a direct method of target identification: immunoaffinity purification of human leukocyte antigen I (HLA-I) peptide complexes from the antigen-presenting immature and mature dendritic cells, and macrophages derived from the myeloid cell lines MUTZ3 and THP1 coupled to liquid chromatographic tandem mass spectrometry (LC-MS/MS). These cell lines were established originally from leukemia cells obtained from patients.…”

Section: Introductionmentioning

confidence: 99%

HLA class I-restricted T cell epitopes isolated and identified from myeloid leukemia cells

Nyambura

Muñoz

Coutre

et al. 2019

Sci Rep

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Leukemia-associated antigens (LAAs) and HLA-I epitopes published previously have shown promise in inducing leukemia-specific T cell responses. However, the clinical responses are limited, and clinical effectiveness is yet to be achieved. Limitations, among others, being the LAAs themselves, the indirect approach to HLA-I epitope identification by reverse immunology, and the use of single or few LAAs and HLA-I epitopes, which limits the spectrum of inducible tumor-specific T cells. Use of a direct approach to identify naturally processed and presented HLA-I epitopes from LAAs, and higher numbers of antigens for T cell-mediated immunotherapy for leukemia may enhance clinical responses and broaden clinical effectiveness. In a prior study we used immunoaffinity purification of HLA-I peptide complexes from the differentiated myeloid tumor cell lines MUTZ3 and THP1 coupled to high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS). From this we identified in the current study seven new HLA-I epitopes and the corresponding LAAs for myeloid leukemia. In comparison, the myeloid HLA-I epitopes reported here were generally stronger HLA-binders that induce stronger T cell responses than those previously published, and their source LAAs had higher immunogenicity, higher expression levels in myeloid tumors cells compared to normal hemopoietin and other major normal tissues, and more protein interaction partners, and they are targeted by CD8 T cells in CML patients. This study analyses and compares the LAAs and HLA-I epitopes based on various immunotherapeutic targets selection criteria, and highlights new targets for T cell-mediated immunotherapy for leukemia.

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Tumor‐Associated Macrophage‐Derived Exosomal LINC01232 Induces the Immune Escape in Glioma by Decreasing Surface MHC‐I Expression

Wang

Yang

et al. 2023

Advanced Science

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Tumor-associated macrophage (TAM) infiltration facilitates glioma malignancy, but the underlying mechanisms remain unclear. Herein, it is reported that TAMs secrete exosomal LINC01232 to induce tumor immune escape. Mechanistically, LINC01232 is found to directly bind E2F2 and promote E2F2 entry into the nucleus; the two synergistically promots the transcription of NBR1. The increase in binding between NBR1 binding and the ubiquitinating MHC-I protein through the ubiquitin domain causes an increase in the degradation of MHC-I in autophagolysosomes and a decrease in the expression of MHC-I on the surface of tumor cells, which in turn led to tumor cell escape from CD8 + CTL immune attack. Disruption of E2F2/NBR1/MHC-I signaling with shRNAs or blockade with the corresponding antibodies largely abolishes the tumor-supportive effects of LINC01232 and inhibits tumor growth driven by M2-type macrophages. Importantly, knockdown of LINC01232 enhances the expression of MHC-I on the surface of tumor cells and improves the response to reinfusion with CD8 + T cells. This study reveals the existence of critical molecular crosstalk between TAMs and glioma mediates through the LINC01232/E2F2/NBR1/MHC-I axis to support malignant tumor growth, indicating that targeting this axis may have therapeutic potential.

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Diverse HLA-I Peptide Repertoires of the APC Lines MUTZ3-Derived Immature and Mature Dendritic Cells and THP1-Derived Macrophages

Cited by 9 publications

References 44 publications

Impact of Leishmania donovani infection on the HLA I self peptide repertoire of human macrophages

Impact of Leishmania donovani infection on the HLA I self peptide repertoire of human macrophages

HLA class I-restricted T cell epitopes isolated and identified from myeloid leukemia cells

Tumor‐Associated Macrophage‐Derived Exosomal LINC01232 Induces the Immune Escape in Glioma by Decreasing Surface MHC‐I Expression

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