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1.
J Virol ; 98(7): e0079124, 2024 Jul 23.
Artículo en Inglés | MEDLINE | ID: mdl-38940584

RESUMEN

Fibrocytes were reported to be host cells for HIV-1, but the immunological recognition of HIV-1-infected fibrocytes has not been studied. Here, we investigated the recognition of HIV-1-infected fibrocytes by HIV-1-specific CD8+ T cells. CD8+ T cells specific for five HIV-1 epitopes (HLA-A*24:02-restricted, HLA-B*52:01-restricted, and HLA-C*12:02-restricted epitopes) produced IFN-γ and expressed CD107a after coculture with HIV-1-infected fibrocytes. HIV-1-infected fibrocytes were effectively killed by HIV-1-specific CD8+ T cells. Although it is well known that HIV-1 Nef-mediated downregulation of HLA-A and HLA-B critically affects the T cell recognition of HIV-1-infected CD4+ T cells and HIV-1-infected macrophages, Nef downregulated HLA-A, but not HLA-B, in HIV-1-infected fibrocytes. These findings suggested that HIV-1-specific CD8+ T cells could recognize HIV-1-infected fibrocytes more strongly than HIV-1-infected CD4+ T cells or HIV-1-infected macrophages. HIV-1-infected fibrocytes were also recognized by HIV-1-specific HLA-DR-restricted T cells, indicating that HIV-1-infected fibrocytes can present HIV-1 epitopes to helper T cells. Collectively, these findings suggest that fibrocytes have an important role as antigen-presenting cells during HIV-1 infection. The present study demonstrates effective recognition of HIV-1-infected fibrocytes by HIV-1-specific T cells and suggests possible roles of fibrocytes in the induction and maintenance of HIV-1-specific T cells. IMPORTANCE: Fibrocytes were identified as unique hematopoietic cells with the features of both macrophages and fibroblasts and were demonstrated to be host cells for HIV-1. However, T cell recognition of HIV-1-infected fibrocytes has not been studied. We investigated the recognition of HIV-1-infected fibrocytes by HIV-1-specific T cells. HIV-1-infected fibrocytes were effectively recognized and killed by CD8+ T cells specific for HIV-1 epitopes presented by HLA-A, HLA-B, or HLA-C and were recognized by HIV-1-specific HLA-DR-restricted CD4+ T cells. HIV-1 Nef-mediated downregulation of HLA-A and HLA-B was found in HIV-1-infected CD4+ T cells, whereas Nef did not downregulate HLA-B in HIV-1-infected fibrocytes. These results suggest that HIV-1-specific CD8+ T cells recognize HIV-1-infected fibrocytes more strongly than HIV-1-infected CD4+ T cells. The present study suggests the importance of fibrocytes in the induction and maintenance of HIV-1-specific T cells.


Asunto(s)
Linfocitos T CD8-positivos , Regulación hacia Abajo , Infecciones por VIH , VIH-1 , Antígenos HLA-B , Productos del Gen nef del Virus de la Inmunodeficiencia Humana , Humanos , VIH-1/inmunología , Productos del Gen nef del Virus de la Inmunodeficiencia Humana/metabolismo , Productos del Gen nef del Virus de la Inmunodeficiencia Humana/inmunología , Linfocitos T CD8-positivos/inmunología , Infecciones por VIH/inmunología , Infecciones por VIH/virología , Antígenos HLA-B/inmunología , Antígenos HLA-B/metabolismo , Fibroblastos/virología , Fibroblastos/inmunología , Fibroblastos/metabolismo , Linfocitos T CD4-Positivos/inmunología , Epítopos de Linfocito T/inmunología , Macrófagos/inmunología , Macrófagos/virología , Macrófagos/metabolismo
2.
Structure ; 32(8): 1121-1136.e5, 2024 Aug 08.
Artículo en Inglés | MEDLINE | ID: mdl-38733995

RESUMEN

Immunogenetic studies have shown that specific HLA-B residues (67, 70, 97, and 156) mediate the impact of HLA class I on HIV infection, but the molecular basis is not well understood. Here we evaluate the function of these residues within the protective HLA-B∗5701 allele. While mutation of Met67, Ser70, and Leu156 disrupt CD8+ T cell recognition, substitution of Val97 had no significant impact. Thermal denaturation of HLA-B∗5701-peptide complexes revealed that Met67 and Leu156 maintain HLA-peptide stability, while Ser70 and Leu156 facilitate T cell receptor (TCR) interactions. Analyses of existing structures and structural models suggested that Val97 mediates HLA-peptide binding to inhibitory KIR3DL1 molecules, which was confirmed by experimental assays. These data thereby demonstrate that the genetic basis by which host immunity impacts HIV outcomes occurs by modulating HLA-B-peptide stability and conformation for interaction with TCR and killer immunoglobulin receptor (KIR) molecules. Moreover, they indicate a key role for epitope specificity and HLA-KIR interactions to HIV control.


Asunto(s)
Antígenos HLA-B , Unión Proteica , Receptores de Antígenos de Linfocitos T , Humanos , Antígenos HLA-B/química , Antígenos HLA-B/metabolismo , Antígenos HLA-B/genética , Antígenos HLA-B/inmunología , Receptores de Antígenos de Linfocitos T/metabolismo , Receptores de Antígenos de Linfocitos T/química , Receptores de Antígenos de Linfocitos T/inmunología , VIH-1/inmunología , VIH-1/metabolismo , Infecciones por VIH/inmunología , Infecciones por VIH/virología , Modelos Moleculares , Receptores KIR3DL1/metabolismo , Receptores KIR3DL1/química , Receptores KIR3DL1/genética , Péptidos/química , Péptidos/metabolismo , Sitios de Unión , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Polimorfismo Genético , Estabilidad Proteica
3.
Eur J Immunol ; 54(6): e2350683, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38549458

RESUMEN

HLA-B*39:06, HLA-B*39:01, and HLA-B*38:01 are closely related HLA allotypes differentially associated with type 1 diabetes (T1D) risk and progression. B*39:06 is highly predisposing, while B*39:01 and B*38:01 are weakly predisposing and protective allotypes, respectively. Here, we aimed to decipher molecular mechanisms underlying the differential association of these allotypes with T1D pathogenesis. We addressed peptide binding and conformational stability of HLA-B allotypes using computational and experimental approaches. Computationally, we found that B*39:06 and B*39:01 allotypes had more rigid peptide-binding grooves and were more promiscuous in binding peptides than B*38:01. Peptidomes of B*39:06 and B*39:01 contained fewer strong binders and were of lower affinity than that of B*38:01. Experimentally, we demonstrated that B*39:06 and B*39:01 had a higher capacity to bind peptides and exit to the cell surface but lower surface levels and were degraded faster than B*38:01. In summary, we propose that promiscuous B*39:06 and B*39:01 may bind suboptimal peptides and transport them the cell surface, where such unstable complexes may contribute to the pathogenesis of T1D.


Asunto(s)
Diabetes Mellitus Tipo 1 , Antígenos HLA-B , Péptidos , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/inmunología , Humanos , Péptidos/química , Péptidos/genética , Péptidos/inmunología , Antígenos HLA-B/genética , Antígenos HLA-B/metabolismo , Polimorfismo Genético , Unión Proteica , Alelos , Estabilidad Proteica , Predisposición Genética a la Enfermedad
4.
Stem Cell Res Ther ; 14(1): 366, 2023 12 13.
Artículo en Inglés | MEDLINE | ID: mdl-38093328

RESUMEN

BACKGROUND: Induced pluripotent stem cell (iPSC)-derived cell therapies are an interesting new area in the field of regenerative medicine. One of the approaches to decrease the costs of iPSC-derived therapies is the use of allogenic homozygous human leukocyte antigen (HLA)-matched donors to generate iPSC lines and to build a clinical-grade iPSC bank covering a high percentage of the Spanish population. METHODS: The Spanish Stem Cell Transplantation Registry was screened for cord blood units (CBUs) homozygous for the most common HLA-A, HLA-B and HLA-DRB1 haplotypes. Seven donors were selected with haplotypes covering 21.37% of the haplotypes of the Spanish population. CD34-positive hematopoietic progenitors were isolated from the mononuclear cell fraction of frozen cord blood units from each donor by density gradient centrifugation and further by immune magnetic labeling and separation using purification columns. Purified CD34 + cells were reprogrammed to iPSCs by transduction with the CTS CytoTune-iPS 2.1 Sendai Reprogramming Kit. RESULTS: The iPSCs generated from the 7 donors were expanded, characterized, banked and registered. Master cell banks (MCBs) and working cell banks (WCBs) from the iPSCs of each donor were produced under GMP conditions in qualified clean rooms. CONCLUSIONS: Here, we present the first clinical-grade, iPSC haplobank in Spain made from CD34 + cells from seven cord blood units homozygous for the most common HLA-A, HLA-B and HLA-DRB1 haplotypes within the Spanish population. We describe their generation by transduction with Sendai viral vectors and their GMP-compliant expansion and banking. These haplolines will constitute starting materials for advanced therapy medicinal product development (ATMP).


Asunto(s)
Células Madre Pluripotentes Inducidas , Humanos , Células Madre Pluripotentes Inducidas/metabolismo , Cadenas HLA-DRB1/metabolismo , Antígenos HLA/metabolismo , Antígenos HLA-B/metabolismo , Antígenos HLA-A/metabolismo
5.
Int J Biol Macromol ; 253(Pt 7): 127199, 2023 Dec 31.
Artículo en Inglés | MEDLINE | ID: mdl-37793526

RESUMEN

The tremendous success of immune checkpoint blockade (ICB) therapy has raised the great demand for the development of predictive biomarkers. A recent cancer genomic study suggested that human leukocyte antigen (HLA)-B*44:02 and HLA-B*15:01 alleles may act as potential biomarkers for ICB therapies, however, the underlying molecular mechanisms remain largely elusive. Here, we investigated the molecular origins of differential responses to ICB therapies for four representative HLA alleles: HLA-B*44:02, HLA-B*15:01, HLA-B*07:02, and HLA-B*53:01, using extensive all-atom molecular dynamics simulations. We first demonstrated that the relatively more rigid peptide-binding groove of HLA-B*15:01, than those in the other three HLA alleles, may result in challenges in its recognition with T-cell receptors. Specifically, the "bridge" structure in HLA-B*15:01 is stabilized through both intramolecular electrostatic interactions between the HLA residues and intermolecular interactions between the HLA and the antigenic peptide. These observations were further confirmed by in silico mutagenesis studies, as well as simulations of several other HLA-B*15:01-peptide complexes. By contrast, the "bridge" structure is either completely absent in HLA-B*44:02 or easily perturbed in HLA-B*07:02 and HLA-B*53:01. Our findings provide detailed structural and mechanistic insights into how HLA genotype influences ICB responses and may have important implications for developing immune markers.


Asunto(s)
Antígenos HLA-B , Neoplasias , Humanos , Antígenos HLA-B/genética , Antígenos HLA-B/metabolismo , Péptidos/química , Inmunidad , Biomarcadores
6.
J Immunol ; 211(9): 1298-1307, 2023 11 01.
Artículo en Inglés | MEDLINE | ID: mdl-37737643

RESUMEN

The extreme polymorphisms of HLA class I proteins result in structural variations in their peptide binding sites to achieve diversity in Ag presentation. External factors could independently constrict or alter HLA class I peptide repertoires. Such effects of the assembly factor tapasin were assessed for HLA-B*44:05 (Y116) and a close variant, HLA-B*44:02 (D116), which have low and high tapasin dependence, respectively, for their cell surface expression. Analyses of the HLA-B*44:05 peptidomes in the presence and absence of tapasin reveal that peptides with C-terminal tryptophans and higher predicted affinities are preferentially selected by tapasin, coincident with reduced frequencies of peptides with other C-terminal amino acids, including leucine. Comparisons of the HLA-B*44:05 and HLA-B*44:02 peptidomes indicate the expected structure-based alterations near the peptide C termini, but also C-terminal amino acid frequency and predicted affinity changes among the unique and shared peptide groups for B*44:02 and B*44:05. Overall, these findings indicate that the presence of tapasin and the tapasin dependence of assembly alter HLA class I peptide-binding preferences at the peptide C terminus. The particular C-terminal amino acid preferences that are altered by tapasin are expected to be determined by the intrinsic peptide-binding specificities of HLA class I allotypes. Additionally, the findings suggest that tapasin deficiency and reduced tapasin dependence expand the permissive affinities of HLA class I-bound peptides, consistent with prior findings that HLA class I allotypes with low tapasin dependence have increased breadth of CD8+ T cell epitope presentation and are more protective in HIV infections.


Asunto(s)
Infecciones por VIH , Triptófano , Humanos , Antígeno HLA-B44/metabolismo , Triptófano/metabolismo , Péptidos/metabolismo , Antígenos de Histocompatibilidad Clase I/genética , Antígenos de Histocompatibilidad Clase I/metabolismo , Inmunoglobulinas/metabolismo , Unión Proteica , Antígenos HLA-B/genética , Antígenos HLA-B/metabolismo
7.
J Biol Chem ; 299(10): 105136, 2023 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-37543367

RESUMEN

Human Tapasin (hTapasin) is the main chaperone of MHC-I molecules, enabling peptide loading and antigen repertoire optimization across HLA allotypes. However, it is restricted to the endoplasmic reticulum (ER) lumen as part of the protein loading complex (PLC), and therefore is highly unstable when expressed in recombinant form. Additional stabilizing co-factors such as ERp57 are required to catalyze peptide exchange in vitro, limiting uses for the generation of pMHC-I molecules of desired antigen specificities. Here, we show that the chicken Tapasin (chTapasin) ortholog can be expressed recombinantly at high yields in a stable form, independent of co-chaperones. chTapasin can bind the human HLA-B∗37:01 with low micromolar-range affinity to form a stable tertiary complex. Biophysical characterization by methyl-based NMR methods reveals that chTapasin recognizes a conserved ß2m epitope on HLA-B∗37:01, consistent with previously solved X-ray structures of hTapasin. Finally, we provide evidence that the B∗37:01/chTapasin complex is peptide-receptive and can be dissociated upon binding of high-affinity peptides. Our results highlight the use of chTapasin as a stable scaffold for protein engineering applications aiming to expand the ligand exchange function on human MHC-I and MHC-like molecules.


Asunto(s)
Presentación de Antígeno , Pollos , Antígenos HLA-B , Proteínas de Transporte de Membrana , Chaperonas Moleculares , Animales , Humanos , Antígenos HLA-B/metabolismo , Inmunoglobulinas/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Chaperonas Moleculares/metabolismo , Péptidos/metabolismo , Proteínas Recombinantes/metabolismo , Epítopos/metabolismo , Ingeniería de Proteínas
8.
Cell Immunol ; 387: 104707, 2023 05.
Artículo en Inglés | MEDLINE | ID: mdl-36933326

RESUMEN

MHC-I molecules of the HLA-B7 supertype preferentially bind peptides with proline at position 2. HLA-B*51:01 and B*51:08 present two predominant subpeptidomes, one with Pro2 and hydrophobic residues at P1, and another with Ala2 and Asp enriched at position 1. Here, we present a meta-analysis of the peptidomes presented by molecules of the B7 supertype to investigate the presence of subpeptidomes across different allotypes. Several allotypes presented subpeptidomes differing in the presence of Pro or another residue at P2. The Ala2 subpeptidomes preferred Asp1 except in HLA-B*54:01, where ligands with Ala2 contained Glu1. Sequence alignment and the analysis of crystal structures allowed us to propose positions 45 and 67 of the MHC heavy chain as relevant for the presence of subpeptidomes. Deciphering the principles behind the presence of subpeptidomes could improve our understanding of antigen presentation in other MHC-I molecules. Running title: HLA-B7 supertype subpeptidomes.


Asunto(s)
Antígeno HLA-B7 , Antígenos de Histocompatibilidad Clase I , Presentación de Antígeno , Antígenos HLA-B/química , Antígenos HLA-B/metabolismo , Antígeno HLA-B7/química , Antígeno HLA-B7/metabolismo , Péptidos/metabolismo , Humanos
9.
Nature ; 612(7941): 771-777, 2022 12.
Artículo en Inglés | MEDLINE | ID: mdl-36477533

RESUMEN

Human leucocyte antigen B*27 (HLA-B*27) is strongly associated with inflammatory diseases of the spine and pelvis (for example, ankylosing spondylitis (AS)) and the eye (that is, acute anterior uveitis (AAU))1. How HLA-B*27 facilitates disease remains unknown, but one possible mechanism could involve presentation of pathogenic peptides to CD8+ T cells. Here we isolated orphan T cell receptors (TCRs) expressing a disease-associated public ß-chain variable region-complementary-determining region 3ß (BV9-CDR3ß) motif2-4 from blood and synovial fluid T cells from individuals with AS and from the eye in individuals with AAU. These TCRs showed consistent α-chain variable region (AV21) chain pairing and were clonally expanded in the joint and eye. We used HLA-B*27:05 yeast display peptide libraries to identify shared self-peptides and microbial peptides that activated the AS- and AAU-derived TCRs. Structural analysis revealed that TCR cross-reactivity for peptide-MHC was rooted in a shared binding motif present in both self-antigens and microbial antigens that engages the BV9-CDR3ß TCRs. These findings support the hypothesis that microbial antigens and self-antigens could play a pathogenic role in HLA-B*27-associated disease.


Asunto(s)
Autoinmunidad , Antígenos HLA-B , Péptidos , Receptores de Antígenos de Linfocitos T , Humanos , Autoantígenos/química , Autoantígenos/inmunología , Autoantígenos/metabolismo , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Antígenos HLA-B/inmunología , Antígenos HLA-B/metabolismo , Péptidos/química , Péptidos/inmunología , Péptidos/metabolismo , Receptores de Antígenos de Linfocitos T/química , Receptores de Antígenos de Linfocitos T/inmunología , Receptores de Antígenos de Linfocitos T/metabolismo , Líquido Sinovial/inmunología , Espondilitis Anquilosante/inmunología , Uveítis Anterior/inmunología , Biblioteca de Péptidos , Reacciones Cruzadas , Secuencias de Aminoácidos
10.
Medicina (Kaunas) ; 58(10)2022 Oct 07.
Artículo en Inglés | MEDLINE | ID: mdl-36295572

RESUMEN

Background and Objectives: Ankylosing spondylitis (AS) is a chronic inflammatory disease and is highly linked with the expression of the human leukocytic antigen-B*27 (HLA-B*27) genotype. HLA-B*27 heavy chain (B*27-HC) has an innate characteristic to slowly fold, resulting in the accumulation of the misfolded B*27-HC and the formation of homo-oligomeric B*27-HC molecules. The homo-oligomeric B*27-HC can act as a ligand of KIR3DL2. Interaction of the homo-oligomeric B*27-HC molecules with KIR3DL2 will trigger the survival and activation of KIR3DL2-positive NK cells. However, the effects of homo-oligomeric B*27-HC molecules associated with KIR3DL2 on the cytotoxic activity of NK cells and their cytokine expressions remain unknown. Materials and Methods: HLA-B*-2704-HC was overexpressed in the HMy2.C1R (C1R) cell line. Western blotting and quantitative RT-PCR were used to analyze the protein expression and cytokine expression, respectively, when C1R-B*-2704 cells that overexpress B*2704-HC were co-cultured with NK-92MI cells. Flow cytometry was used to analyze the cytotoxicity mediated by NK-92MI cells. Results: Our results revealed that NK-92MI cells up-regulated the expression of perforin and enhanced the cytotoxic activity via augmentation of PI3K/AKT signaling after co-culturing with C1R-B*2704 cells. Suppression of the dimerized B*27-HC formation or treatment with an inhibitor of PI3K, LY294002, or with an anti-B*27-HC monoclonal antibody can reduce the perforin expression of NK-92MI after co-culturing with C1R-B*-2704. Co-culturing with C1R-B*-2704 cells suppressed the TNF-α and IL6 expressions of NK-92MI cells. Conclusion: Stimulation of NK cell-mediated cytotoxicity by homo-oligomeric B*27-HC molecules may contribute to the pathogenesis of AS.


Asunto(s)
Fosfatidilinositol 3-Quinasas , Espondilitis Anquilosante , Humanos , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt , Factor de Necrosis Tumoral alfa/metabolismo , Ligandos , Perforina/metabolismo , Interleucina-6/metabolismo , Receptores KIR3DL2/metabolismo , Células Asesinas Naturales/metabolismo , Células Asesinas Naturales/patología , Antígenos HLA-B/genética , Antígenos HLA-B/metabolismo , Anticuerpos Monoclonales
11.
Chem Biol Interact ; 368: 110220, 2022 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-36243146

RESUMEN

BACKGROUND: Recently, Trichloroethylene (TCE) induced TCE hypersensitivity syndrome (THS) has attracted the attention of many researchers in the field of environmental and occupational health. Studies have revealed that Human leukocyte antigen (HLA) polymorphisms were the important genetic determinants of the diseases, but the potential molecular mechanism remains unclear. OBJECTIVE: This study aimed to investigate the association between THS and HLA at the molecular level. METHOD: We chose the human B-lymphoblastoid cell line Hmy2.C1R transfected with cDNA of HLA-B*13:01 and HLA-B*13:02 to analyze the characteristics of HLA-B-binding peptides and investigate the effect of TCE on the binding affinity of peptides to the HLA-B molecules. Further, the mathematical model was used to identify the possible interaction between TCE and HLA-B*13:01 or HLA-B*13:02 molecule. RESULTS: 54 HLA-B*13:01-binding peptides and 85 HLA-B*13:02-binding peptides were identified. Comparing the protein sequences of HLA-B*13:01 and HLA-B*13:02, amino acids were different at positions 94, 95 and 97. The results of the binding affinity of self-peptides to HLA molecules in the presence of TCE showed that TCE significantly decreased the binding affinity of peptides to HLA-B*13:01 only, but did not affect that of HLA-B*13:02. Molecular docking model showed that there was a unique high-affinity binding mode between TCE and HLA-B*13:01 (but not HLA-B*13:02), and the binding site located in the region of F pocket, suggesting that the unique structure of the F pocket of HLA-B*13:01 might provide the possibility of binding TCE. The pathogenesis of interaction between HLA-B*13:01 and TCE might belong to the model of the alteration of the HLA-presented self-peptide repertoire. DISCUSSION: This study explored the molecular mechanism of the association between THS and HLA-B*13:01, and had important implications for understanding the role of gene-environment interaction in the development of complex environment-related diseases.


Asunto(s)
Hipersensibilidad , Salud Laboral , Tricloroetileno , Humanos , Interacción Gen-Ambiente , Antígenos HLA-B/genética , Antígenos HLA-B/metabolismo , Simulación del Acoplamiento Molecular , Péptidos , Tricloroetileno/toxicidad , Hipersensibilidad/epidemiología
12.
Biochem Soc Trans ; 50(5): 1329-1339, 2022 10 31.
Artículo en Inglés | MEDLINE | ID: mdl-36111814

RESUMEN

Understanding the basis of the immune determinants controlling disease outcome is critical to provide better care to patients and could be exploited for therapeutics and vaccine design. The discovery of the human immunodeficiency virus (HIV) virus as the causing agent of acquired immunodeficiency syndrome (AIDS) decades ago, led to a tremendous amount of research. Among the findings, it was discovered that some rare HIV+ individuals, called HIV controllers (HICs), had the ability to control the virus and keep a low viral load without the need of treatment. This ability allows HICs to delay or avoid progression to AIDS. HIV control is strongly associated with the expression of human leukocyte antigen (HLA) alleles in HICs. From the HIV protective HLAs described, HLA-B57 is the most frequent in HIC patients. HLA-B57 can present a large range of highly conserved Gag-derived HIV peptides to CD8+ T cells and natural killer (NK) cells, both the focus of this review. So far there are limited differences in the immune response strength, magnitude, or receptor repertoire towards HIV epitopes that could explain viral control in HICs. Interestingly, some studies revealed that during early infection the large breadth of the immune response towards HIV mutants in HLA-B57+ HIC patients, might in turn influence the disease outcome.


Asunto(s)
Síndrome de Inmunodeficiencia Adquirida , Infecciones por VIH , VIH-1 , Humanos , Antígenos HLA-B/metabolismo , Linfocitos T CD8-positivos/metabolismo , Células Asesinas Naturales/metabolismo
13.
Biomolecules ; 12(8)2022 08 08.
Artículo en Inglés | MEDLINE | ID: mdl-36008984

RESUMEN

The IFITM restriction factors play a role in cancer cell progression through undefined mechanisms. We investigate new protein-protein interactions for IFITM1/3 in the context of cancer that would shed some light on how IFITM1/3 attenuate the expression of targeted proteins such as HLA-B. SBP-tagged IFITM1 protein was used to identify an association of IFITM1 protein with the SRSF1 splicing factor and transporter of mRNA to the ribosome. Using in situ proximity ligation assays, we confirmed a predominant cytosolic protein-protein association for SRSF1 and IFITM1/3. Accordingly, IFITM1/3 interacted with HLA-B mRNA in response to IFNγ stimulation using RNA-protein proximity ligation assays. In addition, RT-qPCR assays in IFITM1/IFITM3 null cells and wt-SiHa cells indicated that HLA-B gene expression at the mRNA level does not account for lowered HLA-B protein synthesis in response to IFNγ. Complementary, shotgun RNA sequencing did not show major transcript differences between IFITM1/IFITM3 null cells and wt-SiHa cells. Furthermore, ribosome profiling using sucrose gradient sedimentation identified a reduction in 80S ribosomal fraction an IFITM1/IFITM3 null cells compared to wild type. It was partially reverted by IFITM1/3 complementation. Our data link IFITM1/3 proteins to HLA-B mRNA and SRSF1 and, all together, our results begin to elucidate how IFITM1/3 catalyze the synthesis of target proteins. IFITMs are widely studied for their role in inhibiting viruses, and multiple studies have associated IFITMs with cancer progression. Our study has identified new proteins associated with IFITMs which support their role in mediating protein expression; a pivotal function that is highly relevant for viral infection and cancer progression. Our results suggest that IFITM1/3 affect the expression of targeted proteins; among them, we identified HLA-B. Changes in HLA-B expression could impact the presentation and recognition of oncogenic antigens on the cell surface by cytotoxic T cells and, ultimately, limit tumor cell eradication. In addition, the role of IFITMs in mediating protein abundance is relevant, as it has the potential for regulating the expression of viral and oncogenic proteins.


Asunto(s)
Antígenos de Diferenciación/metabolismo , Antígenos HLA-B , Neoplasias del Cuello Uterino , Femenino , Antígenos HLA-B/metabolismo , Humanos , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Factores de Empalme de ARN , ARN Mensajero/genética , Proteínas de Unión al ARN/genética , Proteínas de Unión al ARN/metabolismo , Factores de Empalme Serina-Arginina/genética , Neoplasias del Cuello Uterino/genética
14.
Cell Host Microbe ; 30(8): 1173-1185.e8, 2022 08 10.
Artículo en Inglés | MEDLINE | ID: mdl-35841889

RESUMEN

Human leukocyte antigen (HLA) alleles have been linked to HIV disease progression and attributed to differences in cytotoxic T lymphocyte (CTL) epitope representation. These findings are largely based on treatment-naive individuals of European and African ancestry. We assessed HLA associations with HIV-1 outcomes in 1,318 individuals from Thailand and found HLA-B∗46:01 (B∗46) associated with accelerated disease in three independent cohorts. B∗46 had no detectable effect on HIV-specific T cell responses, but this allele is unusual in containing an HLA-C epitope that binds inhibitory receptors on natural killer (NK) cells. Unbiased transcriptomic screens showed increased NK cell activation in people with HIV, without B∗46, and simultaneous single-cell profiling of surface proteins and transcriptomes revealed a NK cell subset primed for increased responses in the absence of B∗46. These findings support a role for NK cells in HIV pathogenesis, revealed by the unique properties of the B∗46 allele common only in Asia.


Asunto(s)
Infecciones por VIH , Antígenos HLA-B , Progresión de la Enfermedad , Epítopos , Infecciones por VIH/metabolismo , Antígenos HLA-B/genética , Antígenos HLA-B/metabolismo , Humanos , Células Asesinas Naturales , Fenotipo
15.
Toxicol In Vitro ; 82: 105383, 2022 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-35568130

RESUMEN

The combination of certain human leukocyte antigen (HLA) polymorphisms with administration of certain drugs shows a strong correlation with developing drug hypersensitivity. Examples of typical combinations are HLA-B*57:01 with abacavir and HLA-B*15:02 with carbamazepine. However, despite belonging to the same serotype, HLA-B*57:03 and HLA-B*15:01 are not associated with drug hypersensitivity. Recent studies have shown that several HLA polymorphisms are associated with multiple drugs rather than a single drug, all resulting in drug hypersensitivity. In this study, we compared the molecular structures and intracellular localization of HLA-B*57:01, HLA-B*58:01, and HLA-B*15:02, which pose risks for developing drug hypersensitivity, as well as HLA-B*57:03 and HLA-B*15:01 that do not present such risks. We found that HLA molecules posing risks have a low affinity for the subunit ß2-microglobulin; notably, the weak hydrogen bond formed via Gln96 of the HLA molecule contributes to this behavior. We also clarified that these HLA molecules are easily accumulated in the endoplasmic reticulum, exhibiting a low expression on the cell surface. Considering that these hypersensitivity risk-associated HLA molecules form complexes with ß2-microglobulin and peptides in the endoplasmic reticulum, we assumed that their low complex formation ability in the endoplasmic reticulum facilitates the interaction with multiple drugs.


Asunto(s)
Hipersensibilidad a las Drogas , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos , Carbamazepina/toxicidad , Hipersensibilidad a las Drogas/genética , Antígenos HLA/genética , Antígenos HLA-B/química , Antígenos HLA-B/metabolismo , Humanos
16.
J Immunol ; 208(1): 3-15, 2022 01 01.
Artículo en Inglés | MEDLINE | ID: mdl-34880106

RESUMEN

CD8+ T cells play an important role in the control of untreated HIV infection. Several studies have suggested a decisive role of TCRs involved in anti-HIV immunity. HLA-B*27 and B*57 are often associated with a delayed HIV disease progression, but the exact correlates that provide superior immunity against HIV are not known. To investigate if the T cell repertoire underlies the protective effect in disease outcome in HLA-B*27 and B*57+ individuals, we analyzed Ag-specific TCR profiles from progressors (n = 13) and slow progressors (n = 11) expressing either B*27 or B*57. Our data showed no differences in TCR diversity between progressors and slow progressors. Both alleles recruit biased T cell repertoires (i.e., TCR populations skewed toward specific TRBV families or CDR3 regions). This bias was unrelated to disease progression and was remarkably profound for HLA-B*57, in which TRBV family usage and CDR3 sequences were shared to some extent even between epitopes. Conclusively, these data suggest that the T cell repertoires recruited by protective HLA alleles are highly similar between progressors and slow progressors in terms of TCR diversity, TCR usage, and cross-reactivity.


Asunto(s)
Regiones Determinantes de Complementariedad/genética , Infecciones por VIH/inmunología , VIH-1/fisiología , Receptores de Antígenos de Linfocitos T alfa-beta/genética , Linfocitos T/fisiología , Alelos , Presentación de Antígeno , Antígenos Virales/metabolismo , Células Cultivadas , Reacciones Cruzadas , Progresión de la Enfermedad , Ensayo de Immunospot Ligado a Enzimas , Epítopos de Linfocito T/metabolismo , Variación Genética , Antígenos HLA-B/genética , Antígenos HLA-B/metabolismo , Antígeno HLA-B27/genética , Antígeno HLA-B27/metabolismo , Humanos , Activación de Linfocitos
17.
Immunobiology ; 227(1): 152127, 2022 01.
Artículo en Inglés | MEDLINE | ID: mdl-34968777

RESUMEN

Head and neck squamous cell carcinoma (HNSCC) arises from the malignant mucosal epithelium of the oral cavity, pharynx, and larynx. Natural killer (NK) cells are fundamental immune cells shaping the anti-HNSCC response. Elucidation of the regulatory mechanisms of NK cell activity is crucial for understanding anti-HNSCC immunity. In this study, we characterized the expression and function of HLA-B-associated transcript 3 (Bat3) in NK cells in a mouse HNSCC model. We found that Bat3 expression was down-regulated in HNSCC-infiltrating NK cells. SCC VII, the mouse HNSCC cell line used in this model, induced Bat3 downregulation through direct cell-to-cell contact. By applying lentivirus-mediated silencing of Bat3, we discovered that Bat3 knockdown impaired the tumoricidal effect of NK cells on SCC VII cells and Hepa1-6RAE1, a genetically modified liver cancer cell line. Furthermore, Bat3 knockdown resulted in a significant decrease in perforin, granzyme B, interferon-γ, and tumor necrosis factor-α in NK cells upon co-culture with SCC VII cells. Further investigations revealed that Bat3 knockdown promoted the binding of T cell immunoglobulin and mucin domain-containing-3 (Tim-3) to Fyn and thus activated the Tim-3 signaling. Blockade of Tim-3 with a neutralizing Tim-3 antibody counteracted the effect of Bat3 knockdown on NK cell cytotoxicity. Taken together, our data suggest that HNSCC might down-regulate Bat3 expression to augment Tim-3 signaling and ultimately suppress the tumoricidal activity of NK cells. This study unveils a novel mechanism by which HNSCC evades NK cell killing, and sheds light on designing novel anti-HNSCC immunotherapy targeting Bat3 and Tim-3 signaling.


Asunto(s)
Neoplasias de Cabeza y Cuello , Receptor 2 Celular del Virus de la Hepatitis A , Chaperonas Moleculares , Proteínas Nucleares , Carcinoma de Células Escamosas de Cabeza y Cuello , Animales , Modelos Animales de Enfermedad , Regulación hacia Abajo , Antígenos HLA-B/metabolismo , Neoplasias de Cabeza y Cuello/genética , Receptor 2 Celular del Virus de la Hepatitis A/genética , Inmunoglobulinas/metabolismo , Células Asesinas Naturales , Ratones , Chaperonas Moleculares/genética , Mucinas/metabolismo , Proteínas Nucleares/genética , Carcinoma de Células Escamosas de Cabeza y Cuello/genética , Linfocitos T
18.
Pharmacogenomics J ; 22(1): 69-74, 2022 02.
Artículo en Inglés | MEDLINE | ID: mdl-34671112

RESUMEN

PURPOSE: The Pharmacogenomics (PGx) Profile Service was a proof-of-concept project to implement PGx in patient care at Mayo Clinic. METHODS: Eighty-two healthy individuals aged 18 and older underwent genotyping of CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4, CYP3A5, SLCO1B1, HLA-B*58:01, and VKORC1. A PGx pharmacist was involved in ordering, meeting with patients, interpreting, reviewing, and documenting results. RESULTS: Ninety three percent were CYP1A2 rapid metabolizers, 92% CYP3A4 normal metabolizers, and 88% CYP3A5 poor metabolizers; phenotype frequencies for CYP2C19 and CYP2D6 varied. Seventy-three percent had normal functioning SLCO1B1 transporter, 4% carried the HLA-B*58:01 risk variant, and 35% carried VKORC1 and CYP2C9 variants that increased warfarin sensitivity. CONCLUSION: Pre-emptive PGx testing offered medication improvement opportunity in 56% of participants for commonly used medications. A collaborative approach involving a PGx pharmacist integrated within a clinical practice with regards to utility of PGx results allowed for implementation of the PGx Profile Service. KEY POINTS: The Mayo Clinic PGx (PGx) Profile Service was a proof-of-concept project to utilize PGx testing as another clinical tool to enhance medication selection and decrease serious adverse reactions or medication failures. Over one-half of participants in the pilot using the PGx Profile Service were predicted to benefit from pre-emptive PGx testing to guide pharmacotherapy. PGx pharmacists played a crucial role in the PGx Profile Service by educating participants, identifying medication-gene interactions, and providing evidence-based (CPIC and DPWG) PGx recommendations for past, current, and future medication us.


Asunto(s)
Farmacogenética/métodos , Pruebas de Farmacogenómica , Adolescente , Adulto , Anciano , Sistema Enzimático del Citocromo P-450/genética , Femenino , Pruebas Genéticas , Genotipo , Antígenos HLA-B/genética , Antígenos HLA-B/metabolismo , Voluntarios Sanos , Heterocigoto , Humanos , Transportador 1 de Anión Orgánico Específico del Hígado/genética , Transportador 1 de Anión Orgánico Específico del Hígado/metabolismo , Masculino , Persona de Mediana Edad , Farmacocinética , Fenotipo , Estudios Retrospectivos , Adulto Joven
20.
Immunohorizons ; 5(8): 687-702, 2021 08 25.
Artículo en Inglés | MEDLINE | ID: mdl-34433624

RESUMEN

Tyrosine kinase inhibitor (TKI)-treated chronic myeloid leukemia (CML) patients with increased NK cell number have a better prognosis, and thus, NK cells may suppress CML. However, the efficacy of TKIs varies for reasons yet to be fully elucidated. As NK cell activity is modulated by interactions between their killer cell Ig-like receptors (KIRs) and HLAs of target cells, the combination of their polymorphisms may have functional significance. We previously showed that allelic polymorphisms of KIR3DL1 and HLAs were associated with the prognosis of TKI-treated CML patients. In this study, we focus on differential NK cell activity modulation through KIR3DL1 allotypes. KIR3DL1 expression levels varied according to their alleles. The combination of KIR3DL1 expression level and HLA-Bw4 motifs defined NK cell activity in response to the CML-derived K562 cell line, and Ab-mediated KIR3DL1 blocking reversed this activity. The TKI dasatinib enhanced NK cell activation and cytotoxicity in a KIR3DL1 allotype-dependent manner but did not significantly decrease effector regulatory T cells, suggesting that it directly activated NK cells. Dasatinib also enhanced NK cell cytotoxicity against K562 bearing the BCR-ABL1 T315I TKI resistance-conferring mutation, depending on KIR3DL1/HLA-Bw4 allotypes. Transduction of KIR3DL1*01502 into the NK cell line NK-92 resulted in KIR3DL1 expression and suppression of NK-92 activity by HLA-B ligation, which was reversed by anti-KIR3DL1 Ab. Finally, KIR3DL1 expression levels also defined activation patterns in CML patient-derived NK cells. Our findings raise the possibility of a novel strategy to enhance antitumor NK cell immunity against CML in a KIR3DL1 allotype-dependent manner.


Asunto(s)
Regulación Leucémica de la Expresión Génica/inmunología , Células Asesinas Naturales/inmunología , Leucemia Mielógena Crónica BCR-ABL Positiva/inmunología , Receptores KIR3DL1/inmunología , Alelos , Línea Celular Tumoral , Citotoxicidad Inmunológica/efectos de los fármacos , Citotoxicidad Inmunológica/genética , Citotoxicidad Inmunológica/inmunología , Dasatinib/farmacología , Proteínas de Fusión bcr-abl/genética , Proteínas de Fusión bcr-abl/inmunología , Regulación Leucémica de la Expresión Génica/efectos de los fármacos , Regulación Leucémica de la Expresión Génica/genética , Antígenos HLA-B/genética , Antígenos HLA-B/inmunología , Antígenos HLA-B/metabolismo , Humanos , Células K562 , Células Asesinas Naturales/efectos de los fármacos , Células Asesinas Naturales/metabolismo , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Activación de Linfocitos/efectos de los fármacos , Activación de Linfocitos/genética , Activación de Linfocitos/inmunología , Mutación , Inhibidores de Proteínas Quinasas/farmacología , Receptores KIR3DL1/genética , Receptores KIR3DL1/metabolismo
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