scholarly journals A molecular approach to the identification of cytotoxic T-lymphocyte epitopes within equine herpesvirus 1

2006 ◽  
Vol 87 (9) ◽  
pp. 2507-2515 ◽  
Author(s):  
Julia H. Kydd ◽  
N. J. Davis-Poynter ◽  
J. Birch ◽  
D. Hannant ◽  
J. Minke ◽  
...  
Keyword(s):  
Mhc Class I ◽  
Cytotoxic T Lymphocyte ◽  
Class I ◽  
Target Cells ◽  
Equine Herpesvirus ◽  
Equine Herpesvirus 1 ◽  
Leukocyte Antigen ◽  
Histocompatibility Complex ◽  
Herpesvirus 1 ◽  
Functional Peptide

Equine herpesvirus 1 (EHV-1) causes respiratory and neurological disease and abortion in horses. Animals with high frequencies of cytotoxic T lymphocytes (CTL) show reduced severity of respiratory disease and frequency of abortion, probably by CTL-mediated control of cell-associated viraemia. This study aimed to identify CTL epitopes restricted by selected major histocompatibility complex (MHC) class I alleles expressed in the equine leukocyte antigen (ELA) A3 haplotype. Effector CTL were induced from EHV-1-primed ponies and thoroughbreds with characterized MHC class I haplotypes and screened against P815 target cells transfected with selected EHV-1 genes and MHC class I genes. Targets that expressed EHV-1 gene 64 and the MHC B2 gene were lysed by effector CTL in a genetically restricted manner. There was no T-cell recognition of targets expressing either the MHC B2 gene and EHV-1 genes 2, 12, 14, 16, 35, 63 or 69, or the MHC C1 gene and EHV-1 genes 12, 14, 16 or 64. A vaccinia virus vector encoding gene 64 (NYVAC-64) was also investigated. Using lymphocytes from ELA-A3 horses, the recombinant NYVAC-64 virus induced effector CTL that lysed EHV-1-infected target cells; the recombinant virus also supplied a functional peptide that was expressed by target cells and recognized in an MHC-restricted fashion by CTL induced with EHV-1. This construct may therefore be used to determine the antigenicity of EHV-1 gene 64 for other MHC haplotypes. These techniques are broadly applicable to the identification of additional CTL target proteins and their presenting MHC alleles, not only for EHV-1, but for other equine viruses.

scholarly journals Exogenous Expression of Equine MHC Class I Molecules in Mice Increases Susceptibility to Equine Herpesvirus 1 Pulmonary Infection

2019 ◽  
Vol 56 (5) ◽  
pp. 703-710 ◽  
Author(s):  
Erina Minato ◽  
Keisuke Aoshima ◽  
Atsushi Kobayashi ◽  
Naomi Ohnishi ◽  
Nobuya Sasaki ◽  
...  
Keyword(s):  
Mhc Class I ◽  
Virus Antigen ◽  
Class I ◽  
Equine Herpesvirus ◽  
Class I Mhc ◽  
Equine Herpesvirus 1 ◽  
Exogenous Expression ◽  
Herpesvirus 1 ◽  
Entry Receptor

Equine herpesvirus 1 (EHV-1) uses equine major histocompatibility complex class I (MHC class I) as an entry receptor. Exogenous expression of equine MHC class I genes in murine cell lines confers susceptibility to EHV-1 infection. To examine the in vivo role of equine MHC class I as an entry receptor for EHV-1, we generated transgenic (Tg) mice expressing equine MHC class I under the control of the CAG promoter. Equine MHC class I protein was expressed in the liver, spleen, lung, and brain of Tg mice, which was confirmed by Western blot. However, equine MHC class I antigen was only detected in bronchiolar epithelium and not in other tissues, using the immunofluorescence method employed in this study. Both Tg and wild-type (WT) mice developed pneumonia 3 days after intranasal infection with EHV-1. The bronchiolar epithelial cells of Tg mice showed more severe necrosis, compared with those in WT mice. In addition, the number of virus antigen-positive cells in the lungs was higher in Tg mice than in WT mice. These results suggest that exogenous expression of equine MHC class I renders mice more susceptible to EHV-1 infection.

scholarly journals Equine Herpesvirus 1 Multiply Inserted Transmembrane Protein pUL43 Cooperates with pUL56 in Downregulation of Cell Surface Major Histocompatibility Complex Class I

2015 ◽  
Vol 89 (12) ◽  
pp. 6251-6263 ◽  
Author(s):  
Teng Huang ◽  
Guanggang Ma ◽  
Nikolaus Osterrieder
Keyword(s):  
Cell Surface ◽  
Class I ◽  
Equine Herpesvirus ◽  
Lysosomal Degradation ◽  
Mhc I ◽  
Equine Herpesvirus 1 ◽  
Transfected Cells ◽  
Histocompatibility Complex ◽  
Herpesvirus 1 ◽  
Late Domains

ABSTRACTHerpesviruses have evolved an array of strategies to counteract antigen presentation by major histocompatibility complex class I (MHC-I). Previously, we identified pUL56 of equine herpesvirus 1 (EHV-1) as one major determinant of the downregulation of cell surface MHC-I (G. Ma, S. Feineis, N. Osterrieder, and G. R. Van de Walle, J. Virol. 86:3554–3563, 2012,http://dx.doi.org/10.1128/JVI.06994-11; T. Huang, M. J. Lehmann, A. Said, G. Ma, and N. Osterrieder, J. Virol. 88:12802–12815, 2014,http://dx.doi.org/10.1128/JVI.02079-14). Since pUL56 was able to exert its function only in the context of virus infection, we hypothesized that pUL56 cooperates with another viral protein. Here, we generated and screened a series of EHV-1 single-gene deletion mutants and found that the pUL43 orthologue was required for downregulation of cell surface MHC-I expression at the same time of infection as when pUL56 exerts its function. We demonstrate that the absence of pUL43 was not deleterious to virus growth and that expression of pUL43 was detectable from 2 h postinfection (p.i.) but decreased after 8 h p.i. due to lysosomal degradation. pUL43 localized within Golgi vesicles and required a unique hydrophilic N-terminal domain to function properly. Finally, coexpression of pUL43 and pUL56 in transfected cells reduced the cell surface expression of MHC-I. This process was dependent on PPxY motifs present in pUL56, suggesting that late domains are required for pUL43- and pUL56-dependent sorting of MHC class I for lysosomal degradation.IMPORTANCEWe describe here that the poorly characterized herpesviral protein pUL43 is involved in downregulation of cell surface MHC-I. pUL43 is an early protein and degraded in lysosomes. pUL43 resides in the Golgi vesicles and needs an intact N terminus to induce MHC-I downregulation in infected cells. Importantly, pUL43 and pUL56 cooperate to reduce MHC-I expression on the surface of transfected cells. Our results suggest a model for MHC-I downregulation in which late domains in pUL56 are required for the rerouting of vesicles containing MHC-I, pUL56, and pUL43 to the lysosomal compartment.

scholarly journals Down-regulation of MHC class I expression by equine herpesvirus-1

2003 ◽  
Vol 84 (2) ◽  
pp. 293-300 ◽  
Author(s):  
Giovanna Rappocciolo ◽  
James Birch ◽  
Shirley A. Ellis
Keyword(s):  
Mhc Class I ◽  
Class I ◽  
Equine Herpesvirus ◽  
Down Regulation ◽  
Equine Herpesvirus 1 ◽  
Herpesvirus 1

Conservation of recognition of antibody and T-cell-defined alloantigens between species of equids

2001 ◽  
Vol 13 (8) ◽  
pp. 635
Author(s):  
Jessica M. Baker ◽  
M. Stidworthy ◽  
Tamara Gull ◽  
Jodi Novak ◽  
Jane M. Miller ◽  
...  
Keyword(s):  
T Cell ◽  
Mhc Class I ◽  
Molecular Techniques ◽  
Class I ◽  
Antigenic Determinants ◽  
Immune Recognition ◽  
Large Measure ◽  
Leukocyte Antigen ◽  
Cellular Assays ◽  
Histocompatibility Complex

Serological and cellular assays and molecular techniques were used to define features of the major histocompatibility complex (MHC) of the donkey. With this information in hand, immune recognition of MHC determinants within and between donkeys and horses was compared. An antibody-mediated, complement-dependent, microcytotoxicity assay using a variety of antisera to donkey histocompatibility antigens, including those induced as a result of intraspecies or interspecies pregnancy in horse mares and jenny donkeys, delineated five donkey leukocyte antigen (DoLA) specificities. Antisera raised across species barriers (horse anti-donkey and donkey anti-horse) recognized polymorphic antigenic determinants in the target species. These determinants were often indistinguishable from polymorphic antigens recognized by alloantisera raised within horses or donkeys. The data indicate that a strong correlation exists between the serological antigenic types identified and the specificity of cytotoxic T lymphocytes raised by lymphocyte co-cultures, either within or between species. Moreover, in an analysis of a small number of donkey MHC class I cDNA gene sequences, no features distinguishing horse or donkey MHC class I molecules were identified. These molecular findings explain in large measure why antibody- and T-cell-defined alloantigen recognition is conserved among these closely related equids.

scholarly journals Escape in One of Two Cytotoxic T-Lymphocyte Epitopes Bound by a High-Frequency Major Histocompatibility Complex Class I Molecule, Mamu-A*02: a Paradigm for Virus Evolution and Persistence?

2002 ◽  
Vol 76 (22) ◽  
pp. 11623-11636 ◽  
Author(s):  
Thorsten U. Vogel ◽  
Thomas C. Friedrich ◽  
David H. O'Connor ◽  
William Rehrauer ◽  
Elizabeth J. Dodds ◽  
...  
Keyword(s):  
Major Histocompatibility Complex ◽  
Mhc Class I ◽  
T Lymphocyte ◽  
Cytotoxic T Lymphocyte ◽  
Class I ◽  
Major Histocompatibility ◽  
Histocompatibility Complex ◽  
Mhc Class I Molecule ◽  
Immunodeficiency Virus ◽  
Ctl Responses

ABSTRACT It is now accepted that an effective vaccine against AIDS must include effective cytotoxic-T-lymphocyte (CTL) responses. The simian immunodeficiency virus (SIV)-infected rhesus macaque is the best available animal model for AIDS, but analysis of macaque CTL responses has hitherto focused mainly on epitopes bound by a single major histocompatibility complex (MHC) class I molecule, Mamu-A*01. The availability of Mamu-A*01-positive macaques for vaccine studies is therefore severely limited. Furthermore, it is becoming clear that different CTL responses are able to control immunodeficiency virus replication with varying success, making it a priority to identify and analyze CTL responses restricted by common MHC class I molecules other than Mamu-A*01. Here we describe two novel epitopes derived from SIV, one from Gag (Gag71-79 GY9), and one from the Nef protein (Nef159-167 YY9). Both epitopes are bound by the common macaque MHC class I molecule, Mamu-A*02. The sequences of these two eptiopes are consistent with the molecule's peptide-binding motif, which we have defined by elution of natural ligands from Mamu-A*02. Strikingly, we found evidence for the selection of escape variant viruses by CTL specific for Nef159-167 YY9 in 6 of 6 Mamu-A*02-positive animals. In contrast, viral sequences encoding the Gag71-79 GY9 epitope remained intact in each animal. This situation is reminiscent of Mamu-A*01-restricted CTL that recognize Tat28-35 SL8, which reproducibly selects for escape variants during acute infection, and Gag181-189 CM9, which does not. Differential selection by CTL may therefore be a paradigm of immunodeficiency virus infection.

scholarly journals Peptide transport activity of the transporter associated with antigen processing (TAP) is inhibited by an early protein of equine herpesvirus-1

2004 ◽  
Vol 85 (2) ◽  
pp. 349-353 ◽  
Author(s):  
Aruna P. N. Ambagala ◽  
Raju S. Gopinath ◽  
S. Srikumaran
Keyword(s):  
Antigen Processing ◽  
Protein S ◽  
Class I ◽  
Peptide Transport ◽  
Equine Herpesvirus ◽  
Equine Herpesvirus 1 ◽  
Early Protein ◽  
Infected Cells ◽  
Herpesvirus 1

Equine herpesvirus-1 (EHV-1) downregulates surface expression of major histocompatibility complex (MHC) class I molecules on infected cells. The objective of this study was to investigate whether EHV-1 interferes with peptide translocation by the transporter associated with antigen processing (TAP) and to identify the proteins responsible. Using an in vitro transport assay, we showed that EHV-1 inhibited transport of peptides by TAP as early as 2 h post-infection (p.i). Complete shutdown of peptide transport was observed by 8 h p.i. Furthermore, pulse–chase experiments revealed that maturation of class I molecules in the endoplasmic reticulum (ER) was delayed in EHV-1-infected cells, which may be due to reduced availability of peptides in the ER as a result of TAP inhibition. Metabolic inhibition studies indicated that an early protein(s) of EHV-1 is responsible for this effect.

scholarly journals Definition of Five New Simian Immunodeficiency Virus Cytotoxic T-Lymphocyte Epitopes and Their Restricting Major Histocompatibility Complex Class I Molecules: Evidence for an Influence on Disease Progression

2000 ◽  
Vol 74 (16) ◽  
pp. 7400-7410 ◽  
Author(s):  
David T. Evans ◽  
Peicheng Jing ◽  
Todd M. Allen ◽  
David H. O'Connor ◽  
Helen Horton ◽  
...  
Keyword(s):  
Disease Progression ◽  
Mhc Class I ◽  
T Lymphocyte ◽  
Cytotoxic T Lymphocyte ◽  
Rhesus Macaques ◽  
Class I ◽  
Ctl Epitopes ◽  
Histocompatibility Complex ◽  
Immunodeficiency Virus ◽  
Mhc Class I Molecules

ABSTRACT Simian immunodeficiency virus (SIV) infection of the rhesus macaque is currently the best animal model for AIDS vaccine development. One limitation of this model, however, has been the small number of cytotoxic T-lymphocyte (CTL) epitopes and restricting major histocompatibility complex (MHC) class I molecules available for investigating virus-specific CTL responses. To identify new MHC class I-restricted CTL epitopes, we infected five members of a family of MHC-defined rhesus macaques intravenously with SIV. Five new CTL epitopes bound by four different MHC class I molecules were defined. These included two Env epitopes bound by Mamu-A*11 and -B*03 and three Nef epitopes bound by Mamu-B*03, -B*04, and -B*17. All four restricting MHC class I molecules were encoded on only two haplotypes (b or c). Interestingly, resistance to disease progression within this family appeared to be associated with the inheritance of one or both of these MHC class I haplotypes. Two individuals that inherited haplotypes b and cseparately survived for 299 and 511 days, respectively, while another individual that inherited both haplotypes survived for 889 days. In contrast, two MHC class I-identical individuals that did not inherit either haplotype rapidly progressed to disease (survived <80 days). Since all five offspring were identical at their Mamu-DRBloci, MHC class II differences are unlikely to account for their patterns of disease progression. These results double the number of SIV CTL epitopes defined in rhesus macaques and provide evidence that allelic differences at the MHC class I loci may influence rates of disease progression among AIDS virus-infected individuals.

scholarly journals Human Histocompatibility Leukocyte Antigen (HLA)-G Molecules Inhibit NKAT3 Expressing Natural Killer Cells

1997 ◽  
Vol 185 (3) ◽  
pp. 385-392 ◽  
Author(s):  
Christian Münz ◽  
Nicholas Holmes ◽  
Ashley King ◽  
Yung Wai Loke ◽  
Marco Colonna ◽  
...  
Keyword(s):  
Natural Killer ◽  
Mhc Class I ◽  
Class I ◽  
Specific Expression ◽  
Human Major Histocompatibility Complex ◽  
Leukocyte Antigen ◽  
Histocompatibility Complex ◽  
Mother And Child ◽  
Secondary Function ◽  
Mhc Class I Molecules

The crucial immunological function of the classical human major histocompatibility complex (MHC) class I molecules, human histocompatibility leukocyte antigen (HLA)-A, -B, and -C, is the presentation of peptides to T cells. A secondary function is the inhibition of natural killer (NK) cells, mediated by binding of class I molecules to NK receptors. In contrast, the function of the nonclassical human MHC class I molecules, HLA-E, -F, and -G, is still a mystery. The specific expression of HLA-G in placental trophoblast suggests an important role for this molecule in the immunological interaction between mother and child. The fetus, semiallograft by its genotype, escapes maternal allorecognition by downregulation of HLA-A and HLA-B molecules at this interface. It has been suggested that the maternal NK recognition of this downregulation is balanced by the expression of HLA-G, thus preventing damage to the placenta. Here, we describe the partial inhibition of NK lysis of the MHC class I negative cell line LCL721.221 upon HLA-G transfection. We present three NK lines that are inhibited via the interaction of their NKAT3 receptor with HLA-G and with HLA-Bw4 molecules. Inhibition can be blocked by the anti-NKAT3 antibody 5.133. In conclusion, NK inhibition by HLA-G via NKAT3 may contribute to the survival of the fetal semiallograft in the mother during pregnancy.

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