Transcriptome analysis of Brucella abortus S19∆per immunized mouse spleen revealed activation of MHC-I and MHC-II pathways

Brucellosis is a disease that can be prevented through vaccination. Yet, the effectiveness of the vaccination to fight this disease is considered weak. Fortunately, attempts to modify brucellosis vaccine is still keep going. Some brucellosis vaccines have been found and developed in the past time such as the vaccine B.abortus strain 19-BA and 104M which was made from weakened microbes which had been widely used in Uni Soviet and China. The other brucellosis vaccine that were used in the past were the phenolinsoluble peptidoglycan vaccine which was made in France and polysaccharideprotein vaccine which was used in Russia. This research attempted to see the determinant of antigenic Outer Membrane Protein (OM) 36 kDa Brucella abortus local isolation which has immunogenic character to be developed as an advanced brucellosis vaccine. The method used in this research was the Omp2 gene of Brucella abortus of local isolate employed the PCR technique. The result of the PCR was then sequenced to analyze the determinant antigenic and the bounding prediction of either the T cell or the B cell which were responsible for immune response. The result of this study showed that the gen Omp2 which encoded the OMP 36 kDa Brucella abortus of local isolation with primary JPF 5’ GCG CTC AGG CTG CCG ACG CAA 3’ and JPR 5’ CAT TGC GGT CGG TAC CGG AG 3’ targeted the gene 162 bp, was then translated into amino acids to be later undergo the in silico test using Kolaskar and Tongaonkar Antigenicity Prediction method. The epitope prediction resulted were MSRVCDAYGAGYFYI and TETCLRVHGYVRYD. The result of the epitope prediction of MSRVCDAYGAGYFYI showed that there was a bond with MHC I in YGAGYFYI of the 8th amino acid series to the 15th series, while the epitope prediction of TETCLRVHGYVRYD showed that there was a bond to the ETCLRVHGY of the series of amino acids number 2 to 10. Bond with MHC II existed in the amino acid series of MSRVCDAYGAGYFYI, while the bond with the B cells existed in BCSAYGA and CLRVHG amino acid series. This research has been successful in predicting the epitope of the OMP 36 kDa Brucella abortus of local isolate which had immunogenic characteristic for its ability to bond with the MHC I, MHC II and B cells.

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RNA-seq liver transcriptome analysis reveals an activated MHC-I pathway and an inhibited MHC-II pathway at the early stage of vaccine immunization in zebrafish

BMC Genomics ◽

10.1186/1471-2164-13-319 ◽

2012 ◽

Vol 13 (1) ◽

pp. 319 ◽

Cited By ~ 50

Author(s):

Dahai Yang ◽

Qin Liu ◽

Minjun Yang ◽

Haizhen Wu ◽

Qiyao Wang ◽

...

Keyword(s):

Transcriptome Analysis ◽

Early Stage ◽

Rna Seq ◽

Mhc I ◽

Mhc Ii ◽

Liver Transcriptome

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Design of Epitope Based Peptide Vaccine against Brucella Abortus OmpW Family Protein using Immunoinformatics

10.1101/2021.12.27.474190 ◽

2021 ◽

Author(s):

Mustafa Elhag ◽

Abdelrahman Hamza Abdelmoneim ◽

Anfal Osama Sati ◽

Moaaz Mohammed Saadaldin ◽

Nagla Mohammad Ahmad ◽

...

Keyword(s):

Brucella Abortus ◽

Peptide Vaccine ◽

Reproductive Organs ◽

Population Coverage ◽

Mhc I ◽

B Cell Epitopes ◽

Mhc Ii ◽

Family Protein ◽

Initial Interaction ◽

Chronic Disorders

Brucella abortus is a small aerobic, non-spore-forming, non-motile intracellular coccobacilli localized in the reproductive organs of host animals and causes acute or chronic disorders. It infects approximately 200 cases per 100,000 of the population and has become endemic in many countries. OmpW family protein is an outer membrane protein involved in the initial interaction between the pathogen and its host. This study predicts an effective epitope-based vaccine against OmpW family protein of Brucella abortus using immunoinformatics tools. Sequences were obtained from NCBI and prediction tests were accomplished to analyze possible epitopes for B and T cells. Seven B cell epitopes passed the antigenicity, accessibility and hydrophilicity tests. Forty-three MHC I epitopes were the most promising, while 438 from MHC II. For the population coverage, the epitopes covered 99.97% of the alleles worldwide excluding certain MHC II alleles. We recommend invivo and invitro studies to prove its effectiveness.

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Transcriptome Analysis Shows That IFN-I Treatment and Concurrent SAV3 Infection Enriches MHC-I Antigen Processing and Presentation Pathways in Atlantic Salmon-Derived Macrophage/Dendritic Cells

Viruses ◽

10.3390/v11050464 ◽

2019 ◽

Vol 11 (5) ◽

pp. 464 ◽

Cited By ~ 4

Author(s):

Cheng Xu ◽

Øystein Evensen ◽

Hetron Mweemba Munang’andu

Keyword(s):

Atlantic Salmon ◽

Immune Responses ◽

Transcriptome Analysis ◽

Antigen Processing ◽

Type I ◽

Adaptive Immune Responses ◽

Mhc I ◽

Mhc Ii ◽

Adaptive Immune ◽

Antigen Processing And Presentation

Type I interferons (IFNs) have been shown to play an important role in shaping adaptive immune responses in addition to their antiviral properties in immune cells. To gain insight into the impact of IFN-I-induced pathways involved in early adaptive immune responses, i.e., antigen-presenting pathways, in an Atlantic salmon-derived (Salmo salar L.) macrophage cell line (TO-cells), we used a comparative de novo transcriptome analysis where cells were treated with IFN-I or kept untreated and concurrently infected with salmonid alphavirus subtype 3 (SAV3). We found that concurrent treatment of TO-cells with IFN-I and SAV3 infection (SAV3/IFN+) significantly enriched the major histocompatibility complex class I (MHC-I) pathway unlike the non-IFN-I treated TO-cells (SAV3/IFN−) that had lower expression levels of MHC-I pathway-related genes. Genes such as the proteasomal activator (PA28) and β-2 microglobulin (β2M) were only differentially expressed in the SAV3/IFN+ cells and not in the SAV3/IFN− cells. MHC-I pathway genes like heat shock protein 90 (Hsp90), transporter of antigen associated proteins (TAPs) and tapasin had higher expression levels in the SAV3/IFN+ cells than in the SAV3/IFN− cells. There were no MHC-II pathway-related genes upregulated in SAV3/IFN+-treated cells, and cathepsin S linked to the degradation of endosomal antigens in the MHC-II pathway was downregulated in the SAV3/IFN− cells. Overall, our findings show that concurrent IFN-I treatment of TO-cells and SAV3 infection enriched gene expression linked to the MHC-I antigen presentation pathway. Data presented indicate a role of type I IFNs in strengthening antigen processing and presentation that may facilitate activation particularly of CD8+ T-cell responses following SAV3 infection, while SAV3 infection alone downplayed MHC-II pathways.

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NetCleave: an open-source algorithm for predicting C-terminal antigen processing for MHC-I and MHC-II

Scientific Reports ◽

10.1038/s41598-021-92632-y ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Pep Amengual-Rigo ◽

Victor Guallar

Keyword(s):

Open Source ◽

Antigen Processing ◽

High Accuracy ◽

Biological Processes ◽

Mhc I ◽

Mhc Ii ◽

Prediction Tools ◽

Immunogenic Peptides ◽

Physicochemical Descriptors ◽

Peptide Presentation

AbstractAntigens presented on the cell surface have been subjected to multiple biological processes. Among them, C-terminal antigen processing constitutes one of the main bottlenecks of the peptide presentation pathways, as it delimits the peptidome that will be subjected downstream. Here, we present NetCleave, an open-source and retrainable algorithm for the prediction of the C-terminal antigen processing for both MHC-I and MHC-II pathways. NetCleave architecture consists of a neural network trained on 46 different physicochemical descriptors of the cleavage site amino acids. Our results demonstrate that prediction of C-terminal antigen processing achieves high accuracy on MHC-I (AUC of 0.91), while it remains challenging for MHC-II (AUC of 0.66). Moreover, we evaluated the performance of NetCleave and other prediction tools for the evaluation of four independent immunogenicity datasets (H2-Db, H2-Kb, HLA-A*02:01 and HLA-B:07:02). Overall, we demonstrate that NetCleave stands out as one of the best algorithms for the prediction of C-terminal processing, and we provide one of the first evidence that C-terminal processing predictions may help in the discovery of immunogenic peptides.

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823 CD4 T cells are essential for an anti-tumor effect in a B78 murine melanoma tumor model

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-sitc2020.0823 ◽

2020 ◽

Vol 8 (Suppl 3) ◽

pp. A873-A873

Author(s):

Arika Feils ◽

Mackenzie Heck ◽

Anna Hoefges ◽

Peter Carlson ◽

Luke Zangl ◽

...

Keyword(s):

T Cells ◽

Tumor Growth ◽

Cd4 T Cells ◽

Tumor Response ◽

Immune Cell ◽

Cd8 T Cell ◽

Mhc I ◽

Mhc Ii ◽

Flow Cytometric

BackgroundMice bearing B78 melanoma tumors can be cured using an in situ vaccine (ISV) regimen that includes radiation (RT) together with immunocytokine (tumor-targeting mAb conjugated to IL-2). B78 melanoma cells, derived from B16 cells, express minimal to no MHC-I but express MHC-II upon IFN-g/TNF-a stimulation. Although B78 cells are primarily MHC-I-deficient, an increased CD8 T cell infiltration into the tumor microenvironment (TME) has been shown following ISV.1 To further investigate the potential role of specific immune cell lineages in the B78 anti-tumor response to ISV, immune subset depletion studies and flow cytometric analyses were performed.MethodsC57BL/6 mice bearing B78 tumors were depleted of immune cell subsets with mAbs (anti-CD4, anti-CD8, anti-NK1.1, or Rat IgG control) for 3 weeks during the course of treatment. Treatment groups included no treatment, RT (12 Gy), or ISV (RT D0 and immunocytokine D5-D9). 6 mice/group (repeated three times) were followed for survival/tumor growth, and flow cytometry studies included 4 mice/group, sacrificed on D8 and D13 following the start of ISV.ResultsMice depleted of CD4 T cells during the course of ISV showed a significant reduction of anti-tumor effect as compared to mice treated with ISV/Rat IgG (pConclusionsThese studies suggest that CD4 T cells are essential for an anti-tumor response in the B78 melanoma model. In vivo depletion data show that CD4 T cells, but not CD8 or NK cells, are required for a decrease in tumor growth via ISV. Flow cytometric analyses suggest an interplay between CD4 and CD8 T cells as indicated by a decrease in CD8/IFN-g expression following ISV in the absence of CD4 T cells. The role that MHC-I and MHC-II expression plays in this CD4/CD8 T cell anti-tumor response is under investigation. In future studies, B78 melanoma may serve as a critical syngeneic model for development of more effective immunotherapy treatment regimens.Ethics ApprovalAll animal experiments were performed in accordance with protocols approved by Animal Care and Use Committees of the University of Wisconsin-Madison.ReferenceMorris Z, Guy E, Francis D, et al. In situ tumor vaccination by combining local radiation and tumor-specific antibody or immunocytokine treatments. Cancer Res 2016;76(13):3929-3941.

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High Levels of a Major Histocompatibility Complex II–Self Peptide Complex on Dendritic Cells from the T Cell Areas of Lymph Nodes

Journal of Experimental Medicine ◽

10.1084/jem.186.5.665 ◽

1997 ◽

Vol 186 (5) ◽

pp. 665-672 ◽

Cited By ~ 205

Author(s):

Kayo Inaba ◽

Maggie Pack ◽

Muneo Inaba ◽

Hiraki Sakuta ◽

Frank Isdell ◽

...

Keyword(s):

Dendritic Cells ◽

T Cell ◽

Lymph Nodes ◽

Mhc I ◽

Peptide Complex ◽

Mhc Ii ◽

Histocompatibility Complex ◽

Peripheral Lymph ◽

Free Media ◽

Very High

T lymphocytes recirculate continually through the T cell areas of peripheral lymph nodes. During each passage, the T cells survey the surface of large dendritic cells (DCs), also known as interdigitating cells. However, these DCs have been difficult to release from the lymph node. By emphasizing the use of calcium-free media, as shown by Vremec et al. (Vremec, D., M. Zorbas, R. Scollay, D.J. Saunders, C.F. Ardavin, L. Wu, and K. Shortman. 1992. J. Exp. Med. 176:47–58.), we have been able to release and enrich DCs from the T cell areas. The DCs express the CD11c leukocyte integrin, the DEC-205 multilectin receptor for antigen presentation, the intracellular granule antigens which are recognized by monoclonal antibodies M342, 2A1, and MIDC-8, very high levels of MHC I and MHC II, and abundant accessory molecules such as CD40, CD54, and CD86. When examined with the Y-Ae monoclonal which recognizes complexes formed between I-Ab and a peptide derived from I-Eα, the T cell area DCs expressed the highest levels. The enriched DCs also stimulated a T-T hybridoma specific for this MHC II–peptide complex, and the hybridoma underwent apoptosis. Therefore DCs within the T cell areas can be isolated. Because they present very high levels of self peptides, these DCs should be considered in the regulation of self reactivity in the periphery.

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Abstract 5702: Oxidative Stress in Patients with Lone Atrial Fibrillation

Circulation ◽

10.1161/circ.118.suppl_18.s_988-b ◽

2008 ◽

Vol 118 (suppl_18) ◽

Author(s):

Kengo Kusano ◽

Kazufumi Nakamura ◽

Tohru Ohe

Keyword(s):

Oxidative Stress ◽

Atrial Fibrillation ◽

Immunohistochemical Analysis ◽

Lipid Peroxidation Product ◽

Lone Atrial Fibrillation ◽

Activated T Cells ◽

Mhc I ◽

Mhc Ii ◽

Peroxidation Product ◽

Ros Formation

Background: Lone atrial fibrillation (LAF) is a common arrhythmia, but its mechanism and the aggravated factor of arrhythmia are poorly understood. Oxidative stress has been implicated in the pathogenesis of heart failure. We have previously demonstrated that the amount of 4-hydroxy-2-nonenal (HNE) modified protein, which is a major lipid peroxidation product and a cytotoxic aldehyde, causes intracellular Ca2+ overload via reactive oxygen species (ROS) formation in cardiomyocytes and leads to develop arrhythmia. Accordingly we examined the levels of HNE and major histocompatibility complex (MHC) with the disease severity in LAF patients. Method: Atrial and ventricular myocardial samples were obtained from twelve patients (11 men and 1 woman, mean 48±14 years old) by endomyocardial biopsy in 10, autopsy sample in 1 and surgical resection sample in 1. Histological assessments and immunohistochemical analysis for HNE modified protein, MHC class-I and -II antigens (grade 0 to 3) were performed and compared with LAF severity. Results: Histological assessment showed that increased number of interstitial cells (mainly activated T cells) was observed only in the atrium but not in the ventricle. Moderate to severe expression of MHC antigens (grade 2 or 3) was more observed in the atrium than the ventricle (MHC-I: seven in atrium and three in ventricle; MHC-II: ten in atrium and four in ventricle). Atrial myocarditis was detected in 6 out of 11 samples. HNE modified protein was also more observed in the atrium than that in the ventricle. In addition, more severe expression of HNE staining was observed in the samples from persistent/chronic LAF than that in paroxysmal LAF. Conclusion: These data indicates that oxidative stress plays an important role as an aggravating factor in LAF patients.

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In silico and in vitro analysis of a multiepitope L1-E7 fusion construct for vaccine development against human papillomaviruses

Letters in Drug Design & Discovery ◽

10.2174/1570180818666210602160150 ◽

2021 ◽

Vol 18 ◽

Author(s):

Elnaz Abbasifarid ◽

Azam Bolhassani ◽

Shiva Irani ◽

Fattah Sotoodehnejadnematalahi

Keyword(s):

Western Blotting ◽

Vaccine Development ◽

Fluorescent Microscopy ◽

Therapeutic Vaccine ◽

Human Papillomaviruses ◽

Mhc I ◽

Fusion Construct ◽

Mhc Ii ◽

Hpv Types

Background: Human papillomavirus (HPV) infection is the major risk factor for cervical cancer. Current prophylactic HPV vaccines provide immunity against most genital and carcinogenic HPV types. However, these vaccines failed to produce immune responses against already established HPV infections. Methods: For the design of a therapeutic vaccine candidate, we utilized immunoinformatics tools to design a potential multiepitope fusion construct based on L1 and E7 genes from different high- and low-risk HPV types. After determination of CD4+ and CD8+ T cell epitopes, the allergenicity, toxicity, immunogenicity, conservancy, and population coverage were analyzed for epitope selection. Then, the hemolytic probability of the selected epitopes, and molecular docking between major histocompatibility complex (MHC) and the chosen epitopes were performed by different web servers. Next, a multiepitope peptide construct consisting of 12 epitopes linked by the AAY proteasomal sequence was designed. After that, physicochemical properties, solubility, secondary and tertiary structures of this construct were evaluated by bioinformatics tools. Finally, after amino acid reverse translation of the multiepitope peptide construct, expression of the L1-E7 DNA construct (pEGFP-L1-E7) was investigated in HEK-293T cells using fluorescent microscopy, flow cytometry, and western blotting. Results: Considering various parameters, the immunodominant peptides such as L1(MHC-I)-DLDQFPLGRKFLLQ, L1(MHC-II)-NQLFVTVVDTTRSTN, E7-HPV16(MHC-I)-AEPDRAHYNIVTF, E7-HPV18(MHC-I)-HGPKATVQDIVLHL, E7-HPV31(MHC-I)-KPDTSNYNIVTF, E7-HPV33(MHC-I)-RPDGQAQPATADYYI, E7-HPV45(MHC-I)- RTLQQLFLSFV, E7-HPV16(MHC-II)-TLHEYMLDLQPETTD, E7-HPV18(MHC-II)-LRAFQQLFLNTLSFV, E7-HPV31(MHC-II)-PTLQDYVLDLQPEAT, E7-HPV33(MHC-II)-LKEYVLDLYPEPTDL and E7-HPV45(MHC-II)-LQQLFLSTLSFVCPW were determined to design the vaccine construct. The results indicated efficient expression of the L1-E7 DNA construct (74 ± 2.19%) in vitro. Moreover, the polyepitope peptide generated in the cells was detected as a clear band of ~ 50 kDa in western blotting. Conclusion: Regarding the favorable transfection efficiency of the designed L1-E7 multiepitope construct, in vivo validation study on its therapeutic potential is underway.

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Efficacy of an experimental drug based on technologically processed antibodies in models of influenza infection and secondary bacterial pneumonia: Results of a preclinical study

Nauchno-prakticheskii zhurnal «Patogenez» ◽

10.25557/2310-0435.2020.04.55-63 ◽

2020 ◽

pp. 55-63

Author(s):

Н.В. Петрова ◽

А.Г. Емельянова ◽

С.А. Тарасов ◽

Н. П. Карташова ◽

Е.А. Глубокова

Keyword(s):

Survival Rate ◽

Bacterial Pneumonia ◽

Influenza Infection ◽

Control Groups ◽

Mhc I ◽

Mhc Ii ◽

Cd4 Receptor ◽

2009 H1n1 ◽

Secondary Bacterial Pneumonia ◽

Complex Drug

Целью исследования была оценка противовирусной активности экспериментальных образцов сверхвысоких разведений антител к различным мишеням, вовлеченным в реакции иммунного ответа (MHC I, MHC II, CD4 рецептор, ИФН-γ). Методы. Исследование противовирусной активности сверхвысоких разведений антител к молекуле MHC I проводили на модели летальной гриппозной инфекции (грипп А/Калифорния/04/2009 (H1N1)pdm09), тогда как протективный эффект комплексного препарата, содержащего сверхвысокие разведения антител к молекулам MHC I, MHC II, ИФН-γ и к CD4 рецептору, оценивали на модели смешанной вирусно-бактериальной пневмонии (последовательное инфицирование вирусом гриппа А/Калифорния/04/2009 (H1N1) и Staphylococcus aureus). Результаты. Показано, что сверхвысокие разведения антител к MHC I увеличивали выживаемость животных в эксперименте на 46,7% и 52,4% по сравнению с группами отрицательного контроля и плацебо (p < 0,05), соответственно. Препарат сравнения Осельтамивир повышал этот же показатель на 60% и 85,7% по сравнению с теми же группами животных (p < 0,05). На модели смешанной вирусно-бактериальной инфекции комплексный препарат повышал выживаемость мышей на 30% и 40% (p < 0,05) относительно контрольных групп. Введение Осельтамивира в комбинации с Цефуроксимом выражалось в увеличении выживаемости животных на 50% и 60%, соответственно. Статистически значимого снижения вирусной или бактериальной нагрузки ни для одной из групп выявлено не было. Заключение. Впервые продемонстрирована эффективность экспериментальных препаратов, содержащих сверхвысокие разведения антител к молекуле MHC I, а также их комплекс к MHC I, MHC II, ИФН-γ и к CD4 рецептору в моделях гриппозной инфекции и вирусно-бактериальной пневмонии у животных. Полученные данные свидетельствуют о перспективности дальнейшего изучения протективных эффектов данных образцов при вирусных и бактериальных инфекциях. The aim of this study was to evaluate the antiviral activity of ultra-highly diluted antibodies to various targets (MHC I, MHС II, INFg, and CD4 receptor) involved in the immune response. Methods. The antiviral activity of ultra-highly diluted antibodies to the MHC I molecule was evaluated in a standard model of the lethal A/California/04/2009 (H1N1)pdm09 influenza infection, and the protective effect of a complex drug containing highly diluted antibodies to MHC I and MHC II molecules, INFg, and CD4 receptor was accessed in a model of secondary bacterial pneumonia (A/California/04/2009 (H1N1) influenza virus challenge followed by Staphylococcus aureus inoculation). Results. The treatment with ultra-highly diluted antibodies to MHC I increased the survival rate of mice by 46.7% and 52.4% vs. the negative control and placebo groups (p < 0.05), respectively. The survival rate was increased in the Oseltamivir group by 60% and 85.7% vs. the same control groups (p < 0.05). In the model of secondary bacterial pneumonia following influenza, the complex drug increased the survival rate of mice by 30% and 40% (p < 0.05) vs. the control groups. The combined application of Oseltamivir and Cefuroxime increased the survival rate by 50% and 60%, respectively. There was no statistically significant decrease in the viral or bacterial load in any of the groups. Conclusion. The study showed for the first time that highly diluted antibodies to the MHC I molecule as well as the complex drug containing highly diluted antibodies to MHC I, MHС II, INFg, and CD4 receptor were effective in animal models of influenza infection and secondary bacterial pneumonia. Further investigation of protective effects of these samples in viral and bacterial infections is promising.

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