Full-length transcriptome sequencing and identification and immune response of TRIM genes in Dabry's sturgeon (Acipenser dabryanus)

ABSTRACT Chemically inactivated tetanus toxoid (CITT) is clinically effective and widely used. However, CITT is a crude nonmalleable vaccine that contains hundreds of Clostridium tetani proteins, and the active component is present in variable and sometimes minor percentages of vaccine mass. Recombinant production of a genetically inactivated tetanus vaccine offers an opportunity to replace and improve the current tetanus vaccine. Previous studies showed the feasibility of engineering full-length tetanus toxin (TT) in Escherichia coli. In the present study, full-length TT was engineered with eight individual amino acid mutations (8MTT) to inactivate catalysis, translocation, and host receptor-binding functions, retaining 99.4% amino acid identity to native tetanus toxin. 8MTT purified as a 150-kDa single-chain protein, which trypsin nicked to a 100-kDa heavy chain and 50-kDa light chain. The 8MTT was not toxic for outbred mice and was >50 million-fold less toxic than native TT. Relative to CITT, 8MTT vaccination elicited a strong immune response and showed good vaccine potency against TT challenge. The strength of the immune response to both vaccines varied among individual outbred mice. These data support 8MTT as a candidate vaccine against tetanus and a malleable candidate conjugate vaccine platform to enhance the immune response to polysaccharides and other macromolecular molecules to facilitate a rapid response to emerging microbial pathogens. IMPORTANCE Chemical inactivation is a clinically effective mechanism to detoxify protein toxins to produce vaccines against microbial infections and to serve as a platform for production of conjugate polysaccharide vaccines. This method is widely used for the production of protein toxin vaccines, including tetanus toxoid. However, chemical modification alters the protein structure with unknown effects on antigenicity. Here, a recombinant full-length tetanus toxin (TT) is engineered with 8 mutations (8MTT) that inactivate three toxin functions: catalysis, translocation, and receptor binding. 8MTT is nontoxic and elicits a potent immune response in outbred mice. 8MTT also represents a malleable platform for the production of conjugate vaccines, which can facilitate a rapid vaccine response against emerging microbial pathogens.

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Full-length transcriptome sequencing and identification of immune-related genes in the critically endangered Hucho bleekeri

Developmental & Comparative Immunology ◽

10.1016/j.dci.2020.103934 ◽

2021 ◽

Vol 116 ◽

pp. 103934

Author(s):

Yeyu Chen ◽

Huanchao Yang ◽

Yanling Chen ◽

Mingjiang Song ◽

Bo Liu ◽

...

Keyword(s):

Transcriptome Sequencing ◽

Full Length ◽

Critically Endangered ◽

Hucho Bleekeri ◽

Immune Related Genes

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Effects of dendritic cells transfected with full length wild type P53 and modified by gastric cancer lysate on immune response

Journal of Huazhong University of Science and Technology [Medical Sciences] ◽

10.1007/bf02831108 ◽

2004 ◽

Vol 24 (5) ◽

pp. 460-463

Author(s):

Sun Huawen ◽

Tang Qibing ◽

Chen Yongjun ◽

Tang Cong ◽

Zou ShengQian ◽

...

Keyword(s):

Gastric Cancer ◽

Immune Response ◽

Dendritic Cells ◽

Full Length ◽

Wild Type ◽

Wild Type P53

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REDIportal: millions of novel A-to-I RNA editing events from thousands of RNAseq experiments

Nucleic Acids Research ◽

10.1093/nar/gkaa916 ◽

2020 ◽

Vol 49 (D1) ◽

pp. D1012-D1019 ◽

Cited By ~ 1

Author(s):

Luigi Mansi ◽

Marco Antonio Tangaro ◽

Claudio Lo Giudice ◽

Tiziano Flati ◽

Eli Kopel ◽

...

Keyword(s):

Gene Expression ◽

Immune Response ◽

Comparative Genomics ◽

Rna Editing ◽

Innate Immune Response ◽

Transcriptome Sequencing ◽

Regulation Of Gene Expression ◽

Innate Immune ◽

Neurotransmitter Receptors ◽

Eukaryotic Organisms

Abstract RNA editing is a relevant epitranscriptome phenomenon able to increase the transcriptome and proteome diversity of eukaryotic organisms. ADAR mediated RNA editing is widespread in humans in which millions of A-to-I changes modify thousands of primary transcripts. RNA editing has pivotal roles in the regulation of gene expression or modulation of the innate immune response or functioning of several neurotransmitter receptors. Massive transcriptome sequencing has fostered the research in this field. Nonetheless, different aspects of the RNA editing biology are still unknown and need to be elucidated. To support the study of A-to-I RNA editing we have updated our REDIportal catalogue raising its content to about 16 millions of events detected in 9642 human RNAseq samples from the GTEx project by using a dedicated pipeline based on the HPC version of the REDItools software. REDIportal now allows searches at sample level, provides overviews of RNA editing profiles per each RNAseq experiment, implements a Gene View module to look at individual events in their genic context and hosts the CLAIRE database. Starting from this novel version, REDIportal will start collecting non-human RNA editing changes for comparative genomics investigations. The database is freely available at http://srv00.recas.ba.infn.it/atlas/index.html.

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Effects of a Sudden Drop in Salinity on Immune Response Mechanisms of Anadara kagoshimensis

International Journal of Molecular Sciences ◽

10.3390/ijms20184365 ◽

2019 ◽

Vol 20 (18) ◽

pp. 4365 ◽

Cited By ~ 2

Author(s):

Mo Zhang ◽

Li Li ◽

Ying Liu ◽

Xiaolong Gao

Keyword(s):

Immune Response ◽

Signaling Pathway ◽

Transcriptome Sequencing ◽

Differential Expression Analysis ◽

Enrichment Analysis ◽

Total Count ◽

Sudden Drop ◽

Blood Lymphocytes ◽

Anadara Kagoshimensis ◽

Comparison Groups

In this experiment, the effects of a sudden drop of salinity on the immune response mechanisms of the ark shell Anadara kagoshimensis were examined by simulating the sudden drop of salinity that occurs in seawater after a rainstorm. Additionally, the differentially expressed genes (DEGs) were identified using transcriptome sequencing. When the salinity dropped from 30‰ (S30) to 14‰ (S14), the phagocytic activity of blood lymphocytes, the O2− levels produced from respiratory burst, the content of reactive oxygen species, and the activities of lysozymes and acid phosphatases increased significantly, whereas the total count of blood lymphocytes did not increase. Total count of blood lymphocytes in 22‰ salinity (S22) was significantly higher than that in any other group. The raw data obtained from sequencing were processed with Trimmomatic (Version 0.36). The expression levels of unigenes were calculated using transcripts per million (TPM) based on the effects of sequencing depth, gene length, and sample on reads. Differential expression analysis was performed using DESeq (Version 1.12.4). Transcriptome sequencing revealed 269 (101 up-regulated, 168 down-regulated), 326 (246 up-regulated, 80 down-regulated), and 185 (132 up-regulated, 53 down-regulated) significant DEGs from comparison of the S14 vs. S22, S22 vs. S30, and S14 vs. S30 groups, respectively. Gene Ontology enrichment analysis of the DEGs in these salinity comparison groups revealed that the cellular amino acid metabolic process, the regulation of protein processing, the regulation of response to stress, and other terms were significantly enriched. Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that nucleotide-binding, oligomerization domain (NOD)-like receptor signaling pathway (ko04621), apoptosis-multiple species (ko04215), Toll and Imd signaling pathway (ko04624), NF-κB signaling pathway (ko04064), apoptosis (ko04210), and focal adhesion (ko04510) were significantly enriched in all salinity comparison groups. qRT-PCR verification of 12 DEGs in the above six pathways was conducted, and the results were consistent with the transcriptome sequencing results in terms of up-regulation and down-regulation, which illustrates that the transcriptome sequencing data are credible. These results were used to preliminarily explore the effects of a sudden drop of salinity on blood physiological and biochemical indexes and immunoregulatory mechanisms of A. kagoshimensis. They also provide a theoretical basis for the selection of bottom areas optimal for release and proliferation of A. kagoshimensis required to restore the declining populations of this species.

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