Wang B, Zhang Z, Wang Y et al.Molecular cloning and characterization of macrophage migration inhibitory factor from small abalone Haliotis diversicolor supertexta. Fish Shellfish Immunol 27:57-64

The Key Laboratory of Science and Technology for Aquaculture and Food Safety, Fisheries College, Jimei University, Yindou Road #43, Xiamen, Fujian, China.
Fish &amp Shellfish Immunology (Impact Factor: 2.67). 06/2009; 27(1):57-64. DOI: 10.1016/j.fsi.2009.04.004
Source: PubMed


The macrophage migration inhibitory factor (mif) cDNA and its genome were cloned from small abalone Haliotis diversicolor supertexta. Small abalone mif (samif) was originally identified from an expressed sequence tag (EST) fragment from a normalized cDNA library. It's 5' untranslated region (UTR) was obtained by 5' rapid amplification of cDNA end (RACE) techniques and its genomic DNA was cloned by PCR. The full-length cDNA of samif was of 535 bp, consisting of a 5'-terminal UTR of 49 bp, an open reading frame of 384 bp and a 3'-terminal UTR of 102 bp. The deduced protein was composed of 128 amino acids, with an estimated molecular mass of 14.0 kDa and a predicted pI of 6.90. The full-length samif genomic DNA comprises 3238 bp, containing three exons and two introns. Real time quantitative PCR analysis revealed that samif gene is constitutively expressed in 6 selected tissues, and its expression level in hepatopancreas is higher than that in the other tissues (p < 0.01). Samif expression level in the hepatopancreas at 24 and 48 h after Vibrio parahaemolyticus injection was upregulated significantly (p < 0.01), but there was no significant change after exposure to tributyltin (TBT) (p > 0.05).

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Available from: Guodong Wang, Sep 01, 2015
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    • "In vertebrates, MIFs are important pro-inflammatory cytokines acting on key cellular processes of the immune response such as cell proliferation and apoptosis [22, 23]. MIFs were identified in a variety of species, including protozoan, nematode, mollusk and crustacean species [5, 24–29]. It was shown that a mollusk MIF (BgMIF1) not only presented the expected activities on cell proliferation and apoptosis but played a major role in the response against parasitic infection [5]. "
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    ABSTRACT: Background The widespread use of genome sequencing provided evidences for the high degree of conservation in innate immunity signalling pathways across animal phyla. However, the functioning and evolutionary history of immune-related genes remains unknown for most invertebrate species. A striking observation coming from the analysis of the pea aphid Acyrthosiphon pisum genome is the absence of important conserved genes known to be involved in the antimicrobial responses of other insects. This reduction in antibacterial immune defences is thought to be related to their long-term association with beneficial symbiotic bacteria and to facilitate symbiont maintenance. An additional possibility to avoid elimination of mutualistic symbionts is a fine-tuning of the host immune response. To explore this hypothesis we investigated the existence and potential involvement of immune regulators in aphid agonistic and antagonistic interactions. Results In contrast to the limited antibacterial arsenal, we showed that the pea aphid Acyrthosiphon pisum expresses 5 members of Macrophage Migration Inhibitory Factors (ApMIF), known to be key regulators of the innate immune response. In silico searches for MIF members in insect genomes followed by phylogenetic reconstruction suggest that evolution of MIF genes in hemipteran species has been shaped both by differential losses and serial duplications, raising the question of the functional importance of these genes in aphid immune responses. Expression analyses of ApMIFs revealed reduced expression levels in the presence, or during the establishment of secondary symbionts. By contrast, ApMIFs expression levels significantly increased upon challenge with a parasitoid or a Gram-negative bacteria. This increased expression in the presence of a pathogen/parasitoid was reduced or missing, in the presence of facultative symbiotic bacteria. Conclusions This work provides evidence that while aphid’s antibacterial arsenal is reduced, other immune genes widely absent from insect genomes are present, diversified and differentially regulated during antagonistic or agonistic interactions. Electronic supplementary material The online version of this article (doi:10.1186/1471-2164-15-762) contains supplementary material, which is available to authorized users.
    BMC Genomics 09/2014; 15(1):762. DOI:10.1186/1471-2164-15-762 · 3.99 Impact Factor
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    • "Another ancient and conserved cytokine of the immune system is the macrophage migration inhibitory factor (MIF), which is reported from cyanobacteria and sea urchins to humans [8], [82] and was also recently discovered in the group of mollusca [83], [84]. Previously found to be a T-cell cytokine of the adaptive immune system, MIF is emerging as an important mediator of the innate immune system and the conservation of MIF across taxa indicates an important biological function. "
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    ABSTRACT: The marine mussel Mytilus edulis and its closely related sister species are distributed world-wide and play an important role in coastal ecology and economy. The diversification in different species and their hybrids, broad ecological distribution, as well as the filter feeding mode of life has made this genus an attractive model to investigate physiological and molecular adaptations and responses to various biotic and abiotic environmental factors. In the present study we investigated the immune system of Mytilus, which may contribute to the ecological plasticity of this species. We generated a large Mytilus transcriptome database from different tissues of immune challenged and stress treated individuals from the Baltic Sea using 454 pyrosequencing. Phylogenetic comparison of orthologous groups of 23 species demonstrated the basal position of lophotrochozoans within protostomes. The investigation of immune related transcripts revealed a complex repertoire of innate recognition receptors and downstream pathway members including transcripts for 27 toll-like receptors and 524 C1q domain containing transcripts. NOD-like receptors on the other hand were absent. We also found evidence for sophisticated TNF, autophagy and apoptosis systems as well as for cytokines. Gill tissue and hemocytes showed highest expression of putative immune related contigs and are promising tissues for further functional studies. Our results partly contrast with findings of a less complex immune repertoire in ecdysozoan and other lophotrochozoan protostomes. We show that bivalves are interesting candidates to investigate the evolution of the immune system from basal metazoans to deuterostomes and protostomes and provide a basis for future molecular work directed to immune system functioning in Mytilus.
    PLoS ONE 03/2012; 7(3):e33091. DOI:10.1371/journal.pone.0033091 · 3.23 Impact Factor
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    • "Only 4 cDNA sequences related to MIF have been reported so far from gastropods (Mitta et al., 2005; Wang et al., 2009a) and 2 from bivalves (Cui et al., 2011; Wang et al., 2009b). In addition , the role of MIF in the immune response against bacteria or parasites has been investigated only in the gastropods, Haliotis diversicolor (Wang et al., 2009a) and Biomphalaria glabrata (Baeza Garcia et al., 2010), and in the bivalve, Pinctada fucata (Cui et al., 2011). "
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    ABSTRACT: Three macrophage migration inhibitory factor (MIF)-related sequences were identified from a Mytilus galloprovincialis EST library. The consensus sequence included a 5'-UTR of 32 nucleotides, the complete ORF of 345 nucleotides, and a 3'-UTR of 349 nucleotides. As for other MIFs, M. galloprovincialis ORF does not include any signal or C-terminus extensions. The translated sequence of 115 amino acids possesses a molecular mass of 12,681.4, a pI of 6.27 and a stability index of 21.48. Its 3D structure resembles human MIF except for one shorter α-helix. Although evolutionary separated from ticks and vertebrates, Mg-MIF appeared to be closely related to Pinctada fucata and Haliotis, but not to Chlamys farreri and Biomphalaria glabrata. Numerous mutation points were observed within the Mg-MIF ORF, defining 11 amino acid variants within the mussels from Palavas-France and 14 amino acid variants within the mussels from Palermo-Italy. The 2 major variants from Palavas were identical to 2 of the 4 major variants from Palermo. In all the 18 Mg-MIF variants, residues involved in tautomerase and in oxidoreductase activities were conserved. Generally, one mussel expressed 2 Mg-MIF amino acid sequences but with different frequencies of occurrence. Mg-MIF is constitutively expressed principally in hemocytes and in the mantle. In contrast to other animal models, Mg-MIF expression was always down regulated following challenge by bacteria and fungi, confirming previous data obtained with microarray. Down regulation started as soon as 1 h and Mg-MIF expression returned to background 9-48 h after the challenge. Exception was regarding the yeast, Candidaalbicans, down-regulation between 9 and 72 h, suggesting yeast and bacteria-filamentous fungi trigger different mechanisms of elimination.
    Developmental and comparative immunology 11/2011; 36(4):688-96. DOI:10.1016/j.dci.2011.10.014 · 2.82 Impact Factor
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