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Identification and immunologic property of macrophage migration inhibitory factor (MIF) in grass carp
Yinghuan Wu1, Danru Yang1, Yaqiu Lin1
1Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization of Ministry of Education, College of Life Science and Technology, Southwest Minzu University, Chengdu, 610041, PR China.
Insights
Macrophage migration inhibitory factor (MIF) in grass carp is predominantly expressed in immune tissues and supports white blood cell function. This study reveals MIF
Area of Science:
- Immunology
- Fish Biology
- Cytokine Research
Background:
- Macrophage migration inhibitory factor (MIF) is a key pro-inflammatory cytokine regulating T cell activation.
- The immunological role of MIF in freshwater fish remains largely unexplored.
Purpose of the Study:
- To identify and characterize the MIF gene in grass carp.
- To investigate the immunologic properties and functions of grass carp MIF.
Main Methods:
- Bioinformatics analysis of grass carp MIF gene and protein.
- Gene expression analysis in various tissues.
- In vitro assays using recombinant MIF and peripheral blood leukocytes.
- Cadmium-induced immunosuppression model.
Main Results:
- Grass carp MIF gene was identified, with the protein predicted to bind CD74.
- MIF expression was highest in immune organs (spleen, head kidney) and correlated with white blood cell counts.
- Grass carp MIF enhanced leukocyte viability, inhibited apoptosis, and promoted pro-inflammatory cytokine secretion (TNF-α, IL1β, IL6).
Conclusions:
- Grass carp MIF possesses significant immunologic activity.
- MIF plays a crucial role in regulating immune responses in grass carp, particularly in immune cell function and cytokine production.
Abstract:
Macrophage migration inhibitory factor (MIF), a pro-inflammatory cytokine, plays an important regulatory role in the activation of T cells induced by mitogenic or antigenic stimuli. However, the immunologic property of MIF in freshwater fish is limitedly known by now. In the present study, MIF gene was identified in grass carp. Bioinformatics analysis revealed that the molecular weight of grass carp MIF protein was 12.377 kDa and it could also bind to CD74. MIF gene was predominantly expressed in immune tissues including spleen and head kidney, then liver, skin, gill, intestine and blood, while a relative low level expression in heart, brain, fat and red muscle. The predicted receptor and tissues distribution of MIF implied the immunologic activity of grass carp MIF. Then grass carp MIF antigen and the polyclonal antibodies against it were prepared. Using cadmium as an immunosuppressive agent, MIF expression in spleen and head kidney was depressed in a dose-dependent manner with cadmium consumption. On the same time, white blood cell count decrease displayed a similar pattern with MIF expression, which suggested a possible positive correlation between MIF and white blood cell count. Thereafter, MIF enhanced the viability of grass carp peripheral blood leukocytes and inhibited cell apoptosis with depressed reactive oxygen species production in vitro. In addition, recombinant grass carp MIF promoted tumor necrosis factor-alpha (TNF-α), interleukin 1β (IL1β) and interleukin 6 (IL6) secretion from peripheral blood leukocytes. These results indicated the immunologic property of grass carp MIF.
