Recombinant Macrophage Migration Inhibitory Factor Derived from Trichinella spiralis Suppresses Obesity by Reducing Body Fat and Inflammation

Abstract

Obesity, an escalating global health crisis, is characterized by adipose tissue hypertrophy and chronic low-grade inflammation. Although anti-obesity drugs can induce weight loss, their use is limited by adverse effects, underscoring the need for safer therapeutic strategies. In this study, we generated a recombinant form of Trichinella spiralis-derived macrophage migration inhibitory factor (rTs-MIF) and investigated its anti-inflammatory and anti-obesity effects via immunometabolic regulation. Male C57BL/6 mice fed a 45% high-fat diet were orally administered rTs-MIF, and its effects were evaluated by measuring fat mass, glucose metabolism, serum cytokines, liver histology, and adipose tissue parameters. In 3T3-L1 cells, we examined the effects of rTs-MIF on adipocyte differentiation, obesity-related gene expression, and intracellular signaling pathways. Oral rTs-MIF suppressed body weight gain, reduced fat mass, improved glucose levels, and decreased the food efficiency ratio. It also lowered pro-inflammatory cytokines and increased markers associated with M2 macrophages. In 3T3-L1 cells, rTs-MIF inhibited adipocyte differentiation and reduced the expression of lipogenic transcription factors and mouse Mif while modulating AKT and p44/42 MAPK signaling. These findings identify rTs-MIF as a potential bioactive candidate that ameliorates obesity by regulating the immune–metabolic axis.

1. Introduction

Obesity is a major risk factor for chronic diseases, including cardiovascular diseases, diabetes, and hypertension [1,2,3]. Since the 1990s, obesity occurrences have more than doubled in children and tripled in adolescents [4]. If this trend continues, it is estimated that 51% of the global population will be obese by 2030 [5]. This trend is evident in both developed and developing regions, and the demand for anti-obesity therapies continues to rise [6,7]. Glucagon-like peptide-1 (GLP-1)-based drugs have recently attracted significant attention as anti-obesity therapeutics [8]; however, their long-term use is associated with pancreatitis, thyroid cancer, and gastrointestinal complications [9,10].

Obesity is not merely a state of energy imbalance, but is also characterized by chronic low-grade inflammation within the adipose tissue [11]. Classically activated M1 macrophages produce pro-inflammatory cytokines such as IL-6, IL-1β, and TNF-α; meanwhile, activated M2 macrophages secrete IL-10 and TGF-β, thereby promoting tissue repair and anti-inflammatory responses [12]. Adipocytes themselves also produce pro-inflammatory cytokines [13,14]. Interestingly, various helminth infections are known to induce Th2 or Treg-biased immune responses, suppress inflammation, and restore tissue homeostasis [15,16]. Previous studies have shown that Trichinella spiralis infection suppressed body weight gain and alleviated adipose tissue inflammation in high-fat diet (HFD)-induced obese mouse models [17]. Moreover, the total lysate of T. spiralis has been reported to reduce body weight and fat mass and improve blood glucose levels [18]. Accordingly, we focused on helminth-derived macrophage migration inhibitory factor (MIF) with immunoregulatory properties.

Host MIF is a well-known pro-inflammatory factor that binds to CD74 and activates inflammatory signaling pathways [19]. At the molecular level, MIF activates the ERK1/2 MAPK pathway via CD74/CD44 [20], inducing the production of pro-inflammatory cytokines such as TNF-α, IL-1β, IL-6, and IFN-γ and amplifying immune responses [21]. In contrast, helminth-derived MIFs have been reported to exert anti-inflammatory effects, including the suppression of allergic inflammation, attenuation of colitis, and modulation of immune homeostasis [22,23,24]. The structural features unique to the T. spiralis MIF (rTs-MIF) suggest functional properties distinct from those of the host MIF, highlighting its potential as a therapeutic target.

Moreover, MIF family proteins are small and characterized by relatively stable molecules with a trimer structure that can be easily mass produced in their recombinant form, making them attractive candidates for biological therapeutics [25]. As rTs-MIF does not directly target the pathways associated with adverse gastrointestinal, neurological, or cardiovascular effects commonly observed with current anti-obesity drugs, it is expected to have a lower risk of long-term side effects [26]. Meanwhile, parasite-derived immunomodulatory proteins have evolutionarily adapted to minimize host tissue damage, suggesting their high degree of safety and substantial potential for use as therapeutic agents [27].

Here, we investigated the potential of T. spiralis-derived MIF as a modulator of obesity and evaluated its feasibility as a novel anti-obesity therapeutic candidate with fewer adverse effects by targeting the immune–metabolic axis.

2. Results

2.1. Structural Differences Between T. spiralis MIF and M. musculus MIF

To analyze the similarity of MIF among these species, we performed multiple sequence alignments of T. spiralis and M. musculus (Figure 1A). The sequence identity between T. spiralis MIF and M. musculus MIF was 40.35%. A superimposition of T. spiralis MIF (green) and M. musculus MIF (yellow) is shown in the Figure (Figure 1B). The root-mean-square deviation (RMSD) between the two structures was 0.647 Å.

2.2. rTs-MIF Reduces Obesity in Mice

The overall experimental scheme is presented in Figure 2A. After 8 weeks of rTs-MIF administration, body fat accumulation was reduced compared with that in the HFD group (Figure 2B). In the 100 µg group, body weight gain was significantly suppressed from week 7 onward (Figure 2C), and FER significantly decreased (Figure 2D). OGTT results showed improved glucose tolerance (Figure 2E). Consistently, the total fat mass measured by MRI and the eWAT weight were reduced in a dose-dependent manner in rTs-MIF-treated mice (Figure 2F,G).

2.3. rTs-MIF Inhibits Hepatic Lipid Accumulation and Lipogenic Gene Expression

H&E staining of liver sections revealed minimal lipid droplet formation in ND mice, whereas HFD mice exhibited a marked increase in intrahepatic lipid droplets. By contrast, mice treated with rTs-MIF (50 or 100 µg) showed a substantial reduction in the size and number of hepatic lipid droplets (Figure 3A). Quantitative analysis of the lipid droplet area confirmed that the pronounced HFD-induced increase in hepatic lipid accumulation was significantly attenuated by rTs-MIF treatment (Figure 3B). Gene expression analysis revealed that rTs-MIF downregulated HFD-induced expression of Acox1, Mcp1, Srebp1c, and Gpat1 (Figure 3C). These findings indicate that rTs-MIF effectively suppressed hepatic lipogenesis and ameliorated HFD-induced metabolic disturbances in the liver.

2.4. rTs-MIF Decreases Th1 Cytokines and Increases Th2 Cytokines

The 45% HFD significantly increased serum levels of the pro-inflammatory cytokines IL-6, IL-1β, and IFN-γ. In contrast, treatment with rTs-MIF (50 or 100 µg) reduced the levels of these pro-inflammatory cytokines. Anti-inflammatory cytokines IL-10 and TGF-β were elevated in the HFD group and further increased in the rTs-MIF-treated groups, suggesting that rTs-MIF enhances anti-inflammatory responses and contributes to the resolution of HFD-induced inflammation (Figure 4).

2.5. rTs-MIF Modulates Macrophage Composition in White Adipose Tissue

Flow cytometric analysis revealed that high-fat diet (HFD) feeding markedly increased the proportion of CD11c⁺ macrophages within the stromal vascular fraction (SVF) of epididymal white adipose tissue (eWAT), indicating an accumulation of pro-inflammatory M1 macrophages. In contrast, mice treated with rTs-MIF at doses of 50 and 100 µg exhibited a reduction in CD11c⁺ cells together with an increase in CD206⁺ cells, suggesting a shift in macrophage composition toward an anti-inflammatory phenotype (Figure 5A). Quantitative assessment confirmed that rTs-MIF treatment significantly reduced the CD11c⁺F4/80⁺ M1 macrophage population, whereas CD206⁺F4/80⁺ M2 macrophages showed an increasing trend that did not reach statistical significance. Accordingly, the M2/M1 macrophage ratio was elevated in rTs-MIF-treated groups compared with HFD controls, indicating a relative enrichment of M2-like macrophages in adipose tissue (Figure 5B). Consistent with these cellular changes, mRNA expression levels of M2-associated markers, including Mrc1 (encoding CD206) and Mgl2, were significantly upregulated in the adipose tissue of rTs-MIF-treated mice. These molecular findings support the notion that rTs-MIF is associated with a shift in adipose tissue macrophage balance toward an anti-inflammatory M2 profile under HFD conditions (Figure 5C).

2.6. rTs-MIF Inhibits Adipocyte Differentiation in 3T3-L1 Cells

Oil Red O staining demonstrated abundant lipid droplet formation in fully differentiated adipocytes in the positive control group, whereas rTs-MIF treatment (1, 5, and 10 µg/mL) reduced the size and number of lipid droplets in a dose-dependent manner (Figure 6A). Quantification of Oil Red O absorbance demonstrated that intracellular lipid accumulation was significantly decreased by rTs-MIF treatment (Figure 6B). During adipocyte differentiation, expression of the adipogenic transcription factors Pparg, Cebpa, Fabp4, and mouse Mif progressively increased in the positive control group, whereas this increase was attenuated in cells treated with 10 µg/mL rTs-MIF. The difference in the expression levels between the positive control and rTs-MIF groups was particularly pronounced at the late differentiation stage (day 10), demonstrating that rTs-MIF inhibited adipocyte differentiation (Figure 6C).

2.7. rTs-MIF Modulates Obesity-Related Signaling Pathways During Early 3T3-L1 Differentiation

Time-course analysis of signaling pathways after rTs-MIF (10 µg/mL) treatment revealed that phosphorylation of AKT and p44/42 MAPK tended to increase at 5 min but decreased at 15–30 min. The total AKT and total p44/42 MAPK levels remained relatively unchanged over time (Figure 7A). Quantification of band intensities confirmed a transient increase in phosphorylation of AKT and p44/42 MAPK at early time points, followed by a decline at later time points (Figure 7B).

3. Discussion

In this study, we comprehensively evaluated the anti-obesity and anti-inflammatory effects of rTs-MIF at systemic, tissue, and cellular levels. We demonstrated that rTs-MIF suppressed body weight gain, reduced fat accumulation, alleviated hepatic lipid deposition, decreased systemic inflammation, promoted macrophage polarization toward M2, inhibited adipocyte differentiation, downregulated lipogenic transcription factors, and modulated AKT and p44/42 MAPK signaling in HFD-induced obese mice and 3T3-L1 cells.

HFD-fed mice exhibited continuous body weight gain compared to ND controls, whereas rTs-MIF-treated mice showed attenuated weight gain. The FER analysis indicated reduced weight gain relative to food intake. Although the food efficiency ratio (FER) can be influenced by factors such as nutrient absorption and intestinal microflora, the observed changes were interpreted in the context of longitudinal analyses of body weight and food intake using repeated-measures statistical approaches. Improvement in glucose tolerance was consistent with previous observations that helminth-derived molecules support metabolic homeostasis [28,29]. These effects extend beyond simple weight reduction and align with an anti-obesity pattern wherein reduced metabolic stress and inflammation contribute to improved insulin sensitivity [30,31,32]. Liver histology showed that rTs-MIF markedly reduced HFD-induced hepatic lipid droplet accumulation, suggesting attenuation of fatty liver and improvement of hepatic lipid metabolism through the suppression of lipogenesis and/or enhancement of fatty acid oxidation. HFD typically upregulates lipogenesis-related genes such as Acox1, Mcp1, Srebp1c, and Gpat1 in the liver, promoting lipid accumulation [33,34,35,36]. Although reductions in hepatic lipid droplet area were accompanied by coordinated changes in lipid metabolism-related gene expression, correlation analyses between gene expression levels and lipid droplet area may help clarify the underlying mechanisms in future studies. In this study, rTs-MIF reversed these gene expression changes, thus mitigating HFD-induced hepatic metabolic stress and facilitating the restoration of lipid homeostasis. In rTs-MIF-treated mice, reductions in IL-6, IL-1β, and IFN-γ, along with further increases in IL-10 and TGF-β, clearly showed a shift from a pro-inflammatory to an anti-inflammatory systemic cytokine profile. Obesity is generally associated with elevated IL-6, IL-1β, and IFN-γ levels due to an increased M1 macrophage population and chronic low-grade inflammation [37]. Meanwhile, IL-10 and TGF-β may increase as part of an adaptive immunoregulatory response aimed at limiting tissue damage and stress under chronic inflammatory conditions [38]. The additional increase in anti-inflammatory cytokines in rTs-MIF-treated mice suggests that helminth-derived MIF directly enhances anti-inflammatory signaling, rather than merely reflecting passive compensation. FACS demonstrated that rTs-MIF reduced the HFD-induced accumulation of CD11c⁺ M1 macrophages in adipose tissue and promoted an increase in CD206⁺ M2 macrophages. The elevated M2/M1 ratio in rTs-MIF-treated mice indicated that rTs-MIF actively reprogrammed macrophage polarization toward an anti-inflammatory phenotype. The increased Mrc1, and Mgl2 expression supports the notion that rTs-MIF promotes M2 polarization. Together, these findings suggest that rTs-MIF alleviates the inflammatory microenvironment in the adipose tissue, contributing to improved adipocyte function and systemic metabolic normalization.

In vitro, rTs-MIF reduced lipid accumulation in 3T3-L1 adipocytes and suppressed Pparg, Cebpa, and Fabp4 expression. Pparg, Cebpa, and Fabp4 are key adipogenic transcription factors and markers of adipocyte differentiation and maturation [39]. Furthermore, rTs-MIF treatment significantly suppressed the expression of mouse Mif, which typically increases during adipocyte differentiation. Because host MIF is known to promote adipogenesis and amplify inflammatory signaling [40], this finding provides important evidence that T. spiralis MIF may counter-regulate host MIF signaling, contributing to its anti-inflammatory and anti-obesity effects. Western blot analysis revealed that rTs-MIF induced a transient increase in AKT and p44/42 MAPK phosphorylation at 5 min, which decreased subsequently. The AKT and p44/42 MAPK signaling pathways are critical early regulators of adipocyte differentiation [41,42]. The observed transient changes in AKT and p44/42 MAPK phosphorylation suggest that rTs-MIF may interfere with the maintenance of sustained signaling required for adipogenesis, rather than inducing prolonged activation of these pathways. Host MIF is known to strongly activate p44/42 MAPK via CD74–CD44 and promote inflammation [43]. In contrast, rTs-MIF in this study induced only transient activation, which was followed by downregulation of these pathways. This suggests that helminth-derived MIF competitively interferes with host MIF or modulates receptor interactions in a manner that attenuates pro-inflammatory signaling, which in turn likely contributes to the inhibition of adipocyte differentiation, regulation of macrophage polarization, and mitigation of inflammation. These findings are in line with previous reports that helminth-derived MIFs exhibit distinct immunoregulatory and metabolic functions, despite their structural similarity to host MIF [43,44]. Differences in the exposed loop regions of helminth MIF may differentially affect its interactions with CD74 or other receptors, thereby conferring anti-inflammatory and anti-obesity properties.

Taken together, these findings suggest that rTs-MIF may act as a molecular regulator involved in both immune modulation and metabolic control, rather than functioning solely as an inhibitor of adipogenesis. Further studies focusing on receptor-level interactions, including CD74–CD44 engagement, as well as structural–functional analyses of helminth-derived MIF, will be crucial in defining the mechanisms underlying its immunometabolic effects more precisely. In addition, long-term and tissue-specific investigations may help clarify how rTs-MIF contributes to the maintenance of immune–metabolic homeostasis in obesity.

4. Materials and Methods

4.1. Protein Expression and Purification

Primers targeting the T. spiralis MIF gene (XM_003378364.1) were designed as follows: forward, 5′-GAATTCATGCCGATTTTTAC-3′; reverse, 5′-CTCGAGTCAGAATGTAGTACC-3′. The target gene was amplified using PCR, cloned into a T-vector, and subcloned into the pET-28a vector (Novagen, Darmstadt, Germany). The recombinant construct was transfected into Escherichia coli BL21 cells. Protein expression was induced with 0.5 mM IPTG (Sigma-Aldrich, St. Louis, MO, USA), and the cells were harvested via centrifugation (4000 rpm, 20 min, 4 °C). The cell pellets were disrupted using an ultrasonic homogenizer, and the supernatants were collected. The recombinant His-tagged protein was purified using a His-tag affinity column (Sigma-Aldrich, St. Louis, MO, USA), followed by endotoxin removal using an LPS depletion column (Thermo Fisher Scientific, Waltham, MA, USA).

4.2. Mice and Diet

Six-week-old, male C57BL/6 mice were purchased from Samtako (Seongnam-si, Republic of Korea). Mice were randomly allocated to groups and fed either a normal diet (ND) or a 45% high-fat diet (HFD; Research Diets, New Brunswick, NJ, USA) until the end of the experiment. After 4 weeks of HFD feeding, a subset of HFD-fed mice received oral rTs-MIF at doses of 50 or 100 µg every other day for 8 weeks. rTs-MIF was administered mixed with corn oil, with corn oil accounting for one-third of the total administration volume. Control HFD mice received an equal volume of vehicle consisting of PBS mixed with corn oil in the same proportion. The body weight and food intake were measured weekly. The food efficiency ratio (FER) was calculated as daily body weight gain (g)/daily food intake (g). All animal procedures were approved by the Institutional Animal Care and Use Committee (IACUC) of Pusan National University (Approval No. PNU-2025-0305).

4.3. Measurement of Body Fat Mass and Adipose Tissue Weight

One day before sacrifice, total body fat mass was quantified using a body composition analyzer (Bruker Minispec LF50, Billerica, MA, USA). After sacrifice, the epididymal white adipose tissue (eWAT) surrounding the testes was dissected and weighed to assess changes in fat mass.

4.4. Oral Glucose Tolerance Test (OGTT)

After 8 weeks of rTs-MIF administration, mice were fasted for 16 h and then orally gavaged with a 20% glucose solution at a dose of 10 µL per gram of body weight. Blood glucose levels were measured at 0, 15, 30, 60, and 120 min after glucose administration.

4.5. Histological Analysis

Liver samples were fixed in 4% formalin, embedded in paraffin, sectioned, and stained with hematoxylin and eosin (H&E) for histological evaluation. Lipid droplets in H&E-stained liver sections were quantified using ImageJ (version 1.54g; NIH, Bethesda, MD, USA). Images were captured at the same magnification, and the droplet area was measured as a percentage of the total hepatic tissue area. For each animal, three to five randomly selected fields were analyzed.

4.6. RNA Extraction and cDNA Synthesis

Total RNA was extracted from the liver tissue, eWAT, and 3T3-L1 cells using TRIzol reagent (Invitrogen, Burlington, ON, Canada). Quantitative PCR was performed using SYBR™ Select Master Mix (Thermo Fisher Scientific, Waltham, MA, USA). Gapdh was used as an endogenous control. The genes analyzed included Acox1, Mcp1, Srebp1c, Gpat1, Pparg, Cebpa, Fabp4, mouse Mif, Mrc1, and Mgl2. The primer sequences used for each gene are listed in

Table 1. Primer sequences for quantitative PCR.

Primer	Sequence
Gapdh-for 1	5′-CAT CAC TGC CAC CCA GAA GAC TG-3′
Gapdh-rev	5′-ATG CCA GTG AGC TTC CCG TTC AG-3′
Acox1-for	5′-GCC ATT CGA TAC AGT GCT GTG AG-3′
Acox1-rev	5′-CCG AGA AAG TGG AAG GCA TAG G-3′
Mcp1-for	5′-GCT ACA AGA GGA TCA CCA GCA G-3′
Mcp1-rev	5′-GTC TGG ACC CAT TCC TTC TTG G-3′
Srebp1c-for	5′-GGA GCC ATG GAT TGC ACA TT-3′
Srebp1c-rev	5′-GGC CCG GGA AGT CAC TGT-3′
Gpat1-for	5′-GCA AGC ACT GTT ACC AGC GAT C-3′
Gpat1-rev	5′-TGC AAT CAG CCT TCG TCG GAA G-3′
Mgl2-for	5′-CGA GAC TTG AGC CAG AAG GTG A-3′
Mgl2-rev	5′-GCC TTC AAG TCT GTC TCC AGC T-3′
Mrc1-for	5′-CAG GTG TGG GCT CAG GTA GT-3′
Mrc1-rev	5′-TGT GGT GAG CTG AAA GGT GA-3′
Pparg-for	5′-GCC TCC TGG TGA CTT TAT GGA-3′
Pparg-rev	5′-GCA GCA GGT TGT CTT GGA TG-3′
Cebpa-for	5′-CAA AGC CAA GAA GTC GGT GGA CAA-3′
Cebpa-rev	5′-TCA TTG TGA CTG GTC AAC TCC AGC-3′
Fabp4-for	5′-TGA AAT CAC CGC AGA CGA CAG G-3′
Fabp4-rev	5′-GCT TGT CAC CAT CTC GTT TTC TC-3′
mouse Mif-for	5′-ATG CCT ATG TTC ATC GTG-3′
mouse Mif-rev	5′-TCA AGC GAA GGT GGA ACC-3′

¹ for: forward; rev: reverse.

.

4.7. Cytokine Analysis Using ELISA

At sacrifice, blood was collected from the heart, and the serum was separated by centrifugation. Serum concentrations of IL-6, IL-1β, IFN-γ, IL-10, and TGF-β were measured using ELISA kits (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s instructions.

4.8. Flow Cytometry (FACS)

eWAT was excised and enzymatically digested with 0.2% collagenase type I (Sigma-Aldrich, St. Louis, MO, USA) to isolate the stromal vascular fraction (SVF). Isolated SVF cells were stained with fluorophore-conjugated antibodies against F4/80, CD11c, and CD206 (eBioscience, San Diego, CA, USA). Macrophages were identified as F4/80⁺ cells, and CD11c and CD206 were used as representative markers of M1 and M2 macrophage phenotypes, respectively. Data acquisition was performed using a BD FACSCanto II flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA), and data analysis was conducted using FlowJo software (version 10; BD Biosciences) according to the manufacturer’s instructions. CD11c and CD206 were selected as representative markers of classically activated (M1) and alternatively activated (M2) macrophages, respectively, as these markers are widely used to assess macrophage polarization in adipose tissue inflammation and metabolic disease models [11,12]. Detailed information regarding antibody specifications, gating strategy, and flow cytometry controls is provided in Supplementary Figure S1 and Supplementary Table S1.

4.9. 3T3-L1 Cell Culture and Differentiation

3T3-L1 preadipocytes were seeded in 24-well plates at a density of 5 × 10⁴ cells/well and cultured in DMEM supplemented with 10% FBS and penicillin/streptomycin until reaching 100% confluence. Upon confluence, adipocyte differentiation was induced for 48 h using DMEM containing 0.5 mM IBMX, 1 μM dexamethasone, and 1 μg/mL insulin with 10% FBS. rTs-MIF (1, 5, or 10 μg/mL) was co-treated throughout the differentiation period. After 48 h, cells were switched to maintenance medium (DMEM containing 1 μg/mL insulin), and the medium was replaced every 48 h until day 10. Lipid accumulation was assessed on day 10 by Oil Red O staining, and the expression of adipocyte differentiation-related genes was evaluated on days 2, 4, 7, and 10.

4.10. Oil Red O Staining

After 10 d of differentiation, the cells were washed with PBS and fixed with 4% formalin. The lipid droplets were stained with Oil Red O solution (0.5 g/L). The dye was then eluted with 100% isopropanol, and the absorbance was measured at 490 nm.

4.11. Western Blot Analysis

3T3-L1 cells were treated with 10 µg/mL rTs-MIF for 5, 15, or 30 min, and total protein was extracted. Proteins were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto PVDF membranes. Membranes were probed with primary antibodies against phospho-AKT, total AKT, phospho-p44/42 MAPK, total p44/42 MAPK, and actin (Cell Signaling Technology, Danvers, MA, USA), followed by incubation with HRP-conjugated secondary antibodies. β-actin was used as a loading control for protein normalization. Signals were detected using WESTSAVE™ (AbFrontier, Seoul, Republic of Korea), and band intensities were quantified using ImageJ software.

4.12. Statistical Analysis

Data are presented as mean ± standard deviation (SD). For comparisons among three or more groups, one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparisons post hoc test was used to compare all treatment groups with the HFD control group. For repeated measurements over time, two-way repeated-measures ANOVA was applied. All statistical analyses were performed using GraphPad Prism 5 (GraphPad Software, San Diego, CA, USA). Statistical significance was set at p < 0.05.

5. Conclusions

In summary, rTs-MIF exerted anti-obesity effects by reducing body fat mass, suppressing hepatic steatosis, attenuating systemic and adipose tissue inflammation, inhibiting adipocyte differentiation, downregulating AKT and p44/42 MAPK signaling, and promoting an M2-dominant macrophage milieu in adipose tissue.

Given that current anti-obesity drugs are limited by adverse effects, high cost, and the need for long-term administration, rTs-MIF represents a promising new class of anti-obesity candidates that target the immune–metabolic axis. This study suggests that helminth-derived proteins may be broadly applicable as therapeutics for metabolic and immune-mediated diseases beyond obesity.

6. Patents

A patent application related to Ts-MIF and its potential therapeutic use has been filed. Seo Yeong Choi, Mi-Kyung Park, Yu Jin Jeong, Shin Ae Kang, and Hak Sun Yu are listed as inventors.