Dermatomyositis (DM) is an immune-mediated myopathy marked by chronic inflammation and heterogeneous clinical trajectories. The molecular determinants driving disease onset and progression remain poorly defined. This study aimed to construct a high-accuracy diagnostic risk model, identify causally relevant genes, and uncover cell-type-specific immune circuits contributing to DM pathogenesis, with emphasis on BTN3A2. We integrated three bulk transcriptomic datasets to identify dysregulated genes and performed functional enrichment analyses. An 18-gene diagnostic risk score was generated using LASSO and validated externally. Summary-based Mendelian randomization with tissue-specific eQTLs was applied to identify causal genes. Single-cell RNA sequencing (GSE190684) was analyzed to map cellular heterogeneity, quantify risk-score and BTN3A2 distributions, and evaluate ligand–receptor communication across clinical states and expression subgroups. Bulk analyses identified 2,051 dysregulated genes enriched in antiviral responses, antigen presentation, and immune activation. Summary-data based Mendelian Randomization (SMR) uncovered 85 causal genes shared across skin and muscle, converging on MHC-related and autoimmune pathways. Integrating causal and transcriptional evidence yielded 16 core DM genes, highlighting BTN3A2 for its potential dual diagnostic and suggestive causal relevance. The 18-gene risk score demonstrated excellent predictive performance (AUC 0.957/0.724). Single-cell profiling revealed that both the risk score and BTN3A2 were predominantly enriched in CD14⁺/CD16⁺ monocytes and T-cell subsets. Cell–cell communication analysis identified a monocyte-derived LGALS9-CD44/CD45 signaling axis as the key interaction pathway distinguishing clinical states and risk groups. This multi-omics study establishes a precise diagnostic risk score and suggests that BTN3A2 may serve as a mechanistically relevant biomarker in DM. The discovery of an LGALS9-CD44/CD45 monocyte-T cell circuit provides new insight into DM immunopathology and highlights actionable targets for precision diagnosis and therapy.

Ischemic stroke is a leading cause of disability and mortality, with neuroinflammation playing a key role in post-stroke injury. The molecular mechanisms remain incompletely defined. This study explored the functions of IRF7 and its downstream target ISG15 in stroke-associated neuroinflammation and prognosis of ischemic stroke. Bioinformatic analysis of transcriptomic datasets from microglia and ischemic brain tissues identified both molecules as hub genes. Their expression was validated in a mouse transient middle cerebral artery occlusion (tMCAO) model and in microglial cultures exposed to oxygen-glucose deprivation/reoxygenation (OGD/R). The effects of these molecules were assessed using siRNA knockdown, conditioned media assays with SY5Y neuronal cells, in vivo overexpression of ISG15, histological and functional assessments. Both IRF7 and ISG15 were significantly upregulated after stroke. IRF7 knockdown reduced ISG15 expression, whereas ISG15 knockdown did not affect IRF7, suggesting a unidirectional regulatory relationship. Conditioned media from microglia treated with siIRF7 or siISG15 increased SY5Y cell mortality, with a stronger effect in the siISG15 group, highlighting neuroprotective role of ISG15. ISG15 knockdown also enhanced microglial migration. Conversely, microglial ISG15 overexpression in vivo promoted a shift toward reduced neuroinflammation, improved neuronal survival, and enhanced functional recovery. Mechanistically, ISG15 stabilized NLRP3 protein but more strongly decreased its mRNA stability through accelerated degradation. These findings demonstrate that the activation of IRF7/ISG15 axis in microglia inhibits NLRP3 Expression and improve the prognosis of ischemia/reperfusion in mice, with ISG15 exerting stronger neuroprotective effects. Targeting microglial ISG15 may offer a promising therapeutic strategy for ischemic stroke. MAIN POINTS ### Competing Interest Statement The authors have declared no competing interest. * List of abbreviations : OGD/R : oxygen-glucose deprivation/reoxygenation BBB : blood-brain barrier NF-κB : nuclear factor-κB TLRs : Toll-like receptors IRFs : interferon regulatory factors LPS : lipopolysaccharide CpG-ODN : CpG oligodeoxynucleotides tMCAO : transient middle cerebral artery occlusion CCA : right common carotid artery ECA : external carotid artery ICA : internal carotid artery MCA : middle cerebral artery TTC : triphenyltetrazolium chloride PFA : paraformaldehyde PMSF : phenylmethylsulfonyl fluoride TBST : Tris-buffered saline with Tween 20 NCM : normal culture medium PS : Penicillin-Streptomycin PPI : Protein-protein interaction ISG15⁻/⁻ : ISG15 knockout HGF : hepatocyte growth factor DMEM : Dulbecco’s Modified Eagle’s Medium FBS : fetal bovine serum DEGs : Differentially expressed genes GO : Gene Ontology KEGG : Kyoto Encyclopedia of Genes and Genomes GSEA : Gene set enrichment analysis SD : standard deviation pIRF7 : phosphorylated IRF7 Il-1βmat : active form of Il-1β GSDMDmat : active form of GSDMD GSDMDpre : precursor of GSDMD NC : negative control CHX : cycloheximide TBI : traumatic brain injury siISG15 : siRNA targeting ISG15 siIRF7 : siRNA targeting IRF7 Sichuan Provincial Science and Technology Program Project, 2023NSFSC0643, 2023YFS0050, 2024YFHZ0009

UFMylation, a novel ubiquitin-like protein modification system, has been recently found to be activated in inflammation. However, the effects of UFMylation activation on inflammation in vivo remains unclear. In the present study, we generated a UFMylation activated mice using transgenic (TG) techniques. Lipopolysaccharide (LPS) was used to induce systemic inflammation in both TG and non-transgenic (NTG) mice. Serum cytokines were detected using a Mouse Cytokine Array, and the proportions of splenic NK, B and T cells were determined by using flow cytometry. We found that TG mice showed increased serum G-CSF, TNF RII and decreased serum TCA-3, CD30L, bFGF, IL-15 and MIG compared with NTG mice at baseline. Furthermore, serum cytokines in TG mice exhibited different responses to LPS compared to NTG mice. LPS up-regulated serum TNF RII, G-CSF, MCP-5, RANTES, KC, BLC, MIG and down-regulated IL-1b, IL-2, IL-3, IL-4, IL-5, IL-7, IL-10, IL-12p40, IL-15, IL-17, IFN-γ, TCA-3, Eotaxin-2, LIX, MCP-1, TNFα, GM-CSF in NTG mice, whereas LPS up-regulated G-CSF, MCP-5, RANTES, KC, BLC, MIG, ICAM-1, PF4, Eotaxin, CD30L, MIP-1a, TNFRI and down-regulated IL-1b, IL-3, LIX, MCP-1, TNFα, GM-CSF in TG mice. Data from flow cytometry indicated that LPS significantly reduced the percentages of NK and NKT cells in NTG mice, whereas UFMylation activation inhibited LPS-induced NKT cell decrease. The proportions of B cells, total CD4+ and total CD8+ T cells were comparable between TG and NTG mice in response to LPS treatment, whereas the percentages of CD4+CD69+ and CD8+CD69+T cells were lower in TG mice. These findings suggest that UFMylation may alter LPS-induced serum cytokine profile and participate in splenic T cell activation in vivo.

Objective:To explore and evaluate a appropriate suitable method for detection of Campylobacter and antibiotic sensitivity test for foodborne diarrhea in clinical laboratories. Methods:Pre-experiment:a total number of 400 fecal samples of patients with foodborne diarrhea were prospectively collected from the intestinal disease clinic of Beijing Tongren Hospital from September 2017 to January 2018. Double-hole filtration culture method and modified cefoperazone charcoal deoxycholate (CCD) agar culture method were used for fecal culture in micro-aerobic environment for 48 hours, and then suspicious colonies were identified by matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. Meanwhile, C. jejuni and C. coli were detected by real-time quantitative polymerase chain reaction(qPCR). Large sample verification: 2 062 fecal samples of patients with foodborne diarrhea in three hospitals of different levels in different areas of Beijing were collected for qPCR detection and culture from April 2018 to March 2019. The antimicrobial sensitivity test (AST) of C. jejuni and C. coli was performed according to the disk diffusion method and agar dilution method recommended by Clinical and Laboratory Standards Institute and National Antimicrobial Resistance Monitoring System for Enteric Bacteria. The results of the three detection methods and the consistency of the two antibiotic sensitivity tests were compared. Results:In the pre-experiment, the positive rates of Campylobacter ( jejuni/coli) detected of qPCR, double-hole filtration culture and modified CCD agar culture were 9.0% (36/400), 5.0% (20/400)and 3.5% (14/400), and the difference was statistically significant ( P<0.01). The samples with negative result of qPCR were negative by both culture methods. The total positive rates of Campylobacter detected by qPCR was 8.1% (168/ 2 062)including 7.0% (144/2 062) for C. jejuni and 1.2% (24/2 062) for C. coli. The samples with positive qPCR results were cultured by double-hole filtration culture method and the positive rate was 61.9%(104/168), among which, the positive rate of C. jejuni and C. coli were 58.3%(84/144) and 83.3%(20/24) respectively, which was not significantly different from the detection rate and culture positive rate in the pre-test ( P>0.1). The resistance rates of C. jejuni and C. coli to ciprofloxacin were 94.0%(94/100) and 100.0%(24/24) and to erythromycin were 6.0%(6/100) and 33.3%(8/24). The results from two antibiotic sensitivity test methods were consistent (Kappa>0.75). Conclusions:qPCR is rapid, sensitive and easy to operate, so it is suitable for routine development in clinical laboratories. The double-hole filtration culture method is beneficial to the acquisition of strains and is essential for the further study of Campylobacter. There was no significant difference between agar dilution method and disk diffusion method in antibiotic sensitivity test. Campylobacter showed a very high resistance rate to quinolones, which was no longer suitable for the treatment of Campylobacter foodborne diarrhea in Beijing area. Macrocyclic lipid antibiotics should be preferred.

目的 对针刺蝶腭神经节治疗变应性鼻炎患者多中心无对照研究资料中相关症状改善的程度、时间、持续性进行观察,分析该针刺法对变应性鼻炎的疗效.方法 2017年4月—2018年6月在北京地区7个分中心对变应性鼻炎患者进行针刺蝶腭神经节治疗(每周2次,共4周),共纳入符合标准患者222例;在治疗前(第0周),治疗第2、3、4周(第2、3、4周)及随访(第8周)共5个时点记录患者的鼻炎症状量表(TNSS)中鼻塞、流涕、鼻痒、喷嚏4个主要症状不同程度例数和鼻炎伴随症状量表(TNNSS)中鼻涕从咽部流过、流泪、鼻或眼部瘙痒、鼻或口腔上颌疼痛、头痛5个伴随症状例数.比较主要症状及伴随症状分布的变化情况,以随访结束时较治疗前症状是否缓解、消失或不变、加重的例数为依据,比较针刺蝶腭神经节对诸症状疗效的差别.结果 治疗前后,鼻塞、流涕、鼻痒、喷嚏症状分数总体分布差异具有统计学意义(P<0.05),且每2个相邻时点比较,后一时点症状更轻(P<0.01);伴随症状中鼻涕从咽部流过、流泪、鼻或眼部瘙痒、鼻或口腔上颌疼痛、头痛症状分数总体分布差异具有统计学意义(P<0.05),其中鼻涕从咽部流过、流泪、鼻或眼部瘙痒、鼻或口腔上颌疼痛后一时点较相邻的前一时点均症状减轻(P<0.01).结论 针刺蝶腭神经节可减轻变应性鼻炎主要症状及伴随症状,疗效确切;而以喷嚏、鼻塞、流涕、鼻痒为主要临床表现的变应性鼻炎患者采用针刺蝶腭神经节治疗可能获得较满意疗效.

Most protein-coding genes in eukaryotes possess at least two poly(A) sites, and alternative polyadenylation is considered a contributing factor to transcriptomic and proteomic diversity. Following transcription, a nascent RNA usually undergoes capping, splicing, cleavage, and polyadenylation, resulting in a mature messenger RNA (mRNA); however, increasing evidence suggests that transcription and RNA processing are coupled. Plants, which must produce rapid responses to environmental changes because of their limited mobility, exhibit such coupling. In this review, we summarize recent advances in our understanding of the coupling of transcription with RNA processing in plants, and we describe the possible spatial environment and important proteins involved. Moreover, we describe how liquid–liquid phase separation, mediated by the C-terminal domain of RNA polymerase II and RNA processing factors with intrinsically disordered regions, enables efficient co-transcriptional mRNA processing in plants.

Cellular protrusion formation capacity is a key feature of developing neurons and many eukaryotic cells. However, the mechanisms underlying membrane growth in protrusion formation are largely unclear. In this study, photo-reactive unnatural amino acid 3-(3-methyl-3H-diazirin-3-yl)-propamino-carbonyl-Nε-l-lysine was incorporated by a genetic code expansion strategy into protrudin, a protein localized in acidic endosomes and in the endoplasmic reticulum, that induces cellular protrusion and neurite formation. The modified protrudin was used for covalent trapping of protrudin-interacting proteins in living cells. Fatty acid synthase (FASN), which synthesizes free fatty acids, was identified to transiently interact with protrudin. Further characterization revealed a unique cooperation mechanism in which protrudin cooperates with FASN to facilitate cellular protrusion formation. This work reveals a novel mechanism involved in protrusion formation that is dependent on transient interaction between FASN and protrudin, and establishes a creative strategy to investigate transient protein-protein interactions in mammalian cells.

留学生教学已成为我国高等教育的重要组成部分.人体寄生虫学是一门医学生的必修课,根据留学生的特点,从加强沟通、教材选择及建设、完善教学内容、运用多种教学方法和将科学研究渗入教学等方面进行探讨,以提高学生的综合素质及教学质量.

基础医学专业是首都医科大学的新设专业,目的在于培养具备自然科学、生命科学和医学科学基本理论知识和实验技能的高级专门人才.人体寄生虫学是一门联系基础医学与临床医学的桥梁学科.本文在总结本教研室近年来从事基础医学专业人体寄生虫学教学体会的同时,发现教学中的有待改进之处,并提出了相应的可行性建议,旨在进一步提高教学质量.

Trichinellosis, a widespread zoonosis, is considered to be an emerging or re-emerging infectious parasitic disease. The development of vaccines to prevent Trichinella infection in domestic animals and humans is important for disease control. In a previous study, we identified Ts14-3-3 as an immunodominant protein from Trichinella spiralis (T. spiralis) adult worms recognized by early infection sera from pigs and mice. In this study, we further confirmed that Ts14-3-3 mRNA is expressed in both adult worms and in the larval stages of T. spiralis. Immunostaining with anti-Ts14-3-3 mouse sera further confirmed that native Ts14-3-3 is highly expressed on the surface of T. spiralis muscle larvae. The immune recognition by infected sera, its expression in both adult and larval stages and its exposure on the surface of the parasite led us to explore Ts14-3-3 as a vaccine antigen. Recombinant Ts14-3-3 formulated with an ISA50v2 adjuvant produced strong total IgG and balanced IgG1 and IgG2a responses in vaccinated mice and stimulated mouse splenocytes to produce high levels of Th1 (INF-γ, IL-2) and Th2 (IL4, IL5) cytokines. These results indicate that Ts14-3-3 is highly immunogenic and is able to induce balanced Th1/Th2 immune responses. These vaccine-induced immune responses resulted in a reduction in muscle larvae of up to 46.2% in vaccinated mice upon subsequent larval challenge relative to the number of larvae in mice received PBS control. The significant reduction in muscle larvae in vaccinated mice suggests that Ts14-3-3 is a promising vaccine target for potential use in domestic pigs to prevent trichinellosis transmission.

慕课是大规模开放在线课程的新型教育模式，它把优秀的教学资源较好地融合起来，对知识点进行最优呈现。人体寄生虫学的教学需要借鉴慕课的优点，转变教学理念，优化教学内容和教学设计，并提供多元化的教学评价方式，提高教学效果。

BACKGROUND:Our previous studies showed that Trichinella spiralis paramyosin (TsPmy) is an immunomodulatory protein that inhibits complement C1q and C8/C9 to evade host complement attack. Vaccination with recombinant TsPmy protein induced protective immunity against T. spiralis larval challenge. Due to the difficulty in producing TsPmy as a soluble recombinant protein, we prepared a DNA vaccine as an alternative approach in order to elicit a robust immunity against Trichinella infection.METHODS AND FINDINGS:The full-length TsPmy coding DNA was cloned into the eukaryotic expression plasmid pVAX1, and the recombinant pVAX1/TsPmy was transformed into attenuated Salmonella typhimurium strain SL7207. Oral vaccination of mice with this attenuated Salmonella-delivered TsPmy DNA vaccine elicited a significant mucosal sIgA response in the intestine and a systemic IgG antibody response with IgG2a as the predominant subclass. Cytokine analysis also showed a significant increase in the Th1 (IFN-γ, IL-2) and Th2 (IL-4, 5, 6, 10) responses in lymphocytes from the spleen and MLNs of immunized mice upon stimulation with TsPmy protein. The expression of the homing receptors CCR9/CCR10 on antibody secreting B cells may be related to the translocation of IgA-secreted B cells to local intestinal mucosa. The mice immunized with Salmonella-delivered TsPmy DNA vaccine produced a significant 44.8% reduction in adult worm and a 46.6% reduction in muscle larvae after challenge with T. spiralis larvae.CONCLUSION:Our results demonstrated that oral vaccination with TsPmy DNA delivered by live attenuated S. typhimurium elicited a significant local IgA response and a mixed Th1/Th2 immune response that elicited a significant protection against T. spiralis infection in mice.

为优化抗旋毛虫病核酸疫苗免疫策略,本文探讨了Ts87 DNA疫苗滴鼻初免与蛋白疫苗皮下注射加强免疫诱导产生的保护效果及免疫应答特征.分别在第0d以旋毛虫Ts87 DNA疫苗SL7207/pVAX1-Ts87滴鼻初免,第14/28 d以蛋白疫苗rTs87皮下注射加强免疫小鼠,在第35 d攻击感染旋毛虫肌幼虫,第84 d剖杀小鼠,计算减虫率观察免疫保护效果;进行小鼠血清中特异性IgG抗体效价、抗体亚型分析及肠道盥洗液sIgA水平分析,检测脾淋巴细胞增殖反应、脾及颈部淋巴结细胞分泌细胞因子水平.结果显示,DNA疫苗滴鼻初免—重组蛋白加强的联合免疫组的保护作用优于单独DNA疫苗滴鼻及单独重组蛋白免疫组,显著提高了减虫率(46.1％);DNA疫苗滴鼻初免和蛋白疫苗加强免疫方式不仅诱导小鼠产生了粘膜免疫应答,同时也诱导了有效的细胞和体液免疫应答,其免疫应答类型是以Th2型为主的混合型Th1/Th2免疫反应.该研究为新型抗旋毛虫病疫苗接种策略的优化提供了实验依据.

Trichinellosis is a worldwide zoonosis and vaccinating swine with a potent vaccine is a practical approach to prevent Trichinella infections in China. Paramyosin of T. spiralis (Ts-Pmy) was shown in our previous work to be a good vaccine candidate against Trichinella infections. Because CD4+ T cells play a crucial role in effective immunity against T. spiralis infection, identifying CD4+ T cell epitopes of paramyosin is crucial for constructing a chimeric subunit epitope vaccine. Twelve CD4+ T cell epitopes of Ts-Pmy with the highest scores were predicted and synthesized as peptides. Five of the twelve peptides, P2, P3, P4, P5 and P12, induced strong splenocyte proliferation and secretion of the Th2 cytokines IL-4 and IL-5 from rTs-Pmy-immunized mouse splenocytes. To assess the immunogenicity of CD4+ T cell epitopes in vivo, splenocytes from mice immunized with individual peptides were stimulated with the corresponding peptides. P2, P3, P4 and P5 induced strong cell proliferation and secretion of both Th1 (INF-γ, IL-2) and Th2 (IL-4, IL-5) cytokines. The results indicate that the peptides P2, P3, P4 and P5 are immunodominant CD4+ T cell epitopes of Ts-Pmy. This study will facilitate the design of an effective epitope-based multivalent subunit vaccine against Trichinella infections.

Background: Trichinellosis, a widespread zoonosis, is regarded as an emerging or reemerging disease. Effective treatment and prognosis of trichinellosis depends on early diagnosis of the infection. The objective of this study was to identify sensitive and specific antigens for early diagnosis or effective vaccine antigens for preventing infection.Methods: The somatic proteins of T. spiralis adult worms were separated by two-dimensional gel electrophoresis (2-DE). The separated proteins were probed with early infection sera from swine or mice infected with T. spiralis for 7 days. The primary immunoreactive spots were characterized by MALDI-TOF/TOF-MS analysis in combination with bioinformatics. The identified proteins were annotated using WEGO based on their functions. The immunodominant protein was chosen for expression as recombinant protein in E. coli and the purified recombinant protein was used to confirm its antigenicity by Western blot with the same infection sera.Results: Approximately 300 spots were separated by 2-DE, with molecular weights ranging from 10 to 130 kDa, and pI values ranging from pH 4 to 10. The sera from swine and mice infected with T. spiralis for 7 days recognized 64 proteins. MALDI-TOF/TOF-MS analysis identified 55 proteins, some with different isoforms. Finally, 40 individual immunoreactive proteins were obtained with a wide range of biological functions. Several proteins, such as heat shock protein 70, 14-3-3 protein, and cysteine protease could be used as immunodiagnostic or vaccine antigens. Among these identified proteins, the highly immunodominant Ts14-3-3 was chosen for expression in E. coli and purified recombinant Ts14-3-3 was able to be strongly recognized by the same T. spiralis infected sera used for identifying these antigens, therefore the most promising antigen for early immunodiagnosis of Trichinella infection.Conclusions: A total of 64 proteins from the adult worm were recognized by early infection sera from swine and mice infected with T. spiralis for 7 days. Several proteins, are of particular interest as immunodiagnostic or vaccine antigens, especially with Ts14-3-3 as most promising due to its highly immunogenicity during early infection, expressed protein can be recognized by Trichinella early infection sera and the native Ts14-3-3 expression in both adult and larval stages.

To construct and express the fusion protein that combines Trichinella spiralis heat shock protein 70 with excretory-secretory antigen Ts87, restriction enzyme EcoRⅠ site was applied to fuse Ts-Hsp70 with Ts87.The target genes were inserted into plasmid pET28-a ( +), and transformed into E.coli.BL21 (DE3).After expression and purification, the fusion protein rTs-Hsp70-Ts87 was characterized by SDS-PAGE and Western blot.The results showed that the fusion gene was 2 721 bp.The results of PCR, enzyme digestion and sequencing analysis showed that the recombinant plasmid was affirmed to be successfully constructed.The fusion protein rTs-Hsp70-Ts87 was expressed with IPTG induction and purified by Ni-affinity chromatography and refolded.The results of SDS-PAGE and Western blot indicated that molecular weight of the protein is about 100 kDa, and it could be recognized by anti His-tag monoclonal antibody, anti-rTs-Hsp70 immune serum and anti-rTs87 immune serum.In conclusion, T.spiralis recombinant fusion protein rTs-Hsp70-Ts87 was constructed and expressed successfully, which had laid the groundwork for further research.

Budding yeast divides asymmetrically, giving rise to a mother cell that progressively ages and a daughter cell with full lifespan. It is generally assumed that mother cells retain damaged, lifespan limiting materials ("aging factors") through asymmetric division. However, the identity of these aging factors and the mechanisms through which they limit lifespan remain poorly understood. Using a flow cytometry-based, high-throughput approach, we quantified the asymmetric partitioning of the yeast proteome between mother and daughter cells during cell division, discovering 74 mother-enriched and 60 daughter-enriched proteins. While daughter-enriched proteins are biased toward those needed for bud construction and genome maintenance, mother-enriched proteins are biased towards those localized in the plasma membrane and vacuole. Deletion of 23 of the 74 mother-enriched proteins leads to lifespan extension, a fraction that is about six times that of the genes picked randomly from the genome. Among these lifespan-extending genes, three are involved in endosomal sorting/endosome to vacuole transport, and three are nitrogen source transporters. Tracking the dynamic expression of specific mother-enriched proteins revealed that their concentration steadily increases in the mother cells as they age, but is kept relatively low in the daughter cells via asymmetric distribution. Our results suggest that some mother-enriched proteins may increase to a concentration that becomes deleterious and lifespan-limiting in aged cells, possibly by upsetting homeostasis or leading to aberrant signaling. Our study provides a comprehensive resource for analyzing asymmetric cell division and aging in yeast, which should also be valuable for understanding similar phenomena in other organisms.

Objective In order to obtain vaccine or diagnostic candidate antigens for trichinellosis, the adult cDNA library of T. spiralis was immunoscreened. Methods The adult cDNA library of T. spiralis was screened using the swine sera infected with T. spiralis. The cDNA sequence of the positive clone was analyzed and the recombinant protein was expressed and purified. The antigenicity and immunogenicity of the recombinant protein was analyzed with ELISA and Western blotting. Results Forty-two positive clones were identified by immunscreened. A novel gene Tmp10 of T. spiralis was sequenced and recombinant plasmid pET-28a (+)/Tmp10 was successfully constructed. The recombinant protein(rTmp10) was expressed and the molecular weight was about 40 000. The results of Western blotting showed that the rTmp10 could be recognized by sera from the mice and swine infected with T. spiralis respectively, which showed that the rTmp10 had the specific antigenicity. The rTmp10 could induce a high level of specific anti-Tmp10 IgG antibodies, which showed that the rTmp10 had the immunogenicity. Conclusion The novel gene Tmp10 of T. spiralis was obtained by immunoscreening and predicted to be a membrane protein. The recombinant protein Tmp10 had the high specific antigenicity and immunogenicity.

BACKGROUND:Trichinella spiralis expresses paramyosin (Ts-Pmy) as a defense mechanism. Ts-Pmy is a functional protein with binding activity to human complement C8 and C9 and thus plays a role in evading the attack of the host's immune system. In the present study, the binding activity of Ts-Pmy to human complement C1q and its ability to inhibit classical complement activation were investigated.METHODS AND FINDINGS:The binding of recombinant and natural Ts-Pmy to human C1q were determined by ELISA, Far Western blotting and immunoprecipitation, respectively. Binding of recombinant Ts-Pmy (rTs-Pmy) to C1q inhibited C1q binding to IgM and consequently inhibited C3 deposition. The lysis of antibody-sensitized erythrocytes (EAs) elicited by the classical complement pathway was also inhibited in the presence of rTs-Pmy. In addition to inhibiting classical complement activation, rTs-Pmy also suppressed C1q binding to THP-1-derived macrophages, thereby reducing C1q-induced macrophages migration.CONCLUSION:Our results suggest that T. spiralis paramyosin plays an important role in immune evasion by interfering with complement activation through binding to C1q in addition to C8 and C9.