Pathogenicity and transcriptomic profiling reveals immunology molecular hallmarks after CA10 virus infection

Abstract

Background: Hand, foot and mouth disease (HFMD) is a common infectious disease caused by viral infection by a variety of enteroviruses, with coxsackievirus A 10 (CA10) having become more prevalent in recent years.

Methods: In this study, models of CA10 infection were established in 7-day-old Institute of Cancer Research (ICR) mice by intraperitoneal injection to analyze the pathogenicity of the virus. RNA sequencing analysis was used to screen the differentially expressed genes (DEGs) after CA10 infection. Coxsackievirus A 16 (CA16) and enterovirus 71 (EV71) infections were also compared with CA10.

Results: After CA10 virus infection, the mice showed paralysis of the hind limbs at 3 days post infection and weight loss at 5 days post infection. We observed viral replication in various tissues and severe inflammatory cell infiltration in skeletal muscle. The RNA-sequencing analysis showed that the DEGs in blood, muscle, thymus and spleen showed heterogeneity after CA10 infection and the most up-regulated DEGs in muscle were enriched in immune-related pathways. Compared with CA16 and EV71 infection, CA10 may have an inhibitory effect on T helper (Th) cell differentiation and cell growth. Additionally, the common DEGs in the three viruses were most enriched in the immune system response, including the Toll-like receptor pathway and the nucleotide-binding and oligomerization domain (NOD)-like pathway.

Conclusions: Our findings revealed a group of genes that coordinate in response to CA10 infection, which increases our understanding of the pathological mechanism of HFMD.

Keywords

CA10 / HFMD / RNA-Seq

References

[1] Yi L, Lu J, Kung HF, He ML. The virology and developments toward control of human enterovirus 71. Crit Rev Microbiol. 2011;37(4):313-327.

[2] Hong J, Liu F, Qi H, et al. Changing epidemiology of hand, foot, and mouth disease in China, 2013–2019: a population-based study. Lancet Reg Health West Pac. 2022;20:100370.

[3] Nguyen TT, Chiu CH, Lin CY, et al. Efficacy, safety, and immunogenicity of an inactivated, adjuvanted enterovirus 71 vaccine in infants and children: a multiregion, double-blind, randomised, placebo-controlled, phase 3 trial. Lancet. 2022;399(10336):1708-1717.

[4] Zhu FC, Meng FY, Li JX, et al. Efficacy, safety, and immunology of an inactivated alum-adjuvant enterovirus 71 vaccine in children in China: a multicentre, randomised, double-blind, placebo-controlled, phase 3 trial. Lancet. 2013;381(9882):2024-2032.

[5] Zhu FC, Liang ZL, Li XL, et al. Immunogenicity and safety of an enterovirus 71 vaccine in healthy Chinese children and infants: a randomised, double-blind, placebo-controlled phase 2 clinical trial. Lancet. 2013;381(9871):1037-1045.

[6] Head JR, Collender PA, Lewnard JA, et al. Early evidence of inactivated enterovirus 71 vaccine impact against hand, foot, and mouth disease in a major center of ongoing transmission in China, 2011–2018: a longitudinal surveillance study. Clin Infect Dis. 2020;71(12):3088-3095.

[7] Wang J, Qi S, Zhang X, et al. Coxsackievirus A 16 infection does not interfere with the specific immune response induced by an enterovirus 71 inactivated vaccine in rhesus monkeys. Vaccine. 2014;32(35):4436-4442.

[8] Yang L, Liu Y, Li S, et al. A novel inactivated enterovirus 71 vaccine can elicit cross-protective immunity against coxsackievirus A16 in mice. Vaccine. 2016;34(48):5938-5945.

[9] Du Z, Huang Y, Bloom MS, et al. Assessing the vaccine effectiveness for hand, foot, and mouth disease in Guangzhou, China: a time-series analysis. Hum Vaccin Immunother. 2021;17(1):217-223.

[10] Aswathyraj S, Arunkumar G, Alidjinou EK, Hober D. Hand, foot and mouth disease (HFMD):emerging epidemiology and the need for a vaccine strategy. Med Microbiol Immunol. 2016;205(5):397-407.

[11] Bian L, Gao F, Mao Q, et al. Hand, foot, and mouth disease associated with coxsackievirus A10: more serious than it seems. Expert Rev Anti Infect Ther. 2019;17(4):233-242.

[12] Cai Y, Ku Z, Liu Q, Leng Q, Huang Z. A combination vaccine comprising of inactivated enterovirus 71 and coxsackievirus A16 elicits balanced protective immunity against both viruses. Vaccine. 2014;32(21):2406-2412.

[13] Fan S, Liao Y, Jiang G, et al. Efficacy of an inactivated bivalent vaccine for enterovirus 71 and coxsackievirus A16 in mice immunized intradermally. Vaccine. 2021;39(3):596-604.

[14] Halfmann PJ, Iida S, Iwatsuki-Horimoto K, et al. SARS-CoV-2 omicron virus causes attenuated disease in mice and hamsters. Nature. 2022;603(7902):687-692.

[15] Ding Z, Chen T, Lan J, Wong G. Application of animal models to compare and contrast the virulence of current and future potential SARS-CoV-2 variants. Biosaf Health. 2022;4:154-160.

[16] Wang YF, Yu CK. Animal models of enterovirus 71 infection: applications and limitations. J Biomed Sci. 2014;21:31.

[17] Solomon T, Lewthwaite P, Perera D, Cardosa MJ, Mcminn P, Ooi MH. Virology, epidemiology, pathogenesis, and control of enterovirus 71. Lancet Infect Dis. 2010;10(11):778-790.

[18] Li S, Zhao H, Yang L, et al. A neonatal mouse model of coxsackievirus A10 infection for anti-viral evaluation. Antiviral Res. 2017;144:247-255.

[19] Chen C, Xia Y, Zhu S, et al. Muscle destruction caused by coxsackievirus A10 in gerbils: construction of a novel animal model for antiviral evaluation. Virus Res. 2020;286:198067.

[20] Westermann AJ, Vogel J. Cross-species RNA-seq for deciphering host-microbe interactions. Nat Rev Genet. 2021;22(6):361-378.

[21] Toro-Moreno M, Sylvester K, Srivastava T, Posfai D, Derbyshire ER. RNA-seq analysis illuminates the early stages of plasmodium liver infection. MBio. 2020;11(1):e03234-19.

[22] Ding Y, Dai X, Bao M, et al. Hepatic transcriptome signatures in mice and humans with nonalcoholic fatty liver disease. Animal Model Exp Med. 2023;6(4):317-328.

[23] Liu J, Yang Y, Xu Y, Ma C, Qin C, Zhang L. Lycorine reduces mortality of human enterovirus 71-infected mice by inhibiting virus replication. Virol J. 2011;8:483.

[24] Luo Z, Su R, Wang W, et al. EV71 infection induces neurodegeneration via activating TLR7 signaling and IL-6 production. PLoS Pathog. 2019;15(11):e1008142.

[25] Cui G, Wang H, Yang C, et al. Berberine prevents lethal EV71 neurological infection in newborn mice. Front Pharmacol. 2022;13:1027566.

[26] Pei Z, Wang H, Zhao Z, Chen X, Huan C, Zhang W. Chemokine PF4 inhibits EV71 and CA16 infections at the entry stage. J Virol. 2022;96(11):e0043522.

[27] Zhang Y, Han J. Differential privacy fuzzy C-means clustering algorithm based on gaussian kernel function. PLoS ONE. 2021;16(3):e0248737.

[28] He Y, Yang J, Zeng G, et al. Risk factors for critical disease and death from hand, foot and mouth disease. Pediatr Infect Dis J. 2014;33(9):966-970.

[29] Gonzalez G, Carr MJ, Kobayashi M, Hanaoka N, Fujimoto T. Enterovirus-associated hand-foot and mouth disease and neurological complications in Japan and the rest of the world. Int J Mol Sci. 2019;20(20):5201.

[30] Wang Y, Zhao H, Ou R, et al. Epidemiological and clinical characteristics of severe hand-foot-and-mouth disease (HFMD) among children: a 6-year population-based study. BMC Public Health. 2020;20(1):801.

[31] Nicot C. RNA-seq reveals novel CircRNAs involved in breast cancer progression and patient therapy response. Mol Cancer. 2020;19(1):76.

[32] Wang C, Li D, Zhang L, et al. RNA sequencing analyses of gene expression during Epstein-Barr virus infection of primary B lymphocytes. J Virol. 2019;93(13):e00226-19.

[33] Konig A, Yang J, Jo E, et al. Efficient long-term amplification of hepatitis B virus isolates after infection of slow proliferating HepG2-NTCP cells. J Hepatol. 2019;71(2):289-300.

[34] Luo W, Hu J, Xu W, Dong J. Distinct spatial and temporal roles for Th1, Th2, and Th17 cells in asthma. Front Immunol. 2022;13:974066.

[35] Muhammad YF, Wong KK, Mohd RN. Th1, Th2, and Th17 cytokines in systemic lupus erythematosus. Autoimmunity. 2020;53(1):8-20.

[36] Arnold PK, Jackson BT, Paras KI, et al. A non-canonical tricarboxylic acid cycle underlies cellular identity. Nature. 2022;603(7901):477-481.

[37] Wang W, Xiao F, Wan P, et al. EV71 3D protein binds with NLRP3 and enhances the assembly of inflammasome complex. PLoS Pathog. 2017;13(1):e1006123.

[38] Platnich JM, Muruve DA. NOD-like receptors and inflammasomes: a review of their canonical and non-canonical signaling pathways. Arch Biochem Biophys. 2019;670:4-14.

[39] Trindade BC, Chen GY. NOD1 and NOD2 in inflammatory and infectious diseases. Immunol Rev. 2020;297(1):139-161.

[40] Xin P, Xu X, Deng C, et al. The role of JAK/STAT signaling pathway and its inhibitors in diseases. Int Immunopharmacol. 2020;80:106210.

[41] Luo W, Li YX, Jiang LJ, Chen Q, Wang T, Ye DW. Targeting JAK-STAT signaling to control cytokine release syndrome in COVID-19. Trends Pharmacol Sci. 2020;41(8):531-543.

[42] Khanmohammadi S, Rezaei N. Role of toll-like receptors in the pathogenesis of COVID-19. J Med Virol. 2021;93(5):2735-2739.

[43] Diamond MS, Kanneganti TD. Innate immunity: the first line of defense against SARS-CoV-2. Nat Immunol. 2022;23(2):165-176.

[44] Zheng C. The emerging roles of NOD-like receptors in antiviral innate immune signaling pathways. Int J Biol Macromol. 2021;169:407-413.

[45] Zhu X, Xie C, Li Y, et al. TMEM2 inhibits hepatitis B virus infection in HepG2 and HepG2.2.15 cells by activating the JAK–STAT signaling pathway. Cell Death Dis. 2016;7(6):e2239.

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2024 The Authors. Animal Models and Experimental Medicine published by John Wiley & Sons Australia, Ltd on behalf of The Chinese Association for Laboratory Animal Sciences.