TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infection
Abstract Background Toxoplasma gondii is an opportunistic protozoan infecting almost one-third of the world’s population. Toxoplasma gondii rhoptry protein 18 (TgROP18) is a key virulence factor determining the parasite’s acute virulence and is secreted into host cells during infection. We previousl...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2020-08-01
|
| Series: | Parasites & Vectors |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13071-020-04251-7 |
| _version_ | 1818406525780099072 |
|---|---|
| author | Ling Kong Dan Jiang Cheng He Jing Xia Haixia Wei Lijuan Zhou Hongjuan Peng |
| author_facet | Ling Kong Dan Jiang Cheng He Jing Xia Haixia Wei Lijuan Zhou Hongjuan Peng |
| author_sort | Ling Kong |
| collection | DOAJ |
| description | Abstract Background Toxoplasma gondii is an opportunistic protozoan infecting almost one-third of the world’s population. Toxoplasma gondii rhoptry protein 18 (TgROP18) is a key virulence factor determining the parasite’s acute virulence and is secreted into host cells during infection. We previously identified the interaction of TgROP18 and host cell immune-related receptor protein IL20RB, and observed the activation of STAT3 in human keratinocytes (HaCaT) cells infected by the rop16 knockout RH strain, though TgROP16 is regarded as being responsible for host STAT3 activation during T. gondii invasion. Therefore, we hypothesize TgROP18 can activate host STAT3 through binding to IL20RB. Methods CRISPR-CAS9 technology was used to generate the ROP16 and ROP18 double knockout RH strain, RH-∆rop16∆rop18. SDS-PAGE and western blot were used to detect STAT3 activation in different HaCaT cells with high endogenous IL20RB expression treated with T. gondii tachyzoites infection, recombinant ROP18, or IL-20. FRET and co-immunoprecipitation (Co-IP) was used to detect the protein-protein interaction. Results We observed that TgROP18 was involved in a synergic activation of the host JAK/STAT3 pathway together with TgROP16 in human HaCaT cells infected with T. gondii or treated with recombinant TgROP18 protein, stimulating host proinflammatory immune responses such as expression of TNF-α. The effect of recombinant ROP18 on STAT3 phosphorylation was presented in a dose-dependent manner. Additionally, TgROP18 was identified to target IL20RB on its extracellular domain. When we treated different cell lines with the recombinant ROP18, STAT3 phosphorylation could only be observed in the cells with endogenous IL20RB expression, such as HaCaT cells. Conclusions These findings indicate that TgROP18-IL20RB interaction upon T. gondii invasion was involved in STAT3 activation, which is associated with host cell defense. |
| first_indexed | 2024-12-14T09:13:20Z |
| format | Article |
| id | doaj.art-e6ad8307f829401eada65dfd887f6bb7 |
| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-12-14T09:13:20Z |
| publishDate | 2020-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Parasites & Vectors |
| spelling | doaj.art-e6ad8307f829401eada65dfd887f6bb72022-12-21T23:08:31ZengBMCParasites & Vectors1756-33052020-08-0113111410.1186/s13071-020-04251-7TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infectionLing Kong0Dan Jiang1Cheng He2Jing Xia3Haixia Wei4Lijuan Zhou5Hongjuan Peng6Department of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical UniversityDepartment of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical UniversityDepartment of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical UniversityDepartment of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical UniversityDepartment of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical UniversityDepartment of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical UniversityDepartment of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical UniversityAbstract Background Toxoplasma gondii is an opportunistic protozoan infecting almost one-third of the world’s population. Toxoplasma gondii rhoptry protein 18 (TgROP18) is a key virulence factor determining the parasite’s acute virulence and is secreted into host cells during infection. We previously identified the interaction of TgROP18 and host cell immune-related receptor protein IL20RB, and observed the activation of STAT3 in human keratinocytes (HaCaT) cells infected by the rop16 knockout RH strain, though TgROP16 is regarded as being responsible for host STAT3 activation during T. gondii invasion. Therefore, we hypothesize TgROP18 can activate host STAT3 through binding to IL20RB. Methods CRISPR-CAS9 technology was used to generate the ROP16 and ROP18 double knockout RH strain, RH-∆rop16∆rop18. SDS-PAGE and western blot were used to detect STAT3 activation in different HaCaT cells with high endogenous IL20RB expression treated with T. gondii tachyzoites infection, recombinant ROP18, or IL-20. FRET and co-immunoprecipitation (Co-IP) was used to detect the protein-protein interaction. Results We observed that TgROP18 was involved in a synergic activation of the host JAK/STAT3 pathway together with TgROP16 in human HaCaT cells infected with T. gondii or treated with recombinant TgROP18 protein, stimulating host proinflammatory immune responses such as expression of TNF-α. The effect of recombinant ROP18 on STAT3 phosphorylation was presented in a dose-dependent manner. Additionally, TgROP18 was identified to target IL20RB on its extracellular domain. When we treated different cell lines with the recombinant ROP18, STAT3 phosphorylation could only be observed in the cells with endogenous IL20RB expression, such as HaCaT cells. Conclusions These findings indicate that TgROP18-IL20RB interaction upon T. gondii invasion was involved in STAT3 activation, which is associated with host cell defense.http://link.springer.com/article/10.1186/s13071-020-04251-7Toxoplasma gondiiROP18IL20RBSTAT3Proinflammatory immunity |
| spellingShingle | Ling Kong Dan Jiang Cheng He Jing Xia Haixia Wei Lijuan Zhou Hongjuan Peng TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infection Parasites & Vectors Toxoplasma gondii ROP18 IL20RB STAT3 Proinflammatory immunity |
| title | TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infection |
| title_full | TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infection |
| title_fullStr | TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infection |
| title_full_unstemmed | TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infection |
| title_short | TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infection |
| title_sort | tgrop18 targets il20rb for host defense related stat3 activation during toxoplasma gondii infection |
| topic | Toxoplasma gondii ROP18 IL20RB STAT3 Proinflammatory immunity |
| url | http://link.springer.com/article/10.1186/s13071-020-04251-7 |
| work_keys_str_mv | AT lingkong tgrop18targetsil20rbforhostdefenserelatedstat3activationduringtoxoplasmagondiiinfection AT danjiang tgrop18targetsil20rbforhostdefenserelatedstat3activationduringtoxoplasmagondiiinfection AT chenghe tgrop18targetsil20rbforhostdefenserelatedstat3activationduringtoxoplasmagondiiinfection AT jingxia tgrop18targetsil20rbforhostdefenserelatedstat3activationduringtoxoplasmagondiiinfection AT haixiawei tgrop18targetsil20rbforhostdefenserelatedstat3activationduringtoxoplasmagondiiinfection AT lijuanzhou tgrop18targetsil20rbforhostdefenserelatedstat3activationduringtoxoplasmagondiiinfection AT hongjuanpeng tgrop18targetsil20rbforhostdefenserelatedstat3activationduringtoxoplasmagondiiinfection |