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Mechanism by which the combination of SjCL3 and SjGAPDH protects against Schistosoma japonicum infection

  • Immunology and Host-Parasite Interactions - Original Paper
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Abstract

A vaccine is an important method to control schistosomiasis. Molecules related to lung-stage schistosomulum are considered potential vaccine candidates. We previously showed that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and cathepsin L3 (CL3) displayed differential expression in the lung-stage schistosomula of Schistosoma japonicum cocultured with host cells. In the present study, we prepared the two proteins and detected the protective effects of SjGAPDH by immunizing mice with this protein alone and in combination with SjCL3 with or without Freund’s adjuvant. Then, we investigated the possible mechanisms underlying S. japonicum infection. The results showed that vaccination of adjuvanted SjGAPDH decreased the worm burden (37.8%) and egg load (38.1%), and the combination of adjuvanted SjGAPDH and SjCL3 further decreased the worm burden (65.6%) and egg load (70.9%) during Schistosoma japonicum infection. However, the immunization of a combination of adjuvant-free SjGAPDH and SjCL3 displayed a lower protective effect (< 15%) than those of the adjuvanted SjCL3, the adjuvanted SjGAPDH, and a combination of adjuvanted SjGAPDH and SjCL3. Flow cytometric results showed that the frequency of regulatory T cells (Tregs) was lower (P < 0.05) in the group with adjuvanted SjGAPDH and SjCL3 (2.61%) than the remaining groups. The enzyme-linked immunosorbent assay (ELISA) results indicated that except for the uninfected and infected control groups, the remaining groups displayed a Th1-type shift in immune responses. These results showed the immunization of SjGAPDH resulted in partial protection (approximately 38%); inoculation with a combination of SjCL3 and SjGAPDH in Freund’s adjuvant resulted in a high immunoprotective effect (> 65%) against Schistosoma japonicum infection in mice, which was possibly caused by the reduced percentage of Tregs and a Th1-type shift in immune responses; and SjCL3 has no adjuvant-like effect, dissimilar to SmCL3.

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Acknowledgments

We thank Yanru Gao for her assistance in this work. We thank Chunlian Tang for guiding the flow cytometric analysis and the modification of this article. We thank the Structural Bioinformatics Group (Imperial College, London) for the free use of Phyre2. We also acknowledge the molecular graphics and analyses generated by UCSF Chimera, developed by the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco, with support from NIH P41-GM103311.

Funding

This study was funded by the National Natural Science Foundation of China (No. 81273010) and the Scientific Research Subject of the Health and Family Planning Commission of Hubei Province (No. XF2012-18).

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Authors and Affiliations

  1. Hubei Province Key Laboratory of Allergy and Immunology, Department of Parasitology, Wuhan University School of Basic Medical Sciences, Wuhan, Hubei Province, China

    Wenling Huang, Mengjie Gu, Wenjun Cheng, Shuhong Wang, Kaijie Li, Qin Ping Zhao, Zhenping Ming & Huifen Dong

  2. Hubei Provincial Center for Disease Control and Prevention, Wuhan, Hubei Province, China

    Kaijie Li

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  1. Wenling Huang
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Correspondence to Huifen Dong.

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All animal experiments were carried out in absolute accordance with the Guideline for the Care and Use of Laboratory Animals of the National Institutes of Health in China and were approved by the Center for Animal Experiment of Wuhan University (approval no. 2019103).

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Fig. S1

The PCR and SDS-PAGE results of SjGAPDH. a PCR of SjGAPDH expression. M, marker; lanes 1 and 2, PCR amplification products of the SjGAPDH partial CDS. b SDS-PAGE of SjGAPDH. M, marker; lanes 1 and lane 2, E. coli BL21 (DE3) cells transformed with plasmid pET-28a(+)/SjGAPDH before and after culture with 1 mM isopropyl-β-thiogalactopyranoside; lanes 3 and 4, soluble and insoluble components after lysis. c Purified SjGAPDH. M, marker; lane 1, solution after Ni-NTA agarose binding; lanes 2, 3, and 4, first, second and third wash solutions; lane 5, purified SjGAPDH. d Kinetic activity assay of rSjGAPDH (PNG 414 kb)

High resolution image (TIF 5121 kb)

Fig. S2

Representative regulatory T cell frequency in each group. a, a' A representative image of the sample from the uninfected control group. b, b' A representative image of the infected control group. c, c' A representative image of the Freund’s adjuvant control group. d, d' A representative image of the adjuvanted SjCL3 group. e, e' A representative image of the adjuvanted SjGAPDH group. f, f' A representative image of the adjuvant-free combination group. g, g' A representative picture of the adjuvanted combination group. The upper row represents all splenic lymphocytes from one mouse, in which the cells in the gate show the CD4+ cell frequency. The lower row represents the CD25+ and/or Foxp3+ frequency derived from the CD4+ cells (cells from the upper row gate), in which Q2 represents the CD25+Foxp3+ cell frequency (% CD4+). (PNG 590 kb)

High resolution image (TIF 6337 kb)

ESM 1

Sequencing results of SjGAPDH (TXT 356 bytes)

ESM 2

All raw data in this study (XLSX 42 kb)

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Huang, W., Gu, M., Cheng, W. et al. Mechanism by which the combination of SjCL3 and SjGAPDH protects against Schistosoma japonicum infection. Parasitol Res 120, 173–185 (2021). https://doi.org/10.1007/s00436-020-06916-9

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