Expression of sphingosine-1-phosphate receptor 1 in the brain of fatal cerebral malaria

Srisook, Charit; Nintasen, Rungrat; Punsawad, Chuchard; Glaharn, Supattra; Techarang, Tachpon; Viriyavejakul, Parnpen

doi:10.1038/s41598-026-36072-6

Download PDF

Article
Open access
Published: 17 January 2026

Expression of sphingosine-1-phosphate receptor 1 in the brain of fatal cerebral malaria

Charit Srisook¹,
Rungrat Nintasen²,
Chuchard Punsawad^3,4,
Supattra Glaharn¹,
Tachpon Techarang¹ &
…
Parnpen Viriyavejakul¹

Scientific Reports , Article number: (2026) Cite this article

784 Accesses
Metrics details

We are providing an unedited version of this manuscript to give early access to its findings. Before final publication, the manuscript will undergo further editing. Please note there may be errors present which affect the content, and all legal disclaimers apply.

Subjects

Abstract

The signaling pathway activated by sphingosine-1-phosphate (S1P) through S1P receptor 1 (S1PR1) plays specific roles in regulating vascular integrity and preventing vascular leakage during inflammatory response. Endothelial cell barrier dysfunction has been implicated in cerebral malaria (CM) pathology. To explore the S1P/S1PR1 signaling pathway in CM, the study investigated the expression of S1PR1 in the brain of fatal malaria and correlated with the level of S1P and malaria severity. Localisation of S1PR1 in brain tissues was evaluated using immunohistochemistry technique in archived brain tissues of fatal P. falciparum malaria. S1P level was determined using enzyme-linked immunosorbent assay (ELISA). S1PR1 expression was intense in cerebral blood vessels and neurons of fatal CM patients compared to brain tissues from control group and non-CM patients (all p < 0.001). S1P level in the blood decreased significantly in CM group and was negatively correlated with S1PR1 expression in blood vessels and neurons. The expression of S1PR1 in cerebral blood vessels and neurons indicates that S1P/S1PR1 signaling pathway is involved in malaria pathogenesis and represents potential targets for S1P/S1PR1 modulators to treat CM. The outcomes can serve as a basis to explore measures to block the expression of S1PR1 which could reduce sequestration.

Mesenchymal stem cells alleviate experimental cerebral malaria disease severity by inducing RoRγt⁺ Foxp3⁺ T regulatory (Tr 17) cells and modulating the dysregulated Th17/Treg axis

Article Open access 30 January 2026

Single-cell atlas of the human brain vasculature across development, adulthood and disease

Article Open access 10 July 2024

Shaping the brain vasculature in development and disease in the single-cell era

Article 20 March 2023

Data availability

All data generated and analysed during the current study are included in this published articles and its supplementary information.

Abbreviations

ABC:: Avidin–biotin complex
BBB:: Blood brain barrier
CM:: Cerebral malaria
°C:: Degree Celsius
DAB:: 3,3'-Diaminobenzidine
ECs:: Endothelial cells
EDG1:: Endothelial differentiation gene 1
ELISA:: Enzyme-linked immunosorbent assay
Fig:: Figure
FTY720:: Fingolimod hydrochloride
Gi:: G protein alpha subunit
h:: Hour
H&E:: Haematoxylin and eosin
HCl:: Hydrochloric acid
HRP:: Horseradish peroxidase
IFN-γ:: Interferon gamma
IL:: Interleukin
LPF:: Low power fields
min:: Minute
N:: Normal
NCM:: Non-cerebral malaria
p :: p-value
pH:: Potential of hydrogen
PRBCs:: Parasitized red blood cells
RBCs:: Red blood cells
r _s :: Spearman’s rank correlation coefficient
RT:: Room temperature
SARS-CoV-2:: Severe acute respiratory syndrome coronavirus 2
SEM:: Standard error of the mean
S1P:: Sphingosine-1-phosphate
S1PR1:: Sphingosine-1-phosphate receptor 1
SPSS:: Statistical Package for the Social Sciences
TBS:: Tris-buffered saline
TMB:: 3, 3’, 5, 5’-Tetramethylbenzidine
TNF:: Tumour necrosis factor
TS:: Total score
µm:: Micrometre
µM:: Micromolar

References

Song, X. et al. Cerebral malaria induced by Plasmodium falciparum: clinical features, pathogenesis, diagnosis, and treatment. Front. Cell. Infect. Microbiol. 12, 939532. https://doi.org/10.3389/fcimb.2022.939532 (2022).
Google Scholar
Solomon, W. et al. Neuregulin-1 attenuates mortality associated with experimental cerebral malaria. J. neuroinflammation. 11, 1–13. https://doi.org/10.1186/1742-2094-11-9 (2014).
Google Scholar
Wiser, M. F. Knobs, adhesion, and severe falciparum malaria. Trop. Med. Infect. Dis. 8, 353. https://doi.org/10.3390/tropicalmed8070353 (2023).
Google Scholar
Nishanth, G. & Schluter, D. Blood-brain barrier in cerebral malaria: Pathogenesis and therapeutic intervention. Trends Parasitol. 35, 516–528. https://doi.org/10.1016/j.pt.2019.04.010 (2019).
Google Scholar
Leao, L. et al. Association of cerebral malaria and TNF-alpha levels: a systematic review. BMC Infect. Dis. 20, 442. https://doi.org/10.1186/s12879-020-05107-2 (2020).
Google Scholar
Dunst, J., Kamena, F. & Matuschewski, K. Cytokines and chemokines in cerebral malaria pathogenesis. Front. Cell. Infect. Microbiol. 7, 324. https://doi.org/10.3389/fcimb.2017.00324 (2017).
Google Scholar
Royo, J. et al. Elevated plasma interleukin-8 as a risk factor for mortality in children presenting with cerebral malaria. Infect. Dis. Poverty. 12, 8. https://doi.org/10.1186/s40249-023-01059-2 (2023).
Google Scholar
Idro, R., Jenkins, N. E. & Newton, C. R. Pathogenesis, clinical features, and neurological outcome of cerebral malaria. Lancet Neurol. 4, 827–840. https://doi.org/10.1186/s40249-023-01059-210.1016/S1474-4422(05)70247-7 (2005).
Google Scholar
Lyke, K. E. et al. Serum levels of the proinflammatory cytokines interleukin-1 beta (IL-1beta), IL-6, IL-8, IL-10, tumor necrosis factor alpha, and IL-12 (p70) in Malian children with severe Plasmodium falciparum malaria and matched uncomplicated malaria or healthy controls. Infect. Immun. 72, 5630–5637. https://doi.org/10.1128/IAI.72.10.5630-5637.2004 (2004).
Google Scholar
Mshana, R. N., Boulandi, J., Mshana, N. M., Mayombo, J. & Mendome, G. Cytokines in the pathogenesis of malaria: levels of IL-I beta, IL-4, IL-6, TNF-alpha and IFN-gamma in plasma of healthy individuals and malaria patients in a holoendemic area. J Clin Lab Immunol. 34, 131–139 (1991).
Google Scholar
Okamoto, Y., Wang, F., Yoshioka, K., Takuwa, N. & Takuwa, Y. Sphingosine-1-phosphate-specific G protein-coupled receptors as novel therapeutic targets for atherosclerosis. Pharmaceuticals. 4(1), 117–137. https://doi.org/10.3390/ph4010117 (2011).
Google Scholar
Weigel, C., Bellaci, J. & Spiegel, S. Sphingosine-1-phosphate and its receptors in vascular endothelial and lymphatic barrier function. J. Biol. Chem. 299, 104775. https://doi.org/10.1016/j.jbc.2023.104775 (2023).
Google Scholar
Chatzikonstantinou, S. et al. Signaling through the S1P–S1PR Axis in the gut, the immune and the central nervous system in multiple sclerosis: Implication for pathogenesis and treatment. Cells 10, 3217. https://doi.org/10.3390/cells10113217 (2021).
Google Scholar
Ono, Y. et al. Sphingosine 1-phosphate release from platelets during clot formation: close correlation between platelet count and serum sphingosine 1-phosphate concentration. Lipids Health Dis. 12, 20. https://doi.org/10.1186/1476-511X-12-20 (2013).
Google Scholar
Lucke, S. & Levkau, B. Endothelial functions of sphingosine-1-phosphate. Cell. Physiol. Biochem. 26, 87–96. https://doi.org/10.1159/000315109 (2010).
Google Scholar
Mendelson, K., Evans, T. & Hla, T. Sphingosine 1-phosphate signalling. Development 141, 5–9. https://doi.org/10.1242/dev.094805 (2014).
Google Scholar
Lee, M. J. et al. Vascular endothelial cell adherens junction assembly and morphogenesis induced by sphingosine-1-phosphate. Cell 99, 301–312. https://doi.org/10.1016/s0092-8674(00)81661-x (1999).
Google Scholar
Garcia, J. G. et al. Sphingosine 1-phosphate promotes endothelial cell barrier integrity by Edg-dependent cytoskeletal rearrangement. J Clin Invest. 108, 689–701. https://doi.org/10.1172/JCI12450 (2001).
Google Scholar
Yun, T. et al. Expression of sphingosine-1-phosphate receptor 1 in neuroinflammation of canine brains. Top. Companion Anim. Med. 60, 100847. https://doi.org/10.1016/j.tcam.2024.100847 (2024).
Google Scholar
Iwasawa, E. et al. Sphingosine-1-phosphate receptor 1 activation enhances leptomeningeal collateral development and improves outcome after stroke in mice. J Stroke Cerebrovasc Dis. 27, 1237–1251. https://doi.org/10.1016/j.jstrokecerebrovasdis.2017.11.040 (2018).
Google Scholar
Robert, P. et al. EDG1 receptor stimulation leads to cardiac hypertrophy in rat neonatal myocytes. J Mol Cell Cardiol. 33, 1589–1606. https://doi.org/10.1006/jmcc.2001.1433 (2001).
Google Scholar
Punsawad, C., Maneerat, Y., Chaisri, U., Nantavisai, K. & Viriyavejakul, P. Nuclear factor kappa B modulates apoptosis in the brain endothelial cells and intravascular leukocytes of fatal cerebral malaria. Malar J. 12, 260. https://doi.org/10.1186/1475-2875-12-260 (2013).
Google Scholar
Srisook, C., Glaharn, S., Punsawad, C. & Viriyavejakul, P. Apoptotic changes and aquaporin-1 expression in the choroid plexus of cerebral malaria patients. Malar J. 21, 43. https://doi.org/10.1186/s12936-022-04044-6 (2022).
Google Scholar
Turner, G. Cerebral malaria. Brain Pathol. 7, 569–582. https://doi.org/10.1111/j.1750-3639.1997.tb01075.x (1997).
Google Scholar
MacPherson, G. G., Warrell, M. J., White, N. J., Looareesuwan, S. & Warrell, D. A. Human cerebral malaria. A quantitative ultrastructural analysis of parasitized erythrocyte sequestration. Am. J. Pathol. 119, 385–401 (1985).
Google Scholar
Prapansilp, P. et al. A clinicopathological correlation of the expression of the angiopoietin-Tie-2 receptor pathway in the brain of adults with Plasmodium falciparum malaria. Malar J. 12, 50. https://doi.org/10.1186/1475-2875-12-50 (2013).
Google Scholar
Xiao, S., Peng, K., Li, C., Long, Y. & Yu, Q. The role of sphingosine-1-phosphate in autophagy and related disorders. Cell Death Discov. 9, 380. https://doi.org/10.1038/s41420-023-01681-x (2023).
Google Scholar
Hashemi, E. et al. Visualizing Sphingosine-1-phosphate receptor 1 (S1PR1) signaling during central nervous system de- and remyelination. Cell. Mol. Neurobiol. 43, 1219–1236. https://doi.org/10.1007/s10571-022-01245-0 (2023).
Google Scholar
Pyne, S. & Pyne, N. J. Sphingosine 1-phosphate signalling in mammalian cells. Biochem. J. 349, 385–402. https://doi.org/10.1042/0264-6021:3490385 (2000).
Google Scholar
Gaire, B. P. & Choi, J. W. Sphingosine 1-phosphate receptors in cerebral ischemia. Neuromolecular Med. 23, 211–223. https://doi.org/10.1007/s12017-020-08614-2 (2021).
Google Scholar
Garcia, J. G. et al. Sphingosine 1-phosphate promotes endothelial cell barrier integrity by Edg-dependent cytoskeletal rearrangement. J. Clin. Invest. 108, 689–701. https://doi.org/10.1172/JCI12450 (2001).
Google Scholar
Modak, A. et al. Fingolimod (FTY720), an FDA-approved sphingosine 1-phosphate (S1P) receptor agonist, restores endothelial hyperpermeability in cellular and animal models of dengue virus serotype 2 infection. IUBMB Life 76, 267–285. https://doi.org/10.1002/iub.2795 (2024).
Google Scholar
Camerer, E. et al. Sphingosine-1-phosphate in the plasma compartment regulates basal and inflammation-induced vascular leak in mice. J. Clin. Invest. 119, 1871–1879. https://doi.org/10.1172/jci38575 (2009).
Google Scholar
Punsawad, C. & Viriyavejakul, P. Reduction in serum sphingosine 1-phosphate concentration in malaria. PLoS ONE 12, e0180631. https://doi.org/10.1371/journal.pone.0180631 (2017).
Google Scholar
Sah, R. K. et al. Reduction of sphingosine kinase 1 phosphorylation and activity in plasmodium-infected erythrocytes. Front. Cell. Dev. Biol. 8, 80. https://doi.org/10.3389/fcell.2020.00080 (2020).
Google Scholar
Finney, C. A. et al. S1P is associated with protection in human and experimental cerebral malaria. Mol Med. 17, 717–725. https://doi.org/10.2119/molmed.2010.00214 (2011).
Google Scholar
Karam, M. & Auclair, C. Sphingosine-1-phosphate as lung and cardiac vasculature protecting agent in SARS-CoV-2 Infection. Int. J. Mol. Sci. 24, 13088. https://doi.org/10.3390/ijms241713088 (2023).
Google Scholar
Winkler, M. S. et al. Erythrocytes increase endogenous sphingosine 1-phosphate levels as an adaptive response to SARS-CoV-2 infection. Clin. Sci (Lond) 135, 2781–2791. https://doi.org/10.1042/CS20210666 (2021).
Google Scholar
Marfia, G. et al. Decreased serum level of sphingosine-1-phosphate: a novel predictor of clinical severity in COVID-19. EMBO Mol. Med. 13, 13424. https://doi.org/10.15252/emmm.202013424 (2021).
Google Scholar
Gomes, L. et al. Sphingosine 1-phosphate in acute dengue infection. PLoS ONE 9, e113394. https://doi.org/10.1371/journal.pone.0113394 (2014).
Google Scholar
Michels, M. et al. Decreased plasma levels of the endothelial protective sphingosine-1-phosphate are associated with dengue-induced plasma leakage. J. Infect. 71, 480–487. https://doi.org/10.1016/j.jinf.2015.06.014 (2015).
Google Scholar
Winkler, M. S. et al. Loss of sphingosine 1-phosphate (S1P) in septic shock is predominantly caused by decreased levels of high-density lipoproteins (HDL). J. Intensive Care. 7, 23. https://doi.org/10.1186/s40560-019-0376-2 (2019).
Google Scholar
Winkler, M. S. et al. Decreased serum concentrations of sphingosine-1-phosphate in sepsis. Crit Care. 19, 372. https://doi.org/10.1186/s13054-015-1089-0 (2015).
Google Scholar
Bai, Q. et al. Serum sphingosine-1-phosphate level and peritonitis in peritoneal dialysis patients. Ren. Fail. 42, 829–835. https://doi.org/10.1080/0886022X.2020.1805763 (2020).
Google Scholar
Yang, X. et al. Association of plasma sphingosine-1-phosphate levels with disease severity and prognosis after intracerebral hemorrhage. Front. Neurol. 15, 1365902. https://doi.org/10.3389/fneur.2024.1365902 (2024).
Google Scholar
Hsu, S. C. et al. Circulating sphingosine-1-phosphate as a prognostic biomarker for community-acquired pneumonia. PLoS ONE 14, e0216963. https://doi.org/10.1371/journal.pone.0216963 (2019).
Google Scholar
Zhao, Y. et al. Pathogenic sphingosine 1-phosphate pathway in psoriasis: a critical review of its pathogenic significance and potential as a therapeutic target. Lipids Health Dis. 22, 52. https://doi.org/10.1186/s12944-023-01813-3 (2023).
Google Scholar
Ding, F., Wang, Z., Wang, J., Ma, Y. & Jin, J. Serum S1P level in interstitial lung disease (ILD) is a potential biomarker reflecting the severity of pulmonary function. BMC Pulm. Med. 24, 266. https://doi.org/10.1186/s12890-024-03081-y (2024).
Google Scholar
Miura, K. et al. Dysregulation of sphingolipid metabolic enzymes leads to high levels of sphingosine-1-phosphate and ceramide in human hepatocellular carcinoma. Hepatol Res. 51, 614–626. https://doi.org/10.1111/hepr.13625 (2021).
Google Scholar
Blachnio-Zabielska, A. U. et al. The Interplay between oxidative stress and sphingolipid metabolism in endometrial cancer. Int. J. Mol. Sci. 25. https://doi.org/10.3390/ijms251910243 (2024).
Oggungwan, K., Glaharn, S., Ampawong, S., Krudsood, S. & Viriyavejakul, P. FTY720 restores endothelial cell permeability induced by malaria sera. Sci. Rep. 8, 10959. https://doi.org/10.1038/s41598-018-28536-1 (2018).
Google Scholar
Thompson, J. et al. Progressive disruption of sphingosine-1-phosphate receptor 1 correlates with blood-brain barrier leakage in a rat model of chronic hypoxic hypoperfusion. Aging Dis. 16, 2. https://doi.org/10.14336/AD.2024.0098 (2024).
Google Scholar
Nacer, A. et al. Neuroimmunological blood brain barrier opening in experimental cerebral malaria. PLoS Pathog. 8, e1002982. https://doi.org/10.1371/journal.ppat.1002982 (2012).
Google Scholar
Yang, L. X. et al. Sphingosine-1-phosphate receptor 1 regulates blood-brain barrier permeability in epileptic mice. Neural Regen. Res. 18, 1763–1769. https://doi.org/10.4103/1673-5374.360263 (2023).
Google Scholar
Groves, A., Kihara, Y. & Chun, J. Fingolimod: direct CNS effects of sphingosine 1-phosphate (S1P) receptor modulation and implications in multiple sclerosis therapy. J. Neurol. Sci. 328, 9–18. https://doi.org/10.1016/j.jns.2013.02.011 (2013).
Google Scholar
Burg, N., Salmon, J. E. & Hla, T. Sphingosine 1-phosphate receptor-targeted therapeutics in rheumatic diseases. Nat Rev Rheumatol. 18, 335–351. https://doi.org/10.1038/s41584-022-00784-6 (2022).
Google Scholar
Patmanathan, S. N. et al. Mechanisms of sphingosine 1-phosphate receptor signalling in cancer. Cell Signal. 34, 66–75. https://doi.org/10.1016/j.cellsig.2017.03.002 (2017).
Google Scholar
Jiang, H. et al. Quantitative Analysis of S1PR1 expression in the postmortem multiple sclerosis central nervous system. ACS Chem. Neurosci. 14, 4039–4050. https://doi.org/10.1021/acschemneuro.3c00581 (2023).
Google Scholar
Nishimura, H., Akiyama, T., Irei, I., Hamazaki, S. & Sadahira, Y. Cellular localization of sphingosine-1-phosphate receptor 1 expression in the human central nervous system. J. Histochem. Cytochem. 58, 847–856. https://doi.org/10.1369/jhc.2010.956409 (2010).
Google Scholar
Gao, P. et al. S1PR1 regulates NDV-induced IL-1beta expression via NLRP3/caspase-1 inflammasome. Vet Res. 53, 58. https://doi.org/10.1186/s13567-022-01078-1 (2022).
Google Scholar
Chavez, A. et al. S1PR1 Tyr143 phosphorylation downregulates endothelial cell surface S1PR1 expression and responsiveness. J Cell Sci. 128, 878–887. https://doi.org/10.1242/jcs.154476 (2015).
Google Scholar

Download references

Acknowledgements

Our gratitude goes to the Central Equipment Unit, Faculty of Tropical Medicine, Mahidol University, Thailand for facilitating the laboratory equipment. We thank all staff at the Department of Tropical Pathology, Faculty of Tropical Medicine, Mahidol University, Thailand for their support throughout this study.

Funding

This research project is supported by Faculty of Tropical Medicine, Mahidol University, Fiscal year 2011. RN is the grant recipient.

Author information

Authors and Affiliations

Department of Tropical Pathology, Faculty of Tropical Medicine, Mahidol University, 420/6 Rajvithi Road, Bangkok, 10400, Thailand
Charit Srisook, Supattra Glaharn, Tachpon Techarang & Parnpen Viriyavejakul
Independent Researcher, Bangkok, Thailand
Rungrat Nintasen
Department of Medical Science, School of Medicine, Walailak University, Nakhon Si Thammarat, 80160, Thailand
Chuchard Punsawad
Research Center in Tropical Pathobiology, Walailak University, Nakhon Si Thammarat, 80160, Thailand
Chuchard Punsawad

Authors

Charit Srisook
View author publications
Search author on:PubMed Google Scholar
Rungrat Nintasen
View author publications
Search author on:PubMed Google Scholar
Chuchard Punsawad
View author publications
Search author on:PubMed Google Scholar
Supattra Glaharn
View author publications
Search author on:PubMed Google Scholar
Tachpon Techarang
View author publications
Search author on:PubMed Google Scholar
Parnpen Viriyavejakul
View author publications
Search author on:PubMed Google Scholar

Contributions

P.V., C.S. and R.N. initiated the research idea, designed the experiments, gathered clinical data and retrieved archived paraffin blocks. P.V., C.S., R.N., C.P. and S.G. performed immunohistochemistry study. P.V., C.S., S.G. and T.T. worked on ELISA. All authors analysed the results, interpreted the data and drafted the manuscript. PV provided technical guidance, gave advices and revised the final manuscript. R.N. obtained funding, was affiliated with Department of Tropical Pathology, Faculty of Tropical Medicine, Mahidol University during the initial part of the research, and is currently an independent researcher. All authors reviewed and approved the final version of the manuscript.

Corresponding author

Correspondence to Parnpen Viriyavejakul.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethical approval

Ethical approval was obtained from the Ethics Committee of the Faculty of Tropical Medicine, Mahidol University (MUTM 2011-027-01, MUTM 2011-27-02: for using tissue specimens and MUTM 2024-033-01: for using left-over human sera).

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Supplementary Information 1.

Supplementary Information 2.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

Reprints and permissions

About this article

Cite this article

Srisook, C., Nintasen, R., Punsawad, C. et al. Expression of sphingosine-1-phosphate receptor 1 in the brain of fatal cerebral malaria. Sci Rep (2026). https://doi.org/10.1038/s41598-026-36072-6

Download citation

Received: 21 February 2025
Accepted: 09 January 2026
Published: 17 January 2026
DOI: https://doi.org/10.1038/s41598-026-36072-6