Modulation of host immune response by carotene supplementation in a COVID-19 vaccination mouse model

Abstract

Carotenoids, known for their immunomodulatory and gut microbiota-modulating effects, have drawn attention as potential dietary adjuvants to enhance vaccine efficacy and maintain gut health. This study aimed to evaluate the effect of carotene supplementation on immune response with insight into gut microbiome using an in vivo animal model. The BALB/c mice were fed daily with CaroGaia (50 mg/kg of body weight), a carotene supplement that contained 33.3% α-carotene and 66.6% β-carotene, by oral gavage for 70 days. Mice fed with the vehicle served as controls. The mice in the vaccinated groups received two doses of the CoronaVac inactivated virus vaccine on days 14 and 42. Flow cytometry revealed no significant modulation of lymphocyte subsets (total T lymphocytes, T-helper cells, cytotoxic T lymphocytes, and B cells) with carotene supplementation. In addition, there were no significant differences in the levels of SARS-CoV-2 immunoglobulin G and interferon-gamma in plasma between treatment and control groups. In contrast, the vaccinated carotene group showed an increased SARS-CoV-2 antigen-specific splenocyte proliferation. In the gut microbiome, carotene supplementation appeared to alter the gut microbiota composition. However, no significant changes were observed in the short-chain fatty acids (SCFA) levels, such as acetic acid, butyric acid, and propionic acid. Furthermore, the differential abundance analysis showed that carotene supplementation reduced the levels of SCFA producers (Odoribacter and Monoglobus genera) in unvaccinated mice compared to the control group, while it enriched the level of SCFA producers (Ruminococcaceae family) and reduced pathobiont levels, commensal bacteria that have pathogenic potential (Mucispirillum genus), in the vaccinated group.

Keywords

COVID-19 / SARS-CoV-2 / Vaccination / Carotenoid / Carotene / Immune-modulation / Gut microbiota / Short-chain fatty acid

Funding

This study was supported by a research grant from the Jeffrey Cheah School of Medicine, Monash University Malaysia (STG-000056).

Conflict of interest

The authors declare that they have no competing interests.

References

[1] Amanat F, Krammer F. SARS-CoV-2 vaccines: Status report. Immunity. 2020; 52:583-589. doi: 10.1016/j.immuni.2020.03.007

[2] Pennisi F, Genovese C, Gianfredi V. Lessons from the COVID-19 pandemic: Promoting vaccination and public health resilience, a narrative review. Vaccines (Basel). 2024;12:891. doi: 10.3390/vaccines12080891

[3] Lim WW, Mak L, Leung GM, Cowling BJ, Peiris M. Comparative immunogenicity of mRNA and inactivated vaccines against COVID-19. Lancet Microbe. 2021; 2:e423 doi: 10.1016/S2666-5247

[4] Fu Z, Liang D, Zhang W, et al. Host protection against Omicron BA.2.2 sublineages by prior vaccination in spring 2022 COVID-19 outbreak in Shanghai. Front Med. 2023;17:562-575. doi: 10.1007/s11684-022-0977-3

[5] Zeng G, Wu Q, Pan H, et al. Immunogenicity and safety of a third dose of CoronaVac, and immune persistence of a two-dose schedule, in healthy adults: Interim results from two single-centre, double-blind, randomised, placebo-controlled phase 2 clinical trials. Lancet Infect Dis. 2022;22:483-495. doi: 10.1016/s1473-3099(21)00681-2

[6] Carr AC, Maggini S.Vitamin C and immune function. Nutrients. 2017;9:1211. doi: 10.3390/nu9111211

[7] Chen Y, Yin S, Tong X, et al. Dynamic SARS-CoV-2-specific B-cell and T-cell responses following immunization with an inactivated COVID-19 vaccine. Clin Microbiol Infect. 2022; 28:410-418. doi: 10.1016/j.cmi.2021.10.006

[8] Yin J, Zhao Y, Huang F, et al. Immune response and homeostasis mechanism following administration of BBIBP-CorV SARS-CoV-2 inactivated vaccine. Innovation (Camb). 2023;4:100359. doi: 10.1016/j.xinn.2022.100359

[9] Wang Y, Liu J, Burrows PD, Wang JY. B cell development and maturation. In: WangJY,editor. B Cells in Immunity and Tolerance. Singapore: Springer Singapore; 2020. p. 1-22.

[10] Ramljak D, Vukoja M, Curlin M, et al. Early response of CD8+ T cells in COVID-19 patients. J Pers Med. 2021;11:1291. doi: 10.3390/jpm11121291

[11] Gombart AF, Pierre A, Maggini S. A review of micronutrients and the immune system-working in harmony to reduce the risk of infection. Nutrients. 2020;12:236. doi: 10.3390/nu12010236

[12] Hussain Y, Abdullah, Alsharif KF, et al. Therapeutic role of carotenoids in blood cancer: Mechanistic insights and therapeutic potential. Nutrients. 2022;14:1949. doi: 10.3390/nu14091949

[13] Loganathan R, Subramaniam KM, Radhakrishnan AK, Choo YM, Teng KT. Health-promoting effects of red palm oil: Evidence from animal and human studies. Nutr Rev. 2017;75:98-113. doi: 10.1093/nutrit/nuw054

[14] Iddir M, Brito A, Dingeo G, et al. Strengthening the immune system and reducing inflammation and oxidative stress through diet and nutrition: Considerations during the COVID-19 crisis. Nutrients. 2020;12:1562. doi: 10.3390/nu12061562

[15] Lewis ED, Meydani SN, Wu D. Regulatory role of vitamin E in the immune system and inflammation. IUBMB Life. 2019; 71:487-494. doi: 10.1002/iub.1976

[16] Ávila-Román J, García-Gil S, Rodríguez-Luna A, Motilva V, Talero E. Anti-Inflammatory and anticancer effects of microalgal carotenoids. Marine Drugs. 2021;19:531. doi: 10.3390/md19100531

[17] Zainal Z, Abdul Rahim A, Khaza’ai H, Chang SK. Effects of palm oil tocotrienol-rich fraction (TRF) and carotenes in ovalbumin (OVA)-challenged asthmatic brown norway rats. Int J Mol Sci. 2019;20:1764. doi: 10.3390/ijms20071764

[18] Elefson SK, Greiner LL. The evaluation of the supplementation of vitamin A, beta-carotene, and oxidized beta-carotene in prepubertal gilts. J Anim Sci. 2023;101:skad103. doi: 10.1093/jas/skad103

[19] Liang Z, Wang N, Fan C, et al. Disturbance of adaptive immunity system was accompanied by a decrease in plasma short-chain fatty acid for patients hospitalized during SARS-CoV-2 infection after COVID-19 vaccination. J Inflamm Res. 2023; 16:5261-5272. doi: 10.2147/jir.S434860

[20] Li Z, Dai Z, Shi E, et al. Study on the interaction between β-carotene and gut microflora using an in vitro fermentation model. Food Sci Hum Wellness. 2023; 12:1369-1378. doi: 10.1016/j.fshw.2022.10.030

[21] Lynn DJ, Benson SC, Lynn MA, Pulendran B. Modulation of immune responses to vaccination by the microbiota: Implications and potential mechanisms. Nat Rev Immunol. 2022;22:33-46. doi: 10.1038/s41577-021-00554-7

[22] Liu XF, Shao JH, Liao YT, et al. Regulation of short-chain fatty acids in the immune system. Front Immunol. 2023;14:1186892. doi: 10.3389/fimmu.2023.1186892

[23] Takabayashi T, Yoshida K, Imoto Y, Schleimer RP, Fujieda S. Regulation of the expression of SARS-CoV-2 receptor angiotensin-converting enzyme 2 in nasal mucosa. Am J Rhinol Allergy. 2022;36:115-122. doi: 10.1177/19458924211027798

[24] Gray J, Guo B, Bearden R, Manka J. A fast, fully validated GC-MS method using a simplified pretreatment for the quantification of short and branched chain fatty acids in human stool. J Mass Spectrom. 2022;57:e4817. doi: 10.1002/jms.4817

[25] Kers JG, Saccenti E. The power of microbiome studies: Some considerations on which alpha and beta metrics to use and how to report results. Front Microbiol. 2022;12:796025. doi: 10.3389/fmicb.2021.796025

[26] Nearing JT, Douglas GM, Hayes MG, et al. Microbiome differential abundance methods produce different results across 38 datasets. Nat Commun. 2022;13:342. doi: 10.1038/s41467-022-28034-z

[27] Santos MS, Leka LS, Ribaya-Mercado JD, et al. Short-and long-term beta-carotene supplementation do not influence T cell-mediated immunity in healthy elderly persons. Am J Clin Nutr. 1997;66:917-924. doi: 10.1093/ajcn/66.4.917

[28] Hughes DA, Wright AJ, Finglas PM, et al. The effect of beta-carotene supplementation on the immune function of blood monocytes from healthy male nonsmokers. J Lab Clin Med. 1997;129:309-317. doi: 10.1016/s0022-2143(97)90179-7

[29] Jee J, Hoet AE, Azevedo MP, et al. Effects of dietary vitamin A content on antibody responses of feedlot calves inoculated intramuscularly with an inactivated bovine coronavirus vaccine. Am J Vet Res. 2013; 74:1353-1362. doi: 10.2460/ajvr.74.10.1353

[30] Cui D, Moldoveanu Z, Stephensen CB. High-level dietary vitamin A enhances T-helper type 2 cytokine production and secretory immunoglobulin A response to influenza A virus infection in BALB/c mice. J Nutr. 2000;130:1132-1139. doi: 10.1093/jn/130.5.1132

[31] Lin KH, Lin KC, Lu WJ, Thomas PA, Jayakumar T, Sheu JR. 2015;17:44. doi: 10.3390/ijms17010044

[32] Dharra R, Kumar Sharma A, Datta S. Emerging aspects of cytokine storm in COVID-19: The role of proinflammatory cytokines and therapeutic prospects. Cytokine. 2023;169:156287. doi: 10.1016/j.cyto.2023.156287

[33] Thakur A, Pedersen LE, Jungersen G. Immune markers and correlates of protection for vaccine induced immune responses. Vaccine. 2012; 30:4907-4920. doi: 10.1016/j.vaccine.2012.05.049

[34] Sato Y, Akiyama H, Suganuma H, et al. The feeding of beta-carotene down-regulates serum IgE levels and inhibits the type I allergic response in mice. Biol Pharm Bull. 2004; 27:978-984. doi: 10.1248/bpb.27.978

[35] Hiippala K, Barreto G, Burrello C, et al. Novel odoribacter splanchnicus strain and its outer membrane vesicles exert immunoregulatory effects in vitro. Front Microbiol. 2020;11:575455. doi: 10.3389/fmicb.2020.575455

[36] Kim CC, Lunken GR, Kelly WJ, et al. Genomic insights from Monoglobus pectinilyticus: A pectin-degrading specialist bacterium in the human colon. ISME J. 2019;13:1437-1456. doi: 10.1038/s41396-019-0363-6

[37] Kim YJ, Jung DH, Park CS. Important roles of Ruminococcaceae in the human intestine for resistant starch utilization. Food Sci Biotechnol. 2024;33:2009-2019. doi: 10.1007/s10068-024-01621-0

[38] Loy A, Pfann C, Steinberger M, et al. Lifestyle and horizontal gene transfer-mediated evolution of Mucispirillum schaedleri, a core member of the murine gut microbiota. mSystems. 2017; 2:e00171-16. doi: 10.1128/msystems.00171-16

[39] Shi E, Nie M, Wang X, et al. Polysaccharides affect the utilization of β-carotene through gut microbiota investigated by in vitro and in vivo experiments. Food Res Int. 2023;174:113592. doi: 10.1016/j.foodres.2023.113592