{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,28]],"date-time":"2026-03-28T03:16:34Z","timestamp":1774667794855,"version":"3.50.1"},"reference-count":42,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2025,5,19]],"date-time":"2025-05-19T00:00:00Z","timestamp":1747612800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Small and Medium-sized Enterprises (SMEs) and Startups, Republic of Korea","award":["RS-2023-00283265"],"award-info":[{"award-number":["RS-2023-00283265"]}]},{"name":"Ministry of Small and Medium-sized Enterprises (SMEs) and Startups, Republic of Korea","award":["GCU-202206080001"],"award-info":[{"award-number":["GCU-202206080001"]}]},{"name":"Gachon University Research Fund of 2022","award":["RS-2023-00283265"],"award-info":[{"award-number":["RS-2023-00283265"]}]},{"name":"Gachon University Research Fund of 2022","award":["GCU-202206080001"],"award-info":[{"award-number":["GCU-202206080001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Deficiencies in immune function increase susceptibility to infections and chronic diseases by impairing immune surveillance and tolerance mechanisms, especially in children with immature immune systems. Chronic inflammation associated with immune dysfunction can impair childhood by suppressing the GH\u2013IGF-1. HT042 is composed of Astragalus mongholicus, Eleutherococcus senticosus, and Phlomis umbrosa, which are medicinal herbs that are traditionally utilized in East Asia to promote growth and enhance immune function; thus, HT042 itself holds potential as an immunomodulator. We evaluated the immunomodulatory effects of HT042 in a cyclophosphamide (CYP)-induced immunosuppressed mouse model, as well as in ex vivo primary splenocytes and RAW 264.7 macrophages. HT042 demonstrated remarkable immune-enhancing effects, including the restoration of weight loss and hematological parameters, as well as enhancing NK cell activity. Primary splenocytes treated with HT042 showed increased expression of CD3, CD4, and CD8, along with Th subset transcription factors (T-bet, GATA3, ROR\u03b3t, Foxp3) and corresponding cytokines (IFN-\u03b3, IL-4, IL-17, IL-10). In RAW 264.7 macrophages, HT042 increased nitric oxide production and upregulated NOS2, COX-2, and inflammatory cytokines (IL-6, IL-1\u03b2, TNF-\u03b1). It is noteworthy that HT042 enhances both innate and adaptive immune pathways, particularly via T cell modulation and macrophage activation, as this study is among the first to demonstrate such effects in the context of CYP-induced immunosuppression.<\/jats:p>","DOI":"10.3390\/ijms26104850","type":"journal-article","created":{"date-parts":[[2025,5,19]],"date-time":"2025-05-19T07:34:03Z","timestamp":1747640043000},"page":"4850","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Astragalus Extract Mixture HT042 Reverses Cyclophosphamide-Induced Immunosuppression Through Dual Modulation of Innate and Adaptive Immunity"],"prefix":"10.3390","volume":"26","author":[{"given":"Se-Young","family":"Kim","sequence":"first","affiliation":[{"name":"Korea Institute of Science and Technology for Eastern Medicine (KISTEM) NeuMed Inc., 88 Imun-ro, Dongdaemun-gu, Seoul 02440, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joohee","family":"Son","sequence":"additional","affiliation":[{"name":"Department of Herbal Pharmacology, College of Korean Medicine, Gachon University, 1342 Seongnamdae-ro, Sujeong-gu, Seongnam-si 13120, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Minju","family":"Kim","sequence":"additional","affiliation":[{"name":"Korea Institute of Science and Technology for Eastern Medicine (KISTEM) NeuMed Inc., 88 Imun-ro, Dongdaemun-gu, Seoul 02440, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-8167-049X","authenticated-orcid":false,"given":"Chae Yun","family":"Baek","sequence":"additional","affiliation":[{"name":"Department of Herbal Pharmacology, College of Korean Medicine, Gachon University, 1342 Seongnamdae-ro, Sujeong-gu, Seongnam-si 13120, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mi-Yeon","family":"Kim","sequence":"additional","affiliation":[{"name":"Korea Institute of Science and Technology for Eastern Medicine (KISTEM) NeuMed Inc., 88 Imun-ro, Dongdaemun-gu, Seoul 02440, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ari","family":"Shin","sequence":"additional","affiliation":[{"name":"Department of Herbal Pharmacology, College of Korean Medicine, Kyung Hee University, 26 Kyungheedae-ro, Dongdaemun-gu, Seoul 02447, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1075-0713","authenticated-orcid":false,"given":"Donghun","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Herbal Pharmacology, College of Korean Medicine, Gachon University, 1342 Seongnamdae-ro, Sujeong-gu, Seongnam-si 13120, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hocheol","family":"Kim","sequence":"additional","affiliation":[{"name":"Korea Institute of Science and Technology for Eastern Medicine (KISTEM) NeuMed Inc., 88 Imun-ro, Dongdaemun-gu, Seoul 02440, Republic of Korea"},{"name":"Department of Herbal Pharmacology, College of Korean Medicine, Kyung Hee University, 26 Kyungheedae-ro, Dongdaemun-gu, Seoul 02447, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1016\/j.anai.2021.08.413","article-title":"Secondary immunodeficiencies","volume":"127","author":"Tuano","year":"2021","journal-title":"Ann. Allergy Asthma Immunol."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Gombart, A.F., Pierre, A., and Maggini, S. (2020). A Review of Micronutrients and the Immune System\u2013Working in Harmony to Reduce the Risk of Infection. Nutrients, 12.","DOI":"10.3390\/nu12010236"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.copsyc.2015.03.007","article-title":"Current directions in stress and human immune function","volume":"5","author":"Morey","year":"2015","journal-title":"Curr. Opin. Psychol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1038\/nri2075","article-title":"Innate immunity of the newborn: Basic mechanisms and clinical correlates","volume":"7","author":"Levy","year":"2007","journal-title":"Nat. Rev. Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.cellimm.2007.09.010","article-title":"Regulation of IGF-I function by proinflammatory cytokines: At the interface of immunology and endocrinology","volume":"252","author":"McCusker","year":"2008","journal-title":"Cell. Immunol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3702","DOI":"10.1002\/ptr.7068","article-title":"Potential therapeutic applications of phytoconstituents as immunomodulators: Pre-clinical and clinical evidences","volume":"35","author":"Ali","year":"2021","journal-title":"Phytother. Res."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Mizuno, M., and Minato, K.-I. (2024). Anti-inflammatory and immunomodulatory properties of polysaccharides in mushrooms. Curr. Opin. Biotechnol., 86.","DOI":"10.1016\/j.copbio.2024.103076"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"106076","DOI":"10.1016\/j.phrs.2022.106076","article-title":"Future directions for the discovery of natural product-derived immunomodulating drugs: An IUPHAR positional review","volume":"177","author":"Wainwright","year":"2022","journal-title":"Pharmacol. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1007\/s12272-022-01393-3","article-title":"Astragalus polysaccharide: A review of its immunomodulatory effect","volume":"45","author":"Li","year":"2022","journal-title":"Arch. Pharm. Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"113586","DOI":"10.1016\/j.jep.2020.113586","article-title":"A review of Acanthopanax senticosus (Rupr and Maxim.) harms: From ethnopharmacological use to modern application","volume":"268","author":"Jia","year":"2021","journal-title":"J. Ethnopharmacol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1186\/s13020-019-0250-0","article-title":"A review on the immunomodulatory activity of Acanthopanax senticosus and its active components","volume":"14","author":"Lau","year":"2019","journal-title":"Chin. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1210\/er.2015-1026","article-title":"Growth and the Growth Hormone-Insulin Like Growth Factor 1 Axis in Children with Chronic Inflammation: Current Evidence, Gaps in Knowledge, and Future Directions","volume":"37","author":"Wong","year":"2016","journal-title":"Endocr. Rev."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1479","DOI":"10.1038\/ncomms2484","article-title":"IGF-1 promotes the development and cytotoxic activity of human NK cells","volume":"4","author":"Ni","year":"2013","journal-title":"Nat. Commun."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1124\/pr.109.002469","article-title":"Insulin-Like Growth Factor-I Regulation of Immune Function: A Potential Therapeutic Target in Autoimmune Diseases?","volume":"62","author":"Smith","year":"2010","journal-title":"Pharmacol. Rev."},{"key":"ref_15","first-page":"1085","article-title":"Growth hormone enhances thymic function in HIV-1\u2013infected adults","volume":"118","author":"Napolitano","year":"2008","journal-title":"J. Clin. Investig."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Baek, C.Y., Lee, J., Lee, D., and Kim, H. (2024). Astragalus Extract Mixture HT042 Alleviates Dexamethasone-Induced Bone Growth Retardation in Rat Metatarsal Bones. Nutrients, 16.","DOI":"10.3390\/nu16142333"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"661","DOI":"10.1007\/s00280-016-3152-1","article-title":"The effect of cyclophosphamide on the immune system: Implications for clinical cancer therapy","volume":"78","author":"Ahlmann","year":"2016","journal-title":"Cancer Chemother. Pharmacol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e350","DOI":"10.1097\/HS9.0000000000000350","article-title":"Beyond DNA Damage: Exploring the Immunomodulatory Effects of Cyclophosphamide in Multiple Myeloma","volume":"4","author":"Swan","year":"2020","journal-title":"HemaSphere"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"111889","DOI":"10.1016\/j.ecoenv.2020.111889","article-title":"Cyclophosphamide induced physiological and biochemical changes in mice with an emphasis on sensitivity analysis","volume":"211","author":"Zhang","year":"2021","journal-title":"Ecotoxicol. Environ. Saf."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.micres.2014.11.002","article-title":"Effects of cyclophosphamide on immune system and gut microbiota in mice","volume":"171","author":"Xu","year":"2015","journal-title":"Microbiol. Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"897","DOI":"10.1038\/29795","article-title":"Leptin modulates the T-cell immune response and reverses starvation-induced immunosuppression","volume":"394","author":"Lord","year":"1998","journal-title":"Nature"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"865","DOI":"10.1016\/j.ijbiomac.2018.12.156","article-title":"pH-responsive Astragalus polysaccharides-loaded poly(lactic-co-glycolic acid) nanoparticles and their in vitro immunogenicity","volume":"125","author":"Xu","year":"2019","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1186\/s13020-021-00440-3","article-title":"Correction to: Immunomodulatory effects of a new whole ingredients extract from Astragalus: A combined evaluation on chemistry and pharmacology","volume":"16","author":"Li","year":"2021","journal-title":"Chin. Med."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Zhou, L., Liu, Z., Wang, Z., Yu, S., Long, T., Zhou, X., and Bao, Y. (2017). Astragalus polysaccharides exerts immunomodulatory effects via TLR4-mediated MyD88-dependent signaling pathway in vitro and in vivo. Sci. Rep., 7.","DOI":"10.1038\/srep44822"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1016\/0192-0561(91)90075-I","article-title":"Immunomopharmacological effects of polysaccharides from Acanthopanax senticosus on experimental animals","volume":"13","author":"Shen","year":"1991","journal-title":"Int. J. Immunopharmacol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/j.jep.2004.03.052","article-title":"Anti-metastatic activity of Acanthopanax senticosus extract and its possible immunological mechanism of action","volume":"93","author":"Yoon","year":"2004","journal-title":"J. Ethnopharmacol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"6935802","DOI":"10.1155\/2017\/6935802","article-title":"Astragalus Extract Mixture HT042 Increases Longitudinal Bone Growth Rate by Upregulating Circulatory IGF-1 in Rats","volume":"2017","author":"Lee","year":"2017","journal-title":"Evid.-Based Complement. Altern. Med."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Chen, X., Chen, X., Gao, J., Yang, H., Duan, Y., Feng, Y., He, X., Gong, X., Wang, H., and Wu, X. (2019). Astragaloside III Enhances Anti-Tumor Response of NK Cells by Elevating NKG2D and IFN-\u03b3. Front. Pharmacol., 10.","DOI":"10.3389\/fphar.2019.00898"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1016\/j.jep.2004.08.027","article-title":"Effects of various Eleutherococcus senticosus cortex on swimming time, natural killer activity and corticosterone level in forced swimming stressed mice","volume":"95","author":"Kimura","year":"2004","journal-title":"J. Ethnopharmacol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"855","DOI":"10.1016\/j.cell.2020.07.018","article-title":"Multiple Signaling Roles of CD3\u03b5 and Its Application in CAR-T Cell Therapy","volume":"182","author":"Wu","year":"2020","journal-title":"Cell"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1146\/annurev.immunol.021908.132706","article-title":"T Cell Activation","volume":"27","author":"Koretzky","year":"2009","journal-title":"Annu. Rev. Immunol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Love, P.E., and Hayes, S.M. (2010). ITAM-mediated Signaling by the T-Cell Antigen Receptor. Cold Spring Harb. Perspect. Biol., 2.","DOI":"10.1101\/cshperspect.a002485"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1038\/s41392-023-01471-y","article-title":"T cells in health and disease","volume":"8","author":"Sun","year":"2023","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1038\/cr.2009.138","article-title":"Heterogeneity and plasticity of T helper cells","volume":"20","author":"Zhu","year":"2010","journal-title":"Cell Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"155149","DOI":"10.1016\/j.phymed.2023.155149","article-title":"Phytochemicals as treatment for allergic asthma: Therapeutic effects and mechanisms of action","volume":"122","author":"Jasemi","year":"2024","journal-title":"Phytomedicine"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1438","DOI":"10.1016\/j.ijbiomac.2018.06.153","article-title":"Synthesis and evaluation of a novel water-soluble high Se-enriched Astragalus polysaccharide nanoparticles","volume":"118","author":"Meng","year":"2018","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"650","DOI":"10.1016\/S2095-3119(14)60971-0","article-title":"Effect of Fuzheng Jiedu granule on immunological function level of immune-related cytokines in immune-suppressed mice","volume":"15","author":"Yu","year":"2016","journal-title":"J. Integr. Agric."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"303","DOI":"10.4103\/0976-500X.119726","article-title":"How to calculate sample size in animal studies?","volume":"4","author":"Charan","year":"2013","journal-title":"J. Pharmacol. Pharmacother."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Lee, D., Kim, B.H., Lee, S.H., Cho, W.Y., Kim, Y.S., and Kim, H. (2021). Effects of Astragalus Extract Mixture HT042 on Circulating IGF-1 Level and Growth Hormone Axis in Rats. Children, 8.","DOI":"10.3390\/children8110975"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1002\/ptr.5886","article-title":"Effects of Astragalus Extract Mixture HT042 on Height Growth in Children with Mild Short Stature: A Multicenter Randomized Controlled Trial","volume":"32","author":"Lee","year":"2018","journal-title":"Phytother. Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"27","DOI":"10.4103\/0976-0105.177703","article-title":"A simple practice guide for dose conversion between animals and human","volume":"7","author":"Nair","year":"2016","journal-title":"J. Basic Clin. Pharma"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.phymed.2017.03.005","article-title":"Safety evaluation of Astragalus extract mixture HT042 and its constituent herbs in Sprague\u2013Dawley rats","volume":"32","author":"Song","year":"2017","journal-title":"Phytomedicine"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/10\/4850\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:35:05Z","timestamp":1760031305000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/10\/4850"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,5,19]]},"references-count":42,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2025,5]]}},"alternative-id":["ijms26104850"],"URL":"https:\/\/doi.org\/10.3390\/ijms26104850","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,5,19]]}}}