{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,5]],"date-time":"2026-06-05T02:36:32Z","timestamp":1780626992581,"version":"3.54.1"},"reference-count":54,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,1,23]],"date-time":"2025-01-23T00:00:00Z","timestamp":1737590400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003627","name":"National Institute of Agricultural Sciences, Rural Development Administration, Republic of Korea","doi-asserted-by":"publisher","award":["RS-2022-RD010366"],"award-info":[{"award-number":["RS-2022-RD010366"]}],"id":[{"id":"10.13039\/501100003627","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Objectives: The immune-enhancing properties of steamed mature silkworm, known as HongJam (HJ), were investigated using human interleukin-2-independent Natural Killer 92 (NK92-MI) cells and a cyclophosphamide intraperitoneal injection-induced immunosuppressed mice model (CPA-IP). White Jade variety mature silkworm HJ (WJ-HJ) was used to prepare WJ-HJ supercritical fluid extracts (WJ-SCE) and WJ-HJ-supplemented feeds. Results: Treatment with WJ-SCE significantly enhanced proliferation, migration, and cytotoxicity of NK92-MI cells against various cancer cells while improving mitochondrial function and ATP production (p &lt; 0.05). In CPA-IP mice, consumption of WJ-HJ-supplemented feeds restored immune function by improving body weight, immune organ indices, immunoglobulin levels, and blood cytokines. Splenocyte proliferation and cytotoxicity were significantly elevated in both saline intraperitoneal injection (Sal-IP) and CPA-IP groups with WJ-HJ supplementation, independent of mitogen activation (p &lt; 0.05). Conclusions: These results suggest that WJ-HJ enhances immune modulation and immune surveillance functions of NK cells by improving mitochondrial and cytotoxic functions. WJ-HJ holds promise as a functional food for immune enhancement, pending clinical validation.<\/jats:p>","DOI":"10.3390\/nu17030417","type":"journal-article","created":{"date-parts":[[2025,1,23]],"date-time":"2025-01-23T12:24:09Z","timestamp":1737635049000},"page":"417","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Enhanced Immune Functions of In Vitro Human Natural Killer Cells and Splenocytes in Immunosuppressed Mice Supplemented with Mature Silkworm Products"],"prefix":"10.3390","volume":"17","author":[{"given":"Thanh Thi Tam","family":"Nguyen","sequence":"first","affiliation":[{"name":"Department of Biomedical Gerontology, Hallym University Graduate School, Chuncheon 24252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Byungki","family":"Jang","sequence":"additional","affiliation":[{"name":"Ilsong Institute of Life Science, Hallym University, Seoul 07441, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seong-Ruyl","family":"Kim","sequence":"additional","affiliation":[{"name":"Division of Industrial Insects and Sericulture, National Institute of Agricultural Sciences, Wanju 55365, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sang-Kuk","family":"Kang","sequence":"additional","affiliation":[{"name":"Division of Industrial Insects and Sericulture, National Institute of Agricultural Sciences, Wanju 55365, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kee-Young","family":"Kim","sequence":"additional","affiliation":[{"name":"Division of Industrial Insects and Sericulture, National Institute of Agricultural Sciences, Wanju 55365, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoo Hee","family":"Kim","sequence":"additional","affiliation":[{"name":"Ilsong Institute of Life Science, Hallym University, Seoul 07441, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3608-4376","authenticated-orcid":false,"given":"Young Ho","family":"Koh","sequence":"additional","affiliation":[{"name":"Department of Biomedical Gerontology, Hallym University Graduate School, Chuncheon 24252, Republic of Korea"},{"name":"Ilsong Institute of Life Science, Hallym University, Seoul 07441, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,1,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Kim, J.-S., Lee, E.-B., Choi, J.-H., Jung, J., Jeong, U.-Y., Bae, U.-J., Jang, H.-H., Park, S.-Y., Cha, Y.-S., and Lee, S.-H. (2023). Antioxidant and immune stimulating effects of Allium cepa skin in the RAW 264.7 cells and in the C57BL\/6 mouse immunosuppressed by cyclophosphamide. Antioxidants, 12.","DOI":"10.3390\/antiox12040892"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"S3","DOI":"10.1016\/j.jaci.2009.12.980","article-title":"Overview of the Immune Response","volume":"125","author":"Chaplin","year":"2010","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1007\/978-981-10-5987-2_11","article-title":"The development and diversity of ILCs, NK cells and their relevance in health and diseases","volume":"Volume 1024","author":"Xu","year":"2017","journal-title":"Regulation of Inflammatory Signaling in Health and Disease"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1016\/j.ceb.2008.05.006","article-title":"The killer\u2019s kiss: The many functions of NK cell immunological synapses","volume":"20","author":"Krzewski","year":"2008","journal-title":"Curr. Opin. Cell Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1051","DOI":"10.4161\/cbt.3.11.1173","article-title":"Death receptor signals to the mitochondria","volume":"3","year":"2004","journal-title":"Cancer Biol. Ther."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1016\/j.molimm.2004.07.034","article-title":"Activation of NK cell cytotoxicity","volume":"42","author":"Smyth","year":"2005","journal-title":"Mol. Immunol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3231","DOI":"10.4049\/jimmunol.1701500","article-title":"Single degranulations in Natural Killer cells can mediate target cell killing","volume":"200","author":"Gwalani","year":"2018","journal-title":"J. Immunol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Bhat, R., and Watzl, C. (2007). Serial killing of tumor cells by human Natural Killer cells\u2014Enhancement by therapeutic antibodies. PLoS ONE, 2.","DOI":"10.1371\/journal.pone.0000326"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"7922","DOI":"10.1128\/JVI.00199-15","article-title":"Cytokine-Mediated Activation of NK Cells during Viral Infection","volume":"89","author":"Freeman","year":"2015","journal-title":"J. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2167","DOI":"10.1182\/blood-2009-08-238469","article-title":"Regulation of human NK-cell cytokine and chemokine production by target cell recognition","volume":"115","author":"Fauriat","year":"2010","journal-title":"Blood"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/S0733-8635(18)30284-5","article-title":"Immunosuppressive agents in dermatology","volume":"11","author":"Ho","year":"1993","journal-title":"Dermatol. Clin."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"30","DOI":"10.4103\/0974-8490.105646","article-title":"Protective effects of Picrorhiza kurroa on cyclophosphamide-induced immunosuppression in mice","volume":"5","author":"Hussain","year":"2013","journal-title":"Pharmacogn. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1620\/tjem.162.147","article-title":"Differential effects of a single dose of cyclophosphamide on T cell subsets of the thymus and spleen in mice: Flow cytofluorometry analysis","volume":"162","author":"Miyauchi","year":"1990","journal-title":"Tohoku J. Exp. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2360","DOI":"10.1182\/blood.V83.8.2360.2360","article-title":"Effect of total body irradiation, busulfan-cyclophosphamide, or cyclophosphamide conditioning on inflammatory cytokine release and development of acute and chronic graft-versus-host disease in H-2-incompatible transplanted SCID mice","volume":"83","author":"Xun","year":"1994","journal-title":"Blood"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"103886","DOI":"10.1016\/j.jff.2020.103886","article-title":"Mature silkworm powders ameliorated scopolamine-induced amnesia by enhancing mitochondrial functions in the brains of mice","volume":"67","author":"Nguyen","year":"2020","journal-title":"J. Funct. Foods"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"657","DOI":"10.1007\/s10068-022-01090-3","article-title":"Sericulture and the edible-insect industry can help humanity survive: Insects are more than just bugs, food, or feed","volume":"31","author":"Park","year":"2022","journal-title":"Food Sci. Biotechnol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"83","DOI":"10.5625\/lar.2012.28.2.83","article-title":"Effects of male silkworm pupa powder on the erectile dysfunction by chronic ethanol consumption in rats","volume":"28","author":"Oh","year":"2012","journal-title":"Lab. Anim. Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"635","DOI":"10.3892\/etm.2022.11572","article-title":"Effects of Brain Factor-7\u00ae against motor deficit and oxidative stress in a mouse model of MPTP-induced Parkinson\u2019s disease","volume":"24","author":"Lee","year":"2022","journal-title":"Exp. Ther. Med."},{"key":"ref_19","first-page":"38","article-title":"Development of processing technology for edible mature silkworm","volume":"53","author":"Ji","year":"2015","journal-title":"J. Seric. Entomol. Sci."},{"key":"ref_20","first-page":"675","article-title":"New processing technology for steamed-mature silkworms (HongJam) to reduce production costs: Employing a high-speed homogenization and spray drying protocol","volume":"61","author":"Kim","year":"2022","journal-title":"Korean J. Appl. Entomol."},{"key":"ref_21","first-page":"59","article-title":"The molecular metabolism of the key ingredients in the steamed and freeze-dried mature silkworm powder: Effects and mechanisms","volume":"48","author":"Kim","year":"2024","journal-title":"Int. J. Indust. Entomol. Biomater."},{"key":"ref_22","first-page":"84","article-title":"A comparative study of nutrient compositions between HongJams prepared from 5 silkworm varieties making white cocoons","volume":"45","author":"Kim","year":"2022","journal-title":"Int. J. Indust. Entomol. Biomater."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"969","DOI":"10.1016\/j.aspen.2019.07.009","article-title":"Contents of nutrients in ultra-fine powders of steamed and lyophilized mature silkworms generated by four silkworm varieties","volume":"22","author":"Ji","year":"2019","journal-title":"J. Asia-Pac. Entomol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1007\/978-1-4939-3684-7_6","article-title":"In vitro assessment of human Natural killer cell migration and invasion","volume":"1441","author":"Tomin","year":"2016","journal-title":"Methods Mol. Biol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Mai, L.X., Kang, S.-K., Jo, Y.-Y., Nguyen, P., Kim, A.-Y., Kim, K.-Y., Kim, N.-S., and Koh, Y.H. (2022). An alkaline protease-digestion of silkworm powder enhances its effects over healthspan, autophagy, and mitochondria function in a rotenone-induced Drosophila Model. Front. Nutr., 9.","DOI":"10.3389\/fnut.2022.808295"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Percie du Sert, N., Hurst, V., Ahluwalia, A., Alam, S., Avey, M.T., Baker, M., Browne, W.J., Clark, A., Cuthill, I.C., and Dirnagl, U. (2020). The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. PLoS Biol., 18.","DOI":"10.1371\/journal.pbio.3000410"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"114520","DOI":"10.1016\/j.jep.2021.114520","article-title":"The additive memory and healthspan enhancement effects by the combined treatment of mature silkworm powders and Korean angelica extracts","volume":"281","author":"Nguyen","year":"2021","journal-title":"J. Ethnopharmacol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Kim, S.H., Kim, M.J., Lim, M., Kim, J., Kim, H., Yun, C.K., Yoo, Y.J., Lee, Y., Min, K., and Choi, Y.S. (2023). Enhancement of the anticancer ability of natural killer cells through allogeneic mitochondrial transfer. Cancers, 15.","DOI":"10.3390\/cancers15123225"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Van Acker, H.H., Capsomidis, A., Smits, E.L., and Van Tendeloo, V.F. (2017). CD56 in the immune system: More than a marker for cytotoxicity?. Front. Immunol., 8.","DOI":"10.3389\/fimmu.2017.00892"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1038\/nri3839","article-title":"Perforin and granzymes: Function, dysfunction and human pathology","volume":"15","author":"Voskoboinik","year":"2015","journal-title":"Nat. Rev. Immunol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1319","DOI":"10.1002\/JLB.MR0718-269R","article-title":"Mechanisms of natural killer cell-mediated cellular cytotoxicity","volume":"105","author":"Prager","year":"2019","journal-title":"J. Leukoc. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1189\/jlb.0611308","article-title":"Natural killer cell-produced IFN-\u03b3 and TNF-\u03b1 induce target cell cytolysis through up-regulation of ICAM-1","volume":"91","author":"Wang","year":"2011","journal-title":"J. Leukoc. Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1038\/sj.emboj.7600019","article-title":"Unravelling natural killer cell function: Triggering and inhibitory human NK receptors","volume":"23","author":"Moretta","year":"2004","journal-title":"EMBO J."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1590\/S1984-82502009000200017","article-title":"Toxic effects of different doses of cyclophosphamide on the reproductive parameters of male mice","volume":"45","author":"Kanno","year":"2009","journal-title":"Braz. J. Pharm. Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"103408","DOI":"10.1016\/j.jff.2019.06.010","article-title":"Immunoenhancement effects of pentadecapeptide derived from Cyclina sinensis on immune-deficient mice induced by Cyclophosphamide","volume":"60","author":"Yu","year":"2019","journal-title":"J. Funct. Foods"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1323","DOI":"10.1016\/S0140-6736(16)32381-9","article-title":"Future life expectancy in 35 industrialised countries: Projections with a Bayesian model ensemble","volume":"389","author":"Kontis","year":"2017","journal-title":"Lancet"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1038\/s41536-021-00169-5","article-title":"Longevity leap: Mind the healthspan gap","volume":"6","author":"Garmany","year":"2021","journal-title":"npj Regen. Med."},{"key":"ref_38","first-page":"22","article-title":"The memory enhancement and healthspan extension effects of HongJam","volume":"5","author":"Koh","year":"2020","journal-title":"Policy Rep. Natl. Inst. Korean Med. Dev."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Cojocaru, K.-A., Luchian, I., Goriuc, A., Antoci, L.-M., Ciobanu, C.-G., Popescu, R., Vlad, C.-E., Blaj, M., and Foia, L.G. (2023). Mitochondrial dysfunction, oxidative stress, and therapeutic strategies in diabetes, obesity, and cardiovascular disease. Antioxidants, 12.","DOI":"10.3390\/antiox12030658"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1038\/s41392-023-01547-9","article-title":"Mitochondrial dynamics in health and disease: Mechanisms and potential targets","volume":"8","author":"Chen","year":"2023","journal-title":"Signal Transduct. Target Ther."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"7471","DOI":"10.1038\/s41467-023-42277-4","article-title":"Inflammatory macrophages reprogram to immunosuppression by reducing mitochondrial translation","volume":"14","author":"Brischetto","year":"2023","journal-title":"Nat. Commun."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1795","DOI":"10.1016\/S0140-6736(00)03231-1","article-title":"Natural cytotoxic activity of peripheral-blood lymphocytes and cancer incidence: An 11-year follow-up study of a general population","volume":"356","author":"Imai","year":"2000","journal-title":"Lancet"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1038\/s41573-019-0052-1","article-title":"NK cells for cancer immunotherapy","volume":"19","author":"Shimasaki","year":"2020","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Liem, N.T., Van Phong, N., Kien, N.T., Anh, B.V., Huyen, T.L., Thao, C.T., Tu, N.D., Hiep, D.T., Hoai Thu, D.T., and Nhung, H.T.M. (2019). Phase I Clinical Trial Using Autologous Ex vivo expanded NK cells and cytotoxic T lymphocytes for cancer treatment in Vietnam. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20133166"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1186\/s13045-020-01014-w","article-title":"NK cell-based cancer immunotherapy: From basic biology to clinical development","volume":"14","author":"Liu","year":"2021","journal-title":"J. Hematol. Oncol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1284","DOI":"10.4049\/jimmunol.126.4.1284","article-title":"Transfer to cyclophosphamide-treated mice of natural killer (NK) cells and in vivo natural reactivity against tumors","volume":"126","author":"Riccardi","year":"1981","journal-title":"J. Immunol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1007\/s002620050607","article-title":"Effects of macrophage-colony-stimulating factor on cyclophosphamide-injected mouse NK1.1+ cell activity","volume":"49","author":"Sakurai","year":"2000","journal-title":"Cancer Immunol. Immunother."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1182\/blood-2017-05-780668","article-title":"NK cell recovery after haploidentical HSCT with posttransplant cyclophosphamide: Dynamics and clinical implications","volume":"131","author":"Russo","year":"2018","journal-title":"Blood"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"e56221","DOI":"10.7554\/eLife.56221","article-title":"Ageing compromises mouse thymus function and remodels epithelial cell differentiation","volume":"9","author":"Morgan","year":"2020","journal-title":"eLife"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"6085","DOI":"10.1126\/sciimmunol.aau6085","article-title":"Structure and function of the immune system in the spleen","volume":"4","author":"Lewis","year":"2019","journal-title":"Sci. Immunol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1007\/s11626-013-9705-2","article-title":"Concanavalin A-mediated T cell proliferation is regulated by herpes virus entry mediator costimulatory molecule","volume":"50","author":"Ando","year":"2014","journal-title":"In Vitro Cell Dev. Biol. Anim."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1016\/S0165-2478(99)00068-1","article-title":"Bacterial lipopolysaccharide induced B cell activation is mediated via a phosphatidylinositol 3-kinase dependent signaling pathway","volume":"69","author":"Venkataraman","year":"1999","journal-title":"Immunol. Lett."},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Qi, Q., Dong, Z., Sun, Y., Li, S., and Zhao, Z. (2018). Protective Effect of Bergenin against Cyclophosphamide-Induced Immunosuppression by Immunomodulatory Effect and Antioxidation in Balb\/c Mice. Molecules, 23.","DOI":"10.3390\/molecules23102668"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"4891399","DOI":"10.1155\/2022\/4891399","article-title":"rhCNB improves cyclophosphamide-induced immunodeficiency in Balb\/c mice","volume":"2022","author":"Zhong","year":"2022","journal-title":"Evid. Based Complement. Altern. Med."}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/17\/3\/417\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T10:34:56Z","timestamp":1759919696000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/17\/3\/417"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,23]]},"references-count":54,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2025,2]]}},"alternative-id":["nu17030417"],"URL":"https:\/\/doi.org\/10.3390\/nu17030417","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,1,23]]}}}