Cyclophosphamide-induced Immunosuppressed Mice Research Articles

Immunostimulatory activity of sea buckthorn polysaccharides via TLR2/4-mediated MAPK and NF-κB signaling pathways in vitro and in vivo

SP0.1–1, derived from Sea buckthorn (Hippophae rhamnoides L.), has been discovered to exhibit unique antioxidant activity. In this study, we investigated the immunomodulatory activity and mechanisms of SP0.1–1 on macrophage RAW 264.7 cells in vitro and immunosuppressive mice induced by cyclophosphamide in vivo. The results indicated SP0.1–1 strengthened the immune functions via promoting the proliferation of RAW264.7 cells and phagocytic activity, along with stimulating the release of NO, ROS and cytokines including TNF-α, IL-6, IL-1β and IFN-γ. Western blot and molecular docking analysis demonstrated that SP0.1–1 attached to the prime receptors TLR2 and TLR4 in RAW264.7 cells, and triggered the activation of MyD88-mediated MAPK and NF-κB signaling pathways, thereby exerting the immune response in RAW264.7 cells. However, the intervention of specific inhibitors against TLR2, TLR4, JNK, ERK, p38 and NF-κB blocked the TLR-mediated MAPK and NF-κB signaling pathways and downregulated the levels of NO and the aforementioned cytokines, thus suppressing the activation of macrophages. Therefore, it can be speculated that SP0.1–1 activated the macrophages principally via the TLR2/4-MyD88-mediated MAPK and NF-κB signaling pathways. Additionally, SP0.1–1 could protect against the cyclophosphamide-induced immunosuppression in mice, manifested by the improvement of body weight, immune organ indices, phagocytic index, and the relievement of spleen damage, along with the enhancement of cytokines TNF-α, IL-6, IFN-γ and immunoglobulin IgG and IgM. These findings will shed light on the molecular mechanism of SP0.1–1 on the immunoregulatory effect, and lay the foundation for exploiting a potential immunostimulatory agent of SP0.1–1.

Immunomodulation of exopolysaccharide produced by Lacticaseibacillus rhamnosus ZFM216 in cyclophosphamide-induced immunosuppressed mice by modulating gut microbiota

This study investigated the immunoregulatory activity of exopolysaccharides (EPS) produced by Lacticaseibacillus rhamnosus ZFM216 in immunosuppressed mice induced by cyclophosphamide (CTX). The results showed that EPS treatment effectively improved the body weight, immune organ index and splenic lymphocyte proliferation. EPS also mitigated the damage of immune organs, restored intestinal morphology, and regulated the levels of serum hemolysin and cytokines (e.g. TNF-α, INF-γ and IL-10). EPS promoted the release of NO, TNF-α, IL-1β, and IL-6 in RAW 264.7 cells, however, such effect was inhibited in the presence of inhibitors of TLR4 and MAPKs signaling pathways-related proteins, confirming that EPS achieved the immunomodulation by activating these two signaling pathways. Additionally, EPS, as a prebiotic, effectively improved the diversity of microbial communities, regulated the relative abundance of dominant microbial communities, restored CTX-induced gut microbiota dysbiosis, and promoted the production of short chain fatty acids (SCFAs) in the gut of mice. Thus, immunoregulatory effect of EPS could be attributed to its good ability to modulate the gut microbiota. EPS produced by L. rhamnosus ZFM216 has promising application as an ingredient of functional foods due to its potent probiotic effect.

Brassica rapa L. (Tibetan Turnip) polysaccharide improves the immune function and regulates intestinal microbiota in immunosuppressive mice.

In this paper, active polysaccharides were extracted from Brassica rapa L. polysaccharide (BRP), and structural characterization was preliminarily investigated. Its immunomodulatory activity and molecular biological mechanisms in cyclophosphamide-induced immunosuppressed mice were also explored, as well as its effects on intestinal microbiota. Results indicate that BRP is an acidic heteropolysaccharide with the main components of Ara, GalA, and GlcA and has α- and β-glycosidic linkages with pyranose bonds. The results of the study showed that BRP could effectively improve the thymus and spleen indices and repair Cy-induced immune tissue damage in immunosuppressed mice. Meanwhile, BRP increased the immune cell activity and antioxidant levels in mice. In addition, BRP increased the secretion of cytokines (IL-1β, IL-6, IL-10, and TNF-α) and immunoglobulins (IgA, IgG) in mouse serum. It also regulates the relative expression of genes related to the TLR4/NF-κB signaling pathways as well as regulates the diversity and composition of mouse intestinal microbiota. In conclusion, BRP was able to regulated the immune function in immunosuppressed mice, providing a theoretical basis for the development of immunomodulators.

A novel exosome-like nanovesicles from Cordyceps militaris potentiate immunomodulatory and anti-tumor effect by reprogramming macrophages

AimsFungi-derived exosome-like nanovesicles (ENs) are emerging as a highly promising class of nanoparticles, particularly noted for their cost-effective production. However, their impact on immune regulation and their potential as anti-tumor agents need further exploration. Our study specifically focused on the investigation of the immunomodulatory and anti-tumor properties of ENs derived from Cordyceps militaris, an edible fungus that had achieved large-scale commercial production, referred to as CMDENs. Main methodsThe ENs of C. militaris were collected through ultra-high-speed centrifugation, followed by characterization of their physicochemical properties and contents. Subsequently, the biological distribution of these vesicles was investigated using in vivo fluorescence imaging experiments. Finally, the immune activation and polarization of macrophages were examined through both in vitro and in vivo experiments. Key findingsHerein, we presented the discovery of CMDENs that were rich in proteins, lipids, flavonoids and alkaloids. Immunomodulatory experiments conducted in vivo demonstrated that CMDENs exhibited protective effects against cyclophosphamide-induced immunosuppression in mice by significantly enhancing macrophage phagocytosis and peripheral blood immune cell counts. Moreover, CMDENs effectively induced the polarization of M0- and M2-like macrophages toward M1-like phenotype by activating MAPKs signaling pathway. Notably, CMDENs exhibited remarkable capabilities in inhibiting tumor growth by reprogramming tumor-associated macrophages and activating tumor-infiltrating T lymphocytes, without any observed toxicity in mice bearing tumors. SignificanceOur research suggested that CMDENs possessed the potential to be explored as a nano-immunomodulatory agent for cancer.

Low molecular weight polysaccharide of Tremella fuciformis exhibits stronger antioxidant and immunomodulatory activities than high molecular weight polysaccharide

Low molecular weight polysaccharides had higher bio-activity and bioavailability compared to ultra-high molecular weight polysaccharides, this study aimed to obtain low molecular weight polysaccharides from Tremella fuciformis (TFLP) by using high-temperature and high-pressure assisted hydrochloric acid method to degrade Tremella fuciformis polysaccharides (TFP), and the structural characteristics, in vivo antioxidant and immune enhancing activities of TFP and TFLP was explored through Caenorhabditis elegans (C. elegans) and mice model. It was found that TFP and TFLP were acidic polysaccharides with molecular weights of 2238 kDa and 3 kDa, respectively. The glycosidic bonding of TFP and TFLP was mainly composed of different configurations of mannopyranose. TFP and TFLP had excellent in vivo antioxidant activity and stress resistance by regulating the mRNA transcription level and metabolites in C. elegans. Results also showed that TFP and TFLP could enhance the antioxidant capacity and immunity of serum, spleen and small intestine tissues in normal mice and cyclophosphamide-induced immunosuppressive mice through regulating the relative transcription and expression levels of anti-inflammatory related signaling factors, and it has found that TFLP showed better immune enhancement and antioxidant activity than TFP. In addition, Akkermansia, Bacteroides and Alloprevotella were characteristic bacteria at the genus level in immunosuppressed mice intervened with TFLP, with a significant increase in relative abundance. The content of SCFAs significantly increased in immunosuppressed mice by TFLP. These results indicated that TFP and TFLP had potential in vivo antioxidant and immune enhancing activities.

Serum metabolism-transcriptomics investigated into the immunity of whey protein isolate-galacto-oligosaccharide conjugates after dynamic high-pressure microfluidics pretreatment

The objective of this study was to investigate the immunomodulatory effects of whey protein isolate (WPI)-galacto-oligosaccharide conjugates following dynamic high-pressure microfluidics pretreatment (DHPM) in cyclophosphamide-induced immunosuppressed mice. DHPM facilitated the conjugation of WPI and galacto-oligosaccharide, and inhibited the generation of fluorescent advanced glycation end products (AGEs) and pentosidine. The conjugates demonstrated a significant immune recovery effect on CTX-induced immunosuppressed mice, as evidenced by the enhancement of IgG antibody levels (from 3.5 to 4.1) and the reduction of the levels of immunosuppressive effector factors TGF-β (from 148.1 to 111.2) and IFN-γ (from 34.4 to 17.9). Furthermore, the conjugates exhibited a notable ability to repair histological lesion in the spleen of CTX-induced immunosuppressed mice. Spleen transcriptomics revealed that the Marco, Klrc3 and Cd209b genes were associated with the immune enhancement activity of the conjugates. Metabolomic analysis identified arginine biosynthesis, sphingolipid metabolism, alanine, aspartate and glutamate metabolism, and phenylalanine, tyrosine and tryptophan biosynthesis as key pathways in the immune enhancement activity of the conjugates. Metabolomics combined with transcriptomics indicated the importance of macrophage activation in the restoration of immunosuppressed mice’s immunity by the conjugates. Therefore, the improvement in immunity observed with WPI-galacto-oligosaccharide conjugates may be related to the activation of macrophages.

A Hirsutella sinensis Alcohol Extract Exerts Bidirectional Immunoregulatory Effects by Regulating Macrophage Polarization.

For background, Hirsutella sinensis, the only anamorphic fungus considered an effective substitute for Cordyceps sinensis, possesses immunoregulatory properties. However, the specific mechanism underlying the immunoregulatory function of Hirsutella sinensis remains unclear. The purpose is to investigate the therapeutic effects of Hirsutella sinensis alcohol extract (HSAE) on immune dysregulation and elucidate the underlying mechanisms involved. For methods, we established inflammatory and immunosuppression models in vitro and in vivo to evaluate the bidirectional immunoregulatory function of HSAE via qRT-PCR and immunoblotting. We also studied its potential mechanism via RNA sequencing and transcriptional analysis. We further established M1 and M2 cell models to explore the effect of HSAE on M1/M2 polarization using qRT-PCR, immunoblotting, and flow cytometry. For results, our data demonstrated enhanced proliferation, phagocytosis, and antipathogenic activities of macrophages. Treatment with HSAE led to increases in the proportions of CD3+ and CD4+ immune cells in cyclophosphamide-induced immunosuppressed mice. Additionally, HSAE reduced the lipopolysaccharide (LPS)-induced expression of Il1b, Il6, Ifnb1, and Cxcl10 by inhibiting the activation of the NF-κB and MAPK pathways in vitro and improved mouse survival by reducing the proportion of M1/M2 macrophages in septic mice. Finally, we found that HSAE inhibited M1 polarization by decreasing the expression of iNOS and CD86 and promoted M2 polarization by increasing the expression of ARG1 and CD206. For conclusions, our study provides evidence that HSAE has the potential to enhance immune responses and suppress excessive inflammation. These effects were realized by modulating macrophage polarization, providing novel insights into the fundamental mechanism underlying the bidirectional immunomodulatory effect of HSAE.

Immunomodulatory effects and mechanisms research of deer antler water extract (DAWE) on cyclophosphamide-induced immunosuppressive mice based on metabolomics and microbiomics

Deer antler has been a staple food for millennia, yet its precise immunomodulatory mechanisms remain unexplored. In our study, we systematically investigated the effects and mechanisms of deer antler water extract (DAWE) on cyclophosphamide-induced immunosuppressive mice. We identified various bioactive components in DAWE, including monosaccharides, nucleosides, phospholipids, amino acids, lipids, and organic acids. Following 14 days of administration of DAWE (260 mg/kg) significantly enhanced immunity in immunosuppressed mice, alleviating spleen injury, boosting immune cell proliferation, and increasing immunoactive substances production. Moreover, DAWE effectively mitigated intestinal barrier damage by promoting goblet cell and IgA-secreting cell proliferation and upregulating the expression of occludin-1 and claudin-1. Additionally, DAWE modulated gut microbiota diversity, composition, and metabolic activity. The metabonomic analysis highlighted the pivotal role of glutamine and glutamate metabolic pathways in DAWE's immune regulatory effects. These findings provide valuable insights into DAWE as a potential immunomodulator and functional food, warranting further exploration in clinical applications.

The comprehensive analysis of gut microbiome and spleen transcriptome revealed the immunomodulatory mechanism of Dendrobium officinale leaf polysaccharide on immunosuppressed mice

In recent years, the immunomodulatory efficacy of Dendrobium officinale leaf polysaccharide (DOLP) has attracted much attention, but its potential immunomodulatory mechanism remains unclear. Therefore, we investigated the molecular mechanism of DOLP to ameliorate cyclophosphamide-induced immunosuppressed mice based on transcriptome profiling technology. The results indicated that DOLP significantly mitigated damage to immune organs, regulated the expression levels of inflammatory factors and immunoglobulins, and restored the balance of gut microbiota. Furthermore, it modulated metabolic pathways associated with the immune system, including antigen processing and presentation, hematopoietic cell line development, and natural killer cell-mediated cytotoxicity. DOLP might promote host hematopoietic function to enhance immune cell proliferation and differentiation by up-regulating Cd19, Cr2 and Il7r but down-regulating Dntt. DOLP also up-regulated the expression of MHC-1 (Gm11127, H2-K1, H2-Q10, H2-Q6, and H2-Q7), thus promoting antigen recognition by NK cells to enhance the innate immunity and helping T cells to deliver antigen and secrete immune factors so that enhancing the adaptive immunity.

Enhancing immunomodulation in cyclophosphamide-induced immunosuppressed mice through targeted modulation of butyrate-producing gut microbiota via oral administration of astragalus polysaccharides.

Astragalus polysaccharide is one of the most extensively studied traditional Chinese medicinal polysaccharides because of its immunomodulatory activity and has attracted considerable attention. Existing evidence suggests that its potential immunomodulatory mechanism is related to the modulation of intestinal microbiota. However, current research methods on the gut microbiota mainly focus on 16S rRNA sequencing, providing limited evidence of specific changes in functional bacterial groups in the intestine. Butyrate is a class of short-chain fatty acids among the microbial metabolites in the gut and is most closely associated with immunomodulatory activity. Thus, in this study, we extracted and purified a polysaccharide from astragalus composed of a main chain of →4)-α-D-Glcp-(1 → and →4,6)-α-D-Glcp-(1→, with side chains of →6)-α-D-Glcp-(1→ and aggregated arabinose, and investigated the changes in butyrate-producing bacterial groups in mice during the immunomodulation process of astragalus polysaccharide, using two butyrate-producing bacterial-specific primers. The results showed that oral administration of astragalus polysaccharide significantly increased butyrate production in the mouse intestine, restoring the disrupted butyrate-producing bacterial abundance and diversity caused by immunosuppression. In conclusion, our study provides the first evidence of the targeted modulation of the butyrate-producing gut microbiota by astragalus polysaccharide, offering insights into its pharmacological activity.

Immunostimulating effects of ulvan type polysaccharide isolated from Korean Ulva pertusa in cyclophosphamide-induced immunosuppressed BALB/c mice

This study aimed to determine the immunostimulatory activities of ulvan type polysaccharides isolated from Ulva pertusa. First, U. pertusa polysaccharide (UPP) mainly consists of rhamnose, glucuronic acid, iduronic acid, and xylose, which are typical ulvan type monosaccharides. UPP induced phosphorylation of the mitogen-activated protein kinase and nuclear factor-kappa B pathways in macrophages, subsequently triggering cytokine release and phagocytosis. The effects were closely associated with pattern recognition receptors such as dectin-1, mannose receptor, CD11b, CD14, and Toll-like receptors 2 and 4. Moreover, prophylactic administration of UPP was found to protect against body weight loss and lymphatic organ damage in cyclophosphamide-induced immunosuppressed mice. In addition, UPP demonstrated significant stimulatory effects on various immunocytes, such as T cells, B cells, macrophages, and natural killer cells derived from the spleen. These effects were closely related to the mitogen-activated protein kinase and nuclear factor-kappa B pathways, and significant secretion of immunostimulatory cytokines such as IL-6, -12, and TNF-α was noted in both blood and spleen samples. Impairment of the short-chain fatty acid balance in the cecum was prevented by UPP administration in a dose-dependent manner. Consequently, these results suggest that the UPP isolated from U. pertusa contributes to immune system activation.

Protective effects of Bacillus amyloliquefaciens-derived nonapeptide in cyclophosphamide-induced immunosuppressed mice

This study aims to explore the effect of Bacillus amyloliquefaciens-derived nonapeptide (BAP) on growth and immune function in cyclophosphamide (Cy)-induced immunosuppressed mice. The results showed that BAP alleviated the reduction of the body weight, daily feed intake and spleen and thymus index caused by Cy, and restored jejunal tissue morphology, goblet cell numbers and tight junction proteins. BAP enhanced the expression of immunoglobulins, including immunoglobulin M (IgM), immunoglobulin G (IgG) and secretory immunoglobulin A (SIgA), jejunal CD3+, CD4+ and CD8+ lymphocytes, and cytokines including interleukin-2 (IL-2) and interferon-γ (IFN-γ), while it inhibited the expression of interleukin-4 (IL-4) and interleukin-10 (IL-10). The oxidative stress and decreased antioxidant enzymes induced by Cy were also reversed by BAP. Moreover, BAP modulated the structure of gut microbiota and promoted the colonization of potential intestinal probiotics. Our findings establish the groundwork for comprehending the function of BAP and its potential utilization as an immunomodulator.

Astragalus membranaceus (Fisch.) Bge. administered by dissolving microneedles achieves systemic therapeutic effects at low doses

ObjectiveTo determine the main components of Astragalus membranaceus (Fisch.) Bge (A. membranaceus, Huang Qi), Astragaloside IV (AIV) and Astragalus polysaccharides (AP), to characterize their properties, evaluate their in vivo efficacy, and to analyze drug diffusion using dissolving microneedle (DMN) technology in vivo. MethodsRespectively, AIV- and AP-loaded DMNs comprising chitosan (CTS) and polyvinyl alcohol (PVA) were prepared via dual-mold forming. Their morphology, mechanical properties, in vivo solubility, and skin irritation characteristics were tested. In vivo efficacy was assessed in cyclophosphamide-induced immunosuppressed mice, in vivo diffusion of AIV and AP by DMNs and conventional methods was compared, and the rheological properties of AIV-CTS-PVA and AP-CTS-PVA mixtures were measured. ResultsSubcutaneous dissolution and absorption of AIV-CTS-PVA and AP-CTS-PVA microneedles (MNs) at low doses (50%–17% of intraperitoneal AIV injection and 12%–4% of intravenous AP injection) reduced the spleen index and acid phosphatase activity in immunosuppressed mouse models, increased the thymus index, and achieved equivalent or better systemic therapeutic effects. Compared with injections, AIV and AP achieved controllable solid-liquid conversion through delivery with CTS-PVA MNs, resulting in highly localized aggregation within 48 h, reducing the initial explosive effect of the drug, and achieving stable and slow drug release. ConclusionThe present study enhances our understanding of the efficacy and remote effects of drug-loaded DMNs from a traditional Chinese medicine (TCM) perspective, thereby promoting the development of precise and efficient delivery of TCM and further expanding the drug-loading range and application scenarios for DMNs.

Immunostimulatory Effects of Korean Mineral-Rich Seawaters on Cyclophosphamide-Induced Immunosuppression in Mice.

Deep seawater (DS), obtained from a depth over 200 m, has health benefits due to its rich nutrients and minerals, and intake of DS has shown diverse immunomodulatory effects in allergies and cancer. Therefore, the immunostimulatory effects of Korean mineral-rich seawaters were examined in a cyclophosphamide (CPA)-induced immunosuppression model. Three samples of Korean seawater, namely DS from the East Sea off the coasts of Pohang (PDS) and Uljin (UDS), and seawater from the West Sea off the coast of Boryeong (BS), were collected. The seawaters were abundant in several minerals (calcium, iron, zinc, selenium, etc.). Mice were orally administered the seawaters for 42 days, followed by CPA-induced immunosuppression. The CPA induction reduced the weight of the spleen and lymph nodes; however, the administration of seawaters increased the weight of the lymphoid organs, accompanied by stimulation of natural killer cells' activity and NF-kB-mediated cytokine production (IFNγ, TNFα, IL1β, IL6, and IL12). The mouse-derived splenocytes showed lymphoproliferation without cytotoxicity in the seawater groups. Histopathological analysis revealed that the seawaters improved the CPA-induced atrophic changes by promoting lymphoproliferation in the spleen and lymph nodes. These results provide useful information for the use of Korean mineral-rich seawaters, particularly PDS and UDS, as alternative immunostimulants under immunosuppressive conditions.

Yogurt Alleviates Cyclophosphamide-Induced Immunosuppression in Mice through D-Lactate.

Numerous studies have investigated the immunomodulatory effects of yogurt, but the underlying mechanism remained elusive. This study aimed to elucidate the alleviating properties of yogurt on immunosuppression and proposed the underlying mechanism was related to the metabolite D-lactate. In the healthy mice, we validated the safety of daily yogurt consumption (600 μL) or D-lactate (300 mg/kg). In immunosuppressed mice induced by cyclophosphamide (CTX), we evaluated the immune regulation of yogurt and D-lactate. The result showed that yogurt restored body weight, boosted immune organ index, repaired splenic tissue, recovered the severity of delayed-type hypersensitivity reactions and increased serum cytokines (IgA, IgG, IL-6, IFN-γ). Additionally, yogurt enhanced intestinal immune function by restoring the intestinal barrier and upregulating the abundance of Bifidobacterium and Lactobacillus. Further studies showed that D-lactate alleviated immunosuppression in mice mainly by promoting cellular immunity. D-lactate recovered body weight and organ development, elevated serum cytokines (IgA, IgG, IL-6, IFN-γ), enhanced splenic lymphocyte proliferation and increased the mRNA level of T-bet in splenic lymphocyte to bolster Th1 differentiation. Finally, CTX is a chemotherapeutic drug, thus, the application of yogurt and D-lactate in the tumor-bearing mouse model was initially explored. The results showed that both yogurt (600 μL) and D-lactate (300 mg/kg) reduced cyclophosphamide-induced immunosuppression without promoting tumor growth. Overall, this study evaluated the safety, immune efficacy and applicability of yogurt and D-lactate in regulating immunosuppression. It emphasized the potential of yogurt as a functional food for immune regulation, with D-lactate playing a crucial role in its immunomodulatory effects.

Polygonatum sibiricum Saponin Prevents Immune Dysfunction and Strengthens Intestinal Mucosal Barrier Function in Cyclophosphamide-Induced Immunosuppressed BALB/c Mice.

The aim of this study was to explore the immunomodulatory effect of Polygonatum sibiricum saponin (PS) in a cyclophosphamide-induced (Cy) immunosuppression mice model. Oral administration of PS by gavage effectively alleviated weight loss caused by Cy and increased the index of immune organs. PS promoted the proliferation of splenic lymphocytes and T cell subsets (CD3+, CD355+, CD4+/CD8+) and relieved the xylene-induced inflammatory response and Cy-induced increase of serum hemolysin. Moreover, PS increased serum levels of lactate dehydrogenase and acid phosphatase. PS elevated serum level of cytokines and immunoglobulins (TNF-α, IFN-γ, IL-4, IL-6, IL-β, SIgA, and IgG) and the expression of mRNA of IL-10, TNF-α, and IL-6 in the spleen. Increased mRNA expression of tight junction protein (ZO-1, Mucin2, Occludin) expression and protein expression of IL-6/MyD88/TLR4 in the small intestine showed that PS exhibited a restorative effect on intestinal mucosal injury caused by cyclophosphamide. Oral PS prevented Cy-induced decline in leukocytes, red blood cells, lymphocytes, hemoglobin concentrations, and neutrophils, providing evidence for alleviating hematopoietic disorders. In addition, PS increased SOD and NO levels, reduced MDA levels, and improved oxidative damage in the liver. These findings demonstrate that PS has the potential to be developed as a supplemental agent for alleviating immunosuppression caused by chemotherapeutic agents.

Immune-boosting effect of Yookgong-dan against cyclophosphamide-induced immunosuppression in mice

Immune responses must be strictly regulated to prevent autoimmune and infectious diseases and to protect against infectious agents. As people age, their immunity wanes, leading to a decrease in lymphocyte production in bone marrow and thymus and a decline in the efficacy of mature lymphocytes in secondary lymphoid organs. This study explores the immune-boosting potential of Yookgong-dan (YGD) in enhancing the immune system by activating immune cells. In our in vitro experiments, cyclophosphamide (Cy) treatment led to a significant decrease in primary splenocyte viability. However, subsequent treatment with YGD significantly improved cell viability, with doses ranging between 1 and 25 μg/mL in Cy-treated splenocytes. Flow cytometry analysis demonstrated that the Cy group exhibited reduced positivity of CD3+ T cells and CD45+ leukocytes compared to the blank group. In contrast, treatment with YGD led to a notable, dose-responsive increase in these immune cell types. In our in vivo experiments, YGD was orally administered to Cy-induced immunosuppressed mice at 20 and 100 mg/kg doses for 10 days. The results indicated a dose-dependent elevation in immunoglobulin (Ig)G and IgM levels in the serum, emphasizing the immunostimulatory effect of YGD. Furthermore, the Cy-treated group showed decreased T cells, B (CD19+) cells, and leukocytes in the total splenocyte population. Yet, YGD treatment resulted in a dose-dependent reversal of this pattern, suggesting its ability to counter immunosuppression. Notably, YGD was found to effectively stimulate T (CD4+ and CD8+) lymphocyte subsets and natural killer cells, along with enhancing Th1/Th2 cytokines in immunosuppressed conditions. These outcomes correlated with the modulation of BCL-2 and BAX expression, which are critical for apoptosis. In conclusion, YGD has the potential to bolster immune functionality through the activation of immune cells, thereby enhancing the immune system's capacity to combat diseases and improve overall health and wellness.

Immune-enhancing effect of Weizmannia coagulans BCG44 and its supernatant on cyclophosphamide-induced immunosuppressed mice and RAW264.7 cells via the modulation of the gut microbiota.

We established a model of cyclophosphamide (CTX)-induced immunosuppressed mice and RAW264.7 cells to assess the effectiveness of W. coagulans BCG44 and its supernatant in enhancing immune function and modulating the gut microbiota. W. coagulans BCG44 and its supernatant restored Th17/Treg balance and alleviated gut inflammation by elevating the expression of interleukin-10 (IL-10) and decreasing IL-6 and toll-like receptor 4 (TLR4). Meanwhile, W. coagulans BCG44 and its supernatant downregulated the levels of lipopolysaccharide and D-lactic acid while increasing the expression of tight junction proteins (ZO-1 and occludin) and goblet cells/crypts to ameliorate mucosal damage. W. coagulans BCG44 and its supernatant may restore the gut microbiota in the immunosuppressed mice by regulating keystone species (Lactobacillus and Lachnospiraceae). PICRUSt2 function prediction and BugBase analysis showed thatW. coagulansBCG44 and its supernatant significantly down-regulated American trypanosomiasis and potentially_pathogenic. In addition, under normal versus inflamed culture conditions, stimulation of RAW246.7 cells with W. coagulans BCG44 supernatant activated immune response with increasing proliferation ability and the gene expression of IL-10 while decreasing TLR4. Metabolites in the W. coagulans BCG44 supernatant included arginine, tyrosine, solamargine, tryptophan, D-mannose, phenyllactic acid, and arachidonic acid. Collectively, these findings suggested that W. coagulans BCG44 and its supernatant possess potential immunomodulatory activity and modulate gut microbiota dysbiosis in the CTX-induced immunosuppressed mice.

Investigation of immunomodulatory activity of hericium erinaceus on cyclophosphamide-induced immunosuppression in mice

The study was conducted to assess the immunostimulatory effect of Hericium Erinaceus on cyclophosphamide-induced immunodeficient mice. Animals were administered Hericium Erinaceus orally at 0.72 g/kg b.w and 1.44 g/kg b.w for seven consecutive days. On the 4th day, CP (200 mg/kg) was intraperitoneally injected into mice. The results illustrated that Hericium Erinaceus at 0.72 g/kg b.w only posed an immunostimulating potency in the cyclophosphamide-induced immunosuppression model. Besides, Hericium Erinaceus at 1.44 g/kg b.w ameliorated the effects of CP on delayed-type hypersensitivity (DTH) response, relative spleen weight, serum IL-2, TNF-α concentration, and micro-histological images of spleen and thymus substantially. This suggested the potential of Hericium Erinaceus to treat immunosuppression diseases in clinical practice.

Structural characterization and immunomodulatory effect of a starch-like Grifola frondosa polysaccharides on cyclophosphamide-induced immunosuppression in mice

In this study, a pure Grifola frondosa polysaccharide (GFP-1) was extracted and purified from Grifola frondosa. By HPLC, GC-MS, FT-IR, and NMR analysis, GFP-1 was determined to be a starch-like polysaccharide with an average molecular weight of 3370 kDa. It included three monosaccharides, i.e., glucose, galactose, and mannose. The backbone of GFP-1 consisted of →4)-α-Glcp-(1→ and →4,6)-α-Glcp-(1 → . The side branches were composed of →6)-α-Galp-(1→, α-Glcp-(1→, and a small amount of α-Manp-(1 → . By using a cyclophosphamide (CTX)-induced immunosuppressed mice model, we evaluated the immunomodulatory activity of GFP-1. The results showed that GFP-1 increased the thymic and spleen indices, promoted the level of IgG and IgA in serum, and activated the mitogen-activated protein kinase (MAPK) pathway in CTX-induced mice. Also, GFP-1 significantly promoted the mRNA expression of intestinal barrier factors and protected intestinal structural integrity in immunosuppressed mice. In conclusion, the data presented here suggested that GFP-1 might be a potential immune-enhancing supplement.