Immunomodulatory activity of argentatins A and B isolated from guayule

Aniela M. Silva-Nolasco; Miguel A. de la Cruz-Morcillo; M. Mercedes García-Martínez; Amaya Zalacain; Beatriz G. Gálvez; Manuel Carmona

doi:10.1371/journal.pone.0304713

Abstract

Argentatins are secondary metabolites synthesized by guayule (Parthenium argentatum A. Gray) with numerous potential medical applications. In addition to inhibiting insect growth, they are endowed with several pharmacological properties including antimicrobial and antitumorigenic activity. However, their potential as immunomodulators remains unexplored. The aim of the present study was to investigate whether argentatins can modulate the function of the immune system. Human mesenchymal stem cells were treated with argentatins and the production of several anti- and proinflammatory cytokines was evaluated. The effect of argentatins on the polarization of CD4+ T-lymphocytes and macrophages was also assessed. Results demonstrated that argentatins can modulate the production of proinflammatory cytokines and the polarization of cellular phenotypes, including Th2 lymphocytes and M1 macrophages. These findings suggest that argentatins are promising therapeutic agents in autoimmune or allergic diseases, and open new perspectives for the investigation of argentatins in immune response and in the development of more targeted and effective immunomodulatory therapies.

Citation: Silva-Nolasco AM, de la Cruz-Morcillo MA, García-Martínez MM, Zalacain A, Gálvez BG, Carmona M (2024) Immunomodulatory activity of argentatins A and B isolated from guayule. PLoS ONE 19(5): e0304713. https://doi.org/10.1371/journal.pone.0304713

Editor: Nisha Singh, HNBGU: Hemvati Nandan Bahuguna Garhwal University, INDIA

Received: December 18, 2023; Accepted: May 16, 2024; Published: May 31, 2024

Copyright: © 2024 Silva-Nolasco et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Data Availability: All relevant data are within the manuscript and its Supporting Information files.

Funding: This research was funded through Project PID2022-1424550B-100 by the Spanish Ministry of Science, Innovation and Universities MCIN /AEI /10.13039/501100011033 MCIN /AEI /10.13039/501100011033 / and by the European Regional Development Fund (ERDF). Aniela M. Silva-Nolasco acknowledges a predoctoral contract (2022-UNIVERS-11373) from Universidad de Castilla-La Mancha and the European Social Fund. M. Mercedes García-Martínez acknowledges a grant for the requalification of the Spanish university system modality Margarita Salas (2022- NACIONA-11053) funded by the European Union-Next Generation EU through the Universidad de Castilla-La Mancha. Miguel A. de la Cruz-Morcillo acknowledges a postdoctoral contract (2022-UNIVERS-11006) for researcher for scientific excellence within the internal UCLM R&D Plan, co-financed by the European Social Fund Plus (ESF+). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Competing interests: The authors have declared that no competing interests exist.

Introduction

For many decades, plants have provided us with natural bioactive compounds that benefit our health, often serving as starting points for the synthesis of new medicines. A good example of this is the guayule shrub (Parthenium argentatum A. Gray), which is native to the arid landscapes of the Chihuahuan desert in México and the southwestern USA [1]. In addition to its use in the manufacture of hypoallergenic latex, several secondary metabolites isolated from guayule have received increasing attention in recent years due to their unique structural characteristics and biological properties. The most important of these are the argentatins, which are the principal secondary metabolites of guayule resin, accounting for approximately 30% of the total resin obtained from this shrub [2]. Argentatins are cycloartane-type triterpene compounds with diverse pharmacological properties, including antimicrobial and anticarcinogenic activity in vitro. For example, used at a concentration of 238 μM, argentatin A was found to inhibit the growth of the pathogenic yeasts Candida albicans and Torulopsis glabrata better than chloramphenicol, which was used as a control, and also showed inhibitory activity against Klebsiella pneumoniae and Pseudomonas aeruginosa when compared with fluconazole [3]. Regarding their anticarcinogenic properties, both argentatin A and B are effective in inhibiting up to 100% of the growth (at 100 μM) of various cancer cell lines [4], thus showing very promising results in terms of activity.

Another interesting property of the argentatins is their potential anti-inflammatory activity. Although studies are scarce, argentatins have been shown to reduce inflammation in in vivo models of edema induced by tetradecanoylphorbol acetate; argentatin B used at 15 μM inhibited COX-2 activity by 77%, which was superior to indomethacin used as a reference [5]. In the same study, the effect of argentatins A and B on nitric oxide (NO) production was evaluated in mouse peritoneal macrophages stimulated with lipopolysaccharide to induce the release of tumor necrosis factor alpha (TNF-α) and NO. Results showed that both compounds at 31 and 100 μM significantly inhibited the release of nitrites, which are the precursors of NO. Moreover, in metabolic assays, argentatin A and B used at 50 μM reduced the cell viability of resting macrophages by 63% and 53%, respectively, suggesting that the reduction in NO is directly related to their influence on cell viability [5].

An area of growing interest in drug development is immunomodulation, which involves reshaping the host response through therapeutic strategies that enhance or suppress the immune system response as needed. A classical example of immunomodulation is in the treatment of autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus and multiple sclerosis, which is applied to suppress the immune response and reduce inflammation and associated tissue damage [6]. Immunomodulatory drugs are also used to prevent organ and tissue rejection after transplantation [7]. A more recent example of immunomodulation is the use of checkpoint inhibitor-based immunotherapies to block the signals that inhibit the immune response against cancer cells [8]. Also, immunodulatory drugs are used in the context of inflammatory and allergic diseases involving an exaggerated immune response, such as asthma and atopic dermatitis [9, 10]. Finally, there has been an interest in targeting immunomodulation in neurodegenerative disorders such as Alzheimer’s and Parkinson’s disease to treat brain inflammation [11].

Inflammation is a biological protective response to injury, infection, irritation or cellular damage that eliminates infectious agents, removes dead and damaged cells and facilitates the tissue repair process. Its mechanism is complex and involves many different cells of the immune system. The inflammatory response consists of the activation of resident immune cells such as mast cells and macrophages, which recruit leukocytes (neutrophils and monocytes) to the site of injury or infection to repair tissue damage and kill pathogens [12]. It is important to note that although the response to acute infection is usually rapid and transient, inflammation can become chronic. This has been linked to several diseases, including autoimmune, cardiovascular and metabolic disorders, as well as certain types of cancer [13].

The effect of argentatins on the development of cellular immune responses and cytokine production has not yet been evaluated. Accordingly, the present study was designed to investigate the potential of argentatin A and B to shape the immunomodulatory properties of mesenchymal stem cells (MSCs), and to assess their direct effects on immune cells to better understand their potential ability to modulate inflammation.

Material and methods

Chemical and reagents

Dulbecco’s modified Eagle’s medium (DMEM)-high glucose, Roswell Park Memorial Institute (RPMI) 1640 medium, fetal bovine serum (FBS), phosphate buffered saline (PBS), dimethyl sulfoxide (DMSO), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), lipopolysaccharide (LPS), interferon gamma (IFN-γ), macrophage colony-stimulating factor (M-CSF) and human IgG were purchased from Sigma-Aldrich (St Louis, MO, USA). Antibodies for lymphocyte differentiation were purchased from Abcam (Cambridge, UK) (IL-4, IL-10, rIL-2, rIL-12, IL-23p40, IL-4, rIL4, IL-12p40, anti-CD3 and anti-CD28). Penicillin and streptomycin were from Gibco (Gaithersburg, MD, USA). MSCs were purchased from Lonza (Basel, Switzerland) and THP-1 monocytes and CD4+ T-lymphocytes were from the ATC (Manassas, VA, USA). Argentatins A and B were isolated from guayule resin as described [14] with a purity of 88% and 87%, respectively, and were dissolved in DMSO at 20 mM as stock solutions, which were stored at -80°C.

Cell culture and growth conditions

Human MSCs (PT-2501) were plated (1 × 10⁶) in a 100-mm plate and cultured for 48 hours in complete DMEM/10% FBS, as described [15]. When the plate was near confluent the culture supernatant (conditioned medium) was collected and centrifuged at 1,200 × g at room temperature for 10 minutes. The upper aqueous solution was aliquoted in 1.5-ml microtubes and kept at -80°C until use.

THP-1 human leukemia monocytes (TIB-202) were first cultured in RPMI 1640 supplemented with 5% FBS and 10 ng/ml M-CSF at 37°C and 5% CO₂ for 72 hours, and then in the same medium without FBS for a further 48 hours. To induce differentiation to an M1 (proinflammatory) phenotype, the medium was supplemented with 100 ng/ml LPS and 25 U/ml IFN-γ. To induce differentiation to an M2 (anti-inflammatory) phenotype, the medium was supplemented with 20 ng/ml IL-4 and 20 ng/ml IL-10.

CD4+ T-lymphocyte cells (PCS-800-016) were grown in complete RPMI medium and their differentiation into Th1 or Th2 subpopulations was performed as described [16, 17]. Briefly, CD4+ T-cells were cultured in complete RPMI medium and activated with 1 μg/ml anti-CD3, 2 μg/ml anti-CD28 and 100 U/ml rIL-2. For differentiation into Th1 cells (Th1- differentiation condition) 2.5 ng/ml of rIL-12/IL-23p40 and 50 ng/ml of anti-IL-4 were added. For differentiation into Th2 cells (Th2- differentiation condition) 12.5 ng/ml of rIL-4 and 1 μg/ml of anti-IL-12p40/70 were added. Differentiation was evaluated after 5 days of culture.

There were no reports that cell line used in the assays was misidentified or cross-contaminated [18].

Proliferation assay

The viability and proliferation of MSCs upon treatment with argentatins was measured with an MTT reduction assay and spectrophotometry [19]. MSCs were seeded into 96-well tissue culture plates at 1 × 10⁵ cells per well in 0.2 ml of DMEM-high glucose supplemented with 10% FBS and 1% penicillin/streptomycin and grown at 37°C in a humidified 5% CO₂ incubator for 24 hours. Cells were then treated for 24 or 48 hours in the presence of different amounts of argentatins (20, 50 and 100 μM). DMSO alone was used in control conditions. Relative cytotoxicity was expressed as a percentage of [ODsample − ODblank]/[ODcontrol − ODblank] × 100. Each experiment was performed in triplicate.

Flow cytometry analysis

Monocytes and differentiated macrophages were harvested by gentle scratching with a cell scraper (Sarstedt, Newton, NC, USA) and T-lymphocytes were collected after centrifugation. In both cases, cells were resuspended in serum-free RPMI (2 × 10⁶ cells/ml) and incubated for 30 minutes on ice with 150 μg/ml of Human IgG to block Fc receptors. Cells were analyzed on a FACsCanto II flow cytometer (Becton Dickinson, Sunnyvale, CA, USA). Sample acquisition was performed on FACSVerse with FACSuite v.1.0.6.5230 (BD Biosciences) and data analysis was done using FlowJo software (all from Becton Dickinson).

Macrophages were stained with 20 μg/ml of the corresponding primary antibody on ice for 30 minutes. The mixtures of monoclonal antibodies used for detection of the different populations are shown in S1 Table. After 5 days of differentiation in the presence or absence of argentatins, T-helper CD4⁺ lymphocytes were gated by positive surface staining for CD4 and were expressed as a percentage of gated lymphocytes; Th1 cells (CD4+IFN-c+) and Th2 cells (CD4+IL-4+) were expressed as a percentage of CD4+ cells. Macrophage analysis and data acquisition was performed as described for monocytes [20].

Cytokine assay

The concentration of several cytokines was determined using commercial ELISA detection kits (Merck Millipore, Darmstadt, Germany) with a fluorescence plate reader (BioTek Instruments, Winooski, VT, USA). The level of cytokines (IL-2, IL-4, IL-6, IL-10, TNF, INF-γ, IL17-A) in supernatants was measured after collection of conditioned medium of cultured cells, as previously reported [21] in the presence or absence of argentatins.

Statistical analysis

The results are represented as the mean and standard error of the mean (SEM); the Shapiro-Wilk test was applied to check whether the distribution was normal. A parametric analysis of variance (ANOVA) with Dunnet’s post test was used to compare each study group with respect to the control group (for cell proliferation and cytokine release assays), and a Tukey’s post test for cell differentiation assays, where a positive and a negative control were compared; a value of p<0.05* was considered statistically significant and a value of <0.0005*** highly significant. Statistical analysis was performed with GraphPad Prism software version 9.5.1, (GraphPad Prism, Software San Diego, CA, USA).

Results and discussion

Cell viability assay

To evaluate the potential toxicity of the argentatin A and B compounds isolated from guayule resin, we performed a cytotoxicity assay based on the reduction of a yellow tetrazolium salt (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT). Because the argentatins are antiproliferative in several in vitro tumor models, including HCT15, HCT116, SW620 and RKO cells [22], it was important to assess whether the compounds were cytotoxic in the MSC model. Accordingly, MSCs were treated with increasing doses of argentatin A and B (20, 50 and 100 μM) based on the literature, for 24 and 48 hours.

No toxicity was detected for MSCs at 20 μM for both argentatins compared with control (DMSO) after 24 hours (Fig 1A). Nevertheless, argentatin A and B were cytotoxic at the higher concentration of 50 μM (p = 0.0481 and p = 0.0160, respectively). At the highest dose of 100 μM, the cytotoxicity against MSCs was very similar (Fig 1A).

Download:

Fig 1. Viability of mesenchymal stem cells (MSCs) in response to argentatin A and B.

(A) Viability at 24 h, (B) Viability at 48 h. Cell viability was determined by MTT reduction and was compared to the corresponding untreated control (DMSO), which was normalized to 100% cell viability. Error bars represent the standard error of the mean (SEM) of three independent experiments. Two-way ANOVA with Dunnet´s post test was used to determine differences between argentatin A and B treatments compared with control (*p<0.05, **p<0.005).

https://doi.org/10.1371/journal.pone.0304713.g001

These data indicate that concentrations exceeding 50 μM should be avoided for MSCs. After 48 hours of treatment, 50 μM argentatin A exerted a slight but significant cytotoxicity compared with the control (p = 0.0113), but no significant toxicity was observed for argentatin B (Fig 1B). The dose of 100 μM for argentatin A and B was discarded for future studies because of its high toxicity (p<0.0031 and p<0.0251, respectively), with argentatin A being slightly more cytotoxic than argentatin B (Fig 1B). Of note, neither compound led to a cell viability lower than 50% at the highest concentration and treatment duration, contrary to what has been observed in tumor cell lines with a 50% inhibitory concentration at doses between 20 and 30 μM [4, 23].

Some anticancer compounds tend to have certain selectivity only for cancer cells. This may be due to several reasons, such as the difference in proliferation rate, as they tend to divide and grow faster than normal or healthy cells. Or the genetic mutations that cancer cells tend to have and differentiate them from normal cells, so, some anticancer compounds are designed precisely for this, to target specific processes related to cell division, affecting cells that divide more frequently such as cancer cells. Or they are designed to target mutated or overexpressed proteins, without affecting normal cells as much. Such is the case of the selectivity of curcumin, where its potent action against cancer cells has been demonstrated and its immunomodulatory effect evidenced by good bone marrow activity after administration with no apparent toxicity in the normal cells and even in a murine model, justified by normal behavior and the absence of death or health deterioration [24–26]. This is potentially significant as it could inform new strategies targeted at treating autoimmune and inflammatory diseases with MSCs, which are involved in key functions such as tissue repair and regeneration due to their capacity to differentiate into different cell types and to release immunomodulatory and anti-inflammatory factors [27].

Cytokine expression

We next questioned whether argentatin treatment of MSCs promoted cytokine production. MSCs were treated with 50 μM argentatin A and B for 24 hours and the levels of six pro- or anti-inflammatory cytokines were quantified from conditioned medium by ELISA. No significant changes were observed for the production of the anti-inflammatory cytokines IL-10 and TFG-β or for the proinflammatory cytokines IL-6, IL-8 and VEGF (Fig 2).

Download:

Fig 2. Production of anti-inflammatory and proinflammatory mediators in argentatin-stimulated MSCs.

Cytokine production by MSCs was evaluated in the absence or presence of stimulation for 24 hours at 50 μM. Error bars represent the standard error of the mean (SEM) of three individual experiments. Two-way ANOVA with Dunnet´s post test was used to determine differences between argentatin A and B treatments compared with control (***p<0.0005).

https://doi.org/10.1371/journal.pone.0304713.g002

However, results revealed a marked increase in the production of the proinflammatory cytokine MCP-1 after treatment with either argentatin (Fig 2). MCP-1 (also called chemokine [CC-motif] ligand 2) has a crucial role in immunomodulation by participating in the attraction and regulation of monocytes and macrophages, which in turn differentiate into M1 or M2 cells [28]. It thus contributes to both acute and chronic inflammation.

Cellular differentiation

CD4+ T lymphocytes play a critical role in regulating the immune response and can differentiate into specific subtypes based on the presence of cytokines and other signals in the microenvironment, in addition to the levels of antigens, the type of antigen-presenting cell and costimulatory molecule [29]. Given our finding of a marked increase in MCP-1 production following argentatin treatment, we next evaluated whether argentatin A and B could influence the differentiation of CD4+ T lymphocytes to Th1 and Th2 cells, which are the most common subtypes. As negative control for this study we used CD4+ T cells without differentiation factors, and as a positive control we used CD4+ T cells with full differentiation factors (see materials and methods). In the presence of argentatin A (50 μM for 5 days), differentiation of the lymphocyte population to the Th1 phenotype, which is known for facilitate humoral responses, was 2% with respect to the negative control, and differentiation to the Th2 phenotype, which mediates delayed type hypersensitivity responses and inflammatory reactions, was about 3% (Fig 3). Both cell types normally function in a balanced manner, however, exposure to pathogens may cause one subset to predominate. The results for argentatin B treatment were very similar with values of 3% and 2% for the Th1 and Th2 phenotypes, respectively.

Download:

Fig 3. Percentage of LT CD4+ differentiated to Th1 and Th2 subtypes by argentatins.

The treatment with both argentatins was at 50 μM and the differentiation process was evaluated by cytometry after 5 days of incubation. Error bars represent the standard error of the mean (SEM) of three individual experiment and the data can be found in S2 Table. Two-way ANOVA with Tukey´s post test was used to determine differences between argentatin A and B treatments compared with positive control (*p<0.05). No statistically significant differences were found with respect to the negative control.

https://doi.org/10.1371/journal.pone.0304713.g003

Chronic inflammation is a consequence of the predominance of Th1 response, whereas allergy and hypergammaglobulinemic states are caused by the predominance of Th2 responses [16]. While the argentatins had only a modest effect on cell differentiation, there was a relative equivalence in the lack of polarization between the two lymphocyte phenotypes, meaning that an unbalanced immune response such as in the case of autoimmune or allergic diseases would not occur. However, it is important to note that the manipulation of differentiation will depend on the context and the state of activation of the immune system, as this process is essential in many situations such as fighting infections.

To further evaluate the immunomodulating role of argentatins, their effect on the phenotypic differentiation of macrophages was studied. The monocytic cell line THP-1 was chosen to examine macrophage differentiation because of the similarities it shares with primary monocytes, such as morphology and physiology, minimizing cell phenotype variation arising from monocyte-to-macrophage differentiation by using a single source of cells [30]. Similar to lymphocytes, THP-1 macrophage polarization to the principal phenotypes M1 and M2 are processes controlled by the microenvironment and associated signals.

Treatment of THP-1 cells with argentatin A (50 μM for three days) stimulated M1-type macrophage differentiation almost 40-fold greater than that observed in the negative control, whereas M2-type differentiation was about 16-fold higher compared to the negative control (Fig 4). Similarly, the equivalent treatment of THP-1 cells with argentatin B stimulated M1-type macrophage differentiation 30-fold higher than the negative control, a lower percentage than argentatin A, whereas M2-type differentiation was 17-fold higher with respect to the negative control (Fig 4). The function of M1-type macrophages is related to inhibiting cell proliferation and causing tissue damage, whereas M2-type macrophages promote cell proliferation and tissue repair [31].

Download:

Fig 4. Percentage of THP-1 macrophages differentiated to M1 and M2 subtypes by argentatins.

The treatment with both argentatins was at 50 μM and the differentiation process was evaluated by cytometry after 3 days of incubation. Error bars represent the standard error of the mean (SEM) of three individual experiments and the data can be found in S2 Table. Two-way ANOVA with Tukey´s post test was used to determine differences between argentatin A and B treatments compared with controls. The letters refer to the significant difference compared to the negative control and the asterisk compared to the positive control (*p<0.05, **P<0.005, (a) P<0.05, (b) P<0.005).

https://doi.org/10.1371/journal.pone.0304713.g004

Most studies performed in the search for natural immunomodulatory compounds involve strategies to resolve inflammation by increasing M2 levels and/or decreasing M1 levels, rather than directly promoting differentiation to M1. Compounds such as polysaccharide isolated from the goji plant (Lycium barbadum) have been tested both in vitro and in vivo at a concentration of 10 mg/kg and 100 μg/ml, respectively, in microglial cells, and have been found to induce polarization to M1 macrophages, and a similar result have been noted for Platycodon grandifloras polysaccharide in the RAW 264.7 macrophage line at concentrations of 0.01 to 10 μM [32, 33].

In the context of cancer, the propensity for macrophages to polarize to the M1 phenotype may have benefits by secreting proinflammatory cytokines such as TNF-α and IFN-γ, thereby helping to activate an appropriate immune response against cancer cells. This is the case for curcumin (8 μM) in the presence of LPS, which was tested in the A549 lung cell line, and was found to promote the secretion of IL-1β and TNF-α by reprogramming M2 macrophages into tumor-killing M1 macrophages [34]. The same effect was observed with astragalus polysaccharide (16 mg/ml) in the NSCLC cell line H441 co-cultured with monocyte-derived macrophages, and with β-glucan (800 μM) in mouse models implanted with Lewis lung carcinoma cells (LLC) or EO77 cells, which significantly reduced IL-10 levels while increasing the mRNA levels of iNOS, IL-12, TNF-α, IL-1β and IL-6 [35, 36]. Another example is the polysaccharide portion of ginseng berry (GBPP), which at 100 μg was found to promote the production of cytokines such as TNF-α and IL1-12, thus inducing M1 polarization in mouse lung cancer models [37].

Although Th1 and Th2 cells can secrete cytokines in certain contexts that influence the polarization of M1 and M2 macrophages, respectively, they are not the only factor that determines polarization, as shown in this study, where the predominant phenotype was M1 after treatment with both argentatins irrespective of the low differentiation capacity for Th1 cells [38].

Beyond the secretion of inflammatory mediators, M1 cells can directly phagocytose and destroy cancer cells through the production of reactive oxygen and nitrogen species, as well as by promoting adaptive immunity by stimulating the response of cytotoxic T cells (CD8+). This favors the presentation of tumor antigens, which would lead to the enhancement of antitumor immunity [39, 40].

Nonetheless, the balance between M1 and M2 phenotypes in the tumor context may change during cancer progression. Therefore, the investigation of targeted therapies to modulate polarization in macrophages is a promising research approach.

Conclusions

Here we examined the effects of argentatins A and B on MSCs and their potential to regulate the immune response. We found that at doses of 20–50 μM both compounds were not toxic to MSC cells, making them safe for future use. Also, argentatin A promoted the proliferation of MSCs, which could be beneficial for tissue regeneration and immune system modulation. This is supported by the finding that both argentatin A and B, increased the release of MCP-1 and also favored the differentiation of immune cells (CD4+ T lymphocytes and THP-1 macrophages). To the best of our knowledge, this is the first demonstration of an immunomodulatory function of argentatins, but further research is needed to fully understand their effects on the regulation of immune response and inflammation, which might open up new therapeutic avenues for the development of treatments for several diseases with an inflammatory component.

Supporting information

S1 Table. Monoclonal antibody mix used for the assessment of macrophage populations by flow cytometry.

https://doi.org/10.1371/journal.pone.0304713.s001

(PDF)

S2 Table. Percentages of differentiated cells by argentatins.

LT CD4+ differentiated to Th1 and Th2 cell subtypes, and THP-1 macrophages differentiated to M1 and M2 cell subtypes respectively.

https://doi.org/10.1371/journal.pone.0304713.s002

(PDF)

S1 Raw data.

https://doi.org/10.1371/journal.pone.0304713.s003

(XLSX)

Acknowledgments

The authors wish to thank Agroservicios Guayule S.L. for its support in this research. We thank Kenneth McCreath for English editing.

References

1. Rasutis D, Soratana K, McMahan C, Landis AE (2015) A sustainability review of domestic rubber from the guayule plant. Ind Crops Prod 70:383–394
- View Article
- Google Scholar
2. Gallego B, Carrión ME, Latorre G, García-Martínez MM, Hurtado de Mendoza J, Zalacain A, et al. (2022) UHPLC-HRMS profile and accumulation of argentatins in stems of 27 accessions of guayule (Parthenium argentatum A. Gray) and its hybrids along the growth cycle. Ind Crops Prod 187:115463
- View Article
- Google Scholar
3. Martínez‐Vázquez M, Martínez R, Espinosa Pérez G, Díaz M, Herrera Sánchez M (1994) Antimicrobial properties of argentatin A, isolated from Parthenium argentatum. Fitoterapia 65:371–372
- View Article
- Google Scholar
4. Parra-Delgado H, Ramírez-Apan T, Martínez-Vázquez M (2005) Synthesis of argentatin A derivatives as growth inhibitors of human cancer cell lines in vitro. Bioorg Med Chem Lett 15:1005–1008 pmid:15686901
- View Article
- PubMed/NCBI
- Google Scholar
5. Romero JC, Martínez-Vázquez A, Herrera MP, Martinez-Mayorga K, Parra-Delgado H, Pérez-Flores FJ, et al. (2014) Synthesis, anti-inflammatory activity and modeling studies of cycloartane-type terpenes derivatives isolated from Parthenium argentatum. Bioorg Med Chem 22:6893–6898
- View Article
- Google Scholar
6. López-Pedrera C, Villalba JM, Patiño-Trives AM, Luque-Tévar M, Barbarroja N, Aguirre MÁ, et al. (2021) Therapeutic Potential and Immunomodulatory Role of Coenzyme Q10 and Its Analogues in Systemic Autoimmune Diseases. Antioxidants 10:600 pmid:33924642
- View Article
- PubMed/NCBI
- Google Scholar
7. Enshasy H El (2011) Immunomodulators. In: Industrial Applications. Springer Berlin Heidelberg, Berlin, Heidelberg, pp 165–194
8. Sadreddini S, Baradaran B, Aghebati‐Maleki A, Sadreddini S, Shanehbandi D, Fotouhi A, et al. (2019) Immune checkpoint blockade opens a new way to cancer immunotherapy. J Cell Physiol 234:8541–8549 pmid:30511409
- View Article
- PubMed/NCBI
- Google Scholar
9. Kandikattu HK, Mishra A. (2018) Immunomodulatory effects of tacrolimus (FK506) for the treatment of allergic diseases. Int J Cell Biol Physiol. pmid:31595270
- View Article
- PubMed/NCBI
- Google Scholar
10. Jaworek AK, Szepietowski JC, Hałubiec P, Wojas-Pelc A, Jaworek J (2021) Melatonin as an Antioxidant and Immunomodulator in Atopic Dermatitis—A New Look on an Old Story: A Review. Antioxidants 10:1179 pmid:34439427
- View Article
- PubMed/NCBI
- Google Scholar
11. Martinez B, Peplow P (2018) Neuroprotection by immunomodulatory agents in animal models of Parkinson’s disease. Neural Regen Res 13:1493 pmid:30127102
- View Article
- PubMed/NCBI
- Google Scholar
12. Singh P, Chazaud B (2021) Benefits and pathologies associated with the inflammatory response. Exp Cell Res 409:112905 pmid:34736921
- View Article
- PubMed/NCBI
- Google Scholar
13. FERNANDES JV, DE MEDEIROS FERNANDES TAA, DE AZEVEDO JCV, COBUCCI RNO, DE CARVALHO MGF, ANDRADE VS, et al. (2015) Link between chronic inflammation and human papillomavirus-induced carcinogenesis (Review). Oncol Lett 9:1015–1026
14. García-Martínez MM, Gallego B, Latorre G, Carrión ME, De la Cruz-Morcillo MÁ, Zalacain A, et al. (2023) Argentatin Content in Guayule Leaves (Parthenium argentatum A. Gray). Plants 12:1–7 pmid:37653938
- View Article
- PubMed/NCBI
- Google Scholar
15. de Lucas B, Pérez LM, Bernal A, Gálvez BG (2021) Application of low-intensity pulsed therapeutic ultrasound on mesenchymal precursors does not affect their cell properties. PLoS One 16:e0246261 pmid:33571276
- View Article
- PubMed/NCBI
- Google Scholar
16. Constant SL, Bottomly K (1997) INDUCTION OF TH1 AND TH2 CD4 ⁺ T CELL RESPONSES:The Alternative Approaches. Annu Rev Immunol 15:297–322
- View Article
- Google Scholar
17. Bonecchi R, Bianchi G, Bordignon PP, et al (1998) Differential Expression of Chemokine Receptors and Chemotactic Responsiveness of Type 1 T Helper Cells (Th1s) and Th2s. J Exp Med 187:129–134 pmid:9419219
- View Article
- PubMed/NCBI
- Google Scholar
18. Database of Cross-Contaminated or Misidentified Cell Lines International Cell Line Authentication Committee (ICLAC) Version 12;2023.
19. Silva-Nolasco AM, Camacho L, Saavedra-Díaz RO, Hernández-Abreu O, León IE, Sánchez-Lombardo I (2020) Kinetic Studies of Sodium and Metforminium Decavanadates Decomposition and In Vitro Cytotoxicity and Insulin- Like Activity. Inorganics (Basel) 8:67
- View Article
- Google Scholar
20. Uchiyama R, Toyoda E, Maehara M, Wasai S, Omura H, Watanabe M, et al. (2021) Effect of Platelet-Rich Plasma on M1/M2 Macrophage Polarization. Int J Mol Sci 22:2336 pmid:33652994
- View Article
- PubMed/NCBI
- Google Scholar
21. Chiswick EL, Duffy E, Japp B, Remick D (2012) Detection and Quantification of Cytokines and Other Biomarkers. pp 15–30
- View Article
- Google Scholar
22. Tavarez-Santamaría Z, Jacobo-Herrera NJ, Rocha-Zavaleta L, Zentella-Dehesa A, del Carmen Couder-García B, Martínez-Vázquez M (2020) A higher frequency administration of the nontoxic cycloartane-type triterpene argentatin A improved its anti-tumor activity. Molecules 25:1780 pmid:32295227
- View Article
- PubMed/NCBI
- Google Scholar
23. Romero-Benavides JC, Bailon-Moscoso N, Parra-Delgado H, et al (2018) Argentatin B derivatives induce cell cycle arrest and DNA damage in human colon cancer cells through p73/p53 regulation. Medicinal Chemistry Research. https://doi.org/10.1007/s00044-017-2106-4
- View Article
- Google Scholar
24. Aravind SR, Krishnan LK (2016) Curcumin-albumin conjugates as an effective anti-cancer agent with immunomodulatory properties. Int Immunopharmacol 34:78–85 pmid:26927614
- View Article
- PubMed/NCBI
- Google Scholar
25. Boroumand N, Samarghandian S, Hashemy SI (2018) Immunomodulatory, anti-inflammatory, and antioxidant effects of curcumin. Journal of Herbmed Pharmacology 7:211–219
- View Article
- Google Scholar
26. Barati N, Momtazi‐Borojeni AA, Majeed M, Sahebkar A (2019) Potential therapeutic effects of curcumin in gastric cancer. J Cell Physiol 234:2317–2328 pmid:30191991
- View Article
- PubMed/NCBI
- Google Scholar
27. Spees JL, Lee RH, Gregory CA (2016) Mechanisms of mesenchymal stem/stromal cell function. Stem Cell Res Ther 7:125 pmid:27581859
- View Article
- PubMed/NCBI
- Google Scholar
28. Deshmane SL, Kremlev S, Amini S, Sawaya BE (2009) Monocyte Chemoattractant Protein-1 (MCP-1): An Overview. Journal of Interferon & Cytokine Research 29:313–326 pmid:19441883
- View Article
- PubMed/NCBI
- Google Scholar
29. Luckheeram RV, Zhou R, Verma AD, Xia B (2012) CD4 ⁺ T Cells: Differentiation and Functions. Clin Dev Immunol 2012:1–12
- View Article
- Google Scholar
30. Mohd Yasin ZN, Mohd Idrus FN, Hoe CH, Yvonne-Tee GB (2022) Macrophage polarization in THP-1 cell line and primary monocytes: A systematic review. Differentiation 128:67–82
- View Article
- Google Scholar
31. Mills C(2012) M1 and M2 Macrophages: Oracles of Health and Disease. Crit Rev Immunol 32:463–488 pmid:23428224
- View Article
- PubMed/NCBI
- Google Scholar
32. Wang Y, Smith W, Hao D, He B, Kong L(2019) M1 and M2 macrophage polarization and potentially therapeutic naturally occurring compounds. Int Immunopharmacol 70:459–466 pmid:30861466
- View Article
- PubMed/NCBI
- Google Scholar
33. Gao Y, Xu X, Zhang X (2023) Targeting different phenotypes of macrophages: A potential strategy for natural products to treat inflammatory bone and joint diseases. Phytomedicine 118:154952 pmid:37506402
- View Article
- PubMed/NCBI
- Google Scholar
34. Somchit N, Kimseng R, Dhar R, Hiransai P, Changtam C, Suksamrarn A, et al. (2018) Curcumin pyrazole blocks lipopolysaccharide-induced inflammation via suppression of JNK activation in RAW 264.7 macrophages. Asian Pac J Allergy Immunol 36:184–190 pmid:29246079
- View Article
- PubMed/NCBI
- Google Scholar
35. Liu M, Luo F, Ding C, et al (2015) Dectin-1 Activation by a Natural Product β-Glucan Converts Immunosuppressive Macrophages into an M1-like Phenotype. The Journal of Immunology 195:5055–5065
- View Article
- Google Scholar
36. Bamodu OA, Kuo K-T, Wang C-H, Huang W-C, Wu ATH, Tsai J-T, et al. (2019) Astragalus polysaccharides (PG2) Enhances the M1 Polarization of Macrophages, Functional Maturation of Dendritic Cells, and T Cell-Mediated Anticancer Immune Responses in Patients with Lung Cancer. Nutrients 11:2264 pmid:31547048
- View Article
- PubMed/NCBI
- Google Scholar
37. Lee D-Y, Park CW, Lee SJ, Park H-R, Kim SH, Son S-U, et al. (2019) Anti-Cancer Effects of Panax ginseng Berry Polysaccharides via Activation of Immune-Related Cells. Front Pharmacol. pmid:32038228
- View Article
- PubMed/NCBI
- Google Scholar
38. Stout RD, Suttles J (2004) Functional plasticity of macrophages: reversible adaptation to changing microenvironments. J Leukoc Biol 76:509–513 pmid:15218057
- View Article
- PubMed/NCBI
- Google Scholar
39. Lendeckel U, Venz S, Wolke C (2022) Macrophages: shapes and functions. ChemTexts 8:12 pmid:35287314
- View Article
- PubMed/NCBI
- Google Scholar
40. Orihuela R, McPherson CA, Harry GJ (2016) Microglial M1/M2 polarization and metabolic states. Br J Pharmacol 173:649–665 pmid:25800044
- View Article
- PubMed/NCBI
- Google Scholar

[ref1] 1. Rasutis D, Soratana K, McMahan C, Landis AE (2015) A sustainability review of domestic rubber from the guayule plant. Ind Crops Prod 70:383–394
View Article
Google Scholar

[2] View Article

[3] Google Scholar

[ref2] 2. Gallego B, Carrión ME, Latorre G, García-Martínez MM, Hurtado de Mendoza J, Zalacain A, et al. (2022) UHPLC-HRMS profile and accumulation of argentatins in stems of 27 accessions of guayule (Parthenium argentatum A. Gray) and its hybrids along the growth cycle. Ind Crops Prod 187:115463
View Article
Google Scholar

[5] View Article

[6] Google Scholar

[ref3] 3. Martínez‐Vázquez M, Martínez R, Espinosa Pérez G, Díaz M, Herrera Sánchez M (1994) Antimicrobial properties of argentatin A, isolated from Parthenium argentatum. Fitoterapia 65:371–372
View Article
Google Scholar

[8] View Article

[9] Google Scholar

[ref4] 4. Parra-Delgado H, Ramírez-Apan T, Martínez-Vázquez M (2005) Synthesis of argentatin A derivatives as growth inhibitors of human cancer cell lines in vitro. Bioorg Med Chem Lett 15:1005–1008 pmid:15686901
View Article
PubMed/NCBI
Google Scholar

[11] View Article

[12] PubMed/NCBI

[13] Google Scholar

[ref5] 5. Romero JC, Martínez-Vázquez A, Herrera MP, Martinez-Mayorga K, Parra-Delgado H, Pérez-Flores FJ, et al. (2014) Synthesis, anti-inflammatory activity and modeling studies of cycloartane-type terpenes derivatives isolated from Parthenium argentatum. Bioorg Med Chem 22:6893–6898
View Article
Google Scholar

[15] View Article

[16] Google Scholar

[ref6] 6. López-Pedrera C, Villalba JM, Patiño-Trives AM, Luque-Tévar M, Barbarroja N, Aguirre MÁ, et al. (2021) Therapeutic Potential and Immunomodulatory Role of Coenzyme Q10 and Its Analogues in Systemic Autoimmune Diseases. Antioxidants 10:600 pmid:33924642
View Article
PubMed/NCBI
Google Scholar

[18] View Article

[19] PubMed/NCBI

[20] Google Scholar

[ref7] 7. Enshasy H El (2011) Immunomodulators. In: Industrial Applications. Springer Berlin Heidelberg, Berlin, Heidelberg, pp 165–194

[ref8] 8. Sadreddini S, Baradaran B, Aghebati‐Maleki A, Sadreddini S, Shanehbandi D, Fotouhi A, et al. (2019) Immune checkpoint blockade opens a new way to cancer immunotherapy. J Cell Physiol 234:8541–8549 pmid:30511409
View Article
PubMed/NCBI
Google Scholar

[23] View Article

[24] PubMed/NCBI

[25] Google Scholar

[ref9] 9. Kandikattu HK, Mishra A. (2018) Immunomodulatory effects of tacrolimus (FK506) for the treatment of allergic diseases. Int J Cell Biol Physiol. pmid:31595270
View Article
PubMed/NCBI
Google Scholar

[27] View Article

[28] PubMed/NCBI

[29] Google Scholar

[ref10] 10. Jaworek AK, Szepietowski JC, Hałubiec P, Wojas-Pelc A, Jaworek J (2021) Melatonin as an Antioxidant and Immunomodulator in Atopic Dermatitis—A New Look on an Old Story: A Review. Antioxidants 10:1179 pmid:34439427
View Article
PubMed/NCBI
Google Scholar

[31] View Article

[32] PubMed/NCBI

[33] Google Scholar

[ref11] 11. Martinez B, Peplow P (2018) Neuroprotection by immunomodulatory agents in animal models of Parkinson’s disease. Neural Regen Res 13:1493 pmid:30127102
View Article
PubMed/NCBI
Google Scholar

[35] View Article

[36] PubMed/NCBI

[37] Google Scholar

[ref12] 12. Singh P, Chazaud B (2021) Benefits and pathologies associated with the inflammatory response. Exp Cell Res 409:112905 pmid:34736921
View Article
PubMed/NCBI
Google Scholar

[39] View Article

[40] PubMed/NCBI

[41] Google Scholar

[ref13] 13. FERNANDES JV, DE MEDEIROS FERNANDES TAA, DE AZEVEDO JCV, COBUCCI RNO, DE CARVALHO MGF, ANDRADE VS, et al. (2015) Link between chronic inflammation and human papillomavirus-induced carcinogenesis (Review). Oncol Lett 9:1015–1026

[ref14] 14. García-Martínez MM, Gallego B, Latorre G, Carrión ME, De la Cruz-Morcillo MÁ, Zalacain A, et al. (2023) Argentatin Content in Guayule Leaves (Parthenium argentatum A. Gray). Plants 12:1–7 pmid:37653938
View Article
PubMed/NCBI
Google Scholar

[44] View Article

[45] PubMed/NCBI

[46] Google Scholar

[ref15] 15. de Lucas B, Pérez LM, Bernal A, Gálvez BG (2021) Application of low-intensity pulsed therapeutic ultrasound on mesenchymal precursors does not affect their cell properties. PLoS One 16:e0246261 pmid:33571276
View Article
PubMed/NCBI
Google Scholar

[48] View Article

[49] PubMed/NCBI

[50] Google Scholar

[ref16] 16. Constant SL, Bottomly K (1997) INDUCTION OF TH1 AND TH2 CD4 ⁺ T CELL RESPONSES:The Alternative Approaches. Annu Rev Immunol 15:297–322
View Article
Google Scholar

[52] View Article

[53] Google Scholar

[ref17] 17. Bonecchi R, Bianchi G, Bordignon PP, et al (1998) Differential Expression of Chemokine Receptors and Chemotactic Responsiveness of Type 1 T Helper Cells (Th1s) and Th2s. J Exp Med 187:129–134 pmid:9419219
View Article
PubMed/NCBI
Google Scholar

[55] View Article

[56] PubMed/NCBI

[57] Google Scholar

[ref18] 18. Database of Cross-Contaminated or Misidentified Cell Lines International Cell Line Authentication Committee (ICLAC) Version 12;2023.

[ref19] 19. Silva-Nolasco AM, Camacho L, Saavedra-Díaz RO, Hernández-Abreu O, León IE, Sánchez-Lombardo I (2020) Kinetic Studies of Sodium and Metforminium Decavanadates Decomposition and In Vitro Cytotoxicity and Insulin- Like Activity. Inorganics (Basel) 8:67
View Article
Google Scholar

[60] View Article

[61] Google Scholar

[ref20] 20. Uchiyama R, Toyoda E, Maehara M, Wasai S, Omura H, Watanabe M, et al. (2021) Effect of Platelet-Rich Plasma on M1/M2 Macrophage Polarization. Int J Mol Sci 22:2336 pmid:33652994
View Article
PubMed/NCBI
Google Scholar

[63] View Article

[64] PubMed/NCBI

[65] Google Scholar

[ref21] 21. Chiswick EL, Duffy E, Japp B, Remick D (2012) Detection and Quantification of Cytokines and Other Biomarkers. pp 15–30
View Article
Google Scholar

[67] View Article

[68] Google Scholar

[ref22] 22. Tavarez-Santamaría Z, Jacobo-Herrera NJ, Rocha-Zavaleta L, Zentella-Dehesa A, del Carmen Couder-García B, Martínez-Vázquez M (2020) A higher frequency administration of the nontoxic cycloartane-type triterpene argentatin A improved its anti-tumor activity. Molecules 25:1780 pmid:32295227
View Article
PubMed/NCBI
Google Scholar

[70] View Article

[71] PubMed/NCBI

[72] Google Scholar

[ref23] 23. Romero-Benavides JC, Bailon-Moscoso N, Parra-Delgado H, et al (2018) Argentatin B derivatives induce cell cycle arrest and DNA damage in human colon cancer cells through p73/p53 regulation. Medicinal Chemistry Research. https://doi.org/10.1007/s00044-017-2106-4
View Article
Google Scholar

[74] View Article

[75] Google Scholar

[ref24] 24. Aravind SR, Krishnan LK (2016) Curcumin-albumin conjugates as an effective anti-cancer agent with immunomodulatory properties. Int Immunopharmacol 34:78–85 pmid:26927614
View Article
PubMed/NCBI
Google Scholar

[77] View Article

[78] PubMed/NCBI

[79] Google Scholar

[ref25] 25. Boroumand N, Samarghandian S, Hashemy SI (2018) Immunomodulatory, anti-inflammatory, and antioxidant effects of curcumin. Journal of Herbmed Pharmacology 7:211–219
View Article
Google Scholar

[81] View Article

[82] Google Scholar

[ref26] 26. Barati N, Momtazi‐Borojeni AA, Majeed M, Sahebkar A (2019) Potential therapeutic effects of curcumin in gastric cancer. J Cell Physiol 234:2317–2328 pmid:30191991
View Article
PubMed/NCBI
Google Scholar

[84] View Article

[85] PubMed/NCBI

[86] Google Scholar

[ref27] 27. Spees JL, Lee RH, Gregory CA (2016) Mechanisms of mesenchymal stem/stromal cell function. Stem Cell Res Ther 7:125 pmid:27581859
View Article
PubMed/NCBI
Google Scholar

[88] View Article

[89] PubMed/NCBI

[90] Google Scholar

[ref28] 28. Deshmane SL, Kremlev S, Amini S, Sawaya BE (2009) Monocyte Chemoattractant Protein-1 (MCP-1): An Overview. Journal of Interferon & Cytokine Research 29:313–326 pmid:19441883
View Article
PubMed/NCBI
Google Scholar

[92] View Article

[93] PubMed/NCBI

[94] Google Scholar

[ref29] 29. Luckheeram RV, Zhou R, Verma AD, Xia B (2012) CD4 ⁺ T Cells: Differentiation and Functions. Clin Dev Immunol 2012:1–12
View Article
Google Scholar

[96] View Article

[97] Google Scholar

[ref30] 30. Mohd Yasin ZN, Mohd Idrus FN, Hoe CH, Yvonne-Tee GB (2022) Macrophage polarization in THP-1 cell line and primary monocytes: A systematic review. Differentiation 128:67–82
View Article
Google Scholar

[99] View Article

[100] Google Scholar

[ref31] 31. Mills C(2012) M1 and M2 Macrophages: Oracles of Health and Disease. Crit Rev Immunol 32:463–488 pmid:23428224
View Article
PubMed/NCBI
Google Scholar

[102] View Article

[103] PubMed/NCBI

[104] Google Scholar

[ref32] 32. Wang Y, Smith W, Hao D, He B, Kong L(2019) M1 and M2 macrophage polarization and potentially therapeutic naturally occurring compounds. Int Immunopharmacol 70:459–466 pmid:30861466
View Article
PubMed/NCBI
Google Scholar

[106] View Article

[107] PubMed/NCBI

[108] Google Scholar

[ref33] 33. Gao Y, Xu X, Zhang X (2023) Targeting different phenotypes of macrophages: A potential strategy for natural products to treat inflammatory bone and joint diseases. Phytomedicine 118:154952 pmid:37506402
View Article
PubMed/NCBI
Google Scholar

[110] View Article

[111] PubMed/NCBI

[112] Google Scholar

[ref34] 34. Somchit N, Kimseng R, Dhar R, Hiransai P, Changtam C, Suksamrarn A, et al. (2018) Curcumin pyrazole blocks lipopolysaccharide-induced inflammation via suppression of JNK activation in RAW 264.7 macrophages. Asian Pac J Allergy Immunol 36:184–190 pmid:29246079
View Article
PubMed/NCBI
Google Scholar

[114] View Article

[115] PubMed/NCBI

[116] Google Scholar

[ref35] 35. Liu M, Luo F, Ding C, et al (2015) Dectin-1 Activation by a Natural Product β-Glucan Converts Immunosuppressive Macrophages into an M1-like Phenotype. The Journal of Immunology 195:5055–5065
View Article
Google Scholar

[118] View Article

[119] Google Scholar

[ref36] 36. Bamodu OA, Kuo K-T, Wang C-H, Huang W-C, Wu ATH, Tsai J-T, et al. (2019) Astragalus polysaccharides (PG2) Enhances the M1 Polarization of Macrophages, Functional Maturation of Dendritic Cells, and T Cell-Mediated Anticancer Immune Responses in Patients with Lung Cancer. Nutrients 11:2264 pmid:31547048
View Article
PubMed/NCBI
Google Scholar

[121] View Article

[122] PubMed/NCBI

[123] Google Scholar

[ref37] 37. Lee D-Y, Park CW, Lee SJ, Park H-R, Kim SH, Son S-U, et al. (2019) Anti-Cancer Effects of Panax ginseng Berry Polysaccharides via Activation of Immune-Related Cells. Front Pharmacol. pmid:32038228
View Article
PubMed/NCBI
Google Scholar

[125] View Article

[126] PubMed/NCBI

[127] Google Scholar

[ref38] 38. Stout RD, Suttles J (2004) Functional plasticity of macrophages: reversible adaptation to changing microenvironments. J Leukoc Biol 76:509–513 pmid:15218057
View Article
PubMed/NCBI
Google Scholar

[129] View Article

[130] PubMed/NCBI

[131] Google Scholar

[ref39] 39. Lendeckel U, Venz S, Wolke C (2022) Macrophages: shapes and functions. ChemTexts 8:12 pmid:35287314
View Article
PubMed/NCBI
Google Scholar

[133] View Article

[134] PubMed/NCBI

[135] Google Scholar

[ref40] 40. Orihuela R, McPherson CA, Harry GJ (2016) Microglial M1/M2 polarization and metabolic states. Br J Pharmacol 173:649–665 pmid:25800044
View Article
PubMed/NCBI
Google Scholar

[137] View Article

[138] PubMed/NCBI

[139] Google Scholar

Figures

Abstract

Introduction

Material and methods

Chemical and reagents

Cell culture and growth conditions

Proliferation assay

Flow cytometry analysis

Cytokine assay

Statistical analysis

Results and discussion

Cell viability assay

Cytokine expression

Cellular differentiation

Conclusions

Supporting information

S1 Table. Monoclonal antibody mix used for the assessment of macrophage populations by flow cytometry.

S2 Table. Percentages of differentiated cells by argentatins.

S1 Raw data.

Acknowledgments

References