T-cell specific immunosuppression by prodigiosin isolated from Serratia marcescens

doi:10.1016/S0192-0561(97)00062-3

International Journal of Immunopharmacology

Volume 20, Issues 1–3, 8 July 1998, Pages 1-13

International Journa...

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Abstract

Prodigiosin was isolated from the culture broth of Serratia marcescens B-1231. This compound inhibited the T-cell mediated immune responses such as concanavalin-A induced proliferation, mixed lymphocyte response, local graft vs host reaction and T-dependent antibody response at non-toxic concentrations. However, prodigiosin did not affect B-cell mediated immune functions such as lipopolysaccharide-induced proliferation and -activated polyclonal antibody production at the same concentrations. Prodigiosin did not cause death in vitro to lymphocytes at effective concentrations (<100 nM) and also did not show toxicity in vivo to lymphoid organs at effective dosages (10 and 30 mg\kg). The pharmacological potencies were comparable to the activities of other T-cell specific immunosuppressants such as cyclosporin A and FK-506. In conclusion, it might be suggested that prodigiosin could be used as an immunosuppressant in clinical and immunological studies.

Section snippets

Recently, immunomodulators, especially suppressants, made a number of valuable contributions in the improvement of immunosuppressive therapy, in the context of organ and tissue transplantation and in our awareness of immunological phenomena which underlay and might be crucial for the acceptance of organ transplantations (Thomson and Strazi, 1993).Cyclosporin A, cyclophosphamide, rapamycin and FK506 were included in this category and clinically studied or used in organ transplantation (Kino et

Materials

Specific pathogen free (SPF) mice B6C3F1, C3H and BDF1 (female, 5–7 weeks old) were obtained from Daehan Experimental Animal Center (Seoul, Korea). They were maintained under SPF conditions until used. Autoclaved food and water were supplied ad libitum. RPMI 1640 medium was purchased from GIBCO BRL (Grand Island, NY, U.S.A.). Medium was supplemented with 10% fetal calf serum (FCS) (HyClone, Logan, UT, U.S.A.) and 50 μM 2-mercaptoethanol (2-ME) (Sigma, St Rouis, MO, U.S.A.).

Prodigiosin

Isolation of prodigiosin from culture broth of Serratia marcescences

During the course of the screening program to find immunomodulators from microbial sources, an active compound was isolated from the culture broth of bacterial strain B-1231, which was isolated from a marine sample from Mokpo, Chunnam Province, Korea. It was taxonomically identified as Serratina marcescences. An active material was purified by ethyl acetate extraction and silica gel column chromatography. By spectroscopic analysis, the compound was proved to be prodigiosin (molecular weight,

Cytotoxicity of prodigiosin on lymphocytes in vitro

Splenic lymphocytes were cultured with prodigiosin (0.3–30,000 nM) from day 0 to day 3. Fig. 2 shows the relationship between the concentration of prodigiosin and the viability of lymphocytes. At below 100 nM, prodigiosin did not induce cell death. However, above 300 nM, the viability of lymphocytes rapidly decreased starting on day 2. In the following experiments, the concentrations of prodigiosin were adjusted to lower than 100 nM.

Suppression of T- and B-cell proliferation by prodigiosin

Fig. 3 shows the effects of prodigiosin on the proliferation of T and B cells induced by various lymphocyte-mitogens. Prodigiosin suppressed T-cell proliferation induced by concanavalin A (Con A, 5 μg\ml) at concentrations higher than 3 nM and a phenomenal suppression was observed at 30 nM. The proliferation of lymphocytes induced by pokeweed mitogen (PWM, 5 μg\ml), known as a T and B cell common mitogen, was also suppressed starting at a concentration of 10 nM and was strongly suppressed at

Suppression of the immune functions of T and B cells by prodigiosin in vitro

We evaluated the effects of prodigiosin on the immune functions of T and B cells (Fig. 4). The antibody production of B cells activated by polyclonal B cell stimulant, LPS (25 μg\ml) which activated B cells to antibody producing cells, was evaluated on day 3 after adding LPS and prodigiosin (0.3–100 nM). The antibody production of B cells was not changed by prodigiosin at the stated concentration ranges.

Then T-dependent antigen, sheep red blood cells, was chosen as another antigen to immunize B

Subset analysis

Even though the viability of total lymphocytes was not influenced by prodigiosin at concentrations from 0.3–100 nM, the selective toxicity of T cells might occur in the context of the selective suppression of T cells. This possibility was examined by flow cytometric analysis. Prodigiosin was treated in vitro for 3 days. Fig. 5 illustrates that the percentages of B- and T-cell subsets in total cell population are not changed by prodigiosin at concentrations up to 100 nM, suggesting that the

Suppression of graft vs host reaction (GvHR) by prodigiosin

GvHR was a T-cell specific response and was induced by the immunorejection between allogeneic strains. The potential of prodigiosin in the in vivo suppression of T cells was examined in the GvHR (Fig. 6A). Prodigiosin inhibited the enlargement of popliteal lymph nodes by 68.4% and 72.3% of control at 10 and 30 mg\kg, respectively. The positive control was cyclophosphamide (100 mk\kg), which strongly inhibited the GvHR. The treated animals did not show any toxicity and their body weight did not

Suppression of in vivo T-dependent antibody response by prodigiosin

T-dependent antibody response to SRBC which required the participation of T, B, and antigen presenting cells were also suppressed by the in vivo treatment of prodigiosin (Fig. 7). The inhibition effects of prodigiosin on in vivo T-dependent antibody response was exactly the same as those of GvHR. Prodigiosin inhibited the production of antibody forming cells by 68.4% and 72.6% of control at 10 and 30 mg\kg, respectively. Contrary to cyclophosphamide showing strong toxicity to mice, prodigiosin

Discussion

Median inhibition doses (ID₅₀) of prodigiosin on T-cell proliferation induced by Con A and T-cell activation induced by MLR were calculated to be 9.7 nM and 59.5 nM, respectively. On the contrary, ID₅₀ of B-cell functions was much higher than 100 nM. The suppression of T-cell functions was not mediated by cell death at concentrations below 100 nM. Also, in in vivo conditions, prodigiosin suppressed T-cell participating immune functions such as local graft vs host reaction and T-dependent

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Cited by (101)

Prodigiosin from Serratia rubidaea MJ 24 impedes Helicoverpa armigera development by the dysregulation of Juvenile hormone-dopamine system
2023, Microbiological Research
Prodigiosin pigment is a secondary metabolite produced by many bacterial species and is known for its medicinal properties. A few of these prodigiosin-producing bacteria are also reported to be entomopathogenic. It is intriguing to unravel the role of prodigiosin in insecticidal activities and its mode of action. In this study, we have shown the production and characterization of prodigiosin from the Serratia rubidaea MJ 24 isolated from the soil of the Western Ghats, India. Further, we assessed the effect of this pigment on the lepidopteran agricultural pest, Helicoverpa armigera. Prodigiosin-fed H. armigera indicated defective development of insect growth upon treatment. Due to defective early development, about 50% mortality and 40% reduction in body weight were observed in insects fed on a 500 ppm prodigiosin-containing diet. The transcriptomic analysis of these insects indicated significant dysregulation of Juvenile hormone synthesis and response related genes. In addition, dopamine related processes and their resultant melanization and sclerotization processes were also found to be affected. The changes in the expression levels of the key transcripts were further validated using real-time quantitative PCR. The metabolome data confirmed the developmental dysregulation of precursors and products of differentially regulated genes due to prodigiosin. Therefore, the corroborated data suggests that prodigiosin majorly affects H. armigera development through dysregulation of the Juvenile hormone-dopamine system and can be considered as a bioactive scaffold to design insect-pest management compounds. This study provides the first report of in-depth analysis of insecticidal system dynamics in H. armigera insects upon prodigiosin feeding via gene expression and metabolic change via omics approach.
Solid-state fermentation of food waste by Serratia marcescens NCHU05 for prodigiosin production
2023, Journal of the Taiwan Institute of Chemical Engineers
Prodigiosin is a red colored pigment with applications in antibacterial, antifungal, anticancer, and immunosuppressive activities. It is produced as a secondary metabolite by bacteria genera such as Serratia, Pseudomonas, and Streptomyces. The high-value prodigiosin offers a promising opportunity for the valorization of local food wastes.
The newly found red pigment-producing colony was isolated, sequenced, and identified. The red pigment was characterized using UV–VIS and liquid chromatography mass spectrum. Two cultivation methods, submerged culture and solid-state fermentation (SSF), were employed to produce prodigiosin. Three food waste sources, including rice bran, fish meal, and soybean dreg, were employed as both nutrition and solid matrices for SSF.
A new red pigment-producing Serratia marcescens NCHU05 was successfully isolated and identified. The red pigment was characterized as prodigiosin. S. marcescens NCHU05 was employed for prodigiosin production. SSF outperformed submerged cultures in prodigiosin production. Specifically, SSF using LB1 agar plate (1 g/L NaCl) yielded 22 mg/kg-dry-solid, while submerged cultures in LB1 medium produced only 3.2 ± 0.2 mg/L. Remarkably, when rice bran, fish meal, and soybean dregs were used as nutrients and solid matrices for SSF, prodigiosin productions of 50, 54, and 79 mg/kg-dry-solid were achieved, respectively. This study illustrates the feasibility of utilizing locally isolated microorganisms to contribute to the valorization of local wastes.
Concerted regulation of OPG/RANKL/ NF‑κB/MMP-13 trajectories contribute to ameliorative capability of prodigiosin and/or low dose γ-radiation against adjuvant- induced arthritis in rats
2022, International Immunopharmacology
Prodigiosin (PDG) is a microbial red dye with antioxidant and anti-inflammatory properties, although its effect on rheumatoid arthritis (RA) remains uncertain. Also, multiple doses of low dose γ- radiation (LDR) have been observed to be as a successful intervention for RA. Thus, the purpose of this study was to investigate the ameliorative potential of PDG and/or LDR on adjuvant-induced arthritis (AIA) in rats.
The anti-inflammatory and anti-arthritic effects of PDG and/or LDR were examined in vitro and in vivo, respectively. In the AIA model, the arthritic indexes, paw swelling degrees, body weight gain, and histopathological assessment in AIA rats were assayed. The impact of PDG (200 µg/kg; p.o) and/or LDR (0.5 Gy) on the levels of pro- and anti-inflammatory cytokines (IL-1β, TNF-α, IL-6, IL-18, IL-17A, and IL-10) as well as the regulation of osteoprotegrin (OPG)/ receptor activator of nuclear factor κB ligand (RANKL)/ nuclear factor-κB (NF-κB)/MMP-13 pathways was determined. Methotrexate (MTX; 0.05 mg/kg; twice/week, i.p) was administered concurrently as a standard anti-arthritic drug.
PDG and/or LDR markedly diminished the arthritic indexes, paw edema, weigh loss in AIA rats, alleviated the pathological alterations in joints, reduced the levels of pro-inflammatory cytokines IL-1β, TNF-α, IL-6, IL-18, IL-17A, and RANKL in serum and synovial tissues, while increasing anti-inflammatory cytokines IL-10 and OPG levels. Moreover, PDG and/or LDR down-regulated the expression of RANKL, NF-κBp65, MMP13, caspase-3, and decreased the RANKL/OPG ratio, whereas OPG and collagen II were enhanced in synovial tissues.
PDG and/or LDR exhibited obvious anti-RA activity on AIA.
Survivin modulation in the antimelanoma activity of prodiginines
2020, European Journal of Pharmacology
Citation Excerpt :
Prodiginines are a family of red-pigmented bacteria-produced alkaloids classified into linear or cyclic structures based on their alkyl chains (Hu et al., 2016). The SAR of prodiginines has only been addressed for antimalarial activity (Castro, 1967; Campos and De Melo, 2014), but these molecules have displayed a wide range of biological properties, including immunosuppressive (Han et al., 1998; Lee et al., 2000), antibacterial (Lapenda et al., 2015; Danevčič et al., 2016; Suryawanshi et al., 2017) and anticancer activities (Williamson et al., 2006). Recently, Obatoclax [2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methylene)]-3-methoxy-2H-pyrrol-5-yl)-1H-indole (GX15-070) entered phase I and II clinical trials to treat leukemia, lung cancer, lymphoma, and myelodysplastic syndrome (https://www.clinicaltrials.gov).
Melanoma is a type of skin cancer with an elevated incidence of metastasis and chemoresistance. Such features hamper treatment success of these neoplasms, demanding the search for new therapeutic options. Using a two-step resin-based approach, we recently demonstrated that cytotoxic prodiginines bind to the inhibitor of apoptosis protein, survivin. Herein, we explore the role of survivin in melanoma and whether its modulation is related to the antimelanoma properties of three cytotoxic prodiginines (prodigiosin, cyclononylprodigiosin, and nonylprodigiosin) isolated from marine bacteria. In melanoma patients and cell lines, survivin is overexpressed, and higher levels negatively impact survival. All three prodiginines caused a decrease in cell growth with reduced cytotoxicity after 24 h compared to 72 h treatment, suggesting that low concentrations promote cytostatic effects in SK-Mel-19 (BRAF mutant) and SK-Mel-28 (BRAF mutant), but not in SK-Mel-147 (NRAS mutant). An increase in G1 population was observed after 24 h treatment with prodigiosin and cyclononylprodigiosin in SK-Mel-19. Further studies indicate that prodigiosin induced apoptosis and DNA damage, as detected by increased caspase-3 cleavage and histone H2AX phosphorylation, further arguing for the downregulation of survivin. Computer simulations suggest that prodigiosin and cyclononylprodigiosin bind to the BIR domain of survivin. Moreover, knockdown of survivin increased long-term toxicity of prodigiosin, as observed by reduced clonogenic capacity, but did not alter short-term cytotoxicity. In summary, prodiginine treatment provoked cytostatic rather than cytotoxic effects, cell cycle arrest at G0/G1 phase, induction of apoptosis and DNA damage, downregulation of survivin, and decreased clonogenic capacity in survivin knockdown cells.
Isolation of anticancer and antimicrobial metabolites from Epicoccum nigrum; endophyte of Ferula sumbul
2017, Microbial Pathogenesis
Owing to the importance of endophytes, current research was aimed to purify the secondary metabolites from targeted source. Ferula sumbul, a lipophilic extract of the endophyte was prepared in 10% methanol and partitioned with ethyl acetate and bioassay guided isolation was carried using standard protocols against bacterial, fungal and cancer cells. The active fractions consisted of three new metabolites (2-methyl-3-nonyl prodiginine, Bis (2-ethylhexyl) phthalate, and a meroterpenoid, Preaustinoid A). Their structures were confirmed with LCMS/MS. The purified metabolites showed valuable results against tested activities which concluded that these compounds have great potential and these may be applicable to textile (dyeing), pharmaceutical (drug, infectious agents) and food (preservatives) industries. This study reveals the potential of E. nigrum as an important source of bioactive compounds including 2-methyl-3-nonyl prodiginine, Bis (2-ethylhexyl) phthalate, and Preaustinoid A. This is first report of isolation of prodiginines as well as meroterpenoid and Bis (2-ethylhexyl) phthalate from Epicoccum nigrum.
Microbial pigments: Sources, current status, future challenges in cosmetics and therapeutic applications
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T-cell specific immunosuppression by prodigiosin isolated from Serratia marcescens

Abstract

Section snippets

Materials

Isolation of prodigiosin from culture broth of Serratia marcescences

Cytotoxicity of prodigiosin on lymphocytes in vitro

Suppression of T- and B-cell proliferation by prodigiosin

Suppression of the immune functions of T and B cells by prodigiosin in vitro

Subset analysis

Suppression of graft vs host reaction (GvHR) by prodigiosin

Suppression of in vivo T-dependent antibody response by prodigiosin

Discussion

J. Pharmacol.

Immunopharmacol

Int. J. Immunopharmacol

J. Org. Chem.

Eur. J. Immunol.

Nature

Bone Marrow Transplant.

J. Antibiotics

Biochem.

Toxicol.

J. Antibiotics

J. Antibiotics

J. Immunol.