POOLED HUMAN IMMUNOGLOBULIN INHIBITS IL-4 BUT NOT IFN-γ OR TNF-α SECRETION FOLLOWING IN VITRO STIMULATION OF MONONUCLEAR CELLS WITH STAPHYLOCOCCAL SUPERANTIGEN

ABSTRACT Bovine tuberculosis in the United States has proven costly to cattle producers as well as to government regulatory agencies. While in vivo responsiveness to mycobacterial antigens is the current standard for the diagnosis of tuberculosis, in vitro assays are gaining acceptance, especially as ancillary or complementary tests. To evaluate in vitro indices of cellular sensitization, antigen-induced gamma interferon (IFN-γ), nitric oxide (NO), and tumor necrosis factor alpha (TNF-α) responses by blood mononuclear cells from Mycobacterium bovis-infected cattle were quantified and compared. Using an aerosol model of infection, two doses of each of two strains of M. bovis (95-1315 and HC-2045T) were used to induce a range of IFN-γ, NO, and TNF-α responses. Infection-specific increases in NO, but not in IFN-γ or TNF-α, were detected in nonstimulated cultures at 48 h, a finding that is indicative of nonspecific activation and spontaneous release of NO. The infective dose of M. bovis organisms also influenced responses. At 34 days postinfection, IFN-γ, NO, and TNF-α responses in antigen-stimulated cells from cattle receiving 105 CFU of M. bovis organisms were greater than responses of cells from cattle infected with 103 CFU of M. bovis organisms. The NO response, but not the IFN-γ and TNF-α responses, was influenced by infective strains of M. bovis. The TNF-α, NO, and IFN-γ responses followed similar kinetics, with strong positive associations among the three readouts. Overall, these findings indicate that NO and TNF-α, like IFN-γ, may prove useful as indices for the diagnosis of bovine tuberculosis.

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Modulation of TNF-α Secretion in Peripheral Blood Mononuclear Cells by Cocoa Flavanols and Procyanidins

Developmental Immunology ◽

10.1080/1044667031000137601 ◽

2002 ◽

Vol 9 (3) ◽

pp. 135-141 ◽

Cited By ~ 66

Author(s):

T. K. Mao ◽

J. van de Water ◽

C. L. Keen ◽

H. H. Schmitz ◽

M. E. Gershwin

Keyword(s):

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Human Peripheral Blood ◽

Resting Cells ◽

Peripheral Blood Mononuclear ◽

Tnf Α ◽

Blood Mononuclear Cells ◽

Stimulation Of

Epidemiological reports have suggested that the consumption of foods rich in flavonoids is associated with a lower incidence of certain degenerative diseases, including cardiovascular disease. Flavanols and their related oligomers, the procyanidins CFP, isolated from cocoa can modulate the production and level of several signaling molecules associated with immune function and inflammationin vitro, including several cytokines and eicosanoids. To further elucidate the potential immuno-modulatory functions of flavanol-rich cocoa, the present investigation examined whether isolated CFP fractions (monomers through decamers) influence the secretion of tumor necrosis factor-α (TNF-α) from resting and phytohemagluttinin (PHA)-stimulated human peripheral blood mononuclear cells (PBMC). We used anin vitroculture system where PBMC from 14 healthy subjects were introduced to individual CFP fractions for 72 h prior to measuring the levels of TNF-α released. The intermediate-sized CFP fractions (tetramers through octamers) were the most active on resting cells, causing a 3–4 fold increase in TNF-α relative to media baseline. The monomers and dimers were the least stimulatory of the fractions tested, displaying a 42 and 31% increase, respectively, over media control, whereas the trimers, nonamers and decamers showed an intermediate stimulation of this cytokine. In the presence of PHA, the intermediate-sized CFP fractions again were the most active, enhancing TNF-α secretion in the range of 48–128% relative to the PHA control. The monomers and dimers were slightly inhibitory (–1.5 and –15%, respectively), while trimers, nonamers and decamers stimulated moderate increases in TNF-α levels (13, 19 and 15%, respectively). The above results lend support to the concept that CFP can be immunomodulatory. The stimulation of TNF-α secretion may contribute to the putative beneficial effects of dietary flavanoids against microbial infection and tumorigenesis.

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A ComparativeIn VitroStudy of the Effects of Separate and Combined Products ofCitrus e fructibusandCydonia e fructibuson Immunological Parameters of Seasonal Allergic Rhinitis

Mediators of Inflammation ◽

10.1155/2012/109829 ◽

2012 ◽

Vol 2012 ◽

pp. 1-10 ◽

Cited By ~ 2

Author(s):

E. W. Baars ◽

M. C. Jong ◽

I. Boers ◽

A. F. M. Nierop ◽

H. F. J. Savelkoul

Keyword(s):

Allergic Rhinitis ◽

Seasonal Allergic Rhinitis ◽

Mononuclear Cells ◽

Immunological Parameters ◽

Peripheral Blood Mononuclear ◽

Tnf Α ◽

Ifn Γ ◽

Specific Stimulation

This paper examined the effects of the combined product,Citrus e fructibus/Cydonia e fructibus(Citrus/Cydonia; Citrus and Cydonia: each 0.01 g/mL), and separate products of Citrus (0.01 g/mL) and Cydonia (0.01 g/mL) on the immunological pathways involved in seasonal allergic rhinitis (SAR). Peripheral blood mononuclear cells (PBMCs) from five healthy and five grass pollen-allergic donors were isolated and analyzedin vitroafter polyclonal and allergen-specific stimulation of T cells in the presence of the three extracts. The analyses demonstrated acceptable cell survival with no signs of toxicity. Citrus mainly had a selective effect on reducing allergen-specific chronic inflammatory (TNF-α; Citrus compared to Cydonia and Citrus/Cydonia: −87.4 (P<0.001) and −68.0 (P<0.05), resp.) and Th2 pathway activity (IL-5; Citrus compared to Cydonia: −217.8 (P<0.01); while, both Cydonia and Citrus/Cydonia mainly affected the induction of the allergen-specific Th1 pathway (IFN-γ; Cydonia and Citrus/Cydonia compared to Citrus: 3.8 (P<0.01) and 3.0 (P<0.01), resp.). Citrus and Cydonia demonstrated different working mechanisms in the treatment of SAR and the combination product did not demonstrate larger effects than the separate preparations. Further effectiveness and efficacy studies comparing the effects of the products on SARin vivoare indicated.

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Immunostimulatory effect of faecal Bifidobacterium species of breast-fed and formula-fed infants in a peripheral blood mononuclear cell/Caco-2 co-culture system

British Journal Of Nutrition ◽

10.1017/s0007114511001656 ◽

2011 ◽

Vol 106 (8) ◽

pp. 1216-1223 ◽

Cited By ~ 27

Author(s):

T. Pozo-Rubio ◽

J. R. Mujico ◽

A. Marcos ◽

E. Puertollano ◽

I. Nadal ◽

...

Keyword(s):

Peripheral Blood ◽

Mononuclear Cells ◽

Interferon Γ ◽

Direct Stimulation ◽

Peripheral Blood Mononuclear ◽

Bifidobacterium Adolescentis ◽

Ifn Γ ◽

Breast Fed ◽

Stimulation Of

Bifidobacterium spp. typical of the human intestinal microbiota are believed to influence the balance of immune responses in the intestinal mucosa. The aim of the present study was to investigate the effect of different bifidobacterial species and their mixtures in in vitro experiments with peripheral blood mononuclear cells (PBMC) and Caco-2 cells. Bifidobacterium adolescentis, B. angulatum, B. breve, B. catenulatum, B. infantis, B. longum and two combinations of these bifidobacteria simulating the species composition found in faecal samples from breast-fed (BF) and formula-fed (FF) infants were used. The levels of several cytokines were measured by direct stimulation of PBMC and by stimulation of a Caco-2/PBMC co-culture with bifidobacteria. B. catenulatum and B. breve were the strongest enhancers of interferon-γ (IFN-γ) production by direct stimulation of PBMC. B. longum was the highest inducer of IL-10 and the lowest TNF-α stimulus. In the Caco-2/PBMC system, B. breve was the highest inducer of IL-8 production by Caco-2 cells, significantly different from B. infantis, B. adolescentis and the FF mixture (P < 0·05). IFN-γ produced by PBMC stimulated with the BF mixture (containing 22 % B. breve, compared with 7 % in the FF mixture) was significantly higher compared with B. adolescentis, B. infantis and B. longum. B. adolescentis also inhibited IFN-γ production compared with the FF mixture and B. longum. The proportion of different Bifidobacterium strains seems to be an important determinant of the cytokine balance in the simulated intestinal environment studied. B. breve and the combination of the Bifidobacterium species typically found in the microbiota of BF infants have shown the most significant effects.

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Decreased Interleukin-15 From Activated Cord Versus Adult Peripheral Blood Mononuclear Cells and the Effect of Interleukin-15 in Upregulating Antitumor Immune Activity and Cytokine Production in Cord Blood

Blood ◽

10.1182/blood.v90.8.3106 ◽

1997 ◽

Vol 90 (8) ◽

pp. 3106-3117 ◽

Cited By ~ 73

Author(s):

John X. Qian ◽

Sun min Lee ◽

Yu Suen ◽

Eva Knoppel ◽

Carmella van de Ven ◽

...

Keyword(s):

Cellular Immunity ◽

Immune Reconstitution ◽

Protein Production ◽

Low Dose ◽

Mononuclear Cells ◽

Tnf Α ◽

Interleukin 15 ◽

Ifn Γ ◽

Adult Peripheral Blood

Abstract Interleukin-15 (IL-15) is an important lymphokine regulating natural killer (NK) activity, T-cell proliferation, and T-cell cytotoxic activities. We hypothesized that the reduced expression and production of IL-15 from cord blood (CB) may contribute to the immaturity of CB immunity and potentially delay immune reconstitution after CB transplantation. We compared the expression and production of IL-15 from activated cord versus adult mononuclear cells (MNCs) and the regulatory mechanisms associated with IL-15 expression in CB MNCs. We have also studied the effect of exogenous IL-15 stimulation on CB and adult peripheral blood (APB) MNCs in terms of NK and lymphokine-activated killer (LAK) activities and cytokine induction. Lipopolysaccharide (LPS)-stimulated CB and APB MNCs were used to determine IL-15 expression and protein production by Northern analysis and Western immunoblot analysis. IL-15 mRNA expression and protein accumulation in CB MNC were 25% ± 2.0% (12 hours, n = 4, P < .05) and 30% ± 2.5% (12 hours, n = 3, P < .05), respectively, when compared with APB MNCs. Nuclear run-on assays showed no differences between CB and APB MNCs during basal levels of transcription and after transcriptional activation. However, the half-life of IL-15 mRNA was approximately twofold lower in activated CB MNCs than in activated APB MNCs (CB: 101 ± 5.8 minutes v APB: 210 ± 8.2 minutes, n = 3, P < .05). Exogenous IL-15 significantly enhanced CB NK and LAK activities up to comparable levels of APB (P < .05). IL-15 also significantly induced interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) protein production (days 1, 3, and 6, P < .05, n = 3) in CB MNCs. IL-15–stimulated LAK cells induced a significant lytic response against two acute lymphoblastic cell lines and two pediatric neuroblastoma cell lines. Both NK and LAK activities were augmented by the combination of IL-12 and IL-15, and the low-dose combination of IL-12 and IL-15 achieved similar levels of in vitro NK and LAK cytotoxicity compared with higher doses of either lymphokine. The present study suggests that IL-15 mRNA and protein expression is decreased in activated CB, secondary, in part, to altered posttranscriptional regulation. The reduced production of IL-15 from CB MNCs in response to stimulation may contribute to the decrease in IFN-γ and TNF-α production and CB cellular immunity. However, exogenous IL-15 enhanced IFN-γ and TNF-α production and NK and LAK cytotoxicities in CB MNCs. The reduced production of IL-15 from activated CB may contribute to the immaturity of CB cellular immunity and delayed immune reconstitution after unrelated CB transplantation. Exogenous IL-15 administration may compensate for the immaturity of CB immunity. The synergistic in vitro effects of low-dose IL-12 and IL-15 also implies the possible use of low doses each of IL-12 and IL-15 for enhancing immune reconstitution and/or possibly as a form of antitumor immunotherapy after CB transplantation.

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Progesterone and 17β-Estradiol Enhance Regulatory Responses to Human Papillomavirus Type 16 Virus-Like Particles in Peripheral Blood Mononuclear Cells from Healthy Women

Clinical and Vaccine Immunology ◽

10.1128/cvi.00441-09 ◽

2010 ◽

Vol 17 (4) ◽

pp. 609-617 ◽

Cited By ~ 19

Author(s):

Morgan A. Marks ◽

Patti E. Gravitt ◽

Robert D. Burk ◽

Yevgeniy Studentsov ◽

Homayoon Farzadegan ◽

...

Keyword(s):

Human Papillomavirus ◽

Peripheral Blood Mononuclear Cells ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Hpv 16 ◽

Tnf Α ◽

Blood Mononuclear Cells ◽

17Β Estradiol ◽

Ifn Γ

ABSTRACT Human papillomavirus (HPV) virus-like particle (VLP) vaccines are highly effective at preventing viral infections and the development of precancerous lesions through the induction of high-titer neutralizing antibodies and strong cell-mediated immune responses. Women taking combined oral contraceptives (COCs), however, show large variabilities in the magnitudes of their antibody responses. The goal of the present study was to determine the effects of 17β-estradiol (E2) and progesterone (P4) alone and in combination on the cellular immune response to HPV type 16 (HPV-16) VLPs in vitro. Peripheral blood mononuclear cells (PBMCs) from healthy donor women were stimulated in vitro with HPV-16 VLPs (2.5 μg/ml) in the presence of E2 and P4 administered either alone or in combination; and lymphoproliferation, cytokine production, transcription factor expression, and steroid hormone receptor expression were analyzed. HPV-16 VLPs significantly increased the levels of lymphoproliferation, proinflammatory cytokine (gamma interferon [IFN-γ], interleukin-1β [IL-1β], IL-2, IL-6, IL-8, IL-12p70, IL-17, tumor necrosis factor alpha [TNF-α]) production, anti-inflammatory cytokine (IL-1ra, IL-10) production, and the expression of Erα and Erβ but decreased the levels of Foxp3 expression and production of transforming growth factor β (TGF-β). Exposure of PBMCs to E2 and P4 either alone or in combination significantly decreased the levels of lymphoproliferation and production of proinflammatory cytokines (IFN-γ, IL-12p70, TNF-α) but increased the levels of production of IL-10 and TGF-β and the expression of Foxp3 in response to HPV-16 VLPs. Treatment of cells with biologically relevant concentrations of sex steroid hormones suppressed the inflammatory response and enhanced the regulatory response to HPV-16 VLPs, which may have implications for predicting the long-term efficacy of HPV vaccines, adverse events, and cross-protection among women taking COCs.

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Multifunctional, TNF-α and IFN-γ-Secreting CD4 and CD8 T Cells and CD8High T Cells Are Associated With the Cure of Human Visceral Leishmaniasis

Frontiers in Immunology ◽

10.3389/fimmu.2021.773983 ◽

2021 ◽

Vol 12 ◽

Author(s):

Lorranny Santana Rodrigues ◽

Aline Silva Barreto ◽

Lays Gisele Santos Bomfim ◽

Marcos Couto Gomes ◽

Nathalia Luisa Carlos Ferreira ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Visceral Leishmaniasis ◽

Mononuclear Cells ◽

Healing Process ◽

T Cell Response ◽

Peripheral Blood Mononuclear ◽

Tnf Α ◽

Ifn Γ

Visceral leishmaniasis (VL) is a chronic and often fatal disease caused by protozoans of the genus Leishmania that affects millions of people worldwide. Patients with symptomatic VL have an impaired anti-Leishmania-specific CD4+ T-cell response, which is reversed after clinical cure. In contrast, the quality of the CD4+ and CD8+ T-cell responses involved in resistance and/or cure of VL relies on the capability of these cells to activate polyfunctional and memory responses, which are associated with the simultaneous production of three cytokines: IFN-γ, IL-2, and TNF-α. Models for the development of CD4 and CD8 T-cell quality in memory and protection to leishmaniasis have been described previously. We aimed to assess the functionality of the T cells involved in the recovery of the immune suppression throughout the VL treatment. Therefore, we cultured peripheral blood mononuclear cells (PBMCs) from VL patients and healthy controls in vitro with soluble Leishmania antigen (SLA). Cell surface markers and intracellular cytokine production were determined on days 7, 14, 21, 30, 60, 90, and 180 after the beginning of chemotherapy. We observed that the frequencies of CD4+TNF-α+IFN-γ+ and the multifunctional CD4+IL-2+TNF-α+IFN-γ+, together with CD4+TNF-α+ and CD4+IFN-γ+ T cells, increased throughout and at the end of the treatment, respectively. In addition, enhanced frequencies of CD8+IL-2+TNF-α+IFN-γ+ and CD8+TNF-α+IFN-γ T cells were also relevant in the healing process. Noteworthy, the frequencies of the CD4+ and CD8 central-memory T cells, which produce IL-2, TNF-α, and IFN-γ and ensure the memory response against parasite reinfection, are significantly enhanced in cured patients. In addition, the subset of the non-functional CD8Low population is predominant in VL untreated patients and decreases along the chemotherapy treatment. In contrast, a CD8High subset increased towards the cure. Furthermore, the cure due to treatment with meglumine antimoniate or with liposomal amphotericin B was associated with the recovery of the T-cell immune responses. We described the evolution and participation of functional T cells during the treatment of patients with VL. Our results disclosed that the clinical improvement of patients is significantly associated with the participation of the CD4+ and CD8+ cytokine-secreting T cells.

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Pentoxifylline Inhibits Superantigen-Induced Toxic Shock and Cytokine Release

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.6.4.594-598.1999 ◽

1999 ◽

Vol 6 (4) ◽

pp. 594-598 ◽

Cited By ~ 44

Author(s):

Teresa Krakauer ◽

Bradley G. Stiles

Keyword(s):

Mononuclear Cells ◽

Human Peripheral Blood ◽

Toxic Shock ◽

Peripheral Blood Mononuclear ◽

Necrosis Factor Alpha ◽

Tnf Α ◽

Ifn Γ ◽

Toxic Shock Syndrome Toxin

ABSTRACT Tumor necrosis factor alpha (TNF-α) is a critical cytokine that mediates the toxic effects of bacterial superantigens like staphylococcal enterotoxin B (SEB) and toxic shock syndrome toxin 1 (TSST-1). Pentoxifylline, an anti-inflammatory agent that inhibits endotoxemia and lipopolysaccharide (LPS)-induced release of TNF-α, was tested for its ability to inhibit SEB- and TSST-1-induced activation of human peripheral blood mononuclear cells (PBMCs) in vitro and toxin-mediated shock in mice. Stimulation of PBMCs by SEB or TSST-1 was effectively blocked by pentoxifylline (10 mM), as evidenced by the inhibition of TNF-α, interleukin 1β (IL-1β), gamma interferon (IFN-γ), and T-cell proliferation. The levels of TNF-α, IL-1α, and IFN-γ in serum after an SEB or TSST-1 injection were significantly lower in mice given pentoxifylline (5.5 mg/animal) versus control mice. Additionally, pentoxifylline diminished the lethal effects and temperature fluctuations elicited by SEB and TSST-1. Thus, in addition to treating endotoxemias, the cumulative in vitro and in vivo data suggest that pentoxifylline may also be useful in abrogating the ill effects of staphylococcal enterotoxins and TSST-1.

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Entamoeba histolytica L220 induces the in vitro activation of macrophages and neutrophils and is modulated by neurotransmitters

Acta Parasitologica ◽

10.1515/ap-2018-0031 ◽

2018 ◽

Vol 63 (2) ◽

pp. 270-279 ◽

Cited By ~ 3

Author(s):

Fabiola del Rocío Villalobos-Gómez ◽

Mario García-Lorenzana ◽

Galileo Escobedo ◽

Patricia Talamás-Rohana ◽

Rogelio Salinas-Gutiérrez ◽

...

Keyword(s):

Entamoeba Histolytica ◽

Immune Cells ◽

Vecuronium Bromide ◽

First Line ◽

Human Macrophages ◽

Tnf Α ◽

In Vitro Activation ◽

Ifn Γ ◽

Stimulation Of

Abstract The neuroimmunoregulation of inflammation has been well characterized. Entamoeba histolytica provokes an inflammatory response in the host in which macrophages and neutrophils are the first line of defense. The aim of this study was to analyze the effect of the 220 kDa lectin of Entamoeba histolytica on stimulation of human macrophages and neutrophils, especially the secretion of cytokines and the relation of these to neurotransmitters. Human cells were interacted with L220, epinephrine, nicotine, esmolol and vecuronium bromide. The concentrations of IL-1β, IFN-γ, TNF-α and IL-10 were determined by ELISA at, 4 h of interaction. L220 has a cytokine stimulating function of macrophages and neutrophils for secretion of IL-1β, and IL-10 only by macrophages, which was modulated by the effect of vecuronium on cholinergic receptors in this immune cells.

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Bacterium-Dependent Induction of Cytokines in Mononuclear Cells and Their Pathologic Consequences In Vivo

Infection and Immunity ◽

10.1128/iai.67.5.2125-2130.1999 ◽

1999 ◽

Vol 67 (5) ◽

pp. 2125-2130 ◽

Cited By ~ 32

Author(s):

Yanling Jiang ◽

Luciano Magli ◽

Michael Russo

Keyword(s):

Inflammatory Response ◽

Mononuclear Cells ◽

Oral Bacteria ◽

Interleukin 12 ◽

Potent Inducer ◽

Tnf Α ◽

Oral Pathogens ◽

Ifn Γ

ABSTRACT Viridans streptococci are a heterogeneous group of gram-positive bacteria that are normal inhabitants of the mouth. These organisms are thought to contribute significantly to the etiology of infective endocarditis, although recently they have been implicated in serious infections in other settings. Another group of oral bacteria, gram-negative anaerobes, is associated with chronic dental infections, such as periodontal diseases or endodontic lesion formation. We evaluated the ability of the oral pathogens Streptococcus mutans and Porphyromonas endodontalis to induce a pathogenic response in vivo, with the goal of quantifying the inflammatory response in soft tissue by measuring leukocyte recruitment and hard tissues by measuring osteoclastogenesis. S. mutansinduced a strong inflammatory response and was a potent inducer of osteoclast formation, while P. endodontalis was not. To further study the mechanisms by which P. endodontalis andS. mutans elicit significantly different levels of inflammatory responses in vivo, we tested the capacity of each to induce production of cytokines by mononuclear cells in vitro. S. mutans stimulated high levels of interleukin-12 (IL-12), gamma interferon (IFN-γ), and tumor necrosis factor alpha (TNF-α), all of which are associated with inflammation, enhanced monocyte function, and generation of a Th1 response. In contrast, P. endodontalisstimulated production of IL-10 but not of TNF-α, IL-12, or IFN-γ. These results demonstrate that oral pathogens differ dramatically in their abilities to induce inflammatory and immunoregulatory cytokines. Moreover, there is a high degree of correlation between the cytokine profile induced by these bacteria in vitro and their pathogenic capacity in vivo.

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