Showing papers in "Immunopharmacology and Immunotoxicology in 2003"

Modulation of inflammatory mediators by ibuprofen and curcumin treatment during chronic inflammation in rat.

Abstract: Inflammation is a protective tissue response occurring in three distinct phases, acute, subacute and a chronic proliferative phase. We undertook the present study to understand the overall immune response of the body during adjuvant induced chronic inflammation in rat and the effect of ibuprofen and curcumin on this response. Inflammatory mediators were estimated on day 21 and day 35 after adjuvant injection. The level of C-reactive protein increased to 200% on day 21 and then reduced to 50% on day 35 compared to control. Curcumin and ibuprofen further reduced the increased levels at both the time intervals. Haptoglobin level decreased to 42% on day 21 but increased to 5 times of control on day 35. Curcumin and ibuprofen reduced the increased levels at day 35. No significant change was observed in Prostaglandin-E2 and Leukotriene-B4 levels and in Lymphocyte proliferation. The level of Tumor Necrosis Factor-alpha increased by three folds on day 21, but came down to 88% on day 35. Ibuprofen treatment decreased the raised level on day 21 and increased the reduced level on day 35. Interleukin-1beta increased to 2 folds on day 21 and 10 folds on day 35 which were significantly brought down by curcumin and ibuprofen. Nitric oxide level was reduced at both the time intervals, which were increased by drug treatment.

The Influence of β‐Glucan on Immune Responses in Broiler Chicks

Abstract: Beta‐1,3/1,6‐glucan (β‐glucan) was tested as a possible immunomodulator. Chicken macrophages from a macrophage cell line MQ‐NCSU and from abdominal exudate of broiler chickens were exposed to vario...

Immunomodulatory activity of naturally occurring monoterpenes carvone, limonene, and perillic acid.

Abstract: The immunomodulatory activity of some naturally occurring monoterpenes were studied in Balb/c mice. Administration of various monoterpenes such as carvone (100 micromoles/Kg body wt/dose/animal), limonene (100 micromoles/Kg body wt/dose/animal) and perillic acid (50 micromoles/Kg body wt/dose/animal) were found to increase the total white blood cells (WBC) count in Balb/c mice. The maximum total WBC count in carvone treated animals was observed on the 12th day (16,560 cells/cmm) while in limonene (13,783 cells/cmm) and perillic acid (14,437 cells/cmm) treated animals the maximum count was observed on the 9th day after the drug treatment. Administration of terpenoids increased the total antibody production, antibody producing cells in spleen, bone marrow cellularity and alpha-esterase positive cells significantly compared to the normal animals indicating its potentiating effect on the immune system.

Minimal effects on immune parameters following chronic anti-TGF-beta monoclonal antibody administration to normal mice.

Abstract: Mice genetically deficient in TGF-beta1 or TGF-beta signaling capacity in T or B cells demonstrate profound immune dysregulation, as evidenced by increased lymph node size, expression of markers of memory/activation on T cells, inflammation in a variety of tissues and development of autoantibodies. However, this constant and complete lack of TGF-beta1 or TGF-betaR signaling may not reflect effects of TGF-beta neutralization using antibodies in mature animals. Thus, the present studies were designed to determine if administration of an anti-TGF-beta monoclonal antibody (neutralizes TGF-beta1, 2 and 3) to mature, normal mice results in evidence of immune dysregulation or immune-mediated pathology. An initial study examined daily administration of 0.25, 0.75 and 2.5 mg/kg of anti-TGF-beta to mice for three weeks, achieving blood levels of as high as 9 mg/ml. Comprehensive hematological and histopathological evaluation showed no evidence of pathology. A second study was designed to extend the antibody treatment period and further examine the functional status of the immune system. Mice were injected with 1 mg/mouse (approximately 50 mg/kg) of anti-TGF-beta (1D11) three times per week achieving circulating blood levels of 1-2 mg/ml. Many parameters of immune status were assessed, including natural killer (NK) cell activity, lymphocyte proliferative responses, phagocytic activity, phenotypic assessment of leukocyte subsets, and serum measurements of proinflammatory cytokines, autoantibodies and immunoglobulin isotypes. In addition, histopathological assessment of heart, lungs, liver, kidney, salivary glands, skin, spleen and lymph nodes was also performed. Very few of the multiple immune parameters examined showed detectable changes in anti-TGF-beta-treated mice. Changes that were observed were primarily restricted to the spleen and included increased spleen cell recoveries, increased percentages of macrophages, decreased percentages of NK cells, decreased phagocytic activity, decreased proliferative responses to mitogens and slight increases in T and B cells displaying an activated phenotype. Many of these same parameters examined in the lymph nodes were not altered by the anti-TGF-beta treatment. The thymus was decreased in size, but altered only slightly in one population of developing T cells. Most of the changes observed were modest and returned to control levels after discontinuation of treatments. The only serological finding was an increase in IgA levels in anti-TGF-beta-treated mice, but not in any other isotype. Finally, there was no evidence of increased inflammation in any of the peripheral tissues examined in the anti-TGF-beta-treated mice. In conclusion, although there were changes in some of the immunological parameters examined in these studies, they were few and typically reversed following discontinuation of treatment. The modest nature of the changes observed in these studies is particularly evident when compared to published data of those same parameters examined in mice genetically deficient in TGF-beta1 or mice having TGF-beta unresponsive T or B cells. Thus, there does not appear to be any significant immune dysregulation detectable after long-term antibody-mediated neutralization of TGF-beta in normal mice.

Hepatoprotective Effect of Baicalin, a Major Flavone from Scutellaria radix, on Acetaminophen‐Induced Liver Injury in Mice

Abstract: The protective effects of baicalin (BA), a major flavone from Scutellaria radix, on acetaminophen (AP)-induced hepatotoxicity and the possible mechanism(s) of its protective action were investigated in mice. Treatment with BA (300 mg/kg, p.o.) 0.5 h after AP administration significantly prevented an increase in plasma alanine aminotransferase and aspartate aminotransferase activities and AP-induced hepatic necrosis, and also reduced AP-induced mortality from 43% to 0%. In addition, oral treatment with BA significantly prevented AP-induced depletion of glutathione (GSH) contents. However, BA treatment, by itself, did not affect hepatic GSH contents. The effect of BA on the cytochrome P450 2E1 (CYP2E1), the major isozyme involved in AP bioactivation, was investigated. Oral treatment of mice with BA resulted in a significant decrease in AP-induced CYP2E1 activity together with its inhibition of AP-induced CYP2EI expression. These results show that the hepatoprotective effects of BA against AP overdose may be due to its ability to block the bioactivation of AP by inhibiting CYP2E1 expression.

Enhancement of the humoral immune response by Echinacea purpurea in female Swiss mice.

Abstract: Various preparations of the plant Echinacea purpurea have been investigated for their potential to enhance immune function, primarily through activation of innate immune responses. Few studies have examined the potential for enhancement of humoral immunity. Using female Swiss mice we administered a volumetric dose of a glycerine extract of E. purpurea by oral gavage, to evaluate effects on the IgM specific antibody forming cell (AFC) response. Four days of treatment following immunization with sheep red blood cells (SRBC) produced a significant enhancement over naive controls at doses of 0.4 and 0.8 mL/kg/day. A few clinical trials and anecdotal reports have suggested that the greatest efficacy for E. purpurea occurs in acute use following onset of illness. A time course study, using the time of SRBC immunization to mimic the onset of illness, examined the effects of 8 and 4 days of E. purpurea treatment at 0.6 mL/kg/day. Only in the 4-day administration, with dosing beginning 1 hour after SRBC immunization, was there an observed enhancement of the antibody forming cell response. This supports the acute use of E. purpurea as suggested by anecdotal reports, and demonstrates the potential for enhancement of humoral immune responses as well as innate immune responses.

Swertia chirayita Mediated Modulation of Interleukin‐1β Interleukin‐6, Interleukin‐10, Interferon‐γ, and Tumor Necrosis Factor‐α in Arthritic Mice

Abstract: The effect of aqueous extract of Swertia chirayita stem on the pro- and anti-inflammatory cytokines balance in primary joint synovium of adjuvant-induced arthritic mice has been studied. The level of pro-inflammatory cytokines was found elevated in the joint synovium of arthritic mice in comparison to normal joints. Administration of S. chirayita extract in varying doses through the oral route did not modulate the proinflammatory cytokines on day 2. In contrast, by day 12, a dose dependent (0, 11.86 and 23.72 mg/kg body weight) reduction of tumor necrosis factor-alpha (INF-alpha) interleukin-1beta, (IL-beta) and interferon-gamma, (IFN-gamma) and elevation of Interleukin-10 (IL-10) was observed in the joint homogenates of arthritic mice. Interleukin-6 (IL-6) was not down regulated in joint homogenate of arthritic mice at the dose 11.86 mg/kg but at higher doses (23.72 and 35.58 mg/kg) significant reduction was observed. The aqueous extract was found to possess two polar compounds, amerogentin and mangiferin but was devoid of swerchirin, chiratol, methyl swetianin, and swertanone. Mangiferin has been reported to possess potent anti-inflammatory property and we presume its presence in the aqueous extract of S. chirayita is responsible for reducing TNF-alpha, IL-1beta, IL-6, and IFN-gamma and/or elevating IL-10 in the joint homogenates of arthritic mice on day 12. This study will help in our understanding of the mechanism of anti-inflammatory action of S. chirayita in the light of proinflammatory and anti-inflammatory cytokine balance.

Effect of Piperine on the Inhibition of Nitric Oxide (NO) and TNF‐α Production

Abstract: Effect of piperine which is an alkaloid present in plants such as Piper nigrum and Piper longum on the production of nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) level was analyzed using in vitro as well as in vivo systems. The level of nitrite in the LPS stimulated Balb/C mice (95.3 microM) was reduced in the piperine treated animals (25 microM) significantly. Nitrite level in the Concanavalin-A (Con-A) treated control animals (83.1 microM) was also significantly reduced to 18.5 microM in the piperine treated mice. The drastically elevated levels of TNF-alpha in the lipopolysaccharide (LPS) stimulated animals (625.8 pg/mL) was lowered in the piperine treated animals (105.8 pg/mL). Piperine also inhibited the Con-A induced TNF-alpha production. Piperine could inhibit the nitrite production by in vitro activated macrophages (116.25 microM) to the normal level (15.67 microM) at concentration of 5 microg/mL. In vitro L929 bioassay also revealed the inhibition of TNF-alpha production by the piperine treatment.

Enhanced Cytokine Synthesis of Leukocytes by a β‐Glucan Preparation, SCG, Extracted from a Medicinal Mushroom, Sparassis crispa

Abstract: Sparassis crispa is edible mushroom recently cultivable in Japan. It contains significantly high content (∼40%) of 6‐branched 1,3‐β‐D‐glucan showing antitumor activity in mice. We recently purified a β‐glucan preparation designated as “SCG.” It was considered worth while to test SCG in vitro with whole blood collected from human volunteers. The present study is focusing on the cytokine productivity of SCG in an in vitro human system. The following results were observed: (i) SCG dose dependently enhanced IL‐8 synthesis of whole blood cell culture of human peripheral blood. (ii) IL‐8 synthesis was enhanced in both PBMC and PMN cultures. (iii) IL‐8 synthesis was induced in the culture with autologous plasma, but significantly reduced after 56°C treatment. (iv) The activity was also weak in heat inactivated fetal calf serum (FCS). (v) A complement fragment, C5a, was released by SCG dependently upon dose and kinetics. (vi) Anti‐SCG natural antibody was detected in human plasma. From these facts, SCG wa...

Anti-inflammatory activity of Korean folk medicine purple bamboo salt.

Abstract: Purple bamboo salt is a specially processed salt according to the traditional recipe using normal salt and bamboo. It has been used for the purpose of prevention and treatment of various diseases in Korea. We investigated the anti-inflammatory activity of purple bamboo salt by using human mast cell line (HMC-1). Purple bamboo salt (1 mg/mL) inhibited phorbol 12-myristate 13-acetate (PMA) plus calcium ionophore A23187-stimulated tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-6 secretion, by 67.04% +/- 0.08%, 68.01% +/- 1.85%, and 69.48% +/- 0.54%, respectively. In addition, the expression of TNF-alpha mRNA in HMC-1 cells was inhibited by purple bamboo salt under the same condition. When NaCl (1 mg/mL) was added, the secretion of TNF-alpha and IL-6 was also inhibited but the effect was markedly lower than purple bamboo salt. Our results suggest that purple bamboo salt importantly contributes to the prevention or treatment of inflammatory diseases.

Salidroside from Rhodiola sachalinensis protects neuronal PC12 cells against cytotoxicity induced by amyloid-beta.

Abstract: The amyloid β‐peptide (Aβ)‐induced oxidative stress is a well‐established pathway of neuronal cell death in Alzheimer's disease (AD). Salidroside, one of the major compounds from the roots of Rhodiola species (Crassulaceae), was investigated in vitro for its cytoprotection against Aβ‐induced toxicity on rat neuronal PC12 cells. Salidroside significantly reduced Aβ‐induced cytotoxicity in a dose‐dependent manner. Salidroside also reduced Aβ‐mediated intracellular accumulation of reactive oxygen species and malondialdehyde (MDA), a product of lipid peroxides, by preventing Aβ‐induced decline of antioxidant enzyme activities. These results suggest that salidroside protects neuronal PC12 cells from Aβ‐induced cytotoxicity via its antioxidant pathway.

Effect of Phlomis umbrosa root on mast cell-dependent immediate-type allergic reactions by anal therapy.

Abstract: The effect of aqueous extract of Phlomis umbrosa Turcz. (Labiatae) root (PUAE) on mast cell-dependent immediate-type allergic reaction by anal therapy was investigated. PUAE (0.01 to 1 g/kg) dose-dependently inhibited systemic anaphylaxis induced by compound 48/80 in mice. When PUAE was pretreated at the same concentrations with systemic anaphylaxis, the plasma histamine levels were reduced in a dose-dependent manner. PUAE (0.1 and 1 g/kg) also significantly inhibited local anaphylaxis activated by anti-DNP IgE. PUAE (0.001 to 1 mg/mL) dose-dependently inhibited the histamine release from rat peritoneal mast cells (RPMC) activated by compound 48/80 or anti-DNP IgE. The level of cyclic AMP (cAMP) in human mast cells (HMC-1 cells) when PUAE (1 mg/mL) was added, transiently and significantly increased compared with that of basal cells. In addition, PUAE (0.1 and 1 mg/mL) inhibited the secretion of tumor necrosis factor-α (TNF-α), interleukin (IL)-6 in phorbol 12-myristate 13-acetate (PMA) plus calcium ionoph...

A toxicokinetic study of nickel-induced immunosuppression in rats.

Abstract: The aim of the present study was to investigate dose‐ and time‐dependent effects of NiCl2 on T‐lymphocyte and macrophage‐derived cytokine production in rats. Moreover we have determined the concentrations of nickel in the plasma that are required to elicit alterations in T‐lymphocyte and macrophage function. NiCl2 suppressed T‐lymphocyte proliferation and Th1 (IFN‐γ) and Th2 (IL‐10) cytokine production in a dose‐ and time‐dependent fashion. In addition, NiCl2 inhibited production of the pro‐inflammatory cytokine TNF‐α and increased production of the anti‐inflammatory cytokine IL‐10 from lipopolysaccharide (LPS) stimulated cultures. We have determined that the minimal plasma concentrations of nickel required to provoke immunosuppression are in the range 209–585 ng/mL. In the time‐course study NiCl2 (3.3 mg/kg) provoked immunological changes that were maximal 1 h following administration, and some of these changes persisted for up to 24 h post administration. Overall these data clearly demonstrate t...

Effect of naturally occurring isothiocyanates on the immune system.

Abstract: Administration of allyl isothiocyanate (AITC) and phenyl isothiocyanate (PITC) was found to stimulate the immunological response in Balb/c mice. Treatment with five doses of AITC and PITC (25 µg/dose/animal, i.p.) was found to enhance the total WBC count (13,100 cells/mm3 for AITC) on 9th day and (10,316 cells/mm3 for PITC) on 12th day respectively. Bone marrow cellularity (22.36 × 106 cells/femur for AITC, 21.3 × 106 cells/femur for PITC) as well as α‐esterase positive cell number also increased after treatment with AITC (1397 cells/4000 cells) and PITC (1238 cells/4000 cells). Treatment with AITC and PITC along with the antigen sheep red blood cells (SRBC) produced an enhancement in the circulating antibody titer and the number of plaque forming cells (PFC) in the spleen. Maximum number of PFC (1329 PFC/106 spleen cells) for AITC and PITC (1218 PFC/106 spleen cells) was obtained on the 5th day. These results indicate the immunomodulatory activities of the two naturally occurring isothiocyanates ...

Dang-Gui-Bu-Xai-Tang Modulated the Immunity of Tumor Bearing Mice

Abstract: Dang-Gui-Bu-Xai-Tang (DGBXT), which includes Radix Angelicae Sinensis and Radix Astragali Membranaceus, is a traditional Chinese medicine used to modulate the lymphocyte activity of cancer patients after chemotherapy and radiotherapy. In the present study, we examined the cytotoxicity of DGBXT on transformed cells and the immunomodulating effects of DGBXT in a tumor-bearing murine model. DGBXT markedly inhibited the growth of the EJ-Ha-ras transformed LZEJ and LZEJ-C2 cells lines. Oral administration of DGBXT for three weeks significantly prevented the tumor development in mice that injected with LZEJ-C2 cells subcutaneously. Moreover, DGBXT effectively increased the population of cytotoxic T lymphocytes (CTLs) and NK cells, and down-regulated activated T helper cells (CD4+/CD25+) in spleen and tumor-draining lymph nodes (TDLN). Furthermore, DGBXT stimulated the production of tumor necrosis factor-alpha in in vitro cultured splenocytes. These results might explain the antitumor effects of DGBXT.

Inhibition of immunologic and nonimmunologic stimulation-mediated anaphylactic reactions by the aqueous extract of Mentha arvensis.

Abstract: The effect of aqueous extract of Mentha arvensis L. var. piperascens Malinv. (Labiatae) (MAAE) on immunologic and nonimmunologic stimulation-mediated anaphylactic reactions was studied. Nonimmunologic anaphylactic reaction was induced by compound 48/80 injection. MAAE (0.005 to 0.5 g/kg) inhibited systemic anaphylactic reaction induced by compound 48/80. Immunologic anaphylactic reaction was generated by sensitizing the skin with anti-dinitrophenyl (DNP) IgE followed 48 h later with an injection of antigen. MAAE (0.001 to 1 g/kg) dose-dependently inhibited passive cutaneous anaphylaxis (PCA) when intraperitoneally, intraveneously and orally administered. MAAE (0.001 to 1 mg/ml) dose-dependently inhibited the histamine release from rat peritoneal mast cells (RPMC) activated by compound 48/80 or anti-DNP IgE. Moreover, MAAE (0.1 mg/ml) had a significant inhibitory effect on anti-DNP IgE-mediated tumor necrosis factor-alpha (TNF-alpha) production. These results indicate that MAAE inhibits immunologic and nonimmunologic stimulation-mediated anaphylactic reactions and TNF-alpha production from RPMC.

Effect of some mycotoxins on superoxide anion production of isolated human neutrophils and in whole blood.

Abstract: Activity of mycotoxins citrinin, ochratoxin B, rubratoxin B, and zearalenol β was studied on human neutrophils with regard to O2− generation. At the doses of 10−8, 10−6, 10−4, or 10−2 mg/mL the mycotoxins stimulated the superoxide anion production in resting neutrophils in whole blood but inhibited it in isolated cells. In neutrophils stimulated with zymosan or PMA, mycotoxins showed inibiting effect both on isolated and in whole blood cells. The behavior of resting isolated neutrophils, identical to that of stimulated cells, was caused by manipulations to isolate cells that activated O2− production. Activity of mycotoxins was no dose‐dependent permitting to consider their effect immunotoxic rather than immunomodulator; the absence of dose‐dependence was connected to high variability of individual sensitivity to these compounds. Since the smaller concentrations by us used (10−8 and 10−6 mg/mL) are easily reachable in blood, it is important to point out the danger represented for immune system by m...

Natural killer cell activity, lymphocyte proliferation, and cytokine profile in tumor-bearing mice treated with MAPA, a magnesium aggregated polymer from Aspergillus oryzae.

Abstract: The present study examined the effects of MAPA, an antitumor aggregated polymer of protein magnesium ammonium phospholinoleate-palmitoleate anhydride, isolated from Aspergillus oryzae, on concanavalin A (Con A)-induced spleen cell proliferation, cytokine production and on natural killer (NK) cell activity in Ehrlich ascites tumor-bearing mice. The Ehrlich ascites tumor (EAT) growth led to diminished mitogen-induced expansion of spleen cell populations and total NK activity. This was accompanied by striking spleen enlargement, with a marked increase in total cell counts. Moreover, a substantial enhancement in IL-10 levels, paralleled by a significant decrease in IL-2 was observed, while production of IL-4 and interferon-gamma (IFN-gamma) was not altered. Treatment of mice with 5 mg/kg MAPA for 7 days promoted spleen cell proliferation, IL-2 production and NK cell activity regardless of tumor outgrowth. In addition, MAPA treatment markedly enhanced IFN-gamma levels and reduced IL-10 production relative to EAT mice. A 35% reduction in splenomegaly with normal number of nucleated cells was also found. Altogether, our results suggest that MAPA directly and/or indirectly modulates immune cell activity, and probably disengages tumor-induced suppression of these responses. Clearly, MAPA has an impact and may delay tumor outgrowth through immunotherapeutic mechanisms.

Bombycis corpus extract (BCE) protects hippocampal neurons against excitatory amino acid-induced neurotoxicity.

Abstract: Bombycis corpus (BC) or Bombyx Batryticatus, a batryticated silkworm and white-stiff silkworm, is a drug consisting of the dried larva of silkworm, Mobyz mori L, dead and stiffened due to the infection of Beauveria (Bals) Vuill In a previous paper (Kim et al, Pharmacol Res, 43, 12–16, 2001), BC was shown to protect amyloid-β-induced cytotoxicity In the present study, we have found that BCE can prevent or reduce the neurotoxic actions in the hippocampus of the glutamate agonists N-methyl-D-aspartic acid (NMDA) in vitro or alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and kainic acid in vitro Pre-treatment with BCE (0, 1, 2, 5, and 10 μg/ml for 6–8 h) protected primary hippocampal cultures from embryonic day 18 (E18) embryos against NMDA-induced toxicity (01, 1, 10, and 50 mM/ml) BCE added either with NMDA (1 mM) or 1 h later had lesser, but still significant, protective actions BCE also reduced NMDA-induced toxicity (1 mM) BCE (10 μg/ml) protected cultured neurons against the

Thymopoiesis Following Chronic Blockade of β‐Adrenoceptors

Abstract: The present study was undertaken in order to further clarify putative role of the adrenergic innervation in the regulation of the intrathymic T‐cell maturation. For this purpose adult male DA rats were subjected to either 4‐day‐ or 16‐day‐long propranolol treatment (0.40 mg propranolol/100 g/day, s.c.) and the expression of CD4/8/TCRαβ on thymocytes, as well as thymocyte proliferative and apoptotic index, was assessed in these animals by flow cytometric analysis. Propranolol treatment, in spite of duration, increased both the thymocyte proliferative and apoptotic index (vs. respective vehicle‐treated controls). In 4‐day‐treated animals the thymus cellularity and thymus weight remained unaltered, while in 16‐day‐treated rats the values of both of these parameters were reduced (since increase in the thymocyte apoptotic index overcame that in the proliferative index). The treatments of both durations affected the thymocyte phenotypic profile in a similar pattern, but the changes were more pronounced ...

The immunostatus of T helper and T cytotoxic cells in the patients ten years after exposure to sulfur mustard.

Abstract: The treatment and cure of patients exposed to sulfur mustard is a remaining challenge despite ongoing research in this field. A severe suppression of the immune system still remains the major cause...

Effect of Bojungikki‐tang on Lipopolysaccharide‐Induced Cytokine Production from Peripheral Blood Mononuclear Cells of Chronic Fatigue Syndrome Patients

Abstract: Bojungikki-tang (BIT) has been widely used to treat patients suffering from chronic fatigue syndrome (CFS). However, its effect has not been yet investigated experimentally. Based upon the clinical presentation of CFS, we hypothesized that cytokines may play a role in the pathogenesis of the disease. We studied the effect of BIT on lipopolysaccharide (LPS)-induced various cytokines production in peripheral blood mononuclear cells (PBMC) of CFS patients. Bojungikki-tang (1 mg/mL) significantly inhibited LPS-induced tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-10, transforming growth factor (TGF)-beta1 production by 63.55% +/- 0.19%, 55.06% +/- 0.27%, 48.23% +/- 0.48%, 54.09% +/- 0.76%, respectively (P < 0.05). Bojungikki-tang showed a slightly lower inhibitory effect of LPS-induced Interferon (IFN)-gamma production. These results suggest that BIT may be useful in treating fatigue associated with chronic diseases.

Interleukin‐1 and Tumor Necrosis Factor‐α Induce Collagenolysis and Bone Resorption by Regulation of Matrix Metalloproteinase‐2 in Mouse Calvarial Bone Cells

Abstract: Interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) greatly induces osteoclast formation and stimulates bone resorption of mouse calvaria in culture. We examined the effects of the two cytokines on the collagenolysis and bone resorption by induction of matrix metalloproteinases (MMPs). The cells were analyzed using zymographic analysis. It was shown that the mouse calvarial osteoblasts constitutively synthesize progelatinase-A (MMP-2). Interleukin-1beta markedly enhanced the messenger RNAs (mRNAs) expression of MMP-2 (gelatinase A), but slightly MMP-9 (gelatinase B), which associated with increases in bone matrix degradation. Both pro- and active-forms of MMP-2 were detected in the conditioned medium collected from calvarial cultures, and IL-1beta markedly stimulated both pro- and active-forms of the MMP-2. The expression of MMP-2 mRNAs could be detected, and they were markedly enhanced by IL-1beta on days 1 and 2. These results demonstrate that the potency of induction of MMP-2 by IL-1beta and TNF-alpha is closely linked to the respective bone-resorbing activity, suggesting that MMP-2-dependent degradation of bone matrix plays a key role in bone resorption induced by these cytokines. On the other hand, when the mouse osteoblasts were stimulated with parathyroid hormone, 1,25(OH)2D3, mononuclear cell conditioned medium (MCM) and IL-1 as bone resorption agents, collagenolysis was increased by producing the active gelatinase. Interleukin-1 in stimulating bone resorption was examined using fetal mouse long bone organ culture. Interleukin-1 stimulated bone resorption and produced marked resorption when present simultaneously. Furthermore, treatment of indomethacin and dexamethasone clearly abolished the responses of IL-1alpha and IL-1beta.

The inhibition of superoxide production in EL4 lymphoma cells overexpressing growth hormone.

Abstract: A substantial body of research exists to support the production of growth hormone by cells of the immune system. However, the function and mechanism of action of lymphocyte-derived growth hormone remain largely unelucidated. Since, it has been found that exogenous growth hormone (GH) primes neutrophils for the production of reactive oxygen intermediates (ROI) and in particular superoxide (O2-), we investigated the role of GH on the production of O2- in T cells. Furthermore, we examined whether endogenous and exogenous GH act similarly. Our studies show that overexpression of GH in EL4, a T-cell lymphoma cell line, results in a decrease in the production of O2- compared to control cells, as detected using the fluorescent dye, dihydroethidium. O2- production in control cells was not affected by treatment with inhibitors of xanthine oxidase or a non-specific NADPH-oxidase inhibitor. However, treatment with diallyl sulfide, an inhibitor of cytochrome P450 2E1 mimicked the reduction in O2- production seen in cells overexpressing GH. Although no significant change could be detected in CYP2E1 protein levels, CYP2E1 activity was found to be greater in control EL4 than in cells overexpressing GH. Both the decrease in O2- production and the lower CYP2E1 activity in GH overexpressing cells could be abrogated by treatment with N(G)-monomethyl-L-arginine, an inhibitor of nitric oxide synthase. The overexpression of GH protects cells from apoptosis induced by isoniazid, a CYP2E1 inducer, suggesting a role for nitric oxide as a mediator in the regulation of xenobiotic metabolism and apoptosis-protection by lymphocyte GH.

Systemic reactions to allergen immunotherapy: a review of the literature.

Abstract: Since its introduction the safety of specific immunotherapy (SIT) was assessed by many well-designed studies. SIT is accepted as an effective treatment of allergic diseases despite the occurrence of side-effects, among which systemic reactions (SRs) are the most dangerous. The reported frequency of SRs after SIT varies among the studies and several factors influence it. Asthma is a particular risk factor for systemic side-effects. Furthermore, SRs occur more often in patients with high allergen sensitivity as determined by skin testing or RAST. Making dosage errors is also considered to be a high risk. It is reported that reactions are more common during rush and clustered induction treatment, whereas a significantly lower incidence of SRs occurred with the use of standardized modified allergen vaccines than with aqueous extracts. On the basis of valuable guidelines, precautions to minimize the risk of SRs from SIT were recommended. Injections should be given or supervised by doctors well-trained in this form of treatment in a clinic where there is the immediate availability of a resuscitative equipment. Consideration should be given to evaluate the patient's conditions and to monitor subjects for a minimum of 30 minutes after the injections. Therefore, if appropriately done, the risk of SIT is negligible.

Subchronic exposure to ellagic acid impairs cytotoxic T-cell function and suppresses humoral immunity in mice.

Abstract: Ellagic acid (EA) is present in a variety of foods such as grapes, strawberries, raspberries, and nuts. It is a dietary plant phenol that has been shown to inhibit oxidative stress and chemical carcinogenesis. Although several studies have examined the protective mechanisms of dietary EA including the induction of detoxifying enzymes, regulation of cell cycle, chelation of nickel, and prevention of DNA methylation, none have addressed the role of EA in immunological surveillance. This study investigates the status of immune function in B6C3F1 mice exposed continuously to EA in drinking water at 0.5, 1.0, or 2.0 mg/kg/day for 28 days. Although this range of exposure is above the estimated human daily intake (≈940 µg/day for 70 kg person or 13.4 µg/kg/day), these levels would not be unreasonable if EA were used as a dietary supplement or as a chemotherapeutic agent. Previous reports have demonstrated the anticarcinogenic effects of EA at levels 10‐ to 250‐fold greater than those applied in this stud...

Tumor necrosis factor alpha and apoptosis in Helicobacter pylori related progressive gastric damage: a possible mechanism of immune system involvement in epithelial turnover regulation.

Abstract: Helicobacter pylori (HP) related inflammation is mediated by tumour necrosis factor alpha (TNFα), which “in vitro” increases epithelial apoptosis in response to infection. In the early stages of HP gastritis, a raised epithelial apoptosis occurs; this phenomenon becomes less evident with progression towards intestinal metaplasia. Aim of our study was to analyze “in vivo” mucosal TNFα in relation to epithelial apoptosis in the progression of HP related histological damage. Antral biopsies from 20 HP positive patients were retrospectively studied: 10 with and 10 without intestinal metaplasia (IM and CG group respectively); samples of 10 dyspeptics with normal HP negative stomach (N) were used as control. The following parameters were evaluated by immunohistochemistry: 85 kDa caspase-cleaved fragment (p85) of human poly (ADP-ribose) polymerase (PARP) labelling index (LI) as marker of apoptosis and TNFα LI in stromal cells as marker of inflammatory response. Both epithelial apoptosis and mucosal TNFα expressi...

Saliva Secretory IgA Antibodies Against Molds and Mycotoxins in Patients Exposed to Toxigenic Fungi

Abstract: Upper respiratory exposure to different environmental antigens results first in the activation of mucosal immunity and production of IgA antibodies in different secretions including saliva Despite this there is no study, which addresses secretory antibodies against molds and mycotoxins The purpose of this study was to evaluate mold-specific salivary IgA in individuals exposed to molds and mycotoxins in a water-damaged building environment Saliva IgA antibody levels against seven different molds and two mycotoxins were studied in 40 patients exposed to molds and in 40 control subjects Mold-exposed patients showed significantly higher levels of salivary IgA antibodies against one or more mold species A majority of patients with high IgA antibodies against molds exhibited elevation in salivary IgA against mycotoxins, as well These IgA antibodies against molds and mycotoxins are specific, since using molds and mycotoxins in immune absorption could reduce antibody levels, significantly Detection of high counts of molds in water-damaged buildings, strongly suggests the existence of a reservoir of mold spores in the environment This viable microbial activity with specific mold and mycotoxin IgA in saliva may assist in the diagnosis of mold exposure Whether mold and mycotoxin specific IgA antibodies detected in saliva are indicative of the role of IgA antibodies in the late phase of type-1 hypersensitivity reaction or in type-2 and type-3 delayed sensitivities is a matter that warrants further investigation

Anti-immunoglobulin responses to IgG, F(ab')2, and Fab botulinum antitoxins in mice

Abstract: Side effects to botulinum antitoxins, including anaphylaxis and serum sickness, are common. This is due to the immunogenicity of the antitoxin, which can be measured by the production of anti-immunoglobulin antibodies. An ideal botulinum antitoxin would elicit a minimal production of anti-immunoglobulin antibodies from a patient, aiding its safety. To investigate the immunogenicity of different immunoglobulin fragments, whole IgG, F(ab')2 and Fab botulinum antitoxins were administered to mice by either the intravenous or intramuscular route. The production of anti-immunoglobulin antibodies was measured over time after a single dose of antitoxin, and the anti-immunoglobulin antibodies isotyped. When administered by the intramuscular route, Fab showed significantly lower immunogenicity than IgG, while F(ab')2 had an immunogenicity that was intermediate between the two. When administered by the intravenous route there was no significant difference in immunogenicity between IgG and F(ab')2 antitoxins, although Fab antitoxin had a significantly lower immunogenicity than either IgG or F(ab')2. IgG antitoxin was significantly more immunogenic by the intramuscular route than by the intravenous route. Sheep IgG had a lower immunogenicity than goat IgG in mice. There was no significant difference in immunogenicity between the two dosing routes for either F(ab')2 or Fab antitoxin. The anti-antibodies were predominantly IgG1, suggesting a strong Th2 bias to the anti-antibody response. In all cases, Fab represents the least immunogenic form of antitoxin.

Inhibition of CCL11, CCL24, and CCL26-induced degranulation in HL-60 eosinophilic cells by specific inhibitors of MEK1/MEK2, p38 MAP kinase, and PI 3-kinase.

Abstract: Eosinophilic leukocytes are the cellular hallmark of allergic inflammation. Apart from being potent eosinophils chemoattractants, the eotaxins CCL11, CCL24 and CCL26 are capable of activating eosinophils to generate reactive oxygen species, lipid mediators of inflammation and degranulation of toxic granule proteins. Due to their central role in eosinophil trafficking and activation, understanding the signal transduction mechanism of the eotaxin-induced eosinophil effector functions may provide an innovative therapeutic strategy for eosinophil-associated diseases. Thus, these investigations were conducted to delineate signal transduction mechanisms of CCL11, CCL24 and CCL26-induced eosinophil peroxidase (EPO) degranulation following pretreatment of cells with or without a specific inhibitor of MEK1/MEK2 (U0126), inhibitor of p38 MAP kinase (SB203580) or a specific inhibitor of PI 3-kinase (LY294002). Results have shown that CCR3-mediated eotaxin-induced eosinophilic degranulation was concentration-dependently reduced by specific inhibitors of ERK1/ERK2, p38 MAP kinase and PI 3-kinase. However, the rank order of U0126 with respect to inhibition of chemokine-induced degranulation was CCL11 = CCL24 > CCL26. Potentiation of eotaxin-induced EPO degranulation by IL-5 was also seen. These investigations have not only confirmed the reported co-operativity between IL-5 and the eotaxins but also showed that the eosinophil-degranulating capabilities of the eotaxin CCL11, CCL24 and CCL26 is a consequence of activation of ERK1/ERK2, p38 MAP kinase and PI 3-kinase. Thus, these signaling molecules may provide the biochemical basis for mechanism-based therapy of allergic inflammatory diseases.