Immunostimulating effect of a well-known Thai folkloric remedy in breast cancer patients.

The study aimed to evaluate immune-stimulating effects of a well-known Thai folkloric remedy when used for adjuvant therapy with conventional chemotherapeutics for treatment of breast cancer. Immunostimulating influence of the remedy (215 mg/kg body weight per day) on NK cell activity and TNF-α release from the monocytes/macrophages were investigated in a total of 15 healthy women and 13 female patients with breast cancer (Group 1). The effect of breast tumor surgery on NK cell activity was further investigated in 18 female patients with breast cancer (Group 2). NK cell cytotoxic activity was determined by chromium release cytotoxic assay using K562, an erythroleukemic cell line. TNF-α release from monocytes/macrophages separated from blood samples was determined through a biological assay using actinomycin D-treated L929 mouse fibroblast cells in the presence and absence of LPS. Baseline NK cell activity of the monocytes/macrophages separated from Group 2 patients expressed as %cytotoxicity was significantly lower than in the healthy subjects at E:T ratios of 100:1 and 25:1. In healthy subjects, there was no change in NK cell cytotoxic activity (%cytotoxicity or LU) following 1 and 2 weeks of treatment with the remedy compared with the baseline at various E:T ratios but the binding activity (%binding) was significantly increased after 2 weeks of treatment. The addition of one or two conventional chemotherapeutic regimens did not significantly reduce the NK cytotoxic activity but did affect release of TNF-α in both unstimulated and LPS-stimulated samples. Surgery produced a significant suppressive effect on NK cell activity. The use of the remedy as an adjunct therapy may improve therapeutic efficacy and safety profiles of conventional chemotherapeutic regimens through stimulation of the immune system in cancer patients.

The difference in IL-1beta , MIP-1alpha, IL-8 and IL-18 production between the infection of PMA activated U937 cells with recombinant vaccinia viruses inserted 2004 H5N1 influenza HA genes and NS genes.

BACKGROUND: The severity of avian influenza H5N1 disease is correlated with the ability of the virus to induce an over production of proinflammatory cytokines from innate immune cells. However, the role of each virus gene is unknown. To elaborate the function of each virus gene, the recombinant vaccinia virus inserted HA and NS gene from the 2004 H5N1 virus were used in the study. METHODS: U937 cells and PMA activated U937 cells were infected with recombinant vaccinia virus inserted with HA or NS gene. The expressions of HA and NS proteins in cells were detected on immunofluorescence stained slides using a confocal microscope. The cytokine productions in the cell supernatant were quantitated by ELISA. RESULTS: The recombinant vaccinia virus inserted with HA genes induces the production of IL-1beta, MIP-1alpha, IL-8 and IL-18 cytokines from PMA activated U937 cells significantly more than cells infected with wild type vaccinia, whereas the recombinant vaccinia virus inserted with NS genes it was similar to that with the wild type vaccinia virus. However, there was no synergistic nor antagonistic effect of HA genes and NS genes in relation to cytokines production. CONCLUSION: Only the HA gene from the 2004 H5N1 virus induces IL-1beta, MIP-lalpha, IL-8 and IL-18 cytokine productions from activated U937 cells. The same HA gene effect may or may not be the same in respiratory epithelial cells and this needs to be explored.

Co-stimulatory molecules on peripheral blood mononuclear cells and tissue infiltrating cells of skin wart and in vitro poke weed mitogen stimulation.

BACKGROUND: Skin wart is a lesion caused by human papilloma viruses (HPVs) that can infect both male and female. OBJECTIVE: Quantify the number of CD28+, CD86+, CD152+ and gammadelta+ in peripheral blood mononuclear cells (PBMCs) of subjects with skin wart. Identify CD86+ and gammagamma+ cells in skin wart cryosections. MATERIAL AND METHOD: Sixteen subjects with skin warts on face, hand, finger, knee, foot or plantar, both male and female, aged between 19-59 years-old, were recruited from Ramathibodi Hospital, Mahidol University, Bangkok. RESULTS: CD86 and CD152, on peripheral blood mononuclear cells (PBMCs) of subjects with skin wart are significantly lower compared to controls. Tissue cryosection staining for CD86+ and gammadelta+ cells showed no difference among subjects with skin wart and control. Proliferative response to poke weed mitogen of subjects with skin wart is significantly lower than control subjects. CONCLUSION: There was no difference in the number of subjects positive for CD28 and CD86 cell between normal and skin wart subject, but an increase in skin wart subjects with gammadelta+ cells.

Suppression by Curcuma comosa Roxb. of pro-inflammatory cytokine secretion in phorbol-12-myristate-13-acetate stimulated human mononuclear cells.

Curcuma comosa Roxb. is a medicinal plant that has traditionally been used in Thailand for treatment of inflammation in postpartum uterine bleeding. The purpose of this study was to evaluate its anti-inflammatory effects using peripheral blood mononuclear cells (PBMC) and human pro-monocytic cell line (U937). Pretreatment with hexane or ethanol extract or two diarylhepatanoids (5-hydroxy-7-(4-hydroxyphenyl)-1-phenyl-(1E)-1-heptene and 7-(3,4-dihydroxyphenyl)-5-hydroxy-1-phenyl-(1E)-1-heptene) of C. comosa significantly decreased the release of pro-inflammatory cytokines, tumor necrosis factor alpha (TNF-alpha) and interleukin-1beta, from phorbol-12-myristate-13-acetate (PMA)-stimulated PBMC and U937 cells. In PMA-stimulated U937 cells, the two C. comosa diarylhepatanoids reduced the expression of TNF-alpha and suppressed expression of IkappaB kinase and activation of nuclear factor kappa B. These results indicated that C. comosa and its diarylheptanoids have anti-inflammatory properties which could be exploited for clinical use.

Anti-Inflammatory and Immunomodulatory Activities of Stevioside and Its Metabolite Steviol on THP-1 Cells.

Stevioside, a natural noncaloric sweetener isolated from Stevia rebaudiana Bertoni, possesses anti-inflammatory and antitumor promoting properties; however, no information is available to explain its activity. The aim of this study was to elucidate the anti-inflammatory and immunomodulatory activities of stevioside and its metabolite, steviol. Stevioside at 1 mM significantly suppressed lipopolysaccharide (LPS)-induced release of TNF-alpha and IL-1beta and slightly suppressed nitric oxide release in THP-1 cells without exerting any direct toxic effect, whereas steviol at 100 microM did not. Activation of IKKbeta and transcription factor NF-kappaB were suppressed by stevioside, as demonstrated by Western blotting. Furthermore, only stevioside induced TNF-alpha, IL-1beta, and nitric oxide release in unstimulated THP-1 cells. Release of TNF-alpha could be partially neutralized by anti-TLR4 antibody. This study suggested that stevioside attenuates synthesis of inflammatory mediators in LPS-stimulated THP-1 cells by interfering with the IKKbeta and NF-kappaB signaling pathway, and stevioside-induced TNF-alpha secretion is partially mediated through TLR4.

Immunomodulatory effect and quantitation of a cytotoxic glycosphingolipid from Murdannia loriformis.

NMR signal reassignments for a cytotoxic glycosphingolipid compound, 2, ß-O-D-glucopyranosyl-2-(2'-hydroxy-Z-6'-enecosamide)sphingosine, isolated from an ethanolic extract of the herb Murdannia loriformis, have been achieved by use of FAB-MS, and 1D and 2D 1H and 13C NMR. The amount of 2 in the herb juice was quantitatively determined by use of a validated HPLC method (RP-18, MeOH-H2O, UV detection at 210 nm). The immunomodulatory effect of the herb juice and of 2 was proved by means of in vitro cellular immunological assays. Compound 2 at a concentration of 13 nmol L-1 stimulated PBMC proliferation and increased the CD 3,4:CD 3,8 ratio in T lymphocytes.

Cyclins D1, E, A expression and synergistic cytotoxicity to a cholangiocarcinoma cell line from recombinant TNF-alpha and PHA supernate.

We studied the cytotoxic effects of recombinant TNF-alpha and supernate of phytohemagglutinin stimulated peripheral blood mononuclear cells individually and in combination against a cholangiocarcinoma cell line. Levels of cyclins D1, E and A in the cell line were detected by immunoblotting, and the cell cycle stage was assayed by propidium iodide staining followed by flow cytometry analysis. Viable and apoptotic cells were assessed by trypan blue dye exclusion, DAPI staining, agarose DNA laddering and propidium iodide staining. At the beginning of each experiment, the majority of cholangiocarcinoma cells expressed cyclin A and were in S phase as determined by propidium iodide staining. Treatment of such cells with recombinant TNF-alpha resulted in cytotoxic effects clearly evident at 36 hours post exposure. There was a synergistic killing effect when recombinant TNF-alpha was combined with PHA supernate and this effect could be partly neutralized by monoclonal anti TNF-alpha, interleukin (IL)-2, IL-12 and IFN-gamma.