The difference in IL-1β, MIP-1α, 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 pro-inflammatory 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-1β, MIP-1α, 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-1β, MIP-1α, 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.

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.