Mitogenicity and Adjuvanticity of a Marine Bacterium, Vibrio anguillarum, in Mice

Abstract

The effects of whole cells of three different 0 serotypes of Vibrio anguillarum on the murine immune response were studied. The addition of different doses (1-100 p,g/ml) of V. anguillarum cells, as well as Salmonella typhimurium lipopolysaccharide, markedly increased the incorporation of [3H]thymidine into in vitro cultured spleen cells of C57BL/6 mice. All three serotype strains of V. anguillarum were able to induce the mitogenic effect at 10 p,g/ml and 100 p,g/m1, but serotype I strains were more potent than the others. Since pretreatment of spleen cells with rabbit anti-mouse thymocyte antiserum did not affect the mitogenic activity of V. anguillarum, Vibrio cells may be a B-lymphocyte mitogen.When sheep or horse erythrocytes and Vibrio cells were injected intraperitoneally into ddY mice, Vibrio cells exhibited an enhancing effect on antibody response in vivo, regardless of the different serotypes. Vibrio cells, when injected intraperitoneally into mice before the antigen, markedly suppressed the antibody response. Several days after the injection of Vibrio cells, these mice showed an enhanced carbon clearance activity. Acid phosphatase activity in their peritoneal cells was also augmented, suggesting that Vibrio cells activated macrophages in the mice.Vibrio anguillarum, a gram-negative and halophilic marine bacterium, causes vibriosis in fish, both wild and cultured (I, 17,25); this is one of the most serious diseases affecting different kinds of fish (25).We previously reported that V. anguillarum has antitumor activity against Ehrlich carcinoma cells in mice, which may be mediated by immune lymphocytes of the host (24). Although numerous physiological, biochemical and serological studies on V. anguillarum have been carried out (1,15,17,21,25), no report has been published on the effect of the bacteria on the host immune systems.In this paper, we describe the following effects of whole V. anguillarum cells of three different 0 serotypes: a) a mitogenic effect on splenocytes (3[H]thymidine uptake), b) an adjuvant effect on antibody response to xenogeneic erythrocyte antigen, c) stimulation of the carbon clearance system, and d) enhancement of acid phosphatase activity in peritoneal exudate cells.929

A comprehensive review of Vibrio (Listonella) anguillarum: ecology, pathology and prevention

“…Upon exposure to the host kidney, spleen, or lymphocytes, V. anguillarum induces mitosis in such host cells due to antigen content in the bacterial cell and the presence of certain lipopolysaccharides among the bacterial cell wall which are inflammatory to such host tissues (Shimizu et al . ; Yui & Kaattari ; Boltaña et al . ).…”

“…Host gills, anterior and posterior intestinal tissue, as well as oral and anal tissue commonly exude vibriosis symptoms within several hours of the onset of infection but V. anguillarum invasion of internal organs such as the kidney or spleen can take as long as 24 h (Ransom et al 1984;Laurencin & Germon 1987). Upon exposure to the host kidney, spleen, or lymphocytes, V. anguillarum induces mitosis in such host cells due to antigen content in the bacterial cell and the presence of certain lipopolysaccharides among the bacterial cell wall which are inflammatory to such host tissues (Shimizu et al 1981;Yui & Kaattari 1987;Boltaña et al 2014). Fish skin mucus often contains concentrations of antimicrobial substances such as lysozyme or specific peptide components which block bacterial access to the epithelial cellular membrane (Ellis 2001).…”

Phototoxicity of Quinolones: Measurement by UV-A Inhibition of Concanavalin A-Stimulated Lymphocyte Mitogenic Response

“…9 The splenocytes were suspended in RPMI-1640 medium (Nissui Pharmaceutical, Tokyo, Japan) supplemented with 5% autologous serum. A total of 0.1 ml of the cell suspension (1 x 106 cells/ml) and 0.1 ml of a drug solution (5 or 50 #g/ml) were placed in 96-well microplates (Falcon 3072, Becton Dickinson Labware, NJ, USA) and then irradiated with two 15-W standard domestic fluorescent lamps (mellow white FL15N; Toshiba, Tokyo, Japan).The lamps have broad emission spectra in the range between 350 and 750 nm, and the plates received a total energy dose of 3.9 J/cm 2 (measured at 310-400 nm by using a Topcon IM-3 illuminometer, Tokyo Optical, Tokyo, Japan).…”

Biological activities of chemically synthesized 2-keto-3-deoxyoctonic acid-(alpha 2----6)-D-glucosamine analogs of lipid A

“…Male C57BL/6 mice, 7 weeks old, were purchased from the Shizuoka Agricultural Cooperative for Laboratory Animals, Hamamatsu, Japan. The mitogenicity of the compounds was determined on the basis of in vitro [3H]thymidine uptake into murine splenocytes as described in a previous report (20). In brief, the splenocytes were suspended in RPMI-1640 medium (Nissui Pharmaceutical Co., Ltd., Tokyo) supplemented with 10% fetal bovine serum (GIBCO Laboratories, Grand Island, N.Y.).…”