Helicobacter cinaedi-associated Carotid Arteritis

A 65-year-old Japanese man with bilateral carotid atherosclerosis presented with right neck pain and fever. Contrast-enhanced computed tomography suggested carotid arteritis, and carotid ultrasonography showed an unstable plaque. The patient developed a cerebral embolism, causing a transient ischemic attack. Helicobacter cinaedi was detected in blood culture, and H. cinaedi-associated carotid arteritis was diagnosed. Empirical antibiotic therapy was administered for 6 weeks. After readmission for recurrent fever, he was treated another 8 weeks. Although the relationship between H. cinaedi infection and atherosclerosis development remains unclear, the atherosclerotic changes in our patient’s carotid artery might have been attributable to H. cinaedi infection

Development of novel DNA vaccine for VEGF in murine cancer model

We developed DNA vaccine for vascular endothelial growth factor (VEGF), which may provide the therapeutic option instead of anti-VEGF antibody, bevacizumab. Plasmid containing VEGF mini-gene was constructed in the insertion of B-cell epitope of Hepatitis B core protein [HBc-VEGF], which was an epitope carrier. High titer of anti-VEGF antibody was observed in BALB/c mice which were intramuscularly immunized with HBc-VEGF by electropolator. In mice inoculated with colon 26 cells, tumor volume and microvessel density was decreased in HBc-VEGF with a significant prolonged survival. Co-treatment of purified IgG from immunized mice with HBc-VEGF showed in vitro neutralizing activity for VEGF-induced ERK phosphorylation and tube formation in cultured endothelial cells. Furthermore, intravitreally injection of this purified IgG reduced the neovessel formation in the mouse oxygen-induced retinopathy and laser-induced choroidal neovascularization models. These results first provided that DNA vaccine against VEGF possessed the anti-angiogenic effect, leading to prolonged survival in mouse cancer model.Y

Effect of Ang II vaccine on Ang II-induced hypertension.

<p>(A) Systolic BP at steady state and under Ang II infusion on day 49. The control mice were immunized with 1 mg KLH. The experimental mice were immunized with 100ng or 1,000 ng Ang II-KLH. All groups contained 6 to 8 mice. The data are expressed as the mean systolic BP ± standard error (SE) of the mean. †<i>P</i><0.01 *<i>P</i><0.05. (B) The correlation between systolic BP under Ang II infusion and the anti-Ang II antibody titers in the sera of immunized mice. The titer is expressed as the dilution of serum giving half-maximal binding (optical density: OD 50%).</p

Neutralizing antibody production induced by Ang II vaccine.

<p>(A) The antibody titers in the sera of mice immunized with saline, Ang II, KLH or Ang II-KLH (dose: 25 ng/mouse) with or without Freund’s adjuvant (FA) are shown. †<i>P</i><0.001 vs. saline. ELISA was performed using the sera of mice on day 42. (B) The serum antibody titers elicited by various doses (10, 25, 100, 300, 1000 ng/mouse) of Ang II-KLH with Freund’s adjuvant (FA). *<i>P</i><0.05 vs. 10 ng. The titer is expressed as the dilution of serum giving half-maximal binding (optical density: OD 50%) ± SE of the mean. (C) Western blot using anti-phosphorylated ERK (P-ERK) and anti-ERK antibodies. The total protein was extracted from HASMCs treated with 10⁻⁷ M Ang II that was previously incubated with 1 % serum. (D) Promoter activity of <i>c-fos</i> was measured using luciferase activity. HASMCs were transfected with <i>c-fos</i> luciferase reporter gene and stimulated by pre-incubated Ang II (10⁻⁷ M) with 1 % serum of the control or immunized mice for 24 hours. The results from 3 samples are expressed as the mean of the ratio to no stimulation ± SE of the mean. ‘‘Control serum’’ indicates serum from mice immunized by KLH (1 mg, with adjuvant), and ‘‘Immunized serum’’ indicates serum from mice immunized by Ang II-KLH (1,000 ng, with adjuvant). All serum was obtained on day 42. †<i>P</i><0.01.</p

Effect of Ang II vaccine on SHR.

<p>A) Experimental protocol is shown. Ang II-KLH (Ang II-KLH group, n = 5) or saline (saline group, n = 5) were injected on days 0, 14, and 21 in twenty-four-week old male SHR. The anti-Ang II antibody titer and systolic BP were measured on days 0, 14, and 28. The Ang II-KLH group rats were immunized with 5 µg Ang II-KLH with CFA on day 0 and with 5 µg Ang II-KLH with IFA on day 14 and 21. B) Anti-Ang II antibody titers in the sera of immunized rats on days 0, 14, 28, and that of saline injected rats on day 28 were examined. *<i>P</i><0.01 vs. day 0. C) Systolic BP on days 0, 14, and 28 were shown. The data are expressed as the mean systolic BP ± standard error (SE) of the mean. *<i>P</i><0.05. vs. Saline.</p