YOSHIIE Kiyotaka Kagoshima University, Graduate School of Medical and Dental Sciences, Associate Professor, 大学院・医歯学総合研究科, 助教授 (70174886)
MAENO Nobuaki Kagoshima University, Graduate School of Medical and Dental Sciences, Research Associate, 大学院・医歯学総合研究科, 助手 (20305113)
又吉 盛健 鹿児島大学, 大学院・医歯学総合研究科, 講師 (00128456)
藤村 剛 鹿児島大学, 医学部・歯学部附属病院, 助手 (10136880)
|Budget Amount *help
¥3,600,000 (Direct Cost : ¥3,600,000)
Fiscal Year 2005 : ¥800,000 (Direct Cost : ¥800,000)
Fiscal Year 2004 : ¥800,000 (Direct Cost : ¥800,000)
Fiscal Year 2003 : ¥2,000,000 (Direct Cost : ¥2,000,000)
1.Analysis by SDS-PAGE of whole bacterial cells of Bartonella henselae revealed more than 35 protein bands. The molecular weight of major proteins were 90, 62, 45 (43), 40, 35, 31, and 25kDa. Molecular weight of the major outer membrane proteins were estimated as 62, 43 and 31kDa.
2.Thirty-two of 108 patients who had swelling of lymph nodes and CSD was suspected showed more than 256x IgG titer against B.henselae by indirect immunofluorescence assay (IFA). These antibody positive sera were used for the following study.
3.Western blotting analysis of whole bacterial cells of B henselae using patient sera revealed 8 bands (90, 65, 43, 40, 35, 33, 30, and 28kDa). Distribution pattern of positive bands were diverse in case to case, however, 65, 43, and 35kDa proteins showed strong antigenicity. The reactivity of IFA increased by co-cultivation of B.henselae with vero cells. This increase of reactivity were striking in 35kDa band, however, some sera did not reacted with this band. Consequently we could not find sole antigen protein useful for sero-diagnosis. Compound proteins, including 65, 43, and 35kDa protein, should be considered to develop new method for the laboratory diagnosis of CSD.
4.B.henselae stimulated the growth of lymphatic endothelial cells as well as vascular endothelial cells. B.henselae induced CCR7 on the surface of peripheral neutrophils. The bacteria also stimulated the production of CCL21 from lymphatic endothelial cells. Then we examined the diagnostic importance of CCL21 in patients with CSD. The result showed no significant difference of serum CCL21 level between CSD and other patients with various lymph node diseases.
5.We developed a new method to cultivate B.henselae, and isolated 16 strains of B.henselae from stray cats by using the method.