Key wards: cyclophosphamide, neutropenic mice, Pseudomonas aeruginosa, experimental pyelonephritis, G-CSF

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Key wards: cyclophosphamide, neutropenic mice, Pseudomonas aeruginosa, experimental pyelonephritis, G-CSF

Pseudomonas aeruginosa Fig. 1 Method of administration of cyclophosphamide and G-CSF CPM group CPM+G-CSF prophylactic group CPM+G-CSF therapeutic group CPM: Cyclophosphamide

Fig. 2 Effect of cyclophosphamide on total number of white blood cells single injection of cyclophosphamide, double injection of cyclophosphamide Fig. 3 Histological grade of pyelonephritis in cyclophosphamide-treated mice after 7days of inoculation of Pseudomonas aeruginosa (2 ~ 103, 2 ~ 104, 2 ~ 105 CFU/ml ~ 0.05ml)

Fig. 4 Number of bacteria/ml in urine after 7days of inoculation of Pseudomonas aeruginosa (2 ~ 104 CFU/ml ~ 0.05ml) CPM: Cyclophosphamide Fig. 5 Number of bacteria/g in kidney after 7days of inoculation of Pseudomonas aeruginosa (2 ~ 104 CFU/ml ~ 0.05ml) CPM: Cyclophosphamide

Fig. 6 Histological grade of cystitis after 7days of inoculation of Pseudomonas aeruginosa (2 ~ 104 CFU/ml ~ 0.05ml) Histological grade: CPM: Cyclophosphamide Fig. 7 Histological grade of pyelonephritis after 7days of inoculation of Pseudomonas aeruginosa (2 ~ 104 CFU/ml ~ 0.05ml) Histological grade: CPM: Cyclophosphamide

Fig. 8 Effect of G-CSF on the reduction of peripheral white blood cells in cyclophosphamide-treated mice Fig. 9 Effect of G-CSF in the differential white blood cells count in cyclophosphamide-treated mice A: CPM group B: CPM+G-CSF therapeutic group C: CPM+G-CSF prophylactic group CPM: Cyclophosphamide

Fig. 10 Peripheral white blood cells count after 7days of inoculation of Pseudomonas aeruginosa (2 ~ 104 CFU/ml ~ 0.05ml)

Fig. 11 Effect of G-CSF on bactericidal capacity of peritoneal exudating neutrophils CPM: Cyclophosphamide Fig. 12 Survival rate after 7days of inoculation of Pseudomonas aeruginosa (2 ~ 104 CFU/ml ~ 0.05ml) CPM: Cyclophosphamlde

3) Nomura, H., Imazeki, I.,Oheda, M., Kubota, M., Tamura, M., Ono, M., Ueyama, Y. & Asano, S.: Purification and characterization of human granulocyte-colony stimulating factor(g-csf). E. M. B. O. J., 5: 871-876, 1986. 4) Tamura, M., Hattori, K., Nomura, H., Oheda, M., Kubota, M., Imazaki, I., Ono, M., Ueyama, Y., Nagata, S., Shirafuji, N. & Asano, S.: Induction of neutropenic granulocytosis in mice by administration of purified human granulocytecolony stimulating factor (G-CSF). Biochem. Biophys. Res. Commun., 142: 454 6) Kitagawa, S., Yuo, A., Lawrence, M. S., Saito, M., Miura, Y. & Takaku, F.: Recombinant human granulocyte colony-stimulating factor e enhances superoxide release in human granulocytes stimulated by the chemotactic peptide. Biochem. Biophys. Res. Commun., 144: 1143-1146, 1987. 7) Welte, K., Bonilla, M. A., Gillio, A. P., Boone, T. C., Potter, G. K., Gabrilove, J. L., Moore, M. A. S., O'Reilly, R.J. & Souza, L.M.: Recombinant human granulocyte-colony stimulating factor. Effect on hemotopiesis in normal and cyclophosphamide-treated primates. J. Exp. Med., 165: 941-948, 1987. 10) Quie, P. G., White, J. G., Holmes, B. & Good, R. A.: In vitro bacteriocidal capacity of human polymorphonuclear leukocytes : Diminished activity in chronic granulomatous disease of

childhood. J. Clin. Invest., 46: 668-679, 1967. 18) Matsumoto, M., Matsubara, S., Matsuno, T., Tamura, T., Hattori, K., Nomura, H., Ono, M. & Yokota, T.: Protective effect of human 13) Buhles, W. C. & Shifrine, M.: Adjuvant protection against bacterial infection in granulocytopenic mice. J. Infect. Disease., 136: 90-95, 1977. 14) Lumish, R. M. & Norden, C. W.: Therapy of neutropenic rats infected with Pseudomonas aeruginosa. J. Infect. Diseases, 133: 538-547, 1976. 15) Nishi, T. & Tsuchiya, K.: Experimental urinary tract infection with Pseudomonas aerugionsa in mice. Infect. Immun., 22: 508-515, 1978. granulocyte-colony stimulating factor on microbial infection in neutropenic mice. Infect. Immun., 55: 2715-2720, 1987. 19) Shimamura, M., Kobayashi, Y., Yuo, A., Urabe, A., Okabe, T., Komatu, Y., Itoh, S. & Takaku, F.: Effect of human recombinant granulocyte colony-stimulating factor on hematopoietic injury in mice induced by 5-fluorouracil. Blood 69: 353-355, 1987. 20) Kobayashi, Y., Okabe, T., Urabe, A., Suzuki, N. & Takaku, F.: Human granulocyte colony-stimulating factor produced by E. coli shortens the period of granulocytopenia induced by irradiation in mice. Jpn. J. Cancer Res. (Gann), 78: 763-766, 1987. 21) Asano, S. & Ono, M.: Human granurocyte colony-stimulating factor: Its biological actions and clinical implication. Acta Haematol. Jpn., 50: 1550-1556, 1987. Study of the Prophylactic and Therapeutic Effect of Human Granulocyte-Colony Stimulating Factor (G-CSF) on Experimental Pyelonephritis Induced by Pseudomonas Aeruginase in Neutropenic Mice Noriaki TANAKA, Yoshiaki KUMAMOTO, Takaoki HIROSE & Akifumi YOKOO Department of Urology, Sapporo Medical College (Director: Prof. Yoshiaki KUMAMOTO) We investigated the prophylactic and therapeutic effect of human granulocyte-colony stimulating factor (G-CSF) on mice with ascending pyelonephritis induced by Pseudomonas aeruginasa (G-group). This experimental model was established by a two course administration of cyclophosphamide, so that it kept the mice in a neutropenic status (around 2000 white blood cells/mm3) from the time of infection to the time of sacrifice. The cyclophosphamide-treated group increased their susceptibility more than the control group. In the cyclophosphamide-treated group, the prophylactic administration of G-CSF (2,u g/day/mouse) yielded a lower incidence of infection and of infection-induced motality than that of saline alone. However, the therapeutic administration of G-CSF did not produce significant decreases of these rates, suggesting that this type of administration had no effect on infection. At the time of sacrifice, the prophylactic administration of G-CSF increased the number of neutrophils, while at the time of induced infection, no increase of neutrophils was found. G-CSF therapeutic administraiton was not able to increase neutrophils during the experiment. An investigation of the bactericial capacity of peritoneal exudating neutrophils revealed that G-CSF prophylactic administration accelerated its capacity, although cyclophosphamide alone did not. These results suggest that G-CSF has a prophylactic effect on bacterial infeciton in neutropenic mice, and that this effect, in part, depends upon both the increase of neutrophils and the acceleration of bactericidal capacity produced by G-CSF.

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