Key words: Phagocytosis, Pseudomonas aeruginosa, OEP, LPS, Anti-Pseudomonas globulin
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1 Key words: Phagocytosis, Pseudomonas aeruginosa, OEP, LPS, Anti-Pseudomonas globulin
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3 Table 1 Effect of heated serum on phagocytosis of Pseudomonas aeruginosa (serotype G) by PMNs Difference between heated-and fresh serum (p0.001) OEP-HA titer of serum : 16 ~(2ME treated 8 ~)
4 Fig. 1 Effect of chelated or nonchelated serum on phagocytosis of Pseudomonas aeruginosa (serotype G) and Staphylococcus aureus (phagetype 80/81) by PMNs P. aeruginosa (serotype G) S. aureus (phagetype 80/81) Incubation time
5 Table 2 Effect of serum absorbed by Pseudmonas aeruginosa (serotype G) on phagocytosis of Pseudomonas aeruginosa (serotype G) by PMNs Difference between absorbed-and fresh-serum (p<0.001) OEP-HA titer fresh serum 16 ~(2ME treated 8 ~) absorbed serum <4X (2ME treated <4X) Fig. 2 Effect of antibody on phagocytosis of Pseudomonas aeruginosa (serotype A) by PMNs Serum from a patient with septicemea due to P. aeruginosa (serotype A) A: Convalescent serum OEP-HA titer: 256X 2ME treated 64X B: Convalescent serum in 1:8 dilution C: Convalescent serum in 1:16 dilution D: Pooled fresh serum OEP-HA titer: 16X 2ME treated 8X E: Serum absorbed with P. aeruginosa (serotype A) OEP-HA titer: <4X F: Non-PMN.
6 Table 3 Effect of anti-pseudomonas globulin on phagocytosis of Pseudomonas aeruginosa (serotype G) by PMNs in the presence of in high OEP-HA titer serum OEP-HA titer of serum: 32X(2 ME treated 16X)
7 Table 4 Effect of anti-pseudomonas globulin on phagocytosis of Pseudomonas aeruginosa (serotype G) by PMNs in the presence of absorbed serum OEP-HA titer of absorbed serum by Pseudomonas aeruginosa (serotype G): < ~(2 ME treated <4X) Difference of anti-pseudomonas globulin from gammaglobulin and absorbed serum (p40.001) Fig. 3 Effect of anti-pseudomonas globulin on phagocytosis of Pseudomonas aeruginosa (serotype A) PMNs+ FAS+ Anti-Ps- globulin (15%) B) PMNs+ FAS+ Anti-Ps- globulin (7.75%) C) PMNs+ FAS+ Anti-Ps- globulin (3.87%) D) PMNs+ FAS+ Anti-Ps- globulin (1.88%) E) PMNs+ FAS F) PMNs+ fresh-pooled serum OEP-HAtiter of fresh pooled serum: 16 ~(2ME treated 8 ~) FAS.:fresh absorbed serum
8 Fig. 4 Effect of lipopolysaccharides from Pseudomonas aeruginosa (serotype G) on phagocytosis of various strains of Pseudomonas aeruginosa by PMNs P. aeruginosa (serotype G) P. aeruginosa (serotype C) P. aeruginosa (serotype E) Fig. 5 Effect of Original Endotoxin Protein on phagocytosis of various strains of Pseudomonas aeruginosa by PMNs P. aeruginosa (serotype G) P. aeruginosa (serotype C) P. aeruginosa (serotype E)
9 Fig. 6 Effect of elastase, protease and slime on phagocytosis of Pseudomonas aeruginosa (serotype G) by PMNs Elastase P rotease s lime
10 Table 5 Depression of complement activity by Original Endotoxin Protein, lipopolysaccharides, elastase, protease and slime of Pseudomonas aeruginosa
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13 31, ) $enneff, J. J.: Nosocomical infections due to Pseudomonos. J. Infect. Dis., 130: Suppl. 4-7, ) McCracken, G. H. & Eichenwald, H. F.: Leukocyte function and the development of opsonic and complement activity in the neonate. Am. J. Dis. Child., 121: , ) Winklstein, J. A.: Mecical progress, Opsonins: Their function, indentity, and clinical significance. J. Pediat., 82: , ) Capo, C., Bongrand, P., Benoliel, A. M. & Depieds, R.: Phagocytosis. J. Theor. Biol., 47: , ) Weisman, R. A. & Korn, E. D.: Phagocytosis of latex beads by Acanthamoeba, I. Biochemical properties. Biochemistry, 6: , ) Peterson, P. K., Verhoef, J. & Quie, P. G.: Influence of temperature on opsonezation and phagocytosis of staphylococci. Infect. Immun., 5) Wesly, J. & Fisher, M 15: , W.: Immunization against Pseudomonas in infection after thermal 21) Solberg, C. O. & Helium, K. B.: Influence injury. J. Infect. Dis. of serum on the bactericidal activity of neutrophil granulocytes. Acta Pathol. Microbiol., 130: Suppl , ) Young, L. S., Meyer, R. D. & Armstrong Scand. Sect. B., 81: , 1973., D.: Pseudomonas aeruginosa vaccine in cancer 22) Quie, P' G., White, J. G., Holmes, B. & Good, R. A. In vitro bactericidal capacity of human patients. Annals Inter. Med., 79: , polymorphonuclear leucocyte: Diminished activity in chronic granulomotous disease of childhood. J. Clin. Invest., 46: , ) Young, L. S. & Armstrong 8) Homma, J. Y., Abe, C., Okada, K., Tanamoto,, D.: Human im munity to Pseudomonas aeruginosa K. & Hirao, Y.; The biological properties of. I. In vitro intraction of bacteria the protein moiety of endotoxin of Pseudomonas, polymorphonuclear leucocytes, and serum factor aeruginosa. Animal, Plant and Microbial Toxins. J. Infect. Dis.,, 126: , : , ) Bjornson, A. B. & Michael, J. G.: Factor in human serum promoting phagocytosis of Pseudomonas aeruginosa. I. Intraction of opsonins with the bacterium. J. Infect. Dis., 130: Suppl , ) Young, L. S.: Human immunity to Pseudomonas 11) Schimpff, S. C., Green, W. H., Young, V. M. aeruginosa. II. Relationship between heatstable opsonins and type-specific lipopolysac - & Wiernink, P.H.: Significance of Pseudononas aeruginosa in the patient with Leukemia or charides. J. Infect. Dis., 126: Lymphoma. J. Infect. Dis., 130: Suppl, ) Nakao, M. & Saito, K.: Protective com-
14 ponents in the cell wall of salmonella typhimurium against bactericidal action of antibody. Japan J. Microbiol., 14: 83-86, ) Rowly, D.: Endotoxins and bacterial virulence. J. Infect. Des., 123: , activity and description of an anaphylatoxin in activator of human serum. J. Exp. Med., 29) Hoshi, A., Kanazawa, F., Kuretani, K., 129: , Homma, J. Y. & Abe, C.: Antitumor activity 37) Tamura, N., Shimada, A. & Chang, S.: of the protein moiety of the endotoxin of Further evidence for immune cytolysis by Pseudomonas aeruginosa. Gann, 63: , ) Kojima, Y., Homma, J. Y. & Abe, C.: Property to induce interferon of Pseudomonas aeru antibody and the first components of com ginosa endotoxin and it's component. Japan J. Exp. Med., 41: , ) Fukui, G. M., Homma, J. Y. & Abe, C.: Nonspecific protective properties of Pseudomonas aeruginosa endotoxin and it's component. Japan J. Exp. Med., 41: , ) Kawaharajo, K., Abe, C., Homma, J. Y., Kawano, M., Gotoh, E., Tanaka, H. & Morihara, K.: Carneal ulcers caused by protease and elastase from Pseudomonas aeruginosa. Japan J. Exp. Med., 44: , ) Liu, P. V., Abe, Y. & Bates, J.L.: Roles of various fraction of Pseudomonas aeruginosa in it's pathogeuesis. J. Infect. Dis., , ) Abe, C., Tanamoto, K. & Homma, J. Y.: Infection protective property of the common antigen (OEP) of Pseudomonas aeruginosa and it's chemical composition. Japan J. Exp. Med., 47: , ) Bjornson, A. B. & Michael, J. G.: Factor in human serum promoting phagocytosis of Pseudomonas aeruginosa. II. Intraction of opsonin with the phagocyte cell. J. Infect. Dis., 130: Suppl , ) Bokish, V. A., Muller Eberhard, H. J. & Cochrane, C. G.: Isolation of a fragment (C3a) of the third component of human complement containing anaphylatoxin and chemotactic plement in the absence of Cq. Immunology, 22: , ) Jensen, J.: Anaphylatoxin in it's relation to the complement system: Science, 155: , ) Gotze, O. & Muller Eherhand, H. J.: The role of properdin in the alternative pathway of complement activation. J. Exp. Med., 139: 44-59, ) Fearon, D. T., Austeu, K. F. & Ruddy, S.: Propendin factor D: characterization of it's active site and isolation of the precurror form. J. Exp. Med., 139: , ) Forsgren, A. & Quie, P. G.: Opsonic activity in human serum chelated with ethylene gly coltitra-acetic acid. Immunology, 26: , ) Forsgren, A. & Quie, P. Q.: Influence of the alternate complenent pathway on opsonization of several bacterial species. Infict. Immun., 10: , ) Messner, R. P. & Jelinek, J.: Receptors for human rg globulin on human neutrophils. J. Clin. Invest., 49: , 1970.
15 Phagocytosis of polymorphonuclear leukocytes against Pseudomonas aeruginosa Toshio TAKASHIMA Department of Pediatrics, Osaka Medical College The defence mechanism against infection with Pseudomonas aeurginosa was investigated from the viewpoint of phagocytosis by human PMNs. A study of the effect of antibody, complement, endotoxins and exsoenzymes on phagocytosis by PMNs was under taken by the Quie's method, as follows: 1) Phagocytosis of Pseudomonas aeurginosa serotype G was inhibited by LPS from the same serotype, while that with different serotypes C and E was not. 2) OEP from Pseudomonas aeruginosa serotype G did not inhibit the phagocytosis of Pseudomonas aeurginosa serotypes G, C and E. 3) Elastase from Pseudomonas aeruginosa IF03455, protease from Pseudomonas aeruginosa IF03080 and slime from Pseudomonas aeruginosa IF03455 inhibited the phagocytosis of Pseudomonas aeruginosa serotype G. Especially, elastase most potentiality inhibited the phagocytosis of Pseudomonas aeruginosa and complement activity. 4) OEP antibody enhanced the phagocytosis. The serum absorbed by Pseudomonas aeruginosa serotype G did not develop the phagocytosis of Pseudomonas aeruginosa serotype G. 5) Pathway of activation of complements on phagocytosis of Pseudomonas aeruginosa as well as Staphylococcus aureus by leukocytes was examined. Phagocytosis of Staphylococcus aureus was inhibited by both EDTA and EGTA, while that of Pseudomonas aeruginosa was not by EGTA. This result suggests that the alternate pathway of activation of complements relates to the phagocytosis of Pseudomonas aeurginosa. 6) When OEP antibody titer of opsonin was low, Anti-pseudomonas globulin (OEP-HA titer 256x) enhanced the phagocytosis of Pseudomonas aeruginosa. However, as OEP antibody titer of opsonin became higher, Anti-pseudomonas globulin did not influence the activity in phagocytosis of Pseudomonas aeruginosa by PMNs.
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