BioMed Research International / 2014 / Article / Tab 3

Research Article

Interactions of Neuropathogenic Escherichia coli K1 (RS218) and Its Derivatives Lacking Genomic Islands with Phagocytic Acanthamoeba castellanii and Nonphagocytic Brain Endothelial Cells

Table 3

RS218-derived islands (RDI’s) mutants and their interactions with human brain microvascular endothelial cells (HBMEC). For associated, invasion, and survival assays, the number of E. coli K1 c.f.u. interacting with HBMEC was considered as 100% and the results of mutant strains are expressed as the relative change. For cytotoxicity assays, E. coli K1-mediated HBMEC death was considered as 100% and the results of mutant strains are expressed as the relative change.

E. coli HBMEC
AssociationInvasionSurvivalCytotoxicity

K1 (RS218)100100100100
Plasmid cured RS218 27.41 ± 4.5*92.52 ± 5.6*70.86 ± 6*104 ± 7.8
Black hole filled with HB101 DNA31 ± 1.6*62.86 ± 13*163.5 ± 28*87 ± 10
RDI 189.97 ± 1.2112.8 ± 10.696.56 ± 897 ± 13
RDI 298.42 ± 2.672.5 ± 8.7*91.6 ± 9.1108 ± 14
RDI 493.6 ± 3.5298.37 ± 0.896.76 ± 1.8124 ± 2.3
RDI 9101.47 ± 2.6109.6 ± 4.4106.5 ± 3.4110 ± 12
RDI 1061.12 ± 1.4*58.5 ± 6.1*55.47 ± 6*92.8 ± 3
RDI 1270.7 ± 8.3*88.3 ± 7.1107.5 ± 1189.9 ± 9
RDI 14105.84 ± 4.1133.7 ± 2296.7 ± 11103 ± 4
RDI 1553.97 ± 7.5*115.9 ± 5.5113.6 ± 3.1118 ± 7
RDI 1667.78 ± 3*111 ± 13156.1 ± 19*127 ± 5.4
RDI 17104 ± 169.77 ± 7.4*60.4 ± 3.4*105 ± 2
RDI 18107.52 ± 1.6159.2 ± 18*108.6 ± 8.888 ± 1.8
RDI 2132.82 ± 4.6*25 ± 3.6*17.95 ± 3*14.5 ± 1.1*
RDI 2277.9 ± 8.9*61.53 ± 8*50.45 ± 7*127 ± 3.4

(i) Association was calculated as follows: (number of bacterial c.f.u. recovered/original inoculum) × 100 = bacterial association of HBMEC.
(ii) Invasion was calculated as follows: (number of bacterial c.f.u. recovered/original inoculum) × 100 = bacterial invasion of HBMEC.
(iii) Survival was calculated as follows: (number of bacterial c.f.u. recovered/original inoculum) × 100 = bacterial survival of HBMEC.
(iv) Cytotoxicity was calculated as follows: (sample value − negative control value)/(total LDH release − negative control value) × 100 = HBMEC cytoxicity.
*indicates a significant difference (P < 0.05 using paired t-test, one-tailed distribution) when data of mutant strains were compared with the wild type E. coli K1 data.

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