Article Figures & Data
Figures
- Fig. 1.
Vacuolation of mammalian cells caused by a cytotoxin secreted by V. cholerae. (A) Control normal Vero cells. (B) Vacuolated cells after treatment with the supernatant of a V. cholerae strain (Amazonia), seen by phase-contrast microscopy (magnification, ×320). (C andD) Vacuolated cells after a neutral red uptake assay, done after 24 h of incubation with the cytotoxin. The weak base neutral red is taken up by the vacuoles.
Tables
- Table 1.
Characteristics and effect on Vero cells of strains used
Strain Descriptiona VcVac Titerb No vacuolation V. cholerae O1 Amazonia 20/20c FG1066, FG1068 and 18 others Amazonas, Brazil, 91-92 + FG1066 S, 1/640; C, 1/10,240 O1 El Tor 5/18 13/18 E7946 Ogawa, Bahrain, 78 S, C ANCO223 Ogawa, Peru S, C 0872 Inaba, Peru + S, 1/120; C, 1/2,560 N16961 Inaba, Bangladesh, 75 + S, 1/30; C, 1/320 N16117 Ogawa, Bangladesh S, C P27459 Inaba, Bangladesh, 76 S, C 4 strains Brazil, 92-94 S, C T19479 Bangladesh, 79 S, C 26-3 Ogawa, Philippines, 61 + S, 1/60; C, 1/480 30167 Bangladesh, 76 S, C 62746 Bangladesh, 76 S, C N.2 Inaba, China, 86 + S, 1/20; C, 1/160 N.5 Ogawa, China, 77 + S, 1/960; C, >1/2,560 62-6-91 Tanzania S, C LA-M-644 Nigeria S, C O1 Gulf of Mexico El Tor 3/3 E506 Texas + S, 1/20; C, 1/80 SGN7277 Sewage, Louisiana, 80 + S, 1/20 4808-78 Patient, Louisiana, 78 + S, 1/20 O1 classical 6/6 0395 India, 64 S, C AMS 20-A-73 (VRL3) China, 45 S, C N32089 Bangladesh, 82 S, C N19073 Bangladesh, 82 S, C N19812 Bangladesh, 82 S, C N22646 Bangladesh, 82 S, C Environmental, non-O1, non-0139 7/10 3/10 FG1130 Water, RJ, Brazil, 97 + S, 1/320 FG1131 Water, RJ, Brazil, 97 + S, 1/1920 FG1132 Mollusk, RJ, Brazil, 97 S, C FG1133 Mollusk, RJ, Brazil, 97 S, C FG1134 Mollusk, RJ, Brazil, 97 + S, 1/20 FG1138 Mollusk, RJ, Brazil, 95 + S, 1/80 FG1139 Crab, RJ, Brazil, 96 S, C FG1140 Crab, RJ, Brazil, 97 + S, 1/30 FG1135 Water, RJ, Brazil, 97 + S, 1/40 FG1137 Crab, RJ, Brazil, 95 + S, 1/240 E. colid DH5α DH5αλpir From A. Camilli, Tufts University EK322 MM294/pRK2013 VJ737 MC4100/pKAS32 from R. Taylor, Dartmouth University FG1155 DH5αλpir/pFG300, this work FG1221 DH5α/pFG350, this work Other O1 V. cholerae strains JBK56 Inaba, derived from N16961, Δctx Apr JBK70 Inaba, derived from N16961, Δctx Hgr CVD104 Inaba, N16961 Δctx ΔhlyA CVD113 Ogawa, E7946 Δvirulence cassette ΔhlyA::mer ctxB, Δlec FG1106 FG1066 Smr, this work FG1153 FG1106 hlyA::pFG300, Apr, this work FG1173 Hemolytic revertant of FG1153, this work FG1167 Smr revertant of FG1153, this work FG1229 FG1153/pFG350, this work ↵a Numerals indicate years (91-92 denotes 1991 to 1992, etc.) of isolation at the location given.
↵b S, filtered supernatant of culture; C, 80-fold volume concentrate prepared from S by precipitating with 50% ammonium sulfate.
↵c Fraction of VcVac-positive strains in each group.
↵d Either strains have not been tested on cells or their results are presented in a separate table.
- Table 2.
Comparison of vacuolating activities of Amazonia and vaccine strains derived from the El Tor strain N16961
Fraction Dilution yielding maximum vacuolating activity FG1066 (Amazonia) N16961 (El Tor, wild type) JBK56 (CT− Apr) JBK70 (CT−Hgr) CVD104 (CT− HlyA−) Supernatant 1/640 1/20 1/20 1/20 <1/20 Concentrate 1/5120 1/320 1/320 1/320 <1/20 - Table 3.
Comparison of vacuolating cytotoxic activities from various strains in hlyA inactivation, reversion, cloning, and complementation experimentsa
Fraction Dilution yielding maximum vacuolating activity FG1106 FG1153 FG1173 FG1167 DH5α FG1221 FG1229 Supernatant 1/212 <1/20 1/162 1/125 <1/20 <1/20 1/853 Concentrate 1/4,096 <1/20 1/7,680 1/5,120 <1/10 1/30 1/40,960 ↵a See Table 1 for descriptions of the strains used.
