TABLE 3.
ELISA of the supernatants from the two-step coculture experiments
| Groupa | Supernatant source and designation | Cytokine concn (pg/ml)b
|
|
|---|---|---|---|
| TNF-α | IL-1β | ||
| I | Apical sup from: | ||
| EC1 | - | - | |
| EC1/PMN | 73.2 | 229 | |
| L2C1 | - | - | |
| L2C1/PMN | 52.8 | 104.6 | |
| UC1/PMN | - | 3.3 | |
| RPMI/PMN | 0.7 | 0.13 | |
| II | Sup from MdM stimulated for 48 h | ||
| UC1→MdM | - | - | |
| UC1/PMN→MdM | 2.7 (2.7) | 0.9 (-) | |
| EC1→MdM | 6.3 (6.3) | 3.3 (3.3) | |
| EC1/PMN→MdM | 55.4 (18.8) | 102.3 (-) | |
| L2C1→MdM | 1.5 (1.5) | 3.2 (3.2) | |
| L2C1/PMN→MdM | 278.5 (252.1) | 369.3 (317) | |
| III | Basal sup from: | ||
| EC2 | - | - | |
| EC1/PMN→EC2 | 13.6 (-) | 101.1 (-) | |
| UC1→MdM→EC2 | - | - | |
| EC1→MdM→EC2 | 8.4 | 8 | |
| RPMI/PMN→MdM→EC2 | 5.5 | - | |
| UC1/PMN→MdM→EC2 | - | - | |
| EC1/PMN→MdM→EC2 | 13.6 | 11.5 | |
| L2C2 | 0.7 | 0.3 | |
| L2C1/PMN→L2C2 | 10.3 (-) | 47 (-) | |
| UC1→MdM→L2C2 | - | - | |
| L2C1→MdM→L2C2 | 326.6 | 559 | |
| RPMI/PMN→MdM→L2C2 | 3.8 | 0.2 | |
| UC1/PMN→MdM→L2C2 | - | 0.2 | |
| L2C1/PMN→MdM→L2C2 | 588.6 | 564.8 | |
Group I includes supernatants (Sup) from representative apical supernatants from culture 1 of HeLa cells uninfected or infected with C. trachomatis serovar E (EC1) or L2 (L2C1) and incubated in some experiments with PMN (UC1/PMN, EC1/PMN, and L2C1/PMN, respectively). Controls using PMN incubated with RPMI alone were also included. Group II represents supernatants from MdM incubated for 48 h with group I supernatants. Group III represents basal supernatants from cultures of serovar E-infected (EC2) or serovar L2-infected (L2C2) HeLa cells alone or in coculture with MdM preactivated with group I supernatants.
In some experiments, group I supernatants were used as cytokine inducers. Since these supernatants already contain known concentrations of TNF-α and IL-1β and since they were used after a 1:1 dilution in fresh medium, the actual cytokine release after induction, shown in parentheses, was calculated as follows: total amount detected − (concentration in group I supernatant/2). -, not detected; (−), no additional cytokine release after induction. The table shows average cytokine concentrations from duplicate wells of a representative experiment. These coculture experiments were repeated only twice because of their complexity, and the cytokine concentrations detected in these independent experiments were not averaged because some differences in the actual numbers were found; these were probably caused by a variation in the MdM responsiveness or in the number/well. However, the patterns of cytokine response were similar in both experiments.