Attempts have been made by in vitro methods to demonstrate the presence of a cell-mediated immune response to the gonococcus in human peripheral leukocytes. Leukocyte-migration tests in the presence of gonococcal antigen were performed in 34 patients with uro-arthritis (Reiter's disease), in 18 patients with newly developed gonorrhoea without joint manifestations, and in 34 controls (blood donors). In the patients with uncomplicated gonorrhoea the mean migration index (MI) was generally the same as in the healthy controls. A MI below 0.75 was noted in 21 of 34 cases of uro-arthritis. The number of cases with inhibited migration rose parallel with the duration of the disease and the presence of prostatis. Antigen-induced lymphocyte transformation was also studied by quantitation of the 14C-thymidine uptake in cell cultures. Studies were made in three groups of patients, one group with uroarthritis, one with gonococcal urethritis, and one with gonococcal septicaemia, and also in healthy controls. The lymphocyte stimulation induced by gonococcal antigen was significantly greater in uro-arthritis patients with known gonococcal infection (0.01less than p greater than 0.05) than in healthy controls. No difference in DNA synthesis was noted between patients with uncomplicated gonorrhoea and healthy controls, while the gonococcal septicaemia patients demonstrated a poor cellular immune response.
Antigen-induced lymphocyte proliferation was studied by quantitation of 14C-thymidine uptake in cell cultures. The induction of DNA synthesis in vitro in lymphocytes from patients with uro-arthritis after stimulation by whole cells of virulent and avirulent N. gonorrhoeae, meningococci group B, and apathogenic Neisseria (N. pharyngis) was compared with the DNA synthesis in lymphocytes from healthy controls after stimulation with the same Neisseria antigens. The difference between patients and the controls was found to be highly significant after stimulation with virulent or avirulent N. gonorrhoeae organisms but not after stimulation with apathogenic Neisseria. An analysis of the correlation of the lymphocyte reactivity to all the Neisseria antigens showed a highly significant correlation between the response of uro-arthritis patients to the two types of gonococcal antigen (0.932*** and 0.859***), a lower correlation coefficient for group B meningococci and virulent or avirulent gonococci (0.724*** and 0.714***) and no correlation at all between apathogenic Neisseria and gonococci. The DNA synthesis in lymphocytes stimulated by N. gonorrhoeae and other Gram-positive and Gram-negative bacteria was also studied in cell cultures obtained from other healthy controls as well as uro-arthritis patients. There was no significant difference between the patients and the controls with regard to the response to apathogenic Neisseria and other non-Neisseria antigens.
In vitro lymphocyte responses to gonococcal antigen (T2), apathogenic Neisseria pharyngis (APN), PPD and PHA were studied in patients with uroarthritis and in healthy controls. Only the T2 antigen induced a significantly higher DNA synthetic response in cells from patients than in controls, whereas no difference was observed with the other substances. Peak responses occurred on day 6 of culture with a concentration of 4 × 107 bacteria/ml. In order to test whether lymphocyte responses were the result of specific sensitization, experiments were carried out with cord blood cells. These cells reponded to both T2 and APN with maximum activity on day 6 of culture. Since specific sensitization was ruled out in these experiments, we concluded that T2 and APN were capable of inducing a non‐specific mitogenic response in cord blood cells. The mitogenic activity of T2 and APN was further elucidated in experiments with mouse lymphocytes. Both substances induced increased DNA synthesis in mouse spleen cells, with peak activity on days 2–3 of culture. Stimulation occurred in spleen cells depleted of macrophages, in anti‐Thy 1.2 treated spleen cells and in Nude spleen cells, but not in thymus cell cultures. It was concluded that T2 and APN contained polyclonal B cell activating (PBA) substances, which was subsequently proved by their ability to induce a polyclonal antibody response in cell culture. The B cell subpopulation that responds to the mitogenic properties of these bacteria is normally absent from blood, present as a minor proportion of cord blood lymphocytes and present as a major cell population in spleen. Therefore, in spite of the mitogenic properties of these bacterial strains, they are useful tools for the elucidation of a specific reactivity in patients with uroarthritis.
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