The techniques of hemagglutination inhibition and in vitro lymphocyte transformation and the assay of migration inhibitory factor were used for study of the development of rubella antibody activity in serum and tonsillar tissue washings and of the appearance of rubella-specific, cell-mediated immunity in circulating and tonsillar lymphocytes in groups of children who underwent tonsillectomy at various intervals after natural rubella infection, subcutaneous immunization with HPV-77 DE/5 vaccine, or intranasal inoculation with RA27/3 live, attenuated rubella virus vaccine. Antibody response in serum and tonsillar tissue washings was detected regularly after natural infection or immunization. Development of specific cell-mediated immunity in circulating lymphocytes was regularly observed after natural infection and frequently after immunization. Natural infection or intranasal immunization with rubella vaccine resulted in the appearance of cell-mediated immunity in the tonsillar lymphoid tissue, and the response was detectable up to several years after natural infection in several cases. It is significant, however, that the level of cell-mediated immunity in tonsils was conspicuously low after subcutaneous immunization. These data suggest the induction of local cell-mediated immunity in tonsillar lymphoid tissue after local mucosal application of rubella virus.

Employing the techniques of complement fixation, immunofluorescence, and in vitro lymphocyte transformation assay, the antibody and cell-mediated immunity to cytomegalovirus (CMV) were studied in the serum, peripheral blood lymphocytes, tonsillar lymphocytes, and cord blood lymphocytes. The study population consisted of 32 children undergoing tonsillectomy and adenoidectomy. In the lymphocyte transformation assay, three strains of CMV (AD-169, ADH-1-41, and Davis), herpes simplex type 1, and phytohemagglutinin were employed as antigens. Sixty-five percent of the subjects were found to have CMV-specific antibody activity. The lymphocyte transformation response to phytohemagglutinin was similar in all subjects. No CMV-specific lymphocyte transformation activity was detected in cultures of cord blood lymphocytes. Significant cell-mediated immunity was observed in the tonsillar lymphocytes of 30% (3/10) of the seronegative individuals and in the peripheral blood lymphocytes obtained from one such subject. Over 75% (16/21) of the seropositive subjects demonstrated cell-mediated immunity against one or more strains of CMV in the peripheral blood lymphocytes and tonsillar lymphocytes. In the lymphocyte transformation assay, no cross-reactivity was apparent between CMV and herpes simplex type 1. These studies demonstrate the presence of strain-specific systemic and mucosal cell-mediated immune response to CMV in humans. The frequency and distribution of lymphocyte transformation responses to the three CMV strains suggest antigenic heterogeneity of CMV.

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In an in vitro direct assay with tissue-type plasminogen activator (tPA), plasminogen and the chromogenic substrate S-2251, the ability of Mycoplasma fermentans KL4 to stimulate tPA-mediated activation of plasminogen to plasmin was studied. Mycoplasma cells markedly enhanced the activation of plasminogen by tPA in a concentration-, temperature- and pH-dependent manner. Nonidet P-40 (0.01%), sonication, and freezing and thawing of the cells substantially increased the stimulatory effect of mycoplasma on tPA activity. In contrast, the activation of plasminogen by urokinase was refractory to mycoplasma cells. The mycoplasma-mediated stimulation of tPA activity was prevented by epsilon-aminocaproic acid (EACA), a lysine analogue known to block lysine-binding sites (LBS) in plasminogen and tPA. Among several Mycoplasma fermentans strains tested, incognitus strain demonstrated the highest stimulation activity. These results suggest that mycoplasma cells interact with LBS in tPA and plasminogen to enhance plasminogen activation.