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[Investigations about the immunobiology of m-proteins of Streptococcus pyogenes. IV. Cell-mediated immunity of rhesus monkeys (Macaca mulatta) against streptococcal antigens (author's transl)].

Cell-mediated immunity was tested in monkeys (Macaca mulatta) immunized with highly purified M-proteins by means of the lymphocyte transformation reaction. Lymphocytes from immunized and nonimmunized monkeys were stimulated with PAL-M-Proteins. IC-M-proteins were completely free of mitogenicity and no lymphocyte stimulation was to be found after immunization with 1.6 mg. Erythrogenic toxins of strain NY5 were able to stimulate lymphocytes of monkeys unspecifically.

Animals↗

Receptors for erythrogenic toxin A of Streptococcus pyogenes on human peripheral blood lymphocytes and other blood cells.

The binding of erythrogenic toxin A (ETA) of group A streptococci on human peripheral blood cells was demonstrated light- and electronmicroscopically by labelling with ETA-gold. Double labelling experiments of lymphocytes and the use of purified T-lymphocytes revealed that the ETA-receptor was confined to T cells. Besides lymphocytes, monocytes and especially granulocytes and thrombocytes showed a significant ETA-gold labelling. The lymphocyte ETA-receptor was found to be not sensitive to trypsin and neuraminidase and was not identical with the receptors for concanavalin A, Helix pomatia-lectin and sheep erythrocytes.

Adult↗

Adjuvant immunotherapy with intrapleural Streptococcus pyogenes (OK-432) in lung cancer patients after resection.

A prospective randomized study to evaluate the effect of adjuvant intrapleural OK-432 immunotherapy after resection of lung tumor was conducted in 93 patients with primary lung cancer. Among them, 46 patients had had intrapleural OK-432 injection, 47 had not. In the meantime, serial measurements of serum immunosuppressive acidic protein, of serum interleukin-2 receptor and of the subpopulation of the peripheral blood cells and lymphocytes were performed in all these patients. Patient characteristics in these two groups (sex, age, histological type, pathological stage, type of operation, and performance status) were compatible. The results showed that adjuvant intrapleural OK-432 injection after resection had no beneficial effect on a patient's survival time. Patients who received intrapleural OK-432, had an increase in blood leukocytes, granulocytes and monocytes and serum immunosuppressive acidic protein level. But the cell numbers of total T cells, suppressor/cytoxic cells, helper/inducer cells and natural killer cells of peripheral blood were decreased in the OK-432 positive group. Over half of the patients had transient 1- or 2-day febrile reactions after intrapleural OK-432 injection. It was concluded that neither clinical observation nor immunological monitoring of peripheral blood could demonstrate a beneficial effect from intrapleural OK-432 immunotherapy after complete resection of the tumor.

Adjuvants, Immunologic↗

Effectiveness of ribosomal fractions of Klebsiella pneumoniae, Streptococcus pneumoniae, Streptococcus pyogenes, Haemophilus influenzae and the membrane fraction of Kp (Ribomunyl) in the prevention of clinical recurrences of infectious rhinitis. Results of a multicenter double-blind placebo-controlled study.

A multicenter, double-blind, placebo-controlled study was conducted to investigate the efficacy of an immunostimulant, Ribomunyl, in the prevention of recurrences of infectious rhinitis in adults. This trial involved 327 patients (168 Ribomunyl treated and 159 placebo cases) with an average of 4.3 +/- 1.8 rhinitis episodes per patient recorded during the year preceding the study. The main criterion of efficacy was the cumulative number of recurrences of infectious rhinitis during a 6-month follow-up period, as analyzed by standard tests. An additional analysis of relative risk of recurrences used multivariate failure for time data. Ribomunyl was effective throughout the study period, starting from the first month of treatment: a mean of 1.0 +/- 1.1 recurrences was recorded in the Ribomunyl group as compared to 1.5 +/- 1.4 recurrences in the placebo group; this indicated one-third fewer infections (P = 0.001). The protective effect of Ribomunyl on the relative risk for recurrences was estimated to be 0.58 by multivariate analysis (95% CI: 0.43-0.78, P = 0.0001). Analysis of secondary criteria also favored Ribomunyl: 38.5% less antibiotic courses per patient (0.8 +/- 1.3 vs 1.3 +/- 1.6; P = 0.002) and the number of days with antibiotics (5.6 +/- 9.3 vs 9.1 +/- 12.1; P = 0.002).

Adjuvants, Immunologic↗

Activation of human peripheral blood-derived monocytes by OK-432 (Streptococcus pyogenes): augmented cytotoxicity and secretion of TNF and synergy with rIFN-gamma.

The biological response modifier OK-432 has been shown both to exert the enhancement of several immunological activities and to have a direct anti-tumor effect. The present study examines the immunopotentiating effect of OK-432 on peripheral blood monocytes (PBM) derived from normal humans. Monocyte activation was assessed by examining direct cell-mediated cytotoxic activity (CMC) and secretion of cytotoxic factors in the supernatant by the 51Cr release assay and secretion of tumor necrosis factor (TNF)-alpha detected by a sensitive radioimmunoassay. The OK-432-augmented activity was compared to that achieved by recombinant interferon-gamma (rIFN-gamma). Coculture of PBM with OK-432 overnight resulted in significant augmentation of CMC and secretion of cytotoxic factors and TNF in the supernatant. The effects observed were dose dependent and the resulting activity was much more pronounced than that achieved with an optimal concentration of IFN-gamma. The monocyte- and supernatant-mediated cytotoxic activities were in a large part attributed to TNF as both activities were inhibited by anti-TNF antibody. Several parameters of monocyte activation by OK-432 were examined. The kinetics of monocyte activation revealed that a short time exposure (2-6 hr) was sufficient for activation but maximal activation was detected after 18 hr. However, the kinetics of the cytotoxic assay were not shortened and 16-20 hr was necessary for optimal cytotoxic activity. Significant synergy was obtained when suboptimal concentrations of OK-432 and IFN-gamma were used. The synergy was noted in CMC, supernatant activity, and TNF concentration. These results demonstrate that OK-432 is a potent activator of monocyte cytotoxicity and also activates secretion of TNF. Also, OK-432 is a much more potent activator than rIFN-gamma. The synergy with OK-432 and IFN-gamma suggests that OK-432-mediated activation of monocytes takes place by a different mechanism than that mediated by rIFN-gamma. Thus, monocytes and products thereof may actively participate in the in vivo anti-tumor effect mediated by OK-432.

Biological Products↗

Induction of interferon by streptococcus pyogenes extracellular products.

Lymphocyte-activating streptococcal exoproteins, which were previously characterized, have been tested for their capacity to induce interferon in vitro. Two out of the 3 different streptococcal fractions, studied on mice splenocytes, were shown to elicit the production of a significant amount of interferon. A large proportion of the interferon detected in the supernatants from mice activated spleen cells was acid-labile interferon. The highest level of interferon titer was obtained with the streptococcal fraction identified as the erythrogenic toxin.

Animals↗

Activation of a 66-kilodalton human endothelial cell matrix metalloprotease by Streptococcus pyogenes extracellular cysteine protease.

Human umbilical vein endothelial cells (HUVECs) were used to gain insight into the molecular mechanism whereby the major extracellular protease from group A streptococci damages host tissue. HUVECs exposed to streptococcal cysteine protease (SCP) for various times exhibited cytopathic effect and cell detachment from the culture vessel. Gelatin substrate zymography showed that a time- and concentration-dependent increase in the level of activity of an approximately 66-kDa gelatinase occurred in culture medium taken from cells exposed to enzymatically active SCP. This gelatinase comigrated in gelatin zymograms with the activated form of purified recombinant matrix metalloprotease 2 (MMP-2) and had type IV collagenase activity. In contrast, medium taken from cells exposed to inactivated (boiled) SCP and cells exposed to SCP inhibited by treatment with N-benzyloxycarbonyl-leucyl-valyl-glycine diazomethyl ketone lacked the 66-kDa gelatinase. Appearance of the 66-kDa gelatinase activity was also prevented by 1,10-phenanthroline, a zinc chelator and MMP inhibitor. Inasmuch as proteolytically active SCP is required for the emergence of this gelatinase and MMP activation occurs by proteolytic processing, the 66-kDa gelatinase may be a proteolytic cleavage product of a latent MMP expressed extracellularly by HUVECs. Direct SCP treatment of culture supernatant taken from HUVECs not exposed to SCP also produced the 66-kDa gelatinase. The data show that SCP activates an MMP produced by human endothelial cells, a process that may contribute to endothelial cell damage, tissue destruction, and hemodynamic derangement observed in some patients with severe, invasive group A streptococcal infection.

Cells, Cultured↗