In vivo effects of aluminum adjuvant on systemic reaction of bacterial lipopolysaccharide (LPS) in piglets were investigated. Intramuscular injection of 0.1 mg kg-1 of LPS added to aluminum hydroxide gel (LPS(+)AL) mitigated the leukopenia, trembling and serum levels of TNF-alpha and cortisol compared with the injection of LPS suspended in LPS-free saline (LPS(+)SALINE). The serum endotoxin levels were reduced remarkably but relatively long-lasting in the LPS(+)AL. The lethality in mice injected with LPS added to aluminum hydroxide gel was significantly reduced. Likewise, the Limulus activity of a test LPS was reduced by the addition of aluminum hydroxide gel or aluminum chloride.
Clostridium chauvoei strain Okinawa produced spontaneous non-motile variants at an unusually high rate (approx. 10(-4) per generation) under normal conditions without mutagen. Revertants of non-motile variants were detected at a rate of approximately 10(-3). Biochemically, every variant corresponded well with the parental strain. By transmission electron microscopy, three of nine non-motile variants of strain Okinawa were found to be flagellate, while the other six were found to be aflagellate. These phenotypes were confirmed by Western blot analysis using monoclonal antibodies directed against the flagella of C. chauvoei. Moreover, the parental flagellate strain and non-motile flagellate variants were significantly more virulent in mice than non-motile, aflagellate variants. Our results demonstrated that phase variation in motility and flagellation occurs in C. chauvoei, and that the flagella are associated with the full expression of virulence.
The in vivo effects of bacterial lipopolysaccharide (LPS) on the immune systems of piglets were investigated. Intravenous injection of 0.5 mg of LPS per kilogram of body weight induced apoptosis, which was characterized by nuclear chromatin condensation and fragmentation and a ladder formation of nucleosomal DNA in lymphocytes both in the cortex of the thymus and in the germinal centers and paracortical areas of mesenteric lymph nodes at 24 h postinjection. The levels of endotoxin, tumor necrosis factor alpha, and cortisol in serum increased, generally according to the dose of LPS. These findings suggest that LPS can induce in vivo apoptosis of lymphocytes in piglets and support the notion that cytokine and endocrine responses may play an important role in LPS-induced apoptosis in the immune system.
The effect of bacterial lipopolysaccharide (LPS) on the lymphoid organs in C3H/HeN and C3H/HeJ mice was investigated. In C3H/HeN mice, LPS induced apoptosis, characterised by morphological nuclear condensation and DNA fragmentation resulting in thymic atrophy. Similar but less severe changes were also observed in the spleen and lymph nodes. In C3H/HeJ mice, only a slight depletion of lymphocyte numbers was observed in the lymphoid organs. The plasma endotoxin levels were dependent on the LPS dose regardless of mouse strain. On the other hand, the plasma TNF-alpha levels were significantly elevated in C3H/HeN mice 1 h post-injection and the time course of plasma corticosterone concentration correlated well with the development of apoptosis. These findings suggest that TNF-alpha and corticosterone may play an important role in LPS-induced apoptosis of lymphocytes.
Polyclonal rabbit anti-idiotypic antibody (anti-Id) against the protective monoclonal antibody specific to the flagella of Clostridium chauvoei was produced, purified, and characterized. Anti-Id inhibited the binding of its related monoclonal antibody to the flagellar antigen, suggesting that the anti-Id bore an internal image of the flagellar antigen. When mice were immunized with anti-Id intraperitoneally, the survival rate increased significantly, compared with mice immunized with normal rabbit IgG (P < 0.01), and specific anti-flagellar antibodies were induced.