Immunization with the C‐Domain of α‐Toxin Prevents Lethal Infection, Localizes Tissue Injury, and Promotes Host Response to Challenge withClostridium perfringens

Abstract

Clostridium perfringens gas gangrene is characterized by rapid tissue destruction, impaired host response, and, often, death. Phospholipase C (alpha -toxin) is the virulence factor most responsible for these pathologies. The present study investigated the efficacy of active immunization with the C-terminal domain of alpha -toxin (Cpa247-370) in a murine model of gas gangrene. Primary end points of the study were survival, progression of infection, and tissue perfusion. Secondary end points, which were based on findings of histologic evaluation of tissues, included the extent of tissue destruction and microvascular thrombosis, as well as the magnitude of the tissue inflammatory response. Survival among C-domain-immunized animals was significantly greater than that among sham-immunized control animals. Furthermore, immunization with the C-domain localized the infection and prevented ischemia of the feet. Histopathologic findings demonstrated limited muscle necrosis, reduced microvascular thrombosis, and enhanced granulocytic influx in C-domain-immunized mice. We conclude that immunization with the C-domain of phospholipase C is a viable strategy for the prevention of morbidity and mortality associated with C. perfringens gas gangrene.

Progress and problems in vaccination against necrotic enteritis in broiler chickens

“…Reports on monoclonal antibodies against alpha toxin led to the finding that monoclonal antibodies, which were capable of neutralizing the phospholipase C activity, were not necessarily effective in neutralizing the haemolytic and lethal activities of the toxin in gangrene models (Sato et al, 1989;Logan et al, 1991). The immune response against the C-domain provided protection against challenge with alpha toxin and also against experimental gas gangrene in mice (Stevens et al, 2004).…”

“…The alpha toxin is the most investigated C. perfringens toxin in terms of vaccine-induced protection, mainly in mouse gangrene models (Stevens et al, 2004;Titball, 2009). The alpha toxin is composed of two domains, which are associated with phospholipase C activity (N-domain) and membrane recognition (C-domain) (Naylor et al, 1998).…”

Clostridium perfringens Antigens Recognized by Broiler Chickens Immune to Necrotic Enteritis

“…Although vaccination offers an alternative approach to antimicrobial drugs in control of the disease, little is known about immunity to NE. However, there is the suggestion that the alpha-toxin, a phospholipase C exoenzyme, is an important immunogen (1,13,21). A recent study showed that alpha-toxin is not essential in the pathogenesis of NE (10).…”

Bacterial Sphingomyelinases and Phospholipases as Virulence Factors

“…perfringens is the most common cause of gas gangrene, an acute infection associated with traumatic or surgical wounds and characterized by intravascular leukocyte accumulation, thrombosis, and severe myonecrosis (217). A C. perfringens mutant strain with an inactivated plc is unable to produce gas gangrene (218), and immunization with the alpha-toxin C-terminal domain protects mice against experimental gas gangrene induced by C. perfringens (219). Similarly, immunization of guinea pigs with the Clostridium haemolyticum PLC elicits a protective immunity from an intramuscular challenge with 50 LD 50 s of this bacterium (220).…”