The improvement of the therapeutic anti- Lachesis muta serum production in horses

The main features associated with pit viper envenomations include the intense local lesions such as oedema, necrosis, acute renal failure and other effects. The severity of these reactions to snakebite depends on the degree of envenomation. Lachesis muta venom (LMV) has weak lethal activity, but due to the large amount often inoculated, the effects are extremely severe and demand anti-venom with a high neutralizing capacity. LMV had the lowest neutralizing antibody induction capacity in horses when compared with that of other venoms. For example, Bothrops anti-venom serum neutralizes 180 times the equivalent LD 50 to Bothrops venom; Crotalus anti-venom neutralizes 250 LD 50 of this venom, while Lachesis anti-venom neutralizes only five LD 50 of the Lachesis toxins. To examine the reasons for this low antibody induction, the H GP mouse line, genetically selected for high antibody production received, at different times during immunization with sheep erythrocytes (SE), whole LMV and isolated venom fractions I–VI eluted by gel-filtration chromatography on Superdex ®75. The specific antibody responsiveness showed a partial, but significant suppression of the anti-SE antibody responses during the kinetics of the primary and even the secondary immunizations, after 50–100 μg of fractions IV and V administration 72–48 h before the first antigen injections. Fraction IV was then applied in a Superose ®12 column and three samples were obtained. The peak IVA containing a component of Mr 27 kDa was liable with the immunosuppressive effect as made evident by its effect on the H mice anti-SE responses. Horses receiving the LMV exempt of fractions IV and V produce highly significant anti- Lachesis sera with a 45 LD 50 neutralizing activity, providing, for the first time, an efficient specific therapeutic heterologous serum for human use.