Biomedical subjects

T V Freitas

Publications and source records attributed to T V Freitas.

7 recordsLinked to original sources

PubMed2002

Comparison of the biodistribution of free or liposome-entrapped Crotalus durissus terrificus (South American rattlesnake) venom in mice.

The local absorption rate, clearance and tissue distribution of Crotalus durissus terrificus venom, (Cdt) were examined using a two-antibody sandwich ELISA assay. We compared the biodistribution of both free or encapsulated Cdt in mice. Following subcutaneous injection of 10 microg/mouse of free Cdt (0.8 LD50), venom was detected in serum after 15 min, showed its highest level at 30 min (45+/-5 ng/ml) and was cleared from the circulation after 6 h. After 2 h of inoculation, venom was detected in the kidney (57+/-9 ng/g of tissue), spleen (18+/-4 ng/g of tissue) and brain (14+/-6 ng/g of tissue). For both subcutaneous or intravenous injection of free Cdt, venom was firstly detected in the kidney. No Cdt appeared either in the kidney, spleen, brain, or other tissues after subcutaneous inoculation of encapsulated venom even though a higher dose was used, 25 microg/mouse (2 LD50). Venom remained at the site of injection for a period of 1 week. Following intravenous injection of encapsulated venom (5 microg/mouse, 2 LD50), venom was detected in liver and spleen tissues. The biodistribution of encapsulated venom is discussed in relation to the effects of reduction of toxicity and increase of adjuvanticity.

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PubMed2000

Immunization with liposome-encapsulated Bothrops jararaca venom.

The venom of Bothrops jararaca (BjV) snake was encapsulated into liposomes. The toxicity and ability of the resultant liposomes to protect mice were evaluated. No acute toxicity was found in mice, when liposomes were subcutaneously injected whereas the same dose of venom emulsified in Freund's adjuvant caused the mice to die. Immunization with the venom containing liposomes and associated immunostimulants, protected 50% of mice after challenge with BjV (4 x LD50). The hemorrhagic activity induced by BjV was reduced after immunization with liposomes associated with immunostimulants, similarly to immunization with BjV emulsified in Freund's adjuvant. However, mice immunized with BjV:Freund's were better protected against the lethal effects of the venom.

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PubMed1997

Encapsulation of native crotoxin in liposomes: a safe approach for the production of antivenom and vaccination against Crotalus durissus terrificus venom.

Crotoxin, the neurotoxic component of Crotalus durissus terrificus (Cdt) venom that displays phospholipase A2 activity, was successfully encapsulated into dehydration-rehydration vesicles (DRV/crotoxin) and reverse-phase evaporation vesicles (REV/crotoxin) made from sphingomyelin and cholesterol. The encapsulation efficiency of native crotoxin was higher in DRV/crotoxin than in REV/crotoxin. DRV/crotoxin was not toxic when i.v. inoculated in mice at a dose of crotoxin as high as 91 times its L.D50 or when s.c. inoculated at 42 times its LD50. On the other hand, crotoxin released from DRV/crotoxin retained its original toxicity. REV/crotoxin was found to be at least 1.9 times more toxic than DRV/crotoxin. The fact that DRV/crotoxin retained crotoxin more efficiently than REV/crotoxin may account for the difference in acute toxicity between the two preparations. DRV/crotoxin, when s.c. inoculated in mice, induced anti-crotoxin antibodies that protected animals against the lethal effect of Cdt venom. Following immunization with three doses of DRV/crotoxin (3 x 20 micrograms of crotoxin/mouse) and challenge with 8 x LD50 of Cdt venom, 75% of mice were protected. The DRV/crotoxin preparation was compared to crotoxin emulsified in Freund's adjuvant (FCA/crotoxin). DRV/crotoxin was found to be less toxic than FCA/crotoxin, and to induce lower levels of anti-crotoxin antibodies but similar levels of protection when inoculated at high doses (20 or 70 micrograms crotoxin/mouse). When DRV/crotoxin was adsorbed to alum at the time of immunization, it induced antibody and protection levels comparable to those produced by FCA/crotoxin.

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PubMed1992

Biological activities of venoms from South American snakes.

Standard assay procedures for the characterization of snake venoms, developed by the World Health Organization (WHO) Collaborating Centre for the Control of Antivenoms (CCCA), were used to analyse 33 snake venoms including eight International Reference Venoms for the assessment of lethal, defibrinogenating, procoagulant, haemorrhagic and necrotizing properties. The International Reference Venoms were assayed as part of an International Collaborative Programme for the evaluation of Venoms and Antivenoms; the results showed a close relationship to those obtained by the CCCA. Twenty-five venoms from 13 different species of medically important snakes from South America were assayed as standardized by the WHO-CCCA. Additionally, evaluation of lethal activity by the i.p. and intra cerebroventricular routes, proteolytic effects and venom-induced edema were also determined. Venom yields from captive snakes are also presented. Among the venoms examined, from the subfamily Crotalinae, the members of the genera Bothrops and Lachesis had strong haemorrhagic, proteolytic and edema-inducing activities, whereas all Crotalus durissus species had none. The Elapinae, Micrurus frontalis showed no procoagulant, defibrinogenating, haemorrhagic, necrotizing or proteolytic activities. The results reflect differences among individual samples of the same species and of different geographical regions. The results suggest that there is little or no relationship between the properties of the different venoms and that the determination of one effect cannot predict the value of the others. Therefore, the characterization of the different activities of snake venoms is necessary if toxicity is to be properly evaluated and neutralized.

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PubMed1991

Immunization of horses with Crotalus durissus terrificus (South American rattlesnake) venom. A comparison of four different procedures.

1. A comparative study was carried out on horses immunized with Crotalus durissus terrificus venom using four different inoculation procedures, which included the use of Freund's adjuvant, A1(OH)3 and liposomes as adjuvants. The antibody titer was assessed by enzyme linked immunosorbent assay (ELISA) and the neutralizing potency by the neutralizing median effective dose (ED50). 2. The inoculation schedule used in horses to obtain antivenom serum consisted of sc injections of a 7.5 mg venom starting dose in 5.0 ml sterile saline emulsified with an equal volume of Freund's complete adjuvant. One week later, 7.5 mg venom in 5.0 ml sterile saline emulsified with an equal volume of Freund's incomplete adjuvant was injected. This was followed by three doses of 2.5 mg venom in 12.0 ml sterile saline at 2-day intervals. This immunization procedure, based on low doses of antigen (37.5 mg/horse) emulsified with Freund's adjuvant, produced a more protective and sustained immune response when compared with other procedures using A1(OH)3 or liposome emulsions with either low (50.0 mg/horse in A1(OH)3 and 5.0 mg/horse in liposome) or high (870.0 mg/horse in A1(OH)3 and 20.0 mg/horse in liposome) antigen doses. The ED50 values evaluated at the end of the procedure were 15.4 microliters serum/20 g mouse when antigen was emulsified with Freund's adjuvant; 21.7 microliters serum/20 g mouse when 870.0 mg antigen/horse was emulsified with A1(OH)3 and 30.0 microliters serum/20 g mouse when 50.0 mg antigen/horse was emulsified with A1(OH)3. When antigen was emulsified with liposome, the immune serum was ineffective against the lethal effects of C. d. terrificus venom. 3. The inoculation schedule used in horses to obtain hyperimmune serum consisted of reimmunization with sc booster injections of 7.5 mg venom in 5.0 ml sterile saline emulsified with an equal volume of Freund's incomplete adjuvant. One week later, 2.5 mg venom in 12.0 ml sterile saline was inoculated at 2-day intervals. This reimmunization schedule, based on low doses of antigen (15.0 mg/horse) emulsified with Freund's incomplete adjuvant or with saline, produced a protective and sustained immune response, regardless of the initial immunization procedure. The ED50 evaluated for each of the animals five days after the reimmunization period was never more than 20 microliters serum/20 g mouse. 4. The liposome inoculation method employed a membrane-stabilized reverse phase evaporation preparation of sphingomyelin/cholesterol 2.5/l (w/w) liposomes. This procedure permits incorporation of 1.0 mg protein per mg of phospholipid.(ABSTRACT TRUNCATED AT 400 WORDS)

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PubMed1990

Protection against the lethal effects of Crotalus durissus terrificus (South American rattlesnake) venom in animals immunized with crotoxin.

Mice and rabbits were immunized against crotoxin (the neurotoxic component isolated from Crotalus durissus terrificus venom) using small amounts of antigen in a water-in-oil emulsion. Following boosting (three times at 21-day intervals) a high titre of antibodies against crotoxin was obtained. Crotoxin immunoglobulin G antibody recognizes whole venom antigen at a level comparable with that of crotoxin antigen, using the ELISA method for antibody detection. The antibodies generated by crotoxin were capable of providing 100% protection against challenge with 11 and 50 i.p. LD50 doses of whole venom in mice. When 100 i.p. LD50 doses of whole venom were injected survival was 77.8%.

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PubMed1989

Use of liposomes for protective immunisation against Crotalus durissus (tropical rattlesnake) venom.

Crotalus durissus venom has been described as a weak antigen when injected in combination with Freund's complete adjuvant during the course of traditional methods of equine immunisation. Antibody production is slow and unpredictable, with a wide variation in individual responses. In this experimental study, C. durissus venom was incorporated into stabilised sphingomyelin-cholesterol liposomes both in the presence and absence of lipopolysaccharide immunostimulant and injected by both i.v. and s.c. routes into mice and rabbits. A rapid, sustained and protective immune response was obtained following a single injection of these preparations in mice. Antibody levels were estimated using enzyme-linked immunosorbent assay (ELISA), and the protective effect was evaluated by subsequent challenge with a subcutaneous minimum lethal dose of the venom. Results indicated that the immune response was significantly potentiated by the presence of immunostimulant in the venom liposomes. The use of C. durissus venom liposomes should be a useful tool for the immunisation of animals both in experimental and commercial procedures.

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