TY  - JOUR
T1  - Crotalic Venom Fraction as Promoter of the Transfection Mechanism of a Genic Vaccine (Naked DNA) Against Rabies
AU - , Arcelia Alvarado Islas AU - , Angel Horacio Sandoval Trujillo AU - , Victor R. Tenorio Gutierrez AU - , Rogelio Alonso Morales AU - , Octavio de Paz Villafan AU - , Alvaro Aguilar 
JO  - Journal of Animal and Veterinary Advances
VL  - 7
IS  - 9
SP  - 1045
EP  - 1055
PY  - 2008
DA  - 2001/08/19
SN  - 1680-5593
DO  - javaa.2008.1045.1055
UR  - https://makhillpublications.co/view-article.php?doi=javaa.2008.1045.1055
KW  - Hemoagglutinins
KW  -lectins
KW  -plasmids
KW  -Agkistrodon piscivorus
AB  - The AL27 crotalic fraction is a lectin recognized by cell receptors and then introduced into the cytoplasm. In the present research a new transfection promoter agent for a non viral expression vector was used, using as a model a plasmid expressing the gene that codifies for the G glycoprotein of the rabies virus. The  objective  was to evaluate the ability of AL27 to increase the rabies plasmid transfection in <I>in vitro</I> and <I>in vivo</I> conditions. The AL27 fraction was isolated from the venom of <I>Agkistrodon piscivorus</I> by HPLC-RP. AL27 maintained its recognition by sialic acid receptors up to a titer of 64 HU. The pC38 plasmid was selected from a panel of 18 clones constructed by inserting the gene that codifies for glycoprotein G of rabies strain HQIMSS99 into the commercial pCI-neo expression vector. <I>In vitro</I> results showed that pC38, with or without AL27, transfected CHO cells under the calcium-phosphate system and using Confocal Microscopy (CM) increased fluorescence (+++) was observed with the mixture pC38/AL27, than with pC38 alone (++). Flow cytometry (FC) revealed expression percentages of 27.78 and 38.66% for pC38 and pC38/AL27, respectively. The <I>in vivo</I> experiment was performed using three groups of BALB/C mice: group 1 was inoculated with pC38 in the tibial muscle; group 2 with pC38 and AL27 and group 3, negative control, with only PBS. Tissues showed the highest expression by CM (+++) when using pC38/AL27. Antibodies detected ranged from 0.75-1.2 international units (IU) mL <SUP>1</SUP> for pC38 and from 1.1-1.27 IU mL <SUP>1</SUP> for pC38/AL27. Results showed that AL27 promoted cell entrance mechanisms, generating a better expression level of the gene that codifies for the G glycoprotein of the rabies virus and consequently enhancing the stimulation of the immune response.
ER  -