3.1 Expression of wE7, uE7 and IL-12 proteins in Cos7
To study the expression
of wE7, uE7 and IL-12 proteins in cos-7 cells, crude cell lysates were
separated by electrophoresis on SDS-polyacrylamide gel and were identified
using of mouse anti-E7 and mouse anti-IL12 antibodies. Western blot analysis of
cell lysates showed proteins with two size of approximately 35 KDa and 82KDa for
wE7/ uE7 and IL-12 recombinant proteins, respectively. These results confirmed
expression of the proteins as it migrated according to its corresponding size (Fig.1).
Fig.1. Detection of
recombinant proteins in Cos7 cells by Western blot analysis. Lane 1, a 82KDa
band was observed in pcDNA3.1+-IL12 transfected cells using mouse anti-IL12,
Lane 2, molecular weight protein marker (Sinaclon, Iran); lane 3, negative
results in pcDNA3.1+ transfected cells and lane 4 and 5, two recombinant bands
with size of approximately 35KDa was detected in pcDNA3.1+- wE7/ uE7
transfected cells using mouse anti-E7. The blots were developed with ECL.
3.2 LDH Cytolytic responses
LDH release results
show significantly enhanced CTLs responses against wE7&IL12 and uE7&IL12.
As shown in figure 2, lymphocytes from the mice vaccinated with uE7&IL12 (67±
2.6) wE7&IL12 (67± 2.6) had a significantly increased specific
cytolytic activity at an E/T ratio of 50:1 than those vaccinated with PBS (12.1%±0.70%),
pcDNA3.1+ (19.1%±0.99%), pcDNA3.1+wE7 (37.4%±2.09%), or
pcDNA3.1+uE7 (41.3%±2.2%) (P