Construction of exogenous multiple epitopes of helper T lymphocytes and DNA immunization of its chimeric plasmid with HBV pre-S2/S gene

Table 1 Primers and oligonucleotide fragments used in mte5 splicing and construction of eukaryotic expression plasmids

	Sequences (5’→3’)
F1	ATGGCTAAAA CCATCGCCTA TGATGAAGAA GCTCGTCGTG GTCTGGAACG TGGTCTGAAT GCCTCCGAT
F2	GTCCAGGCAG CAACGAATTT AGCTCCCTGC AGCCAGTGTC CAGGCAGAAC ATCGGAGGCA TTCAGACCA
F3	AATTCGTTGC TGCCTGGACC CTGAAAGCTG CCGCTGGAAG ACACGTTGTT ATCGATAAGA GCTTCGGAA
F4	TTCGGTGATT CCGATAAACT TGGAATTAGC TTTGATGTAC TGCTGAGGGC TTCCAAGGCT CTTATCGAT M ` T E -
MTE-P1	TCGGATCCCA TGGCTAAAAC CATCGCC
MTE-P2	GCTCTAGACT TTCGGTGATT CCGATAA
HBV-P1	CCTAAGCTTA TGCAGTGGAA CTCCACT
HBV-P2	TGGAATTCCT TAAATGTATA CCCAGAG
Bam-1	TCGGATCCCA TGGCTAAAAC CATCGCC
Bam-2	GCGGATCCCT TTCGGTGATT CCGATAA
Hinc-1	TCGTTGACAA TGGCTAAAAC CATCGCC
Hinc-2	GCGTTGACAA TTCGGTGATT CCGATAA
Xba-1	TCTCTAGACA TGGCTAAAAC CATCGCC
Xba-2	GCTCTAGACT TTCGGTGATT CCGATAA
Acc-1	TCGTATACAT GGCTAAAACC ATCGCCT
Acc-2	CGCGTATACT TCGGTGATTC CGATAAA

Table 2 Effect of MTE5 on anti-HBs response of HBV pre-S2/S gene

Group	Anti-HBs		Enhancement
	Positive ratio	MIU/L (GMT ± SE)
Empty control	0/5	-	-
PcDNA3	0/5	-	-
PcHB	6/10	27.60 ± 17.3	-
PcHB/pcMTE5	8/10	136.70 ± 69.1b	4.95
PcHB-MTEB	7/10	57.540 ± 7.68	2.09
PcHB-MTEX	0/10	-	-
PcHB-MTEH	1/10	1.50	-
PcHB-MTEA	0/10	-	-

GMT: Geometric mean titer; Enhancement: GMT ratio of given group to pcHB, immune enhancement was considered when the ratio was larger than 1. “-” means ant-HBs negative or the enhancement could not be detected.

^bP < 0.01, t = -6.56 vs pcHB group.