The effects of anti-lung cancer in nude mice by a fully human single-chain antibody against associated antigen Ts7TMR between A549 cells and Trichinella spiralis

Figures & data

Figure 1. The effect of anti-Ts7TMR serum on A549 lung cancer.

Table 1. Effect of anti-Ts7TMR serum on the growth of A549 lung cancer in nude mice.

Groups (n = 6)	Tumour weight (g) (Inhibition rate)
Negative serum	0.174 ± 0.031
anti-Ts7TMR serum	0.061 ± 0.017** (64.77%)
Anti-T. spiralis serum	0.110 ± 0.024* (36.44%)

**p < 0.01 vs Negative serum group.

Figure 2. The effect of anti-Ts7TMR scFv on A549 lung cancer and its preliminary mechanism. A. The effect of anti-Ts7TMR scFv on the proliferation of A549 cells in vitro. A549 cells were co-incubated with different doses of anti-Ts7TMR scFv (5, 25, and 45 μg) for 48h to detect cell proliferation by CCK-8 assay. B. The effect of anti-Ts7TMR scFv on A549 lung cancer growth in nude mice. Nude mice were injected subcutaneously with A549 cells and then treated subcutaneously with PBS, negative serum (Ns), anti-T. spiralis serum (Ts), and different doses of anti-Ts7TMR scFv (25, 50, and 100 μg) around the tumour for 10 consecutive days. C. Scoring of PCNA, BCL-2, and VEGF expression in tumour tissues by immunohistochemistry (400 ×). Expression of PCNA, BCL-2, and VEGF in tumours treated with PBS, negative serum (Ns), anti-T. spiralis serum (Ts), and different doses of anti-Ts7TMR scFv (25, 50, and 100 μg). Scale bar = 20 μm. *p < 0.05, **p < 0.01.

Table 2. Effect of anti-Ts7TMR scFv on the growth of A549 lung cancer in nude mice.

Groups (n = 6)	Tumour volume (mm^3) (Inhibition rate)	Tumour weight (g) (Inhibition rate)
PBS	615.0 ± 30.4	0.511 ± 0.022
Negative serum	593.3 ± 26.0 (3.52%)	0.501 ± 0.021 (1.91%)
25 μg scFv	533.51 ± 22.3** (13.25%)	0.470 ± 0.012** (8.02%)
50 μg scFv	376.81 ± 16.4** (38.73)	0.304 ± 0.006** (40.56%)
100 μg scFv	300.12 ± 21.6** (51.2%)	0.209 ± 0.025** (59.10%)
Anti-T. spiralis serum	352.5 ± 17.6** (42.68%)	0.272 ± 0.010** (46.73%)

**p < 0.01 vs PBS group.

Figure 3. Pathological observations in nude mice (100 ×). Pathological observation of heart, liver, spleen, lung, and kidney in tumour-bearing nude mice treated with PBS, negative serum (Ns), anti-T. spiralis serum (Ts), and different doses of anti-Ts7TMR scFv (25, 50, and 100 μg). Scale bar = 100 μm.

Figure B1. Construction of the Ts7TMR prokaryotic expression vector. A. Using T. spiralis cDNA as a template, the Ts7TMR gene was amplified by PCR and identified. B. The recombinant pET-32a- Ts7TMR plasmid was characterised by restriction digestion Lane 1: pET-32a- Ts7TMR. Lane 2: pET-32a. C. The purified fusion protein Ts7TMR was identified by SDS-PAGE. Lanes 1: Crude purification of Ts7TMR. Lane 2: Refined purification of Ts7TMR.

Figure B2. SDS-PAGE and western blot analysis of anti-Ts7TMR scFv. A. SDS-PAGE analysis of molecular weight and purity of anti-Ts7TMR scFv. B. Western blot identification of binding of anti-Ts7TMR scFv to Ts7TMR.

Figure B3. Anti-Ts7TMR scFv binding to A549 cells (× 800). A: anti-Ts7TMR scFv. B: negative control.

Data availability statement

The datasets generated during the current study are available from the corresponding author upon reasonable request.