Transfer of Invitro CD4 + T Cells with Hypomethylation of Perforin Promoter into Rats’ Abdomens Causes Autoimmune Emphysema

Figures & data

Table 1. The methylation levels of the perforin gene promoter in CD4 + T cells of the two Cell groups.

Groups	N	Methylation level
Cell group 1	10	0.8186 ± 0.0482^a
Cell group 2	10	0.9316 ± 0.0234
T value		−6.66
P value		<0.05

Notes: DNA extraction of rats’ spleens was performed and NGS was carried out. Methylation levels of the perforin gene promoter. ap < 0.05, as compared with the Cell group 2.

NGS, next-generation sequencing.

Figure 1. Histology of lung tissue. Notes: Lung tissue section was stained with HE. (A) Control rats receiving normal CD4 + T cells, adjuvant and PBS; (B) Rats receiving adjuvant and PBS; (C) Rats receiving CD4 + T cells hypomethylated, adjuvant and PBS. Data presented were one representative image data. Magnification: 400×. Abbreviations: HE, hematoxylin-eosin; PBS, phosphate buffered saline.

Figure 2. Expression of VEGF in rat lungs Notes: Expression of VEGF in rat lungs was detected by immunohistochemistry. (A) Control rats receiving normal CD4 + T cells, adjuvant and PBS; (B) Rats receiving adjuvant and PBS; (C) Rats receiving CD4 + T cells hypomethylated, adjuvant and PBS. Data presented were one representative image data. Magnification: 400×. Abbreviations: VEGF, Vascular endothelial growth factor; PBS, phosphate buffered saline.

Table 2. The difference of MLI and MAN in the three groups.

Groups	n	MLI (×10^-6m)	MAN (×106/m^2)
Control group	10	34.75 ± 6.90	467.28 ± 59.97
Sham operation group	10	32.69 ± 6.65	470.36 ± 60.06
Model group	10	47.79 ± 16.83a,b	322.31 ± 90.05a,b
F value		5.359	14.02
P value		<0.05	<0.05

Notes: Lung tissue section was stained with HE. MLI and MAN. ^ap < 0.05, as compared with the control group, ^bp < 0.05, as compared with the sham operation group.

MLI, mean linear intercept; MAN, mean alveolar numbers; HE, hematoxylin-eosin.

Table 3. The difference of AECA in the three groups.

Groups	n	AECA in serum (ng/ml)	AECA in BALF (ng/ml)
Control group	10	27.27 ± 6.41	19.52 ± 9.80
Sham operation group	10	23.82 ± 6.25	16.11 ± 7.83
Model group	10	60.93 ± 12.58a,b	41.57 ± 13.49a,b
F value		52.89	16.89
P value		<0.05	<0.05

Notes: Serum and BALF were collected and ELISA was performed. AECA. ^ap < 0.05, as compared with the control group, ^bp < 0.05, as compared with the sham operation group.

BALF, Bronchoalveolar lavage fluid; ELISA, enzyme linked immunosorbent assay; AECA, anti-endothelial cell antibodies.

Table 4. The difference of VEGF OD in lung tissues in the three groups.

Groups	n	VEGF OD
Control group	10	14.09 ± 2.67
Sham operation group	10	13.99 ± 3.47
Model group	10	20.43 ± 5.90a,b
F value		7.56
P value		<0.05

Notes: Lung tissue was sectioned and immunohistochemistry was carried out. VEGF.

^ap < 0.05, as compared with the control group, ^bp < 0.05, as compared with the sham operation group.

VEGF, Vascular endothelial growth factor; OD, optical density.

Figure 3. Apoptosis of alveolar septal cells in rat lungs. Notes: Apoptosis of alveolar septal cells in rat lungs was detected by immunohistochemistry. (A) Rats receiving CD4 + T cells hypomethylated, adjuvant and PBS; (B) Control rats receiving normal CD4 + T cells, adjuvant and PBS; (C) Rats receiving adjuvant and PBS. Data presented were one representative image data. Magnification: 400×. PBS, phosphate buffered saline.

Table 5. The difference of AI of alveolar septal cells in the three groups.

Groups	n	AI (%)
Control group	10	8.62 ± 3.56
Sham operation group	10	7.82 ± 2.80
Model group	10	21.91 ± 8.83a,b
F value		19.08
P value		<0.05

Notes: Lung tissue was sectioned and TUNEL was carried out. AI. ^ap < 0.05 as compared with the control group, ^bp < 0.05, as compared with the sham operation group.

AI, apoptotic index; TUNEL, Terminal deoxynucleotidyl transferase- mediated dUTP nick end labeling.

Table 6. The methylation levels of the perforin gene promoter in CD4 + T cells in the three groups.

Groups	N	methylation levels
Control group	10	0.9346 ± 0.0192
Sham operation group	10	0.9364 ± 0.0217
Model group	10	0.8610 ± 0.0243a,b
F value		38.77
P value		<0.05

Notes: DNA extraction of rats’ spleens was performed and NGS was carried out. Methylation levels of the perforin gene promoter. ^ap < 0.05, as compared with the control group, ^bp < 0.05, as compared with the sham operation group.

NGS, next-generation sequencing.