The influence of intrauterine exposure to immunosuppressive treatment on changes in the immune system in juvenile Wistar rats

Figures & data

Table 1 The medication concentration in whole blood and weight of rats in study groups treated with full and half dose of immunosuppressive drugs (additional study groups)

Drug and dose	CMG group (n=3×2)	TMG group (n=4×2)	CEG group (n=4×2)	Control group (n=3×2)
Cyclosporin A (ng/mL)	69.37±45.61	–	50.35±8.80	–
Dose 5 mg/kg
Cyclosporin A (ng/mL)	33.57±41.04	–	33.53±12.84	–
Dose 2.5 mg/kg
Tacrolimus (ng/mL)	–	7±6.61	–	–
Dose 4 mg/kg
Tacrolimus (ng/mL)	–	0.77±0.57	–	–
Dose 2 mg/kg
Everolimus (ng/mL)	–	–	1.43±0.17	–
Dose 0.5 mg/kg
Everolimus (ng/mL)	–	–	0.83±0.22	–
Dose 0.25 mg/kg
Body mass (g)	240±21	255±12.5	245±22.5	260±16
Full-dose group
Body mass (g)	250±50.1	235±20	225±11.5	255±15
Half-dose group

Notes: Results are presented as arithmetic mean ± standard deviation. –, Not applicable.

Abbreviations: CMG, CsA + MMF + prednisone; TMG, Tc + MMF + prednisone; CEG, CsA + everolimus + prednisone; CsA, cyclosporine A; MMF, mycophenolate mofetil; Tc, tacrolimus.

Table 2 Immunophenotyping (cytometry results) of 8-week-old rats from control and CMG group (full dose of drugs) – material from spleen

Parameter/group	Control group n=10	CMG group n=5	P-value (Mann–Whitney U test)
CD3^−CD45^+ (B lymphocytes) %
AM ± SD	14.07±6.085	18.41±1.59	0.019
Med	12.825	18.35
Range	7.76–28.98	16.58–20.73
CD3^+CD4^+ (T lymphocytes) %
AM ± SD	34.06±8.34	32.18±2.19	NS
Med	33.4	31.9
Range	21.8–53.2	29.5–34.9
CD3^+CD8A^+ %
AM ± SD	11.56±3.7	16.5±2.16	0.019
Med	11.1	16.9
Range	6.1–19.7	13.3–19.3
CD3^−CD8A^+ (NK cells) %
AM ± SD	3.52±1.69	5.88±1.905	0.028
Med	2.85	5.9
Range	2.3–7.5	3.5–8.8

Abbreviations: AM, arithmetic mean; SD, standard deviation; Med, median; NS, nonsignificant difference; CMG, CsA + MMF + prednisone; NK, natural killer; CsA, cyclosporine A; MMF, mycophenolate mofetil.

Table 3 Immunophenotyping (cytometry results) of 8-week-old rats in the treatment groups (half dose of drugs) – material from spleen

Parameter/group	Control group n=10	CMG group 0.5 n=10	TMG group 0.5 n=10	CEG group 0.5 n=7	P-value (Kruskal–Wallis test)
CD3^−CD45^+ (B lymphocytes) %
AM ± SD	14.07±6.085	14.49±6.71	18.365±3.16	17.56±2.71	0.0665
Med	12.825	13.005	18.58*	18.21*
Range	7.76–28.98	5.41–27.29	11.7–24.16	13.0–21.51
CD3^+CD4^+ (T lymphocytes) %
AM ± SD	34.06±8.34	18.35±9.84	36.42±3.16	26.13±3.89	0.0010
Med	33.4	14.75**	36.6	24.7*
Range	21.8–53.2	4.6–34.0	29.6–42.3	21.6–33.6
CD3^+CD8A^+ %
AM ± SD	11.56±3.7	15.54±8.28	19.44±2.4	11.07±2.18	0.0060
Med	11.1	16.35	19.3**	12.2
Range	6.1–19.7	2.8–26.5	15.6–22.9	7.3–13.0
CD3^−CD8A^+ (NK cells) %
AM ± SD	3.52±1.69	11.57±3.94	4.22±0.975	9.71±3.76	0.0000
Med	2.85	10.8**	4.25	9.4**
Range	2.3–7.5	7.3–19.6	2.8–6.0	4.7–16.6

Notes:

* P<0.05 versus control group,

** P<0.001 versus control group, Mann–Whitney U test.

Abbreviations: AM, arithmetic mean; SD, standard deviation; Med, median; CMG, CsA + MMF + prednisone; TMG, Tc + MMF + prednisone; CEG, CsA + everolimus + prednisone; CsA, cyclosporine A; MMF, mycophenolate mofetil; Tc, tacrolimus; NK, natural killer.

Table 4 IL-17 profile after incubation of lymphocytes from spleen with different doses of ConA (0, 2, and 5 µg/mL)

Parameter/group	Control group n=12	CMG group n=5	CMG group 0.5 n=10	TMG group 0.5 n=10	CEG group 0.5 n=7	P-value (Kruskal–Wallis test)
IL-17
ConA0	29.923	45.569	2.404	0	0	0.1416 (NS)
ConA2	143.78	0*	87.876	135.775	66.951	0.0283
ConA5	230.701	11.034**	204.287	335.653	301.49	0.0185

Notes: Results are presented as arithmetic mean for all groups (control group; full-dose CMG group, and all half-dose groups).

* P<0.01,

** P<0.06 versus control group, Mann–Whitney U test.

Abbreviations: ConA, concanavaline A; IL, interleukine; NS, difference nonsignificant; CMG, CsA + MMF + prednisone; TMG, Tc + MMF + prednisone; CEG, CsA + everolimus + prednisone; CsA, cyclosporine; MMF, mycophenolate mofetil; Tc, tacrolimus.