Safety and immunogenicity of a novel therapeutic DNA vaccine encoding chicken type II collagen for rheumatoid arthritis in normal rats

Figures & data

Table 1. Vaccination effect of normal rats with pcDNA-CCOL2A1 vaccine at the maximum doses of 3 mg/kg on general health status and behavior on 14 days after vaccination (n=6)

	Control		Vaccine
Clinical symptoms	male	female	male	female
Fur shedding and pilo-erection	−	−	−	−
Injection site swelling	−	−	−	−
Sneeze and cough	−	−	−	−
Ecphysesis	−	−	−	−
Tears and eyes purulent sectretion	−	−	−	−
Instability of gait	−	−	−	−
Diarrhea	−	−	−	−
Skin ulcer	−	−	−	−
Pain upon palpation	−	−	−	−
Convulsive	−	−	−	−
Mortality	−	−	−	−

* The vaccinated rats and the control rats were observed for changes in clinical appearance. No significant changes were detected at 0 h, 1 h, 2 h, 3 h, 5 h, 1 day, 2 day, 4 day, 6 day, 8 day, 10 day, 12 day after vaccination(data not shown). This data is representative of 3 experiments. Three separate experiments yielded similar results. −: No corresponding clinical symptoms, +: Presenting corresponding clinical symptoms.

Table 2. Vaccination effect of normal rats with pcDNA-CCOL2A1 vaccine at the maximum doses of 3 mg/kg on hematological clinical rountine parameters on 2 nd week after vaccination (n = 6, ${\displaystyle \overline{x}}$ ± s)

parameters	Vaccine	Control
WBC(×10^9 cells/L)	18.16±4.91	17.68±4.84
RBC(×10^9 cells/L)	7.94±0.40	7.54±0.35
MCHC(g/L)	162.83±5.81	157±9.36
Platelets(×10^9 cells/L)	634.00±61.78	539.8±82.80
Neutrophil(×10^9 cells/L)	1.65±0.58	1.80±0.77
Lymphocytes(×10^9 cells/L)	15.86±4.49	15.33±4.10
Monocytes(×10^9 cells/L)	0.55±0.21	0.38±0.17
Eosinophils(×10^9 cells/L)	0.11±0.02	0.17±0.17
Basophils(×10^9 cells/L)	0.00±0.00	0.00±0.00

Parameters are listed in SI units; WBC: white blood cells, RBC: red blood cell, MCHC: mean corpuscular.

*P < 0.05 compared with control. These data are representative of 3 experiments. Three separate experiments yielded similar results.

Table 3. Vaccination effect of normal rats with pcDNA-CCOL2A1 vaccine at the maximum doses of 3 mg/kg on serum biochemistry rountine parameters on 2nd week after vaccination (n = 6, ${\displaystyle \overline{x}}$ ± s)

	Parameters	Vaccine	Control
	GLU(mmol/L)	8.02±2.30	10.71±5.42
Renal function	BUN(mmol/L)	8.91±1.14	9.19±1.38
	Cre(μmol/L)	37.58±7.47	42.17±9.29
	UA(μmol/L)	317.67±82.74	278.5±93.10
Liver function	ALT(IU/L)	75.17±34.53	57.83±9.20
	TP(g/L)	80.52±3.24	77.30±4.78
	Albumin(g/L)	33.42±3.25	30.93±3.10
	Glubulin(g/L)	47.10±5.15	46.37±2.40
	TBIL(μmol/L)	2.82±0.46	3.13±0.80
	DBIL(μmol/L)	0.57±0.33	0.47±0.18
	TBA(μmol/L)	26.10±9.29	16.66±3.40
Heart function	ALP(IU/L)	344.50±130.96	291.8±105.36
	AST(mmol/L)	252.17±127.47	197.17±64.40
Blood lipids	triglyceride(mmol/L)	2.79±1.03	3.53±0.94
	TC(mmol/L)	2.07±0.11	1.92±0.09
	HDL(mmol/L)	1.52±0.13	1.36±0.05
	LDL(mmol/L)	0.21±0.07	0.20±0.07

Parameters are listed in SI units; GLU: blood glucose; BUN: blood urea nitrogen; Cre: creatinine; UA: uric acid; ALT: alanine aminotransferase; TP: total protein; TBIL: total bilirubin; DBIL: direct bilirubin; TBA: total bile acid; ALP: alkaline phosphatase; AST: aspartate aminotransferase; TC: total cholesterol. HDL: high-density lipoprotein; LDL: low-density lipoprotein.

*P < 0.05 compared with control. These data are representative of 3 experiments. Three separate experiments yielded similar results.

Figure 1. Histopathological analysis of various tissues from normal rats vaccinated with pcDNA-CCOL2A1 at a maximum dosage of 3 mg/kg on day 14 after a single intramuscular injection into the hind leg. H&E staining, original magnification: 200×. (A) Samples from the control rats. (B) Samples from the vaccinated normal rats. These data are representative of 3 experiments. Three separate experiments yielded similar results.

Table 4. Vaccination effect of normal rats with pcDNA-CCOL2A1 vaccine at the maximum doses of 3mg/kg on plasma production of anti-CII IgG antibodies on 2nd week after vaccination

Anti-CII antibodies	Rat-IgG Anti-Rat	Rat-IgG Anti-chicken
NC	0.1077±0.0565	<0.01
DNA vaccine	0.07355 ±0.0558	<0.01

Each plasma was not diluted, and the values were expressed as mean units of activity (units/ml, n = 6, ${\displaystyle \overline{x}}$ +/−s). Statistical analysis was compared with control. P > 0.05 compared with control. These data are representative of 3 experiments. Three separate experiments yielded similar results.