A multiepitope vaccine encoding four Eimeria epitopes with PLGA nanospheres: a novel vaccine candidate against coccidiosis in laying chickens

Yu, ZhengQing; Chen, SiYing; Huang, JianMei; Ding, WenXi; Chen, YuFeng; Su, JunZhi; Yan, RuoFeng; Xu, LiXin; Song, XiaoKai; Li, XiangRui

doi:10.1186/s13567-022-01045-w

Table 2 Antibody and cytokine determination in the sera from immunized chickens at 7, 14, and 21 days of age.

From: A multiepitope vaccine encoding four Eimeria epitopes with PLGA nanospheres: a novel vaccine candidate against coccidiosis in laying chickens

Age	Group	Chicken IgY			Chicken IFN-γ			Chicken IL-4
Age	Group	Titre (OD450)	p value^a	p value^b	Value (pg/mL)	p value^a	p value^b	Value (pg/mL)	p value^a	p value^b
14 days	Blank (PBS)	0.180 ± 0.049	–	0.9721	29.541 ± 8.341	–	0.9968	83.088 ± 10.484	–	0.9977
	Control	0.202 ± 0.073	0.9721	–	27.372 ± 6.812	0.9968	–	79.879 ± 8.845	0.9977	–
	NSLC	0.158 ± 0.032	0.9721	0.5188	33.011 ± 8.623	0.9536	0.6451	79.365 ± 9.826	0.9969	>0.9999
	NSLC-FA	0.167 ± 0.030	0.9977	0.7529	29.564 ± 3.874	> 0.9999	0.9967	87.676 ± 11.376	0.9914	0.8428
	NSLC-PLGA	0.177 ± 0.051	0.9999	0.9427	31.261 ± 4.860	0.9994	0.9162	82.738 ± 12.697	>0.9999	0.9994
	NSLC-CS	0.187 ± 0.023	0.9997	0.9966	32.340 ± 3.372	0.9877	0.7639	80.648 ± 8.978	0.9995	0.9999
	NSLC-ISCOM	0.161 ± 0.033	0.9888	0.5913	29.492 ± 4.459	>0.9999	0.9969	85.182 ± 7.827	0.9996	0.9784
	NSLC-RG1	0.198 ± 0.040	0.9910	0.9998	30.208 ± 7.356	0.9998	0.9865	85.408 ± 9.470	0.9995	0.9721
	NSLC-201VG	0.204 ± 0.051	0.9542	>0.9999	32.229 ± 8.160	0.9903	0.7825	82.859 ± 9.487	>0.9999	0.9994
	NSLC-71VR	0.167 ± 0.029	0.9976	0.7478	31.224 ± 4.402	0.9994	0.9199	85.351 ± 17.905	0.9996	0.9738
21 days	Blank (PBS)	0.184 ± 0.041	–	0.9967	41.568 ± 8.879	–	0.9995	81.744 ± 8.473	–	0.9997
	Control	0.207 ± 0.067	0.9967	–	43.170 ± 6.579	0.9995	–	82.971 ± 9.361	0.9997	–
	NSLC	0.313 ± 0.036	0.0237	0.0877	55.939 ± 7.119	0.0295	0.0662	90.166 ± 13.679	0.7933	0.8931
	NSLC-FA	0.415 ± 0.047	<0.0001	<0.0001	59.452 ± 9.585	0.0041	0.0104	107.073 ± 8.736	0.0065	0.0105
	NSLC-PLGA	0.504 ± 0.055	<0.0001	<0.0001	64.417 ± 5.931	0.0002	0.0005	119.392 ± 12.301	<0.0001	<0.0001
	NSLC-CS	0.401 ± 0.068	<0.0001	0.0003	53.756 ± 4.372	0.0872	0.1763	125.150 ± 13.758	<0.0001	< 0.0001
	NSLC-ISCOM	0.306 ± 0.053	0.0374	0.1298	48.591 ± 3.741	0.5935	0.8279	139.137 ± 5.669	<0.0001	<0.0001
	NSLC-RG1	0.277 ± 0.041	0.1706	0.4424	61.762 ± 11.139	0.0010	0.0026	107.105 ± 11.632	0.0064	0.0104
	NSLC-201VG	0.366 ± 0.104	0.0007	0.0033	58.666 ± 6.583	0.0065	0.0161	131.842 ± 13.803	<0.0001	<0.0001
	NSLC-71VR	0.562 ± 0.102	<0.0001	<0.0001	62.424 ± 7.896	0.0007	0.0018	111.104 ± 10.736	0.0012	0.0021
28 days	Blank (PBS)	0.332 ± 0.051	–	0.9994	45.890 ± 4.798	–	0.9996	85.873 ± 9.740	–	0.9996
	Control	0.310 ± 0.125	0.9994	–	47.480 ± 6.860	0.9996	–	84.185 ± 6.600	0.9996	–
	NSLC	0.465 ± 0.069	0.1196	0.0497	63.593 ± 11.494	0.0271	0.0529	102.648 ± 8.300	0.0929	0.0519
	NSLC-FA	0.830 ± 0.065	<0.0001	<0.0001	65.113 ± 6.952	0.0137	0.0279	119.938 ± 12.526	<0.0001	<0.0001
	NSLC-PLGA	0.865 ± 0.089	<0.0001	<0.0001	71.442 ± 9.612	0.0006	0.0014	138.027 ± 12.043	<0.0001	<0.0001
	NSLC-CS	0.984 ± 0.082	<0.0001	<0.0001	64.268 ± 11.098	0.0201	0.0400	132.733 ± 15.030	< 0.0001	<0.0001
	NSLC-ISCOM	0.706 ± 0.095	<0.0001	<0.0001	64.909 ± 8.996	0.0151	0.0305	140.731 ± 8.930	<0.0001	<0.0001
	NSLC-RG1	0.635 ± 0.097	<0.0001	<0.0001	70.112 ± 7.111	0.0012	0.0027	116.959 ± 11.750	0.0003	0.0001
	NSLC-201VG	0.846 ± 0.104	<0.0001	<0.0001	67.100 ± 11.976	0.0054	0.0114	138.381 ± 8.845	<0.0001	<0.0001
	NSLC-71VR	1.097 ± 0.067	<0.0001	<0.0001	71.827 ± 9.695	0.0005	0.0011	131.663 ± 7.681	<0.0001	<0.0001

Age	Group	Chicken TGF-β			Chicken IL-17
Age	Group	Value (pg/mL)	p value^a	p value^b	Value (pg/mL)	p value^a	p value^b
14 days	Blank (PBS)	123.173 ± 12.248	–	0.9994	19.394 ± 5.137	–	0.9998
	Control	119.752 ± 8.148	0.9994	–	20.002 ± 7.549	0.9998	–
	NSLC	120.828 ± 15.092	0.9996	0.9998	20.369 ± 5.989	0.9996	0.9999
	NSLC-FA	114.260 ± 9.223	0.8371	0.9883	20.572 ± 6.462	0.9996	0.9998
	NSLC-PLGA	117.337 ± 13.643	0.9822	0.9996	20.533 ± 7.125	0.9996	0.9998
	NSLC-CS	110.500 ± 11.116	0.5066	0.8109	19.591 ± 5.885	>0.9999	0.9999
	NSLC-ISCOM	118.102 ± 12.177	0.9922	0.9997	21.239 ± 4.849	0.9975	0.9996
	NSLC-RG1	121.418 ± 11.854	0.9997	0.9997	19.883 ± 5.432	0.9998	>0.9999
	NSLC-201VG	117.947 ± 14.370	0.9911	0.9997	19.466 ± 4.633	>0.9999	0.9998
	NSLC-71VR	120.903 ± 15.142	0.9996	0.9998	18.501 ± 7.115	0.9997	0.9994
21 days	Blank (PBS)	121.005 ± 10.895	–	0.9997	23.460 ± 6.226	–	>0.9999
	Control	118.695 ± 12.262	0.9997	–	23.171 ± 4.923	>0.9999	–
	NSLC	127.328 ± 9.292	0.9902	0.9361	26.059 ± 4.507	0.9899	0.9794
	NSLC-FA	145.266 ± 12.469	0.0836	0.0475	32.456 ± 6.320	0.1396	0.1193
	NSLC-PLGA	135.244 ± 16.664	0.5657	0.3999	34.643 ± 6.006	0.0387	0.0322
	NSLC-CS	142.659 ± 22.132	0.1508	0.0897	32.020 ± 8.837	0.1752	0.1508
	NSLC-ISCOM	161.718 ± 16.989	0.0008	0.0004	29.559 ± 6.695	0.5142	0.4628
	NSLC-RG1	150.956 ± 8.537	0.0194	0.0101	36.107 ± 5.085	0.0147	0.0120
	NSLC-201VG	153.113 ± 17.809	0.0106	0.0054	31.193 ± 5.205	0.2625	0.2289
	NSLC-71VR	151.481 ± 15.511	0.0168	0.0087	26.425 ± 5.641	0.9758	0.9586
28 days	Blank (PBS)	124.590 ± 9.492	–	0.9999	26.130 ± 6.397	–	0.9993
	Control	123.421 ± 9.944	0.9999	–	28.280 ± 7.843	0.9993	–
	NSLC	139.386 ± 17.307	0.5903	0.5071	37.045 ± 6.409	0.1780	0.3860
	NSLC-FA	155.312 ± 15.596	0.0258	0.0191	42.596 ± 8.476	0.0126	0.0385
	NSLC-PLGA	158.200 ± 14.714	0.0122	0.0089	47.206 ± 8.857	0.0009	0.0031
	NSLC-CS	144.567 ± 10.419	0.2691	0.2173	49.501 ± 10.837	0.0002	0.0008
	NSLC-ISCOM	169.032 ± 15.898	0.0006	0.0004	44.582 ± 3.946	0.0041	0.0138
	NSLC-RG1	157.629 ± 16.096	0.0141	0.0103	49.309 ± 10.099	0.0003	0.0009
	NSLC-201VG	161.536 ± 22.182	0.0048	0.0035	40.471 ± 6.735	0.0381	0.1043
	NSLC-71VR	175.787 ± 20.563	<0.0001	<0.0001	42.661 ± 5.175	0.0122	0.0374

At 14 (before the first immunization), 21 (seven days after the first immunization), and 28 (seven days after the second immunization) days of age, sera were collected from five animals in each group. A standard ELISA procedure was carried out to investigate the titres of chicken IgY, and each sample was assessed once. Commercially available ELISA kits were used to determine the levels of IFN-γ, IL-4, TGF-β, and IL-17 in sera, and each sample was analysed once. The results were estimated using one-way ANOVA followed by Dunnett’s test, and the values are shown as the mean ± SD (n = 5).
^{a, b}Were compared with the blank and control groups, respectively.

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