Effect of immunomodulation in turkeys infected with haemorrhagic enteritis virus on the percentage of CD4+ and CD8α+ T lymphocyte subpopulations synthesising IFN-γ

1	2–27	28–38	39–41	42/0	43/1	46/4
I-M/HEV⁺					EI
I-M/ HEV⁻				Meth	PBS
I-B/HEV⁺			β-Glu + MOS		EI
I-B/HEV⁻					PBS
II-M/HEV⁺	Random				EI
II-M/ HEV⁻	division of				PBS	Meth
II-B/HEV⁺	chicks into	Rearing			EI
II-B/ HEV⁻	14 groups of	under			PBS	β-Glu + MOS
	20 birds	standard			EI
HEV⁺ III-M/	each and	techniques			Meth
	introduction	and		Meth	PBS	Meth
III-M/HEV⁻	into	procedures			Meth
	experimental				EI
HEV⁺ III-B/	boxes				β-Glu+MOS
⁻				β-Glu + MOS	PBS	β-Glu + MOS
III-B/HEV					β-Glu+MOS
C/HEV⁺					EI
C/ HEV⁻					PBS

[i] I-M – turkeys immunomodulated with methisoprinol prior to infection; I-B – turkeys immunomodulated with β-glucans and mannan oligosaccharides (MOS) prior to infection; II-M – turkeys immunomodulated with methisoprinol after infection; II-B – turkeys immunomodulated with β-glucans and MOS after infection; III-M – turkeys immunomodulated with methisoprinol prior to, at the time of, and after infection; III-B – turkeys immunomodulated with β-glucans and MOS prior to, at the time of, and after infection; C – control turkeys not receiving immunomodulators; HEV⁺ – turkeys infected with haemorrhagic enteritis adenovirus at 42 days of life; HEV⁻ – uninfected turkeys; EI – experimental infection with HEV at a dose of 10^4.3 50% egg infectious dose (EID⁵⁰); PBS – uninfected turkeys receiving 1 mL of sterile phosphate buffered saline; Meth – methisoprinol administered in drinking water at 200 mg/kg b.w.; β-Glu+MOS – β-glucans and MOS administered at 500 g of Alphamune G per tonne of feed; S – sampling of the spleen from four turkeys of each group

Gating hierarchy chart in the analysis of the percentage of cells synthesising IFN-γ in response to *in vitro* mitogen stimulation. The cells were in the CD4⁺ (A) and (B) CD8α⁺ T lymphocyte subpopulations isolated from turkey spleen tissue

Table 2

Mean percentage of IFN-γ⁺ cells (±SD) in splenic CD4⁺ T lymphocytes collected from the examined turkeys receiving immunomodulators before the day of infection with HEV

3 dpi	5 dpi	7 dpi
I-M/HEV⁺	3.64^c (±0.45)	10.58^a (±2.44)	11.29^a (±1.15)
I-M/HEV⁻	3.04^c (±1.36)	3.69^c (±0.38)	4.75^cd (±0.90)
I-B/HEV⁺	2.31^c (±0.32)	9.12^ab (±0.87)	6.38^bc (±1.85)
I-B/HEV⁻	7.22^a (±2.50)	5.77^cd (±0.92)	8.74^b (±0.94)
C/HEV⁺	2.26^c (±0.41)	7.20^bd (±1.32)	3.54^cd (±0.45)
C/HEV⁻	5.47^b (±1.45)	7.03^bd (±0.98)	4.52^cd (±0.17)

[i] IFN-γ⁺ – interferon gamma–positive; SD – standard deviation; dpi – days post infection; I-M – turkeys immunomodulated with methisoprinol prior to infection; I-B – turkeys immunomodulated with β-glucans and MOS prior to infection; C – control turkeys not receiving immunomodulators; HEV⁺ – turkeys infected with haemorrhagic enteritis adenovirus at 42 days of life; HEV⁻ – uninfected turkeys. Values in the same column with different superscripts (a–d) differ significantly at P < 0.05 in Duncan’s multiple-range test

Table 3

Mean percentage of IFN-γ⁺ cells (±SD) in splenic CD8α⁺ T lymphocytes collected from the examined turkeys receiving immunomodulators before the day of infection with HEV

3 dpi	5 dpi	7 dpi
I-M/HEV⁺	2.78^cd (±0.88)	12.00^b (±0.94)	15.99^a (±0.86)
I-M/HEV⁻	5.33^bd (±0.91)	6.01^d (±0.50)	4.37^c (±0.95)
I-B/HEV⁺	2.73^c (±0.61)	9.34^cd (±1.24)	10.42^a (±0.89)
I-B/HEV⁻	10.54^a (±3.55)	9.94^bc (±1.09)	7.69^b (±1.11)
C/HEV⁺	2.66^c (±0.33)	15.17^a (±0.88)	5.78^c (±0.87)
C/HEV⁻	6.43^b (±1.33)	14.47^a (±2.11)	11.20^a (±0.93)

Table 4

Mean percentage of IFN-γ⁺ cells (±SD) in splenic CD4⁺ T lymphocytes collected from the examined turkeys receiving immunomodulators after the day of infection with HEV

3 dpi	5 dpi	7 dpi
II-M/HEV⁺	2.33^b (±0.48)	10.67^a (±1.12)	5.73^b (±0.93)
II-M/HEV⁻	6.13^a (±0.43)	8.06^ab (±2.46)	7.94^ab (±0.91)
II-B/HEV⁺	1.83^b (±0.23)	8.81^ab (±0.44)	3.52^c (±0.45)
II-B/HEV⁻	4.69^a (±0.49)	10.41^a (±2.08)	7.86^ab (±3.29)
C/HEV⁺	2.26^b (±0.41)	7.20^b (±1.32)	3.54^c (±0.45)
C/HEV⁻	5.47^a (±1.45)	7.03^b (±0.98)	4.52^c (±0.17)

[i] IFN-γ⁺ – interferon gamma–positive; SD – standard deviation; dpi – days post infection; II-M – turkeys immunomodulated with methisoprinol after infection; II-B – turkeys immunomodulated with β-glucans and MOS after infection; C – control turkeys not receiving immunomodulators; HEV⁺ – turkeys infected with haemorrhagic enteritis adenovirus at 42 days of life; HEV⁻ – uninfected turkeys. Values in the same column with different superscripts (a–d) differ significantly at P < 0.05 in Duncan’s multiple-range test

Table 5

Mean percentage of IFN-γ⁺ cells (±SD) within splenic CD8α⁺ T lymphocytes collected from the examined turkeys receiving immunomodulators after the day of infection with HEV

3 dpi	5 dpi	7 dpi
II-M/HEV⁺	2.74^b (±0.35)	17.12^a (±0.96)	10.60^a (±1.16)
II-M/HEV⁻	7.16^a (±0.49)	13.02^bc (±2.49)	7.61^bc (±0.87)
II-B/HEV⁺	3.57^b (±0.24)	15.38^ac (±0.90)	8.32^b (±1.44)
II-B/HEV⁻	5.73^a (±0.82)	7.21^b (±2.40)	5.70^c (±1.32)
C/HEV⁺	2.66^b (±0.33)	15.17^ac (±0.88)	5.78^c (±0.87)
C/HEV⁻	6.43^a (±1.33)	14.47^ac (±2.11)	11.20^a (±0.93)

Table 6

Mean percentage of IFN-γ⁺ cells (±SD) in splenic CD4⁺ T lymphocytes collected from the examined turkeys receiving immunomodulators prior to, at the time of, and after infection with HEV

3 dpi	5 dpi	7 dpi
III-M/HEV⁺	3.56^cd (±0.72)	8.19^cd (±0.95)	10.17^a (±0.80)
III-M/HEV⁻	8.36^a (±0.74)	7.67^cd (±0.99)	6.69^bc (±0.19)
III-B/HEV⁺	2.43^c (±0.36)	9.91^abd (±0.76)	8.37^abc (±0.87)
III-B/HEV⁻	4.71^bd (±0.88)	10.98^a (±0.51)	8.02^ac (±1.73)
C/HEV⁺	2.26^c (±0.41)	7.20^c (±1.32)	3.54^d (±0.45)
C/HEV⁻	5.47^b (±1.45)	7.03^c (±0.98)	4.52^d (±0.17)

[i] IFN-γ⁺ – interferon gamma–positive; SD – standard deviation; dpi – days post infection; III-M – turkeys immunomodulated with methisoprinol prior to, at the time of, and after infection; III-B – turkeys immunomodulated with β-glucans and MOS prior to, at the time of, and after infection; C – control turkeys not receiving immunomodulators; HEV⁺ – turkeys infected with haemorrhagic enteritis adenovirus at 42 days of life; HEV⁻ – uninfected turkeys. Values in the same column with different superscripts (a–d) differ significantly at P < 0.05 in Duncan’s multiple-range test

Table 7

Mean percentage of IFN-γ⁺ cells (±SD) within splenic CD8α⁺ T lymphocytes collected from the examined turkeys receiving immunomodulators prior to, at the time of, and after infection with HEV

3 dpi	5 dpi	7 dpi
III-M/HEV⁺	5.60^bd (±1.94)	12.55^bc (±1.08)	11.42^a (±0.89)
III-M/HEV⁻	9.99^a (±1.93)	11.84^c (±1.07)	6.95^b (±1.30)
III-B/HEV⁺	3.22^cd (±0.87)	16.42^a (±0.88)	11.46^a (±0.84)
III-B/HEV⁻	4.35^bcd (±0.84)	7.51^d (±1.91)	6.87^b (±0.84)
C/HEV⁺	2.66^c (±0.33)	15.17^ab (±0.88)	5.78^b (±0.87)
C/HEV⁻	6.43^b (±1.33)	14.47^ab (±2.11)	11.20^a (±0.93)

Table 8

Results of PCR amplification of the HE virus hexon gene fragment in spleen samples taken from four turkeys from each group

45/3	47/5	49/7
I-M/HEV⁺	0	50	25
I-M/HEV⁻	0	0	0
I-B/HEV⁺	25	75	25
I-B/HEV⁻	0	0	0
II-M/HEV⁺	25	75	50
II-M/HEV⁻	0	0	0
II-B/HEV⁺	50	100	75
II-B/HEV⁻	0	0	0
III-M/HEV⁺	0	75	25
III-M/HEV⁻	0	0	0
III-B/HEV⁺	0	75	50
III-B/HEV⁻	0	0	0
C/HEV⁺	50	100	100
C/HEV⁻	0	0	0

[i] dol – day of life; dpi – days post infection; I-M – turkeys immunomodulated with methisoprinol prior to infection; I-B – turkeys immunomodulated with β-glucans and mannan oligosaccharides (MOS) prior to infection; II-M – turkeys immunomodulated with methisoprinol after infection; II-B – turkeys immunomodulated with β-glucans and MOS after infection; III-M – turkeys immunomodulated with methisoprinol prior to, at the time of, and after infection; III-B – III-B – turkeys immunomodulated with β-glucans and MOS prior to, at the time of, and after infection; C – control turkeys not receiving immunomodulators; HEV⁺ – turkeys infected with haemorrhagic enteritis adenovirus at 42 days of life; HEV⁻ – uninfected turkeys; 25%, 50%, 75% and 100% mean that HEV genetic material was respectively detected in the 1st, in the 1st and 2nd, in the 1st, 2nd, and 3rd, and in all 4 spleens collected from four turkeys in a group on a given day

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DOI: https://doi.org/10.2478/jvetres-2022-0068 | Journal eISSN: 2450-8608 (formerly 2300-3235)

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Language: English

Page range: 537 - 547

Submitted on: Sep 21, 2022

Accepted on: Nov 29, 2022

Published on: Dec 10, 2022

Published by: National Veterinary Research Institute in Pulawy

In partnership with: Paradigm Publishing Services

Keywords:

haemorrhagic enteritis virus,

Related subjects:

Microbiology and virology,

Life sciences, other,

Medicine,

Veterinary medicine

© 2022 Bartłomiej Tykałowski, Andrzej Koncicki, published by National Veterinary Research Institute in Pulawy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Volume 66 (2022): Issue 4 (December 2022)

Effect of immunomodulation in turkeys infected with haemorrhagic enteritis virus on the percentage of CD4+ and CD8α+ T lymphocyte subpopulations synthesising IFN-γ

Figures & Tables

Table 1

Fig. 1