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. 2023 Apr;87(2):110-119.

Response of lymphocytes from pigs naturally infected with porcine respiratory disease complex at 3 different stages of development

Rosa Elvira Nuñez-Anita #  1 , Fernando Calderón-Rico #  1 , Francisco Pérez-Duran  1 , María Concepción Arenas-Arrocena  1 , Alicia Gabriela Zamora-Avilés  1 , Luis Enrique Franco-Correa  1 , Alejandro Bravo-Patiño  1 , Ilane Hernández-Morales  1
Affiliations

Response of lymphocytes from pigs naturally infected with porcine respiratory disease complex at 3 different stages of development

Rosa Elvira Nuñez-Anita et al. Can J Vet Res. 2023 Apr.

Abstract
in English, French

The objective of this study was to analyze the response of lymphocytes from pigs naturally infected with porcine respiratory disease complex (PRDC) at 3 different stages of development. Porcine respiratory disease complexes were isolated from 2 groups: The infected group, consisting of pigs with PRDC and no vaccination against any virus (n = 24), and the control group, consisting of vaccinated and noninfected piglets (n = 24). Both groups were sampled at 3 stages of development: Weaning (WEA) (n = 8), initiation (INI) (n = 8), and growth (GRO) (n = 8). The PRDC status was confirmed by serological testing against porcine circovirus type 2 (PCV-2), porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (H1N1), and Mycoplasma hyopneumoniae. PCV-2+ cells were quantified by flow cytometry. Weight gain was registered at each stage. PCV-2+ cells, CD4+ cells, monocytes and lymphocytes populations were measured. Gene expression in CD4+ cells was quantified for interferon-γ (IFN-γ), GATA binding protein 3 (GATA3), T-box transcription factor (T-bet), interleukin-10 (IL-10), and IL-4. Control piglets gained approximately 35% more weight than those infected with PRDC. Specifically, PCV-2+ cells were detected in piglets from the infected group in the following proportions: WEA ≤ INI ≤ GRO. In infected piglets, the CD4+ count increased at WEA and decreased at GRO, CD4+ expression profile showed an overexpression of T-bet at INI and GRO, and the expression of IFN-γ was lower at WEA and GRO. In contrast, IL-4 was overexpressed at all 3 stages. GATA3 was overexpressed at INI and GRO. The infected piglets showed lymphopenia and less CD4+ cells. CD4+ cells showed a different expression profile than the control group, in which IFN-γ was less expressed, whereas IL-4 and T-bet were overexpressed.

L’objectif de cette étude était d’analyser la réponse des lymphocytes de porcs naturellement infectés par le complexe respiratoire porcin (PRDC) à trois stades de développement différents. Des PRDC ont été isolés à partir de deux groupes : le groupe infecté, composé de porcs atteints de PRDC et non vaccinés contre un virus (n = 24), et le groupe témoin, composé de porcelets vaccinés et non infectés (n = 24). Les deux groupes ont été échantillonnés à trois stades de développement : sevrage (WEA) (n = 8), initiation (INI) (n = 8) et croissance (GRO) (n = 8). Le statut de PRDC a été confirmé par des tests sérologiques contre le circovirus porcin de type 2 (PCV-2), le virus du syndrome reproducteur et respiratoire porcin (PRRSV), le virus de la grippe porcine (H1N1) et Mycoplasma hyopneumoniae. Les cellules PCV-2+ ont été quantifiées par cytométrie en flux. Un gain de poids a été enregistré à chaque étape. Les populations de cellules PCV-2+, de cellules CD4+, de monocytes et de lymphocytes ont été mesurées. L’expression génique dans les cellules CD4+ a été quantifiée pour l’interféron-γ (IFN-γ), la protéine de liaison GATA 3 (GATA3), le facteur de transcription T-box (T-bet), l’interleukine-10 (IL-10) et l’IL-4. Les porcelets témoins ont pris environ 35 % de poids en plus que ceux infectés par le PRDC. Plus précisément, des cellules PCV-2+ ont été détectées chez les porcelets du groupe infecté dans les proportions suivantes : WEA ≤ INI ≤ GRO. Chez les porcelets infectés, le nombre de CD4+ a augmenté à WEA et diminué à GRO, le profil d’expression de CD4+ a montré une surexpression de T-bet à INI et GRO, et l’expression d’IFN-γ était plus faible à WEA et GRO. En revanche, l’IL-4 était surexprimée aux trois stades. GATA3 était surexprimé à INI et GRO. Les porcelets infectés présentaient une lymphopénie et moins de cellules CD4+. Les cellules CD4+ ont montré un profil d’expression différent de celui du groupe témoin, dans lequel l’IFN-γ était moins exprimé, tandis que l’IL-4 et le T-bet étaient surexprimés.(Traduit par Docteur Serge Messier).

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Figures

Figure 1
Figure 1
Weight of pigs (N = 54) from the control farm (n = 27) and the infected farm (n = 27) divided according to the following 3 stages of development (n = 9): weaning (WEA), initiation (INI), and growth (GRO). Bars indicate the mean of 9 individual subjects and the error bars represent the standard error of the mean (SEM). Data are presented as mean + SEM. Statistically significant differences were determined by Student’s t-tests. * P < 0.05.
Figure 2
Figure 2
Detection of coinfections related to PRCD at the infected farm. ELISA and hemaglutination antibody detection from serum of pigs (N = 12) divided (n = 4) according to the following 3 stages of development: weaning (WEA), initiation (INI), and growth (GRO). (A) PCV-2, (B) PRRSV, (C) H1N1, (D) M. hyopneumoniae, and (E) H3N2. PCV-2, PRRSV, and M. hyopneumoniae were determined by ELISA. H1N1 and H3N2 were determined by hemagglutination inhibition. The dotted line represents the limit of detection for positive samples (below the dotted line was considered a negative result). Data are presented as mean + SEM. Statistically significant differences were determined by individual Student’s t-tests. * P < 0.05.
Figure 3
Figure 3
Quantification of PCV-2+ cells at different stages of development. PCV-2+ cells from samples of peripheral blood mononuclear cells (PBMCs) were determined by flow cytometry using an anti-PCV-2-FITC antibody. PBMCs isolated from pigs (N = 48) from the control farm (n = 24) and the infected farm (n = 24) were divided according to the following 3 stages of development (n = 8): weaning (WEA), initiation (INI), and growth (GRO). Data are presented as mean and error bands represent the + SEM. Statistically significant differences were determined by Student’s t-tests. * P < 0.05.
Figure 4
Figure 4
Peripheral blood mononuclear cell (PBMC) populations isolated from blood. Different cell populations were characterized by flow cytometry separation, according to size and complexity. (A) Monocytes, (B) Lymphocytes. Data are presented as the mean of 8 animals (n = 8) and error bars represent the + SEM. Statistically significant differences were determined by Student’s t-tests. * P < 0.05.
Figure 5
Figure 5
CD4+ cell population at different stages of development at the infected farm and the control farm. CD4+ cells from peripheral blood mononuclear cells (PBMCs) were quantified by flow cytometry using an anti-CD4-FITC antibody. Data are presented as mean + SEM and statistically significant differences were determined by Student’s t-tests. * P < 0.05.
Figure 6
Figure 6
Gene expression of CD4+ cells. The gene expression profile of CD4+ cells was evaluated by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). CD4+ cells were isolated from pigs (N = 48) from the control farm (n = 24) and the infected farm (n = 24) and divided according to the following 3 stages of development (n = 8): Weaning (WEA), initiation (INI), and growth (GRO). (A) T-BET, (B) IL-4, (C) INF-γ, (D) GATA3, (E) IL-10. Data are presented as mean and error bands represent the + SEM. Dotted line represents the limit of overexpression gene. Statistically significant differences were determined by Student’s t-tests. * P < 0.05.
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References

    1. Lv QZ, Guo KK, Zhang YM. Current understanding of genomic DNA of porcine circovirus type 2. Virus Genes. 2014;49:1–10. - PubMed
    1. Nauwynck HJ, Sanchez R, Meerts P, et al. Cell tropism and entry of porcine circovirus 2. Virus Res. 2012;164:43–45. - PubMed
    1. Tischer I, Rasch R, Tochtermann G. Characterization of papovavirus- and picornavirus-like particles in permanent pig kidney cell lines. Zentralbl Bakteriol Orig A. 1974;226:153–167. - PubMed
    1. Firth C, Charleston MA, Duffy S, Shapiro B, Holmes EC. Insights into the evolutionary history of an emerging livestock pathogen: Porcine circovirus 2. J Virol. 2009;83:12813–12821. - PMC - PubMed
    1. Noriega J, Reyes P, Bucarey S. Circovirus Porcino : Un virus pequeño que genera un gran problema Porcine Circovirus : A small virus that causes a great problem. Av en Ciencias Vet. 2007;22:62–71.

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