Peripheral blood lymphocytes from food-and-mouth disease virus resistant guinea pigs show increased cytotoxicity to cells with simulative viral infection
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Laboratory Animal Centre, Zhejiang University, Hangzhou 310013, China

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    Abstract:

    Objective To investigate the genetic and immunologic mechanisms of resistance to foot-and-mouth disease virus ( FMDV) in guinea pigs and to assess the possibility of using pseudo-FMDV in the pathogenetic study of FMVD. Methods FMDV-VP1 peptide was expressed in primary renal cells derived from two different strains of guinea pigs using lentivirus. Peripheral blood lymphocytes (PBLC, without antigen processing) derived from two different strains of guinea pigs were isolated and cultured with lentivirus infected primary renal cells. PBLC cytotoxicity was determined using the lactate dehydrogenase ( LDH ) assay, and supernatant cytokines were determined by enzyme-linked immunosorbent assay (ELISA). Moreover, the proportions of CD3+ / CD4+ and CD3+ / CD8+ cells were measured via flow cytometry. Results The cytotoxicity of PBLCs in experimental groups was significantly higher than in corresponding control groups (P < 0. 01). The cytotoxicity of PBLCs to target cells was significantly higher in the FMDV-resistant strain than in the FMDV-sensitive strain (P < 0. 01). Levels of PF, GRA, GRB, and Fas in serum derived from the FMDV-resistant strain were higher than in serum derived from the FMDV-sensitive strain (P < 0. 01). However, the expression of FasL showed no significant difference (P > 0. 05). Similarly, levels of IFN-γ, TNF-α, IL-2, and IL-12 were also higher (P < 0. 05). However, the expression of cell-surface MHC class I and II showed no significant difference (P > 0. 05). The FMDV-VP1 peptide was expressed on the surfaces of primary renal cells derived from guinea pigs when cells were infected by lentiviral-vector containing the DNA sequence of the FMDV-VP1 fragment. The quantity of CD4+ cells in PBLCs derived from the FMDV-resistant strain was significantly lower than in PBLCs derived from the FMDV-sensitive strain (P < 0. 05). The opposite trend was seen for the quantity of CD8+ cells ( P < 0. 05). Conclusions FMDV-VP1 peptides stimulate immunoreactions of effector cells and enhance the cytotoxicity of effector cells to target cells. PBLCs derived from the FMDV-resistant strain exerted strong cytotoxicity to FMDV infected cells. Cytokines secreted by effector cells participate in the cytotoxicity reaction. Primary renal cells derived from guinea pigs infected by lentiviral-vector containing the DNA sequence of the FMDV-VP1 fragment can be used to simulate FMDV infected cells, which could help to reduce risks associated with FMDV research. Finally, the genetic characteristics of different strains of guinea pigs determine their sensitivities to FMDV.

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History
  • Received:August 11,2019
  • Revised:
  • Adopted:
  • Online: April 29,2020
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