reference strain vr2332  (ATCC)

Journal: Frontiers in Immunology

Article Title: Mitochondrial dysfunction in PRRSV-2-infected macrophages

doi: 10.3389/fimmu.2025.1670488

Figure Lengend Snippet: Experimental layout. Lungs were collected from influenza- and PRRSV-negative 8–12-week-old pigs. Mononuclear phagocytes (MNPs) from bronchoalveolar lavage (BAL) and parenchyma (PAR) were enriched using OptiPrep gradient. MNPs were infected with North Carolina (NC) PRRSV-2 strains NC134 and NC174 (MOI = 1) and control media for 12 h. Infected and control porcine alveolar macrophages (PAMs), pulmonary intravascular macrophages (PIMs), monocyte-derived dendritic cells (moDCs), and classical DCs (cDCs) were sorted, and RNA-seq was performed on each cell subset. Additional PAM and PIM sorted cells were used for qPCR and NanoString validations. For other assays, enriched MNPs were plated to allow macrophage adhesion for 2 h. PAMs and PIMs were subsequently infected with different NC PRRSV-2 strains (NC134, NC174, and NC144) or VR2332 prototype strain; uninfected cells were used as negative control. Infected macrophages were used in different assays: mitochondrial function assay (Seahorse Agilent technology) and ROS and NO production assays. Created in BioRender. Crisci, E (2025). https://BioRender.com/65x4hji . PRRSV, porcine reproductive and respiratory syndrome virus; MOI, multiplicity of infection; ROS, reactive oxygen species; NO, nitric oxide.

Article Snippet: Additionally, the reference strain VR2332 (ATCC strain BIAH-001, GenBank accession ID U87392.3 , L5A.1) was used as the PRRSV-2 prototype strain.

Techniques: Infection, Control, Derivative Assay, RNA Sequencing, Negative Control, Functional Assay, Virus

Journal: Vaccines

Article Title: Broad Protection of Pigs against Heterologous PRRSV Strains by a GP5-Mosaic DNA Vaccine Prime/GP5-Mosaic rVaccinia (VACV) Vaccine Boost

doi: 10.3390/vaccines8010106

Figure Lengend Snippet: Experimental design.

Article Snippet: The viruses used in the study included the VR2332 NA reference strain (ATCC VR-2332) and the MN184C strain provided kindly by Drs. Kelly Lager and Kay Faaberg at U.S. Department of Agriculture-Agriculture Research Service (USDA-ARS).

Techniques:

Journal: Vaccines

Article Title: Broad Protection of Pigs against Heterologous PRRSV Strains by a GP5-Mosaic DNA Vaccine Prime/GP5-Mosaic rVaccinia (VACV) Vaccine Boost

doi: 10.3390/vaccines8010106

Figure Lengend Snippet: Phylogenetic analysis of the GP5 Amino acid sequence of the two Mosaic sequences, MN184C and VR2332. The analysis was done via the neighbor-joining method using MEGA7.0.

Article Snippet: The viruses used in the study included the VR2332 NA reference strain (ATCC VR-2332) and the MN184C strain provided kindly by Drs. Kelly Lager and Kay Faaberg at U.S. Department of Agriculture-Agriculture Research Service (USDA-ARS).

Techniques: Sequencing

Journal: Vaccines

Article Title: Broad Protection of Pigs against Heterologous PRRSV Strains by a GP5-Mosaic DNA Vaccine Prime/GP5-Mosaic rVaccinia (VACV) Vaccine Boost

doi: 10.3390/vaccines8010106

Figure Lengend Snippet: Vaccination-induced humoral and cellular responses. ( A ) The virus neutralization was expressed as viral copy numbers (log10 scale), as measured by RT-qPCR in cell supernatants after the infection of MARC-145 cells with pre-incubated serum–virus mixtures. ( B ) The expression of IFN-γ mRNA as fold changes, by either VR2332 or MN184C-stimulated PBMCs collected on the challenge day. Each dot represents the value of one animal. The variation is expressed as standard error of the means. There were three independent replications. Significant differences were calculated by a two-way ANOVA or Student’s t test (* p < 0.05, *** p < 0.0001).

Article Snippet: The viruses used in the study included the VR2332 NA reference strain (ATCC VR-2332) and the MN184C strain provided kindly by Drs. Kelly Lager and Kay Faaberg at U.S. Department of Agriculture-Agriculture Research Service (USDA-ARS).

Techniques: Virus, Neutralization, Quantitative RT-PCR, Infection, Incubation, Expressing

Journal: Vaccines

Article Title: Broad Protection of Pigs against Heterologous PRRSV Strains by a GP5-Mosaic DNA Vaccine Prime/GP5-Mosaic rVaccinia (VACV) Vaccine Boost

doi: 10.3390/vaccines8010106

Figure Lengend Snippet: Virus clearance in sera and tissues. ( A ) Viral copy numbers in serum from 0 to 14 DPC upon VR2332 challenge. ( B ) The viral copy numbers in serum from 0 to 14 DPC upon MN184C challenge. Each dot represents the mean value of each group. The variation bars are expressed as the standard error of the mean. Three separate experiments were performed for each. A two-way ANOVA or Student’s t test (* p < 0.05, ** p < 0.01, was used to calculate significant differences; days post-challenge (DPC). ( C ) The viral copy numbers in tissues at necropsy upon VR2332 challenge. ( D ) The viral copy numbers in tissues at necropsy upon MN184C challenge. Each bar represents the mean value of each group. The bars are the standard error of the mean. Three separate experiments were performed for each. Significant differences were calculated by Student’s t test (* p < 0.05).

Article Snippet: The viruses used in the study included the VR2332 NA reference strain (ATCC VR-2332) and the MN184C strain provided kindly by Drs. Kelly Lager and Kay Faaberg at U.S. Department of Agriculture-Agriculture Research Service (USDA-ARS).

Techniques: Virus

Journal: Vaccines

Article Title: Broad Protection of Pigs against Heterologous PRRSV Strains by a GP5-Mosaic DNA Vaccine Prime/GP5-Mosaic rVaccinia (VACV) Vaccine Boost

doi: 10.3390/vaccines8010106

Figure Lengend Snippet: Virus clearance in bronchioalveolar lavage fluids (BAL) and PAMs. ( A ) The viral copy numbers in BAL fluids at necropsy upon challenge with VR2332. ( B ) The viral copy numbers in BAL fluids at necropsy upon challenge with MN184C. Each bar represents the mean value of each group. Bars represent the standard error of the mean. Three independent experiments were performed for each. Significant differences were calculated by Student’s t test (* p < 0.05). ( C ) The viral copy numbers in PAMs at necropsy upon challenge with VR2332. ( D ) The viral copy numbers in PAMs at necropsy upon challenge with MN184C. Each dot represents the mean value of each animal. The bars represent the standard error of the mean. Three independent experiments were performed for each. Student’s t test was used to calculate significant differences (* p < 0.05, ** p < 0.01).

Article Snippet: The viruses used in the study included the VR2332 NA reference strain (ATCC VR-2332) and the MN184C strain provided kindly by Drs. Kelly Lager and Kay Faaberg at U.S. Department of Agriculture-Agriculture Research Service (USDA-ARS).

Techniques: Virus