Dataset for longitudinal profiling of T-cell response elicited by PCV2 subunit vaccine and upon heterologous PCV2d challenge

At 28 days post-vaccination (dpv), the Vacc-PCV2d and PCV2d groups were challenged both intranasally and intramuscularly with the PCV2d strain CQ2302 (GenBank accession no. PQ511330.1); the Mock group received PK-15 cell culture supernatant via the same routes. Key procedures included: clinical monitoring and rectal temperature recording; serial blood collection for serum and peripheral blood mononuclear cell (PBMC) isolation; PCV2-specific IFNγ-secreting cell enumeration by IFNγ-ELISPOT; high-throughput T-cell response profiling using intracellular cytokine staining (ICS) and flow cytometry; memory T-cell immunophenotyping by surface marker staining; CD8 T-cell degranulation assessment by CD107a mobilization; T-cell proliferation analysis by Ki-67 staining; PCV2-specific antibody detection by indirect ELISA; and viral load quantification in serum and swab samples by real-time quantitative PCR (qPCR).Data ProcessingData processing involved flow cytometry analysis using FlowJo software (version 10, Tree Star Inc.), ELISPOT plate reading using an S6-Entry ImmunoSpot Analyzer (CTL), viral load calculation by interpolation from a standard curve using LightCycler 480 software (Roche), and statistical analysis using SPSS Statistics (version 23.0, IBM Corp.).

For ICS and degranulation assays, antigen-specific T-cell frequencies were calculated by subtracting background values from unstimulated controls, with positivity defined as exceeding the limit of detection (LOD; mean plus three standard deviations of the mock group).Temporal ScopeThe data were generated from laboratory-based animal studies conducted at the experimental animal center of Zhongchong Sino Biotech Co., LTD (Taizhou, China).

The temporal scope covers the experimental timeline: vaccination phase from 0 to 28 days post-vaccination (dpv), and challenge phase from 0 to 14 days post-challenge (dpc). PBMC samples for immunological assays were collected at 0, 7, 14, 21, and 28 dpv, and at 0, 7, and 14 dpc. Serum, nasal swab, and fecal swab samples for virological analysis were collected at 0, 3, 7, 10, and 14 dpc.Data Structure and ContentThe dataset is primarily tabular.

In each table, rows correspond to time points (e.g., 0, 7, 14, 21, 28 dpv or 0, 3, 7, 10, 14 dpc), and columns correspond to individual pigs. Cell values represent the measured response for a specific animal at a specific time point. When multiple experimental groups are included, they are indicated by column grouping, column prefixes, or separate worksheets within the same file.Key data tables include:ELISPOT Data: Rows represent time points (dpv and dpc).

Columns represent individual pigs. Cell values are PCV2-specific IFNγ-secreting cells per million PBMCs (after background subtraction).Intracellular Cytokine Staining (ICS) Data: Rows represent time points (dpv or dpc). Columns represent individual pigs.

Cell values are the frequency of cells positive for each cytokine combination (IFNγ/IL-2/TNFα) within CD4 or CD8 T cells. Immunophenotyping Data: Rows represent time points (dpv). Columns represent individual pigs.

Cell values are the frequency of naïve (TN), central memory (TCM), effector memory (TEM), and terminally differentiated effector memory (TEMRA) cells within CD4 or CD8 T cells.Degranulation Data: Rows represent time points (dpv or 28 dpv). Columns represent individual pigs. Cell values are the frequency of CD107a⁺ cells within CD8 T cells.Proliferation Data: Rows represent time points post-challenge (0, 7, 14 dpc).

Columns represent individual pigs. Cell values are the frequency of Ki-67⁺ cells within CD4 or CD8 T cells.Serology Data: Rows represent time points (0 and 28 dpv). Columns represent individual pigs.

Cell values are PCV2-specific antibody titers determined by indirect ELISA.Clinical and Virological Data: Rows represent time points post-challenge (0, 3, 7, 10, 14 dpc). Columns represent individual pigs. Cell values include rectal temperature, body weight, average daily weight gain (ADWG), and PCV2 DNA copy numbers in serum, nasal swabs, and rectal swabs (log10 copies/mL).The total number of data entries varies by table, corresponding to the number of animals (n=5 per group), time points, and experimental replicates described in the methods.Missing DataNo systematic missing data is reported.

All planned measurements and samples from the described experimental cohorts are accounted for in the results.Data Errors and UncertaintyMeasurement uncertainties are inherent to the techniques used. These include: the limit of detection for real-time qPCR; the error range associated with serial dilution in ELISA and virus titration; and the variability intrinsic to flow cytometry and ELISPOT assays, mitigated by the use of replicate wells, fluorescence-minus-one (FMO) controls, and standardized gating/analysis protocols.

For ELISPOT assays, the limit of detection for a positive response was 30 IFNγ-secreting cells per 10⁶ PBMCs. For ICS and degranulation assays, a response was considered positive only if the background-subtracted value exceeded the mean plus three standard deviations of the mock group. Statistical significance (p-values) and measures of variance (means ± standard deviations) are reported for quantitative comparisons, indicating the precision and reliability of the observed differences.