Table 1.
Comparisons of 28S-gB-CAR-T, BBL-gB-CAR-T, and irrelevant control gp350-CAR-T cells after a single coculture with mesenchymal stem cells infected with human cytomegalovirus
| 28S |
BBL |
gp350 |
p |
|||
|---|---|---|---|---|---|---|
| Mean (SD) | Mean (SD) | Mean (SD) | 28S × BBL | 28S × gp350 | BBL × gp350 | |
| AB (n = 3) | ||||||
| IFN-γ (pg/mL) | 9.3E+02 (5.78E+02) | 1.43E+03 (8.0E+02) | 0 (0) | ns | ** | *** |
| % CD107a+/CD8+ | 7.97E-01 (4.1E-01) | 3.3E+01 (9.5E+00) | 3.23E-01 (2.03E-01) | *** | ns | *** |
| % Dead MSC | 8.4E+01 (5.25E+00) | 7.77E+01 (1.79E+01) | 5.0E+01 (1.41E+01) | ns | * | ns |
| % Proliferated T cells (CD4+) | 6.65E+01 (9.09E+00) | 1.9E+01 (9.57E+00) | 6.87E-01 (3.7E-01) | *** | *** | * |
| % Proliferated T cells (CD8+) | 6.02E+01 (1.04E+00) | 4.21E+01 (1.05E+01) | 2.4E-01 (2.6E-02) | * | *** | *** |
| CB (n = 3) | ||||||
| IFN-γ (pg/mL) | 4.96 + 00 (9.90E+00) | 1.84E+02 (1.56E+02) | 5.52E+00 (7.95E+00) | *** | ns | *** |
| % CD107a+/CD8+ | 3.65E+00 (1.06E+00) | 1.02E+01 (1.10E+00) | 1.97E+00 (2.65E-01) | *** | ns | *** |
| % Dead MSC | 1.62E+01 (1.47E+00) | 2.87E+01 (7.11E+00) | 1.63E+01 (2.66E+00) | *** | ns | *** |
| % Proliferated T cells (CD4+) | 4.31E+00 (2.00E+00) | 3.59E+00 (2.17E+00) | 1.16E+00 (3.5E-01) | ns | ** | * |
| % Proliferated T cells (CD8+) | 9.72E+00 (2.38E+00) | 8.33E+00 (2.8E+00) | 3.66E+00 (1.71E+00) | ns | ns | ns |
CAR-T cells were generated from three AB donors and from three CB donors and cultured with MSC-HCMV or MSC-MOCK at E:T 1:3; 1:1; and 3:1 for 72 h. Values are shown for 3:1 cocultures of CAR-T cells with MSC/HCMV. Triplicate cultures for each donor were analyzed individually for IFN-γ and bioluminescence detection. Cultures were pooled for flow cytometry analyses. Statistical significance was determined by two-way ANOVA and Bonferroni's post hoc test between the indicated groups and p-values are shown.
*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.
AB, adult blood; ANOVA, analysis of variance; CAR, chimeric antigen receptor; CB, cord blood; gB, glycoprotein B; HCMV, human cytomegalovirus; IFN, interferon; MSC, mesenchymal stem cell; ns, not significant (p > 0.05); SD, standard deviation.