Results of head ultrasonography were normal. Because of persistent lethargy, hypotonia, desaturation episodes, and hypothermia, another sepsis evaluation was performed on DOL 11 to evaluate for viral caused. ...neonatal tissues possess high proportions of regulatory T cells that can suppress endogenous T-cell activation,7 potentially increasing susceptibility to disseminated viral infection. Quantitative (plasma) and qualitative (nasopharyngeal aspirates, endotracheal tube aspirates, urine, and stool) adenovirus real-time PCR testing (Molecular Laboratory, Nationwide Children's Hospital) and standardized GvHD assessments were also performed at least weekly for 50 days after VST infusion.Detection of maternal monocytes and lymphocytes Low-resolution HLA typing was performed by using standard PCR-based methodology at the Ohio State University Clinical Histocompatibility Laboratory. DOL Interventions Adenovirus PCR∗ Laboratory studies Antiviral therapy Supportive care Plasma copies/mL (log10/mL) NP/ETT Stool Urine Cr (mg/dL) ALT (U/L) AST (U/L) WBC (× 103/μL) ALC HGB (g/dL) PLT (× 103/μL) LDH (U/L) 14 IVIG, cidofovir† Nasal CPAP >2M (>6.3) Positive Positive — 0.54 92 326 10.4 3,120 11 202 — 16 Cidofovir 17 Intubated, MV started >2M (>6.3) Positive — — 0.42 77 424 10.9 3,488 12 103 — 18 Cidofovir 19 MV→HFOV, iNO started 21 Cidofovir >2M (>6.3) Positive — Positive 0.50 41 141 16.1 4,699 8.9‡ 109 3,435 22 VST infusion 23 Cidofovir iNO stopped — Positive — 24 545,050 (5.7) Positive — Positive 0.41 25 45 23.1 8,547 9.6‡ 174 1,589 25 Cidofovir HFOV→MV 26 Extubated→ BiPAP 28 Cidofovir 29 16,155 (4.2) Positive Negative — 0.49 16 19 36 10,440 9.6 496 968 30 Cidofovir — — — Negative — — — 23.7 9,717 9.0‡ 571 — 31 BiPAP→CPAP 1,951 (3.3) — — — 32 Cidofovir 0.2L O2 NC 34 1,214 (3.1) — — — 0.40 21 23 17.9 9,487 11.3 570 806 35 Cidofovir Negative — — 36 424 (2.6) — — — 37 Cidofovir 43 Negative — — — 0.34 24 26 12.8 7,680 9.1 420 — 57 — — — 0.32 29 23 8.3 5,063 7.5 399 623 64 Negative — — — 0.27 36 31 8.8 6,952 7.2 472 806 69 — — — — 0.28 36 22 8.2 6,314 6.9‡ 483 612 76 — — — — 0.30 35 36 10.8 7,949 11.9 353 697 Table I Clinical course, laboratory evaluations, and administered supportive care and antiviral therapies Antibody/probe Fluorochrome Clone Vendor Anti-human CD3 Allophycocyanin (APC)-Vio 770 REA613 Miltenyi Biotech Anti-human CD4 Allophycocyanin (APC) REA623 Miltenyi Biotech Mouse anti-human CD8 BB515 RPA-T8 BD Biosciences (Franklin Lakes, NJ) Anti-human HLA-A9 Phycoerythrin (PE) REA127 Miltenyi Biotech Viability Ghost Dye Violet 510 — Tonbo Bioscience (San Diego, Calif) Table E1 Conjugated mAbs used in flow cytometric analyses DOL 1 DOL 2 DOL 8 DOL 9 DOL 11 DOL 12 Complete blood count WBC (× 103/μL) 16.8 20.6 22.2 17 Segmented neutrophils (%) 57 79 79 68 Band neutrophils (%) 2 0 3 3 Absolute neutrophil count 9,912 16,274 18,204 12,070 Lymphocytes (%) 24 16 11 24 ALC 4,032 3,296 2,442 4,080 Hemoglobin (g/dL) 15.4 13.6 12.2 13 Platelet (× 103/μL) 305 194 288 309 Inflammatory marker C-reactive protein (mg/L) 3.8 16.8 CSF Glucose (mg/dL) 68 50 Protein (mg/dL) 165 107 RBCs (cells/μL) 2,095 2,420 WBCs (cells/μL) 4 14 Neutrophils (%) 67 47 Lymphocytes (%) 14 33 Monocytes/macrophages (%) 17 13 Eosinophils (%) 1 1 Basophils (%) 0 1 Bacterial cultures Eye including Chlamydia species No growth Blood No growth No growth No growth CSF No growth No growth Urine No growth No growth PCR testing Viral nasopharyngeal panel ADV C NP influenza A/B Negative NP respiratory syncytial virus Negative Herpes simplex virus Blood Negative CSF Negative Pooled surface swabs Negative Meningitis/encephalitis array∗ CSF Negative Antimicrobial therapy Ampicillin, 100 mg/kg every 12 h X X X X Gentamicin, 4 mg/kg daily X X X X X X Nafcillin, 50 mg/kg every 12 h X X Acyclovir, 20 mg/kg every 8 h X X Table E2 Diagnostic evaluations performed and antimicrobial therapies administered at an outside hospital PBMC leukapheresis Adenovirus-specific T cells Cell counts Total volume 196 mL 5.13 mL† Total nucleated cell count 93.79 × 108 cells/kg 1.15 × 106 cells/kg Fresh viability (≥90% expected) 97% — Volume processed on CliniMACS Prodigy 11 mL — Total nucleated cells processed on CliniMACS Prodigy 1 × 109 cells — Postprocessing results Acceptability criteria Result IFN-γ+ T-cell purity (%) — ≥50% CD3+CD4+ cells 90.0% IFN-γ+ T-cell purity (%) — ≥50% CD3+CD8+ cells 86.4% Fold enrichment, CD3+CD4+IFN-γ+ T cells — ≥20-fold 310-fold Fold enrichment, CD3+CD8+IFN-γ+ T cells — ≥20-fold 131-fold Percentage recovery, CD3+CD4+IFN-γ+ T cells — Report only 153% Percentage recovery, CD3+CD8+IFN-γ+ T cells — Report only 55% Release testing results Test method Target specification Result WBC viability (%) — Flow cytometry, 7-AAD ≥45% 47.2% CD3+IFN-γ+ T-cell viability (%) — Flow cytometry, 7-AAD ≥70% 91.7% Microbial contamination through Gram staining — Gram stain No organisms seen No organisms seen Endotoxin (EU/kg) — Limulus amebocyte lysate ≤5.0 EU/kg/h <0.04 EU/kg CD3+IFN-γ+ T-cell dose — Flow cytometry ≤2.4 × 104 cells/kg 1.00 × 105 CD3+ T-cell dose — Flow cytometry ≤1 × 105 cells/kg 1.29 × 105 Microbial contamination (after release) Test method Target specification Result Aerobic bacteria culture No growth Aerobic bacteria culture No growth No growth Anaerobic bacteria culture No growth Anaerobic bacteria culture No growth No growth Table E3 Cell processing and product information∗