(A) Three RhCMV-naïve RMs (T1A-1, T1A-2, and T1A-3) were co-inoculated with 107 PFU each of 68-1/SIVgag (left arm) and ΔRh110/SIVrtni (right arm). One RM each was necropsied at 14, 21, or 28 dpi, and viral genome copy numbers per 107 cell equivalents were determined in the indicated tissues using ultrasensitive nested qPCR specific for SIVgag (68-1) or SIVrtni (ΔRh110). (B) Three naturally RhCMV-infected RMs (T1B-1, T1B-2, and T1B-3) were co-inoculated with 107 PFU each of 68-1/SIVrtni (left arm) and ΔRh110/SIVgag (right arm). One RM each was necropsied at 14, 21, or 28 dpi, and viral genome copy numbers per 107 cell equivalents in the indicated tissues were determined using ultrasensitive nested qPCR specific for SIVgag (ΔRh110) or SIVrtni (68-1). (C) Two naturally RhCMV-infected RMs (T1C-1 and T1C-2) were co-inoculated with 107 PFU each of complemented ΔRh110 and uncomplemented ΔRh110 in different arms, with the SIVgag and SIVrtni inserts used to mark the complemented and uncomplemented vectors, respectively, in RM1 and the reverse in RM2. Both RMs were necropsied at 14 dpi, and viral genome copy numbers were determined using ultrasensitive nested qPCR specific for SIVgag versus SIVrtni. Normalized to 1 × 107 cell equivalents. WT, wild type; GI, gastrointestinal; LN, lymph node; PLN, parietal lymph node.
| A. Primary infection: 68-1 versus ΔRh110 | ||||||
| RM T1A-1 | RM T1A-2 | RM T1A-3 | ||||
| Tissue type | 14 dpi | 21 dpi | 28 dpi | |||
| 68-1/SIVgag | ΔRh110/SIVrtni | 681/SIVgag | ΔRh110/ SIVrtni | 68-1/SIVgag | ΔRh110/ SIVrtni | |
| Skin injection site—right (ΔRh110) | 19,975,462 | <1 | 13,227 | 2 | <1 | <1 |
| Skin injection site—left (WT) | 1,013,192,441 | 3 | 14,453 | <1 | <1 | <1 |
| Axillary LN—right (ΔRh110 draining) | 4,547 | <1 | <1 | <1 | 7 | <1 |
| Axillary LN—left (WT draining) | 17,687,789 | <1 | 29,484 | <1 | 5,771 | <1 |
| PLN (except draining LN) | 265,298 | 49 | 3 | <1 | 40 | <1 |
| Mesenteric LN | <1 | 1 | 5 | <1 | 5 | <1 |
| GI tract | 34 | 5 | 8 | <1 | 64 | <1 |
| Liver/gallbladder | 11 | <1 | <1 | <1 | 41 | <1 |
| Heart/lung/kidney/BAL | 66 | 4 | 77 | <1 | <1 | <1 |
| BM/spleen/tonsil | 4 | 3 | 9,736 | <1 | <1 | <1 |
| Neuro/endocrine | 14 | <1 | 8 | <1 | <1 | <1 |
| Genitourinary tract | 125 | 14 | 14,075 | <1 | 17 | <1 |
| Salivary glands | 13,262 | <1 | 1,242 | <1 | 11 | <1 |
| PBMCs | <1 | 3 | <1 | <1 | <1 | <1 |
| B. Superinfection: 68-1 versus ΔRh110 | ||||||
| RM T1B-1 | RM T1B-2 | RM T1B-3 | ||||
| Tissue type | 14 dpi | 21 dpi | 28 dpi | |||
| 68-1/SIVrtni | ΔRh110/SIVgag | 68-1/SIVrtni | ΔRh110/ SIVgag | 68-1/SIVrtni | ΔRh110/ SIVgag | |
| Skin injection site—right (WT) | 47 | 5 | 16,502 | <1 | 376 | <1 |
| Skin injection site—left (ΔRh110) | 11 | 354 | 7 | <1 | 355 | <1 |
| Axillary LN—right (WT draining) | 117 | <1 | 7 | <1 | 509 | <1 |
| Axillary LN—left (ΔRh110 draining) | <1 | <1 | <1 | <1 | <1 | <1 |
| Peripheral LN | 1,434 | <1 | 41 | <1 | 2 | <1 |
| Mesenteric LN | <1 | 1 | 1 | <1 | <1 | <1 |
| GI tract | 3 | <1 | <1 | <1 | 12 | <1 |
| Liver/gallbladder | 1 | <1 | 9 | <1 | 9 | <1 |
| Heart/lung/kidney/BAL | 5 | <1 | <1 | <1 | 8 | <1 |
| BM/spleen/tonsil | 15 | 5 | <1 | <1 | <1 | <1 |
| Neuro/endocrine | <1 | <1 | 3 | <1 | 3 | <1 |
| Genitourinary tract | 3 | <1 | 4 | <1 | 1,047 | <1 |
| Salivary gland | 23 | 1 | <1 | <1 | <1 | <1 |
| C. ΔRh110: Complemented versus uncomplemented | ||||||
| RM T1C-1 | ←14 dpi→ | RM T1C-2 | ||||
| Tissue type | Uncomplemented ΔRh110/SIVrtni | Complemented ΔRh110/SIVgag | Tissue type | Uncomplemented ΔRh110/SIVgag | Complemented ΔRh110/SIVrtni | |
| Skin injection site—right (complemented) | <1 | <1 | Skin injection site—right (uncomplemented) | <1 | <1 | |
| Skin injection site—left (uncomplemented) | 13 | <1 | Skin injection site—left (complemented) | <1 | <1 | |
| Axillary LN—right (complemented draining) | <1 | <1 | Axillary LN—right (uncomplemented draining) | <1 | <1 | |
| Axillary LN—left (uncomplemented draining) | 17 | <1 | Axillary LN—left (complemented draining) | <1 | <1 | |
| PLN (except draining LN) | <1 | <1 | PLN (except draining LN) | <1 | <1 | |
| Mesenteric LN | <1 | <1 | Mesenteric LN | <1 | <1 | |
| GI tract | <1 | <1 | GI tract | <1 | <1 | |
| Liver/gallbladder | <1 | <1 | Liver/gallbladder | <1 | <1 | |
| Heart/lung/kidney/BAL | <1 | <1 | Heart/lung/kidney/BAL | <1 | <1 | |
| BM/spleen/tonsil | <1 | <1 | BM/spleen/tonsil | <1 | <1 | |
| Neuro/endocrine | <1 | <1 | Neuro/endocrine | <1 | <1 | |
| Genitourinary tract | 5 | <1 | Genitourinary tract | <1 | 2 | |
| Salivary glands | <1 | <1 | Salivary glands | <1 | <1 | |
| PBMCs | <1 | <1 | PBMCs | <1 | <1 | |