Journal:

Article Title: Increased Macrophage Infection upon Subcutaneous Inoculation of Rhesus Macaques with Simian Immunodeficiency Virus-Loaded Dendritic Cells or T Cells but Not with Cell-Free Virus

doi: 10.1128/JVI.76.19.9787-9797.2002

Figure Lengend Snippet: Immature DCs exposed to SIVmac239 in vitro transmit infection after s.c. reinjection. (A and B) Immature monocyte-derived DCs from two healthy macaques (F154 and L780) were exposed to SIVmac239 (MOI of 0.1), washed, and cultured overnight in the presence of 10 μg of TT per ml. The next day, the cells were washed again, recounted, and monitored for their ability to transmit infection in vitro (A) and in vivo (B). (A) Virus-loaded DCs were mixed with 174xCEM cells (104 DCs with 105 174xCEM [CEM] cells [F154 DC-SIV+CEM and L780 DC-SIV+CEM]) or plated alone (F154 DC-SIV and L780 DC-SIV) and cultured for approximately 2 weeks. Starting at day 3 of culture, supernatants were sampled every other day and monitored for the presence of RTase activity. The results are expressed as cpm (103) of RTase activity per μl of culture supernatant. (B) SIV and TT-bearing DCs (L780, 2 × 106 DC-SIV; F154, 1 × 106 DC-SIV) were s.c. reinjected into the donor animals at three sites proximal to an inguinal LN. Blood samples collected over the ensuing weeks were analyzed for virus RNA levels by bDNA analysis. The numbers of RNA copies per milliliter of plasma are shown. (C) LN and spleen (Sp) cell suspensions isolated from tissues obtained at necropsy of animal F154 were assessed for virus levels by SIV gag PCR. Aliquots of 105 cells were sampled, and the SIV gag DNA was amplified. Standard curve controls representing 0 (lane −), 101 (lane 1), 102 (lane 2), 103 (lane 3), and 104 (lane 4) SIV gag copies were included to estimate SIV gag copy numbers in each test sample.

Article Snippet: The sections were hybridized to a 35 S-labeled, single-stranded antisense RNA probe of SIVmac239 (Lofstrand Labs, Gaithersburg, Md.) overnight at 45°C.

Techniques: In Vitro, Infection, Derivative Assay, Cell Culture, In Vivo, Activity Assay, Isolation, Amplification

Journal:

Article Title: Increased Macrophage Infection upon Subcutaneous Inoculation of Rhesus Macaques with Simian Immunodeficiency Virus-Loaded Dendritic Cells or T Cells but Not with Cell-Free Virus

doi: 10.1128/JVI.76.19.9787-9797.2002

Figure Lengend Snippet: Transmission of infection by cell-associated versus cell-free virus. (A) Immature monocyte-derived DCs or CD4+ T cells were isolated from 17 healthy macaques (as indicated) and exposed to an MOI of 0.1 of SIVmac239 wild-type or SIVmac239 delta nef virus. After an overnight incubation, the cells were collected, washed, and recounted, and 3 × 105 to 1.2 × 106 SIV-bearing DCs or T cells were reinjected into the donor animals proximal to the inguinal or axillary LNs (Table ​(Table2).2). Individual animal numbers are listed in each graph. Four other animals were s.c. injected with the indicated doses of cell-free virus. (B) Six healthy macaques were infected i.v. with either 102 TCID50 of wild-type SIVmac239 or 2.35 × 104 TCID50 of delta nef SIVmac239. Plasma viremia was measured by bDNA analysis for up to 100 days after injection of cell-associated or cell-free virus to reveal the number of SIV RNA copies per milliliter of plasma.

Article Snippet: The sections were hybridized to a 35 S-labeled, single-stranded antisense RNA probe of SIVmac239 (Lofstrand Labs, Gaithersburg, Md.) overnight at 45°C.

Techniques: Transmission Assay, Infection, Derivative Assay, Isolation, Incubation, Injection

Journal: Scandinavian Journal of Medicine & Science in Sports

Article Title: Alpha-2 adrenergic stimulation triggers Achilles tenocyte hypercellularity: Comparison between two model systems

doi: 10.1111/j.1600-0838.2011.01442.x

Figure Lengend Snippet: Section of rabbit Achilles tendon tissue processed with immunohistochemistry (a, b; tetramethylrhodamine isothiocyanate staining) and in situ hybridization (inset in b; anti-digoxigenin alkaline phosphatase immunostaining detection), for detection of tyrosine hydroxylase (TH; a), alpha-2A-adrenoreceptor (α 2A AR; b) and TH mRNA (inset in b). (a) TH-positive nerve structures are seen in the paratendinous tissue in the form of perivascular nerves (arrows) and in a nerve fascicle (N). Asterisk marks lumen of vessel. (b) Tenocytes of the tendon tissue proper display immunoreactions for α 2A AR (arrows). (Inset) A tenocyte is positive for TH mRNA (arrow). Antisense staining.

Article Snippet: For control purposes, a negative control using the corresponding sense DIG-hyperlabeled ssDNA probe was used, and positive controls in the form of a Poly(dT) probe (GD4000-OP; GeneDetect) and a β-actin antisense probe (GD5000-OP, GeneDetect) were used.

Techniques: Immunohistochemistry, Staining, In Situ Hybridization, Immunostaining