Journal: Japanese journal of ophthalmology

Article Title: Infection of endotheliotropic human cytomegalovirus of trabecular meshwork cells.

doi: 10.1007/s10384-018-0618-1

Figure Lengend Snippet: Fig. 1 Induction of the mRNA of the genes of human cytomegalovirus (HCMV) in human trabecular meshwork cells (HTMC) after HCMV infection. HTMCs were infected with the Towne strain or the TB40/E strain of HCMV, and the induction of the mRNA of IE1 (a), pp65 (b), and glycopro- tein B (gB) (c) was determined by RT-real-time PCR. Viral genes, IE1, pp65, and gB, were sequentially induced after infection by the Towne strain or the TB40/E strain of HCMV. TB40/E induced significantly higher amounts of IE1, pp65, and gB. *P <0.05, **P <0.05; n = 8

Article Snippet: The transferred membranes were stained with antibodies for HCMV IE1(NEA-9221, Perkin Elmer), pp65 (CA003, EastCoast Bio), or UL44 (CH167, Santa Cruz) [5].

Techniques: Infection, Real-time Polymerase Chain Reaction

Journal: Japanese journal of ophthalmology

Article Title: Infection of endotheliotropic human cytomegalovirus of trabecular meshwork cells.

doi: 10.1007/s10384-018-0618-1

Figure Lengend Snippet: Fig. 6 Induction of viral gene proteins and remodeling of actin cytoskeleton after HCMV infection. HTMCs were infected with the TB40/E strain (a) or the Towne strain (b) of HCMV at MOI of 5 and immunostained for IE1, pp65, and glycoprotein B (Green). TB40/E infection induced significant remodeling of the actin cytoskeleton (c, pp65, green). Actin was stained using phalloidin (red). Nucleus was labelled by DAPI (blue). *Viral assembly compartment

Article Snippet: The transferred membranes were stained with antibodies for HCMV IE1(NEA-9221, Perkin Elmer), pp65 (CA003, EastCoast Bio), or UL44 (CH167, Santa Cruz) [5].

Techniques: Infection, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: 4-1BB is superior to CD28 costimulation for generating CD8+ cytotoxic lymphocytes for adoptive immunotherapy.

doi: 10.4049/jimmunol.179.7.4910

Figure Lengend Snippet: FIGURE 2. PBL costimulated with 4-1BBL, but not CD28, are enriched for memory CTL. A, CD8 PBL were unstimulated (top panel) or stimulated with K32/anti-CD3/4-1BBL (middle panel) or K32/anti-CD3/anti-CD28 (lower panel) for 7 days, then expanded cells were assayed using HLA A*0201 tetramers loaded with CMV pp65, Flu M1 peptide, and ERBB2 peptides. Anti-CD3/4-1BBL-expanded cells, but not anti-CD3/anti-CD28-expanded cells show increased frequencies of cells responding to viral recall Ags. Percentages reflect the percent of tetramer binding of CD8 cells. B, CD8 PBL stimulated with K32/anti-CD3/4-1BBL, but not K32/anti-CD3/anti-CD28, demonstrate cytotoxicity to FluM1- and pp65-pulsed T2 targets on day 7. Results were representative of five different experiments, using cells from different individual donors. C, 4-1BB costimulation preferentially expands memory CD8 cells but anti-CD28 costimulation preferentially expands naive CD8 cells. CD8 T cells were separated into naive (CD45RO) and memory (CD45RO) subsets using anti-CD45RO microbeads, then placed into a 96-well flat-bottom plate coated with anti-CD3 (1 g/ml) and anti-CD28 or anti-4-1BB at the concentrations indicated. Proliferation was measured by [3H]thymidine incorporation during the last 1618 h of a 5-day culture. This is a representative result of three different experiments.

Article Snippet: Where designated, cultures were harvested, cells were counted, and lytic activities were measured using conventional 4-h 51Cr-release assays using T2 target cells that were first pulsed with either 10 M Flu M1 peptide (GILGFVFTL), CMV pp65 peptide (NLVPMVATV), or the carcinoembryonic Ag-derived HLA-A2-binding peptide CAP1 (GVLVGVALI; New England Peptide), then labeled with 51Cr (PerkinElmer).

Techniques: Binding Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: 4-1BB is superior to CD28 costimulation for generating CD8+ cytotoxic lymphocytes for adoptive immunotherapy.

doi: 10.4049/jimmunol.179.7.4910

Figure Lengend Snippet: FIGURE 4. Selective expansion of Ag-specific CTL using K32/4-1BBL/HLA-A2-Fc aAPCs. A, CD8 cells were stimulated using K32/4-1BBL/pp65- HLA-A2-Fc, K32/anti-CD28/pp65-HLA-A2-Fc, or K32/pp65-HLA-A2-Fc. On day 22 after three stimulations, the cells were harvested, counted, and analyzed by FACS. K32/4-1BBL/pp65 aAPCs expanded Ag-specific cells but not cells binding an irrelevant tetramer while K32/anti-CD28pp65 and K32/pp65 did not expand Ag-specific cells. B, K32/4-1BBL/pp65, but not K32/anti-CD28/pp65, expanded CTL demonstrate Ag-specific cytotoxicity as assessed by CD107a expression. CD8 cells stimulated with K32/pp65 did not produce sufficient cell numbers to perform the assay. This is a representative result of three experiments using Ag-specific expansion from different HLA-A2 donors.

Article Snippet: Where designated, cultures were harvested, cells were counted, and lytic activities were measured using conventional 4-h 51Cr-release assays using T2 target cells that were first pulsed with either 10 M Flu M1 peptide (GILGFVFTL), CMV pp65 peptide (NLVPMVATV), or the carcinoembryonic Ag-derived HLA-A2-binding peptide CAP1 (GVLVGVALI; New England Peptide), then labeled with 51Cr (PerkinElmer).

Techniques: Binding Assay, Expressing