Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Transient activation of tumor-associated T-effector/memory cells promotes tumor eradication via NK-cell recruitment: minimal role for long-term T-cell immunity in cure of metastatic disease

doi: 10.1007/s00262-007-0430-0

Figure Lengend Snippet: Effect of NK-cell depletion on post-therapy intra-tumoral CD8+ T-cell activity

Article Snippet: In vivo lymphocyte subset depletions Purified anti-mouse CD4 monoclonal antibody (mAb) GK1.5 and anti-mouse CD8 mAb 53.6.72 were purchased from Bio express.

Techniques: Activation Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Transient activation of tumor-associated T-effector/memory cells promotes tumor eradication via NK-cell recruitment: minimal role for long-term T-cell immunity in cure of metastatic disease

doi: 10.1007/s00262-007-0430-0

Figure Lengend Snippet: CD8+ T- and NK-cell numbers and activity in post-therapy tumors, TDLN and lungs. Mice bearing advanced (500–600 mm3) primary tumors were treated with a single injection of IL-12+GM-CSF microspheres (day 0). Primary tumors, TDLN and lungs were analyzed for CD8+ T- and NK-cell quantity (cells/gram tumor) and activity (% IFNγ and perforin-positive cells) on indicated days. Day 0 data were obtained prior to treatment. a Primary tumors. Relative fold-change in tumor-infiltrating CD8+ T- and CD3- CD49b + NKG2D + NK-cell numbers is shown on the left ordinate. Percent cells positive for IFNγ or perforin are shown on the right ordinate. The differences between day 0 versus day 1 or 3 IFNγ + CD8+ T-cells were significant (P < 0.005). The differences between day 0 versus day 1 or 3 perforin + CD8+ T-cells were also significant (P < 0.05). The difference between day 0 versus day 3 perforin + NK-cells was significant (P = 0.007). Error bars = S.E., n = 6–13 per group. b TDLN. Cell quantity and activity were determined as in a. Two TDLN were analyzed per mouse. Error bars = S.E., n = 6–13 mice per group. c Lungs. CD8, CD94 exon 1A, IFNγ and perforin mRNA levels in whole lung extracts were quantified by real-time PCR. Relative fold-change compared to pre-therapy (day 0) levels is shown. The increase in CD94 exon 1A mRNA on day 3 was highly significant compared to other days (P < 0.0014). The increases in IFNγ and perforin mRNA (days 1–3) were also significant compared to other time points (P < 0.006). Error bars = S.E., n = 4 mice per group

Article Snippet: In vivo lymphocyte subset depletions Purified anti-mouse CD4 monoclonal antibody (mAb) GK1.5 and anti-mouse CD8 mAb 53.6.72 were purchased from Bio express.

Techniques: Activity Assay, Injection, Real-time Polymerase Chain Reaction

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Transient activation of tumor-associated T-effector/memory cells promotes tumor eradication via NK-cell recruitment: minimal role for long-term T-cell immunity in cure of metastatic disease

doi: 10.1007/s00262-007-0430-0

Figure Lengend Snippet: Effect of T-cell depletion on post-therapy NK-cell infiltration. Mice bearing advanced (500–600 mm3) primary tumors were treated with IL-12 + GM-CSF microspheres. The depletion groups received the relevant antibodies on days -4 and -1 prior to treatment. Relative fold-changes in CD3- CD49b + NKG2D + NK-cell numbers were quantified in pre-therapy (day 0) and post-therapy (day 3) primary tumors (cells/gram tumor) and TDLN (cells/lymph node) by flow cytometry. Relative fold-changes in lung CD94 exon 1A mRNA was quantified by real-time PCR analysis. The difference between days 0 and 3 NK-cell numbers in primary tumors was significant in the no depletion group (P = 0.021). The differences between days 0 and 3 in CD8 or CD8+ CD4 depletion groups were not significant (P > 0.7). The differences between days 0 and 3 in no depletion and CD8+ depletion groups were significant in the TDLN and lungs (P < 0.05). Error bars = S.E., n = 5–6 mice per group

Article Snippet: In vivo lymphocyte subset depletions Purified anti-mouse CD4 monoclonal antibody (mAb) GK1.5 and anti-mouse CD8 mAb 53.6.72 were purchased from Bio express.

Techniques: Flow Cytometry, Real-time Polymerase Chain Reaction

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Transient activation of tumor-associated T-effector/memory cells promotes tumor eradication via NK-cell recruitment: minimal role for long-term T-cell immunity in cure of metastatic disease

doi: 10.1007/s00262-007-0430-0

Figure Lengend Snippet: Time-dependent depletion of CD8+ T- and NK-cells in IL-12+GM-CSF microsphere-treated mice. Mice bearing advanced (500–600 mm3) subcutaneous primary tumors were treated with IL-12+GM-CSF microspheres (day 0) and primary tumors were surgically excised 7–8 days after treatment. Mice were monitored for 3 weeks after surgery and lung tumor burden was determined as described in “Materials and methods”. In some groups CD8+ T- or NK-cells were depleted starting on days -1, 4 or 10 via antibody administration. Tumors of control mice (untreated) were excised upon reaching 500–600 mm3 without treatment and lung tumor burden was analyzed 3 weeks after surgery. a CD8+ T-cell panel. Effect of CD8+ T-cell depletion on lung tumor burden. The differences between untreated control and all other groups were significant (P < 0.005). The difference between no depletion and day −1 groups was significant (P = 0.00034). The differences between no depletion and day 4 or day 10 depletion groups were not significant (P > 0.13). Error bars = S.E., n = 21 for no depletion group and eight for all other groups. b NK-cell panel. Effect of NK-cell depletion on lung tumor burden. The differences between control untreated and all other groups were significant (P < 0.03). The difference between no depletion and days -1, 4 or 10 depletion groups was significant (P < 0.004). Error bars = S.E., n = 20 for no depletion group and 8 for all others

Article Snippet: In vivo lymphocyte subset depletions Purified anti-mouse CD4 monoclonal antibody (mAb) GK1.5 and anti-mouse CD8 mAb 53.6.72 were purchased from Bio express.

Techniques:

Journal: Acta Neuropathologica

Article Title: Oxidative tissue injury in multiple sclerosis is only partly reflected in experimental disease models

doi: 10.1007/s00401-014-1263-5

Figure Lengend Snippet: Primary antibodies and antigen retrieval procedures

Article Snippet: Ox8 , Mouse (mAb) , Anti-rat CD8 , 1:400 , 0 , Seralab; MAS041.

Techniques: Binding Assay, Virus

Journal: Acta Neuropathologica

Article Title: Oxidative tissue injury in multiple sclerosis is only partly reflected in experimental disease models

doi: 10.1007/s00401-014-1263-5

Figure Lengend Snippet: Quantification of markers for inflammation and oxidative injury in different experimental models for CNS inflammation and multiple sclerosis

Article Snippet: Ox8 , Mouse (mAb) , Anti-rat CD8 , 1:400 , 0 , Seralab; MAS041.

Techniques: Injection, Virus

Journal: Acta Neuropathologica

Article Title: Oxidative tissue injury in multiple sclerosis is only partly reflected in experimental disease models

doi: 10.1007/s00401-014-1263-5

Figure Lengend Snippet: Oxidative burst, oxidative injury and iron accumulation in CD8 + T cell-mediated EAE, in innate immunity-driven inflammation and in toxic cuprizone-mediated demyelination. a – f Brain inflammation induced by CD8 + T cells. Perivascular inflammatory infiltrates were associated with massive microglial activation in the adjacent tissue ( a ; Iba-1). Perivascular inflammatory cells and to lower degree also the surrounding microglia highly expressed p22phox ( b ), while iNOS expression was minor and restricted to a few perivascular cells ( c ). The immunohistochemistry for oxidised phospholipids (E06; d ) was negative. Iron deposition was lacking in most lesions ( e ) but occurred in few lesions within perivascular macrophages ( f ). g – m Inflammatory demyelinating lesions induced by local injection of LPS into the spinal cord dorsal column. In the early inflammatory stage (1 day after LPS injection), p22phox + granulocytes ( g ) and iNOS + macrophages ( h ) were present in the lesions. At the peak of active demyelination (9 days after LPS injection), lesions were densely infiltrated by macrophages and showed microglial activation at the lesion edge ( i ; Iba-1, asterisk ). Macrophages in these lesions and, to a lower degree, microglia at the lesion edges showed expression of p22phox ( j ). Despite a very strong expression of p22phox in active lesions ( j ), the expression of iNOS was very low ( k ). There was a weak reactivity for oxidised phospholipids in the lesions ( l ; E06). Individual perivascular macrophages were iron positive ( m ). n – s Actively demyelinating lesion in the corpus callosum induced by cuprizone diet. Extensive loss of myelin was seen in the corpus callosum ( n ), which was densely packed with macrophages and microglia ( o ), although their majority did not express p22phox ( p ) and iNOS only very weakly ( q ). Additionally, there was no evidence for oxidised phospholipids ( r ; E06) or iron deposition ( s ) in the lesions. Scale bar 50 μm

Article Snippet: Ox8 , Mouse (mAb) , Anti-rat CD8 , 1:400 , 0 , Seralab; MAS041.

Techniques: Activation Assay, Expressing, Immunohistochemistry, Injection

Journal: Acta Neuropathologica

Article Title: Oxidative tissue injury in multiple sclerosis is only partly reflected in experimental disease models

doi: 10.1007/s00401-014-1263-5

Figure Lengend Snippet: Oxidative burst, oxidative injury and iron accumulation in inflammatory demyelinating lesions in MHV-JHM coronavirus-induced encephalomyelitis. a – i Basic characterisation of inflammatory demyelinating lesions in the spinal cord of an infected Lewis rat with demyelinating encephalomyelitis after infection with MHV-JHM coronavirus. There was extensive plaque-like demyelination ( a ; MBP) with relative preservation of axons ( b and f ; e NAWM; Bodian silver impregnation). Virus antigen was present in the periplaque white and grey matter ( c ; immunohistochemistry for nucleocapsid protein N) in neurons ( g ) as well as in glia cells in the white matter ( h ). Active demyelination was associated with profound T cell-mediated inflammation ( d ), the majority of inflammatory cells being CD8 + T cells ( blue ; i ) and ED1 + macrophages ( brown ; i ). j – n Determination of microglial activation in MHV-JHM coronavirus-induced lesions. Activated microglia formed microglial nodules in the NAWM reactive for Iba-1 ( j ) and p22phox ( l ). The edge of an active lesion is shown with immunocytochemistry for macrophages/microglia (Iba-1; k ), for myelin (MBP; m ) and for expression of NADPH oxidase (p22phox; n ). Active demyelination was associated with profound microglial activation ( k ; Iba-1). Their numbers increased in the periplaque white matter (PPWM) towards the lesion edge ( k ; left side ) showing the highest density at the zone of initial demyelination ( k , middle ) and decreasing towards the centre of the lesion, where many cells exhibited a macrophage phenotype ( k ; right side ). Immunohistochemistry for p22phox in serial-cut adjacent sections from the lesion ( n ) showed a staining pattern similar to that of Iba-1. iNOS expression was sparse in the lesions ( o ). p – u Accumulation of oxidative damage and iron in MHV-JHM coronavirus-induced lesions. Lesions showed abundant reactivity for oxidised phospholipids ( p , q , s E06) in myelin ( p ), in cells with apoptotic nuclei ( q ), in macrophage granules ( q ) and in axonal spheroids ( s ). In addition, nuclei within the lesions contained oxidised DNA (8OHdH reactivity; r ). Most lesions did not show iron deposition ( t ), although iron-positive perivascular macrophages accumulated in individual lesions ( u ). Scale bar 50 μm except for a – d = 0.5 mm

Article Snippet: Ox8 , Mouse (mAb) , Anti-rat CD8 , 1:400 , 0 , Seralab; MAS041.

Techniques: Infection, Preserving, Virus, Immunohistochemistry, Activation Assay, Immunocytochemistry, Expressing, Staining