LacZ inducible, antigen/MHC-specific T cell hybrids

Sanderson, Sarah; Shastri, Nilabh

doi:10.1093/intimm/6.3.369

Cited by 352 publications

(371 citation statements)

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“…27 Thus we were able to confirm the results obtained by IL-2 ELISA by assaying ␤-galactosidase activity following exposure to treated BMDC. A 4.8-fold increase over background in ␤-galactosidase-expressing T cells was observed in PLG-CTAB-pCMVKm2.GagMod.SF2-treated BMDC; the average number of ␤-gal positive cells per 800 m 2 field was 37 ± 12 versus 180 ± 64, respectively (n = 4; P р 0.01).…”

Section: Antigen Processing and Presentation To An H-2k D Restricted supporting

confidence: 74%

“…The hybridoma used in this assay was generated using the lacZ-inducible BWZ.36 fusion partner that contains the Escherichia coli lacZ reporter gene under the control of the nuclear factor of activated T cells (NFAT) enhancer element of the IL-2 gene. 27 Following removal of culture supernatants, remaining cells were stained for ␤-galactosidase activity using the Invitrogen ␤-gal staining kit according to the manufacturer's instructions (Invitrogen, Carlsbad, CA, USA). Relative numbers of stimulated hybridoma cells were determined by averaging the numbers of ␤-galactosidase positive cells counted under an 800 m 2 field of an inverted light microscope.…”

Section: Il-2 Production By T Cell Hybridomamentioning

confidence: 99%

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Plasmid DNA adsorbed onto cationic microparticles mediates target gene expression and antigen presentation by dendritic cells

et al. 2000

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Dendritic cells (DC) play a key role in antigen presentation and activation of specific immunity. Much current research focuses on harnessing the potency of DC for vaccines, gene therapy, and cancer immunotherapy applications. However, DC are not readily transfected in vitro by traditional nonviral techniques. A novel DNA vaccine formulation was used to determine if DC are transfected in vitro. The formulation consists of plasmid DNA adsorbed on to cationic microparticles composed of the biodegradable polymer polylactide-co-glycolide (PLG) and the cationic surfactant, cetyltrimethylammonium bromide (CTAB). Using preparations of fluorescentlabeled plasmid DNA formulated on PLG-CTAB microparticles to study internalization by macrophages and dendritic cells in vitro and in vivo, we found that most, but not all, of

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Section: Antigen Processing and Presentation To An H-2k D Restricted supporting

confidence: 74%

Section: Il-2 Production By T Cell Hybridomamentioning

confidence: 99%

Plasmid DNA adsorbed onto cationic microparticles mediates target gene expression and antigen presentation by dendritic cells

et al. 2000

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“…DCs were enriched from tumor-draining lymph nodes 48 h after ablation and cocultured with B3Z cells for 24 h at 37jC in 96-well plates. The presentation by DCs of SIINFEKL in H-2K b results in production of h-galactosidase by B3Z cells (23), which can be detected by adding 0.15 mmol/L chlorophenolred-h-D-galactopyranoside (Calbiochem), 9 mmol/L MgCl 2 , 0.125% NP40, and 100 mmol/L h-ME in PBS. Plates were incubated for 3 h at 37jC and absorbance values were measured using a photo spectrometer (595 nm).…”

Section: Methodsmentioning

confidence: 99%

In vivo Colocalization of Antigen and CpG within Dendritic Cells Is Associated with the Efficacy of Cancer Immunotherapy

Nierkens¹,

Brok²,

Sutmuller³

et al. 2008

Cancer Research

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Immunostimulatory cytidyl guanosyl (CpG) motifs are of great interest as cancer vaccine adjuvants. They act as potent inducers of Th1 responses, including the activation of cytotoxic CD8 + T lymphocytes (CTL). Whereas animal models have provided clear evidence that CpG enhances antitumor immunity, clinical trials in humans have thus far been less successful. Applying cryosurgery as an instant in situ tumor destruction technique, we now show that timing of CpG administration crucially affects colocalization of antigen and CpG within EEA-1 + and LAMP-1 + compartments within dendritic cells in vivo. Moreover, antigen/CpG colocalization is directly correlated with antigen cross-presentation, the presence of CTL, and protective antitumor immunity. Thus, failure or success of CpG as a vaccine adjuvant may depend on colocalization of antigen/CpG inside DCs and hence on the timing of CpG administration. These data might aid in the design of future immunotherapeutic strategies for cancer patients. [Cancer Res 2008;68(13):5390-6]

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“…The BWZ cell line was derived from BW5147 T cells and contained a lacZ reporter gene as described previously (35). The cells were transfected to surface-express TREM-2A or TREM-3 linked to a CD3ζ cytoplasmic domain.…”

Section: Reporter Cell Assaymentioning

confidence: 99%

TREM-2 binds to lipooligosaccharides of Neisseria gonorrhoeae and is expressed on reproductive tract epithelial cells

et al. 2008

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Triggering receptor expressed on myeloid cells-2 (TREM-2) is an innate immune receptor that initiates cellular activation upon ligation. In this study, we examined the interaction of TREM-2 with Neisseria gonorrhoeae using murine TREM-2A as it has been reported to recognize bacterial ligands. Using a whole bacteria ELISA, TREM-2A bound to all 6 strains in variable degrees. Far western blots of gonococcal outer membranes revealed TREM-2A binding to lipooligosaccharide (LOS) and Opa protein with predominant binding to LOS. Binding of TREM-2A to LOS was confirmed by ELISA and surface plasmon resonance. O-deacylation of the lipid A significantly reduced binding. Flow cytometry and reporter cell assays showed that gonococci bound to TREM-2A-transfected cells and induced transmembrane signaling. In humans, TREM-2 was constitutively expressed by genitourinary and fallopian tube epithelial cells, both of which are primary targets of gonococcal invasion. Ligation of TREM-2 by LOS induced IL-6 production in HeLa cervical carcinoma cells. To our knowledge, this is the first report of the expression of human TREM-2 by cells deriving from a non-myeloid lineage. We conclude that gonococci can interact with TREM-2 receptors through binding to LOS and Opa protein and initiate cell signaling and cytokine production.

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LacZ inducible, antigen/MHC-specific T cell hybrids

Cited by 352 publications

References 0 publications

Plasmid DNA adsorbed onto cationic microparticles mediates target gene expression and antigen presentation by dendritic cells

Plasmid DNA adsorbed onto cationic microparticles mediates target gene expression and antigen presentation by dendritic cells

In vivo Colocalization of Antigen and CpG within Dendritic Cells Is Associated with the Efficacy of Cancer Immunotherapy

TREM-2 binds to lipooligosaccharides of Neisseria gonorrhoeae and is expressed on reproductive tract epithelial cells

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