Journal: Frontiers in Immunology

Article Title: LPA 5 Is an Inhibitory Receptor That Suppresses CD8 T-Cell Cytotoxic Function via Disruption of Early TCR Signaling

doi: 10.3389/fimmu.2019.01159

Figure Lengend Snippet: LPA signaling via LPA 5 suppresses in vivo antigen-specific CD8 T cell cytotoxicity. (A) Frequency of OVA-reactive endogenous CD8 T cells (CD44+H2-K b -OVA tetramer+) in the spleens of C57BL/6 (open) or Enpp2 +/− (filled) mice 4 days after immunization with pI:C, anti-CD40 and OVA. (B) Panels show histograms of representative target cell killing 1 day after the transfer of target cells pulsed either with an irrelevant (HSV; left panel) or antigen-specific (OVA; right panel) peptide. Peptide-pulsed splenocytes were stained with a relatively high concentration of eFluor670 whereas unpulsed splenocytes are stained with a relatively low eFluor670 concentration. (C) Percent specific killing of irrelevant (HSV) peptide-pulsed splenocytes (open circles) and SIINFEKL (N4) peptide-pulsed splenocytes (filled circles) by endogenous SIINFEKL-specific CD8 T cells 4 days after immunization with anti-CD40, pI:C, and OVA. Percent specific killing was calculated as: (1– experimental ratio/pre-transfer ratio) × 100 and where the Ratio = %peptide–pulsed cells/% non-pulsed cells. *** p < 0.0005 using a one-tailed paired Student's t- test. (D) Percent antigen-specific killing of SIINFEKL-pulsed (left) or SIIGFEKL-pulsed (right) target cells 24 h after i.v. transfer of target cells into immunized mice. Data are cumulative of two experiments with 4–10 mice per group. * p < 0.05; *** p < 0.0005 using a one-tailed paired Student's t- test. (E) EG7 lymphoma cells were implanted into either WT, Lpar5 −/− , or Rag2 −/− hosts for 12 days after which they were isolated and weighed. Data are cumulative of 3 independent experiments reflecting tumors recovered from 13 WT hosts, 11 Lpar5 −/− hosts, and 5 Rag2 −/− hosts. * p < 0.05; ** p < 0.005 using a one-tailed paired Student's t- test. (F) B16.cOVA tumor cells were implanted into the flank of B6 mice for 5 days after which either wild type or Lpar5 −/− OT-I CD8 T cells were transferred i.v. into tumor bearing hosts. Fifteen days later recipients were sacrificed and tumors resected and weighed. * p < 0.05; data represents two independent experiments with 12–13 mice per group.

Article Snippet: The EG7 lymphoma cell line ( ) was obtained from ATCC (CRL-2113).

Techniques: In Vivo, Staining, Concentration Assay, One-tailed Test, Isolation

Journal: Leukemia

Article Title: Effect of ligand-activated estrogen receptor β on lymphoma growth in vitro and in vivo.

doi: 10.1038/leu.2011.68

Figure Lengend Snippet: Figure 1 Expression of ERb in murine lymphoma cells as shown by immunofluorescence. ERb expression was detected in the murine EG7 lymphoma cells using the anti-ERb 503 antibody (a). Staining for ERa was negative in EG7 cells (anti-ERa antibody MC-20) (b). To confirm the ERb specificity of the anti-ERb 503 antibody, ERb-negative murine Hepa1c1c7 cells were stained and showed to be negative (c), unless the cells were transfected with a murine ERb-expressing vector (d). Hepa1c1c7 cell expression of ERa was detected by using the anti-ERa MC-20 antibody (e). ERb protein levels were detected in both human Raji (f) and Ramos (g) lymphoma cells using the anti-ERb 503 antibody.

Article Snippet: Murine mouse T lymphoma cell line EG7,25 murine hepatoma Hepa1c1c7, human B-cell Burkitt’s lymphoma cell lines Raji and Ramos and HEK293 cells were obtained from ATCC.

Techniques: Expressing, Staining, Transfection, Plasmid Preparation

Journal: Frontiers in Immunology

Article Title: LTX-315 triggers anticancer immunity by inducing MyD88-dependent maturation of dendritic cells

doi: 10.3389/fimmu.2024.1332922

Figure Lengend Snippet: MyD88 knockout compromised the generation of anti-B16 immunity in response to LTX-315 (LTX) treatment. (A) Schedule of the experiment. C57BL/6J mice and MyD88 -/- mice (female, 8 wks, n = 8-10) were s.c. inoculated into the right flank with 1.5 x 10 5 B16F10 cells on day 0. Mice were i.t. treated with 3 consecutive daily injection of 1 mg LTX-315 or equal volume of PBS. Tumor-free mice were s.c. re-challenged with 1.5 x 10 5 B16F10 and 4 x 10 5 EG7 cells into the right flank and left flank, respectively. (B, C) , B16 tumor growth (B) and survival (C) . (D, E) , Incidence of EG7 and B16 tumors on Day 15 after rechallenge (D) and growth of re-challenged B16 tumors in LTX-315-cured WT and MyD88-/- (E) mice. *p < 0.05 and #p < 0.05 by Repeated Measures Analysis of Variance (ANOVA) and Log-rank test, respectively.

Article Snippet: Human melanoma A375, mouse melanoma B16F10, and mouse lymphoma EG7 cell lines were originally obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Knock-Out, Injection