Additional file 2 of A four-stage model for murine natural killer cell development in vivo

Additional file 2. Materials and Methods

Additional file 1 of A four-stage model for murine natural killer cell development in vivo

Additional file 1. Figure S1. Functional characteristics of four developmental stages of murine NK cells

Endosomal pH-Activatable Poly(ethylene oxide)-<i>graft</i>-Doxorubicin Prodrugs: Synthesis, Drug Release, and Biodistribution in Tumor-Bearing Mice

Novel poly(ethylene oxide)-graft-doxorubicin (PEO-g-DOX) prodrugs with DOX covalently conjugated to PEO via a pH-sensitive hydrazone bond were developed. PEO-g-DOX conjugates could be readily prepared in the following steps: (i) anionic ring-opening copolymerization of ethylene oxide (EO) and allyl glycidyl ether (AGE) afforded functional PEO with controlled molecular weights, low polydispersities, and multiple pendant double bonds (PEO-g-allyl); (ii) conjugation of PEO-g-allyl with methyl mercaptoacetate, followed by treating with hydrazine hydrate, quantitatively transformed allyl into hydrazide groups (PEO-g-hydrazide); and (iii) DOX was covalently immobilized to PEO-g-hydrazide via acid-labile hydrazone bonds (PEO-g-DOX). Here on the basis of PEO-g-allyl4.4 (Mn GPC = 22 400, PDI = 1.19) and PEO-g-allyl7.1 (Mn GPC = 15 300, PDI = 1.16, the subscription refers to number of allyl groups per chain) two freely water-soluble PEO-g-DOX prodrugs with 2.9 and 3.6 DOX per molecule (denoted as PEO-g-DOX2.9 and PEO-g-DOX3.6, corresponding to drug loading content of 5.6 and 9.0 wt %, respectively) were obtained. The in vitro release studies confirmed much faster release of DOX at pH 5.0 and 6.0 than at pH 7.4. For example, approximately 16, 52, and 61% of drug were released in 22 h, and 23, 83, and 92% of drug were released in 120 h from PEO-g-DOX2.9 at pH 7.4, 6.0 and 5.0, respectively. Notably, confocal laser scanning microscope (CLSM) observations revealed that DOX was released and delivered into the nuclei of RAW 264.7 cells following 24 h of incubation. MTT assays demonstrated that PEO-g-DOX2.9 had pronounced cytotoxic effects to RAW 264.7, HeLa, and 4T1 breast tumor cells with IC50 values of about 26.5, 42.5, and 32.0 μg DOX equiv/mL, whereas the corresponding polymer carrier PEO-g-hydrazide4.4 was nontoxic. The In Vivo pharmacokinetics and biodistribution studies in mice showed that PEO-g-DOX2.9 prodrugs had significantly prolonged circulation time and enhanced drug accumulation in the tumor as compared with free DOX. We are convinced that endosomal pH-activatable PEO-g-DOX prodrugs have tremendous potential for targeted cancer therapy

Supplemental Figure Legend from Combining DNA Vaccine and AIDA-1 in Attenuated <i>Salmonella</i> Activates Tumor-Specific CD4⁺ and CD8⁺ T-cell Responses

Supplemental Figure Legend</p

Supplementary Figure 1 from Combining DNA Vaccine and AIDA-1 in Attenuated <i>Salmonella</i> Activates Tumor-Specific CD4⁺ and CD8⁺ T-cell Responses

Representative flow cytometry plots for intracellular staining of TNF-alpha, IFN-gamma and IL-2 by gating on CD4+ or CD8+ T cells from splenocytes stimulated with B16-F10 lysates on day 14 in subcutaneous melanoma model.</p

Supplementary Data from The Mechanism of Anti–PD-L1 Antibody Efficacy against PD-L1–Negative Tumors Identifies NK Cells Expressing PD-L1 as a Cytolytic Effector

Supplementary figures and figure legends as well as supplementary methods and materials</p

Supplementary Figure 2 from Combining DNA Vaccine and AIDA-1 in Attenuated <i>Salmonella</i> Activates Tumor-Specific CD4⁺ and CD8⁺ T-cell Responses

Representative flow cytometry plots for intracellular staining of TNF-alpha, IFN-gamma and IL-2 of CD4+ and CD8+ T cells from splenocytes on day 14 in lung metastasis model stimulated with B16-F10 lysates.</p

Supplementary Data from An Oncolytic Virus Expressing IL15/IL15Rα Combined with Off-the-Shelf EGFR-CAR NK Cells Targets Glioblastoma

Supplementary Materials and Methods</p

DataSheet_1_B7H6 Serves as a Negative Prognostic Marker and an Immune Modulator in Human Pancreatic Cancer.pdf

Pancreatic cancer (PC), the third leading cause of cancer-related death in the U.S., is frequently found too late to be cured by traditional chemotherapy. Expression of B7 homolog 6 (B7H6), a member of the B7 family of immunoreceptors, has been found in PC and several other cancers. B7H6 is a ligand for cytotoxicity triggering receptor 3 (NKp30), which is expressed on NK cells. Here, we demonstrate that B7H6 can be detected in PC tissues but not normal organs. Its expression in patients associated significantly with tumor differentiation grade and lymphatic metastasis. The soluble form of B7H6 was detected in the PC patients’ sera, and its concentration associated with tumor differentiation grade and tumor, node, metastasis (TNM) stages. Also, higher levels of B7H6 in PC patients’ malignant tissues or serum correlated with shorter overall survival. In vitro, downregulation of B7H6 by CRISPR/Cas9 or siRNA technology had no significant impact on the viability or mobility of PC cells. Instead, knocking out B7H6 sensitized PC cells to NK-mediated cytotoxicity and cytokine production. These results indicate that B7H6 not only serves as a negative prognostic marker but also acts as an immune modulator in PC.</p

DataSheet_2_B7H6 Serves as a Negative Prognostic Marker and an Immune Modulator in Human Pancreatic Cancer.xlsx

Pancreatic cancer (PC), the third leading cause of cancer-related death in the U.S., is frequently found too late to be cured by traditional chemotherapy. Expression of B7 homolog 6 (B7H6), a member of the B7 family of immunoreceptors, has been found in PC and several other cancers. B7H6 is a ligand for cytotoxicity triggering receptor 3 (NKp30), which is expressed on NK cells. Here, we demonstrate that B7H6 can be detected in PC tissues but not normal organs. Its expression in patients associated significantly with tumor differentiation grade and lymphatic metastasis. The soluble form of B7H6 was detected in the PC patients’ sera, and its concentration associated with tumor differentiation grade and tumor, node, metastasis (TNM) stages. Also, higher levels of B7H6 in PC patients’ malignant tissues or serum correlated with shorter overall survival. In vitro, downregulation of B7H6 by CRISPR/Cas9 or siRNA technology had no significant impact on the viability or mobility of PC cells. Instead, knocking out B7H6 sensitized PC cells to NK-mediated cytotoxicity and cytokine production. These results indicate that B7H6 not only serves as a negative prognostic marker but also acts as an immune modulator in PC.</p