Chimeric antigen receptor T cells to target CD79b in B-cell lymphomas.

BACKGROUND: Chimeric antigen receptor (CAR) T cells targeting CD19 mediate potent and durable effects in B-cell malignancies. However, antigen loss or downregulation is a frequent cause of resistance. Here, we report development of a novel CAR T-cell therapy product to target CD79b, a pan B-cell antigen, widely expressed in most B-cell lymphomas. METHODS: We generated a novel anti-CD79b monoclonal antibody by hybridoma method. The specificity of the antibody was determined by testing against isogenic cell lines with human CD79b knock-in or knock-out. A single-chain variable fragment derived from the monoclonal antibody was used to make a panel of CD79b-targeting CAR molecules containing various hinge, transmembrane, and co-stimulatory domains. These were lentivirally transduced into primary T cells and tested for antitumor activity in in vitro and in vivo B-cell lymphoma models. RESULTS: We found that the novel anti-CD79b monoclonal antibody was highly specific and bound only to human CD79b and no other cell surface protein. In testing the various CD79b-targeting CAR molecules, superior antitumor efficacy in vitro and in vivo was found for a CAR consisting CD8α hinge and transmembrane domains, an OX40 co-stimulatory domain, and a CD3ζ signaling domain. This CD79b CAR specifically recognized human CD79b-expressing lymphoma cell lines but not CD79b knock-out cell lines. CD79b CAR T cells, generated from T cells from either healthy donors or patients with lymphoma, proliferated, produced cytokines, degranulated, and exhibited robust cytotoxic activity in vitro against CD19+ and CD19- lymphoma cell lines and patient-derived lymphoma tumors relapsing after prior CD19 CAR T-cell therapy. Furthermore, CD79b CAR T cells were highly efficient at eradicating pre-established lymphoma tumors in vivo in three aggressive lymphoma xenograft models, including two cell line-derived xenografts and one patient-derived xenograft. Notably, these CAR T cells did not demonstrate any significant tonic signaling activity or markers of exhaustion. CONCLUSION: Our results indicated that this novel CD79b CAR T-cell therapy product has robust antitumor activity against B-cell lymphomas. These results supported initiation of a phase 1 clinical trial to evaluate this product in patients with relapsed or refractory B-cell lymphomas.

Thin film notch filters as platforms for biological image processing.

Many image processing operations involve the modification of the spatial frequency content of images. Here we demonstrate object-plane spatial frequency filtering utilizing the angular sensitivity of a commercial spectral bandstop filter. This approach to all-optical image processing is shown to generate real-time pseudo-3D images of transparent biological and other samples, such as human cervical cancer cells. This work demonstrates the potential of non-local, non-interferometric approaches to image processing for uses in label-free biological cell imaging and dynamical monitoring.

Nanophotonics enhanced coverslip for phase imaging in biology.

The ability to visualise transparent objects such as live cells is central to understanding biological processes. Here we experimentally demonstrate a novel nanostructured coverslip that converts phase information to high-contrast intensity images. This compact device enables real-time, all-optical generation of pseudo three-dimensional images of phase objects on transmission. We show that by placing unstained human cancer cells on the device, the internal structure within the cells can be clearly seen. Our research demonstrates the significant potential of nanophotonic devices for integration into compact imaging and medical diagnostic devices. The nanophotonics enhanced coverslip (NEC) enables ultra-compact phase imaging of samples placed directly on top of the device. Visualisation of artificial phase objects and unstained biological cells is demonstrated.

Evidence for the expansion of pediatric cochlear implant candidacy.

OBJECTIVE: To test the hypothesis that children who are non-traditional cochlear implant candidates, but are not making progress with appropriately fitted hearing aids and intervention, will demonstrate significant benefit from cochlear implantation as defined by improvement in (1) speech perception, (2) auditory skills development, and/or (3) progress on standardized measures of receptive and expressive language. STUDY DESIGN: Retrospective case series. SETTING: Two tertiary academic cochlear implant centers. PATIENTS: All pediatric patients that underwent cochlear implantation were reviewed. Only those meeting one or both of the following criteria were included: (1) less severe hearing loss than specified in the current indications and (2) open-set word and/or sentence recognition scores greater than 30% for children who are able to participate in speech perception testing. Patients with auditory neuropathy were excluded. INTERVENTION(S): Cochlear implantation. MAIN OUTCOME MEASURES: Pre- and postoperative results of age appropriate speech recognition tests, auditory questionnaires, and standardized norm-referenced estimates of speech and language development. RESULTS: A total of 51 patients met study criteria. The mean age at time of surgery was 8.3 years and 24% underwent bilateral sequential implantation. Overall, the mean speech recognition improvement was 63 percentage points in the implanted ear (p < 0.001) and 40 percentage points in the bimodal condition (p < 0.001). Results of auditory and language development measures revealed significant improvement after implantation (p < 0.05). CONCLUSION: Non-traditional pediatric implant recipients derive significant benefit from cochlear implantation. A large-scale reassessment of pediatric cochlear implant candidacy, including less severe hearing losses and higher preoperative speech recognition, is warranted to allow more children access to the benefits of cochlear implantation.