Direct CD32 T-cell cytotoxicity: implications for breast cancer prognosis and treatment

The FcγRII (CD32) ligands are IgFc fragments and pentraxins. The existence of additional ligands is unknown. We engineered T cells with human chimeric receptors resulting from the fusion between CD32 extracellular portion and transmembrane CD8α linked toCD28/ζ chain intracellular moiety (CD32-CR). Transduced T cells recognized three breast cancer (BC) and one colon cancer cell line among 15 tested in the absence of targeting antibodies. Sensitive BC cell conjugation with CD32-CR T cells induced CD32 polarization and down-regulation, CD107a release, mutual elimination, and proinflammatory cytokine production unaffected by human IgGs but enhanced by cetuximab. CD32-CR T cells protected immunodeficient mice from subcutaneous growth of MDA-MB-468 BC cells. RNAseq analysis identified a 42 gene fingerprint predicting BC cell sensitivity and favorable outcomes in advanced BC. ICAM1 was a major regulator of CD32-CR T cell–mediated cytotoxicity. CD32-CR T cells may help identify cell surface CD32 ligand(s) and novel prognostically relevant transcriptomic signatures and develop innovative BC treatments

Enhancement of anti-leukemia activity of NK cells in vitro and in vivo by inhibition of leukemia cell-induced NK cell damage

Acute myeloid leukemia (AML) cells induce, in vitro, NK cell abnormalities (NKCAs) including apoptosis and activating receptor down-regulation. The potential negative impact of AML cells on the therapeutic efficacy of NK cell-based strategies prompted us to analyze the mechanisms underlying NKCAs and to develop approaches to protect NK cells from NKCAs. NKCA induction by the AML leukemia cells target a subpopulation of peripheral blood NK cells and is interleukin-2 independent but is abrogated by a long-term culture of NK (LTNK) cells at 37°C. LTNK cells displayed a significantly enhanced ability to damage AML cells in vitro and inhibited the subcutaneous growth of ML-2 cells grafted into CB17 SCID mice. Actinomycin D restored the susceptibility of LTNK cells to NKCAs while TAPI-0, a functional analog of the tissue inhibitor of metalloproteinase (TIMP) 3, inhibits ML-2 cell-induced NKCAs suggesting that the generation of NK cell resistance to NKCAs involves RNA transcription and metalloproteinase (MPP) inactivation. This conclusion is supported by the reduced susceptibility to AML cell-induced NKCAs of LTNK cells in which TIMP3 gene and protein are over-expressed. This information may contribute to the rational design of targeted strategies to enhance the efficacy of NK cell-based-immunotherapy of AML with haploidentical NK cells

Natural humoral immune response to ribosomal P0 protein in colorectal cancer patients

Tumor associated antigens are useful in colorectal cancer (CRC) management. The ribosomal P proteins (P0, P1, P2) play an important role in protein synthesis and tumor formation. The immunogenicity of the ribosomal P0 protein in head and neck, in breast and prostate cancer patients and the overexpression of the carboxyl-terminal P0 epitope (C-22 P0) in some tumors were reported

The FLUXNET2015 dataset and the ONEFlux processing pipeline for eddy covariance data

The FLUXNET2015 dataset provides ecosystem-scale data on CO2, water, and energy exchange between the biosphere and the atmosphere, and other meteorological and biological measurements, from 212 sites around the globe (over 1500 site-years, up to and including year 2014). These sites, independently managed and operated, voluntarily contributed their data to create global datasets. Data were quality controlled and processed using uniform methods, to improve consistency and intercomparability across sites. The dataset is already being used in a number of applications, including ecophysiology studies, remote sensing studies, and development of ecosystem and Earth system models. FLUXNET2015 includes derived-data products, such as gap-filled time series, ecosystem respiration and photosynthetic uptake estimates, estimation of uncertainties, and metadata about the measurements, presented for the first time in this paper. In addition, 206 of these sites are for the first time distributed under a Creative Commons (CC-BY 4.0) license. This paper details this enhanced dataset and the processing methods, now made available as open-source codes, making the dataset more accessible, transparent, and reproducible.Peer reviewe

Potenziamento dell’efficacia del cetuximab mediante combinazione con cellule T ingegnerizzate con recettori Fc chimerici per il trattamento del carcinoma del colon retto K-ras mutato.

INTRODUZIONE: L’impiego di anticorpi monoclonali (AM) diretti contro il recettore per il fattore di crescita epidermico (EGFR), tra cui il cetuximab, ha rappresentato un significativo avanzamento nel trattamento del carcinoma colorettale metastatico (mCRC). L’effetto anti-tumorale del cetuximab è dovuto a: i) blocco della proliferazione neoplastica; ii) attivazione di processi apoptotici; iii) induzione di citotossicità cellulare dipendente da anticorpo (ADCC) mediante il legame con i recettori Fc. Tuttavia, diversi fattori limitano l’efficacia terapeutica del cetuximab; tra questi la presenza di mutazioni nel gene Kras (Kras-mut) e la scarsa presenza di cellule Natural Killer, capaci di mediare ADCC, nel microambiente tumorale. In questo studio ipotizziamo di superare i limiti terapeutici del cetuximab attraverso la sua combinazione con cellule T ingegnerizzate con recettori chimerici Fc (Fc-CRs). MATERIALI E METODI: Due Fc-CRs, denominati CD32-CR e CD16-CR, sono stati generati dalla fusione della regione extracellulare del FcRIIA (CD32) o del FcRIIIA (CD16) con il dominio transmembrana del CD8a e con i domini intracitoplasmatici del CD28 e del CD3. Linfociti T attivati sono stati trasdotti con le chimere e la corretta espressione è stata valutata mediante citofluorimetria e western blot. I recettori sono stati caratterizzati funzionalmente in vitro attraverso test di binding e saggi ELISA. L’attività anti-tumorale delle cellule T trasdotte in associazione al cetuximab è stata valutata in test di deplezione in vitro e in topi SCID xenotrapiantati con la linea cellulare di CRC Kras-mut HCT116. RISULTATI: Entrambi i Fc-CRs sono espressi correttamente sulla superficie delle cellule T. Il CD32-CR, ma non il CD16-CR, lega in maniera specifica il frammento Fc degli AM anti-EGFR cetuximab e panitumumab. La dose massima di cetuximab necessaria per saturare tutti i CD32-CR che è pari a 10g/ml ed il binding è mantenuto anche in presenza di quantità crescenti di plasma umano. Sia il CD32-CR che il CD16-CR inducono rilascio di citochine in risposta a stimolazione con cetuximab o panitumumab. La combinazione del cetuximab con cellule T CD32-CR+: i) induce l’eliminazione di HCT116 in vitro; ii) riduce significativamente la crescita tumorale in vivo. CONCLUSIONI: In questo studio preclinico dimostriamo che cellule T armate con CD32-CR potenziano l’efficacia del cetuximab contro CRC Kras-mutato

Treatment with Human Placental Lactogen (hPL-A) Improves Glucose Homeostasis One Year after Pancreatic Islets Transplantation in Mice Anterior Eye Chamber

Type 1 diabetes (T1D) is a chronic disease characterized by destruction of insulin producing beta cells in the pancreas. In addition to conventional insulin treatment, islets transplantation represents an alternative and promising therapeutic approach to treat patients with T1D. Increasing evidences suggest that human placental lactogen (hPL), a polypeptide placental hormone, is able to promote islets survival and proliferation in vitro, however it is unknown its implication in vivo models. Therefore, in the present study, we used hPL isoform A (hPL-A) to evaluate the role of this hormone in murine engrafted islets. T1D was induced in C57BL6/J male mice by a single intraperitoneal high dose (180 mg/kg) of streptozotocin (STZ). Collagenase p and a Histopaque 1110 gradient solution were used to isolate pancreatic islets. The anterior chamber of the eye was used to inject isolated islets. The engraftment was evaluated by confocal microscopy and immunohistochemical staining on section of the right and left eye. Glucose tolerance test (GTT) was used to assess glucose tolerance in mice. First, we evaluated the engraftment of freshly isolated pancreatic islets. Iris dissections showed a positive staining for insulin and glucagon. Second, since 2 months after transplantation, STZ-treated mice injected with islets stimulated with hPL-A, showed an improvement in fed glycaemia compared to STZ-treated mice transplanted with islets alone, and non-transplanted STZ-treated mice, for a follow-up of 1 year. Furthermore, mice treated with hPL-A, showed GTT with a similar pattern to untreated control mice; islets vascularization and angiogenesis was showed using CD31. Taken together, present results provide evidence that hPL-A could have a long-term effect on islet graft function and survival through enhancing glucose homeostasis