Biology subjects

Puccio, I.

Publications and source records attributed to Puccio, I..

2 recordsLinked to original sources

bioRxiv2025

In depth characterisation of the tumour microenvironment reveals HHV-8 dependent immune regulation in HIV-associated and classic Kaposi Sarcoma.

BackgroundKaposis sarcoma (KS) is the most common malignancy occurring in people living with HIV (PLWH). The clinical course of KS in PLWH established on combined anti-retroviral therapy (cART) resembles that of classic KS. However, no studies have so far been conducted to compare clinical and immunologic characteristics of these subtypes. ObjectivesTo compare the clinical and biological characteristics of HIV-associated KS in patients with well-controlled viral infection (N=21) and non-HIV associated KS (N=19). MethodsClinical data were prospectively collected from patients treated at the National Centre for HIV malignancies at Chelsea & Westminster Hospital. Archival tumour samples were retrieved to perform targeted transcriptomic analysis using the Nanostring nCounter platform. The results were validated by multiplex immune-fluorescence. ResultsMedian progression free survival (mPFS) was comparable between groups. The tumour microenvironment (TME) of non-HIV associated KS was characterized by upregulation of pathways associated with adaptive and innate immunity and angiogenesis. Transcripts relating to glycolysis and endothelial to mesenchymal transition were upregulated in the TME of HIV-associated cases. Furthermore, the TME of HIV-associated cases was associated with lower infiltration of CD4 activated cell. In both cohorts, we found the expression of HHV-8 genes to positively correlate with activated CD4, CD8, NK and immune checkpoints gene expression. ConclusionsIn conclusion, we showed that the TME of KS in the presence of well-controlled HIV infection is characterized by a lower degree of inflammation and more pronounced epithelial to mesenchymal transition. We also identified intra-tumoral HHV-8 gene expression as a driver of TME composition.

immunology↗

bioRxiv2019

Megakaryocytes, erythropoietic and granulopoietic cells express CAL2 antibody in myeloproliferative neoplasms carrying CALR gene mutations

The discovery of mutated Calreticulin (CALR) in myeloproliferative neoplasms (MPN) has provided proof of clonality, diagnostic importance, and influence on prognosis of this pathology. The identification of this MPN-associated driver mutation -currently based on molecular assays- is represented as a major diagnostic criterion for essential thrombocythaemia (ET), pre-fibrotic myelofibrosis and primary myelofibrosis (PMF) in the updated World Health Organization (WHO) 2008 classification. In the present study, we validated by immunohistochemistry the diagnostic usefulness of the monoclonal CAL2 antibody. Cases of acute myeloid leukaemia (AML) and myelodysplastic/ myeloproliferative neoplasms (MDS/MPN) have been also investigated to assess the specificity of CAL2 antibody. For this purpose, the result of the CAL2 immunostaining was compared with the result of molecular assays. Additionally, we investigated by double staining whether expression of mutated CALR can also be demonstrated on cells of the erythroid and myeloid lineage. We confirmed the usefulness of the CAL2 monoclonal antibody in successfully detecting mutant CALR in bone marrow biopsies. We showed that the immune-reactivity of CAL2 was absolutely restricted to the presence of CALR mutations, which were seen only in ET and MDS/MPN biopsies, but not in AML biopsies (14/14). There was 100% concordance in biopsy specimens with the concomitant molecular results. We applied double staining technique and confirmed that a subpopulation of granulopoietic and erythropoietic cells express mutated CALR as demonstrated with the CAL2 antibody in cases of MPNs. This supports the suggestion that the CALR mutations occur in a multipotent progenitor capable of generating both myeloid and erythroid progeny with preferential expansion of megakaryocytic cell lineage as a result of CALR mutation in an immature hematopoietic stem cell.

pathology↗