Immune checkpoint inhibitor therapy increases systemic SDF-1, cardiac DAMPs Fibronectin-EDA, S100/Calgranulin, galectine-3, and NLRP3-MyD88-chemokine pathways.

Background: Immune checkpoint inhibitors (ICIs) have significantly changed the oncology clinic in recent years, improving survival expectations in cancer patients. ICI therapy have a broad spectrum of side effects from endocrinopathies to cardiovascular diseases. In this study, pro-inflammatory and pro-fibrotic effects of short-term ICIs therapy in preclinical models were analyzed. Methods: Firstly, in a human in vitro model, human cardiomyocytes co-cultured with hPBMC were exposed to ICIs (with CTLA-4 or PD-1 blocking agents, at 200 nM) for 72 h. After treatment, production of DAMPs and 12 cytokines were analyzed in the supernatant through colorimetric and enzymatic assays. C57/Bl6 mice were treated with CTLA-4 or PD-1 blocking agents (15 mg/kg) for 10 days. Before (T0), after three days (T3) and after treatments (T10), ejection fraction, fractional shortening, radial and longitudinal strain were calculated by using bidimensional echocardiography (Vevo 2100, Fujfilm). Fibrosis, necrosis, hypertrophy and vascular NF-kB expression were analyzed through Immunohistochemistry. Myocardial expression of DAMPs (S100- Calgranulin, Fibronectin and Galectine-3), MyD88, NLRP3 and twelve cytokines have been analyzed. Systemic levels of SDF-1, IL-1ß, and IL-6 were analyzed before, during and after ICIs therapy. Results: Radial and longitudinal strain were decreased after 10 days of ICIs therapy. Histological analysis of NF-kB expression shows that short-term anti-CTLA-4 or anti-PD-1 treatment increased vascular and myocardial inflammation. No myocardial hypertrophy was seen with the exception of the pembrolizumab group. Myocardial fibrosis and expression of galectin-3, pro-collagen 1-α and MMP-9 were increased after treatment with all ICIs. Both anti-CTLA-4 or anti-PD-1 treatments increased the expression of DAMPs, NLRP3 inflammasome and MyD88 and induced both in vitro and in vivo the secretion of IL-1ß, TNF-α and IL-6. Systemic levels of SDF-1, IL-1ß and IL-6 were increased during and after treatment with ICIs. Conclusions: Short therapy with PD-1 and CTLA-4 blocking agents increases vascular expression of NF-kB, systemic SDF-1, IL-1ß, IL-6 levels and myocardial NLRP3, MyD88 and DAMPs expression in preclinical models. A pro-inflammatory cytokine storm was induced in myocardial tissues and in cultured cardiac cells after ICIs therapy. The overall picture of the study suggests new putative biomarkers of ICIs-mediated systemic and myocardial damages potentially useful in clinical cardioncology.

Lymph node fine needle cytology in the diagnosis of infectious diseases and reactive unspecific processes.

Infectious diseases are one of the main causes of lymph node enlargement both in children and adults and represent a benign and reversible process. Clinical evaluation, serological data, microbiological and molecular tests and imaging techniques are generally used in the diagnosis of reactive lymph nodes determined by infectious diseases but, in some cases, do not assess their origin and nature. Surgical excision and histological control represent the gold standard in the diagnosis of lymphadenopaties, but they might be unnecessary procedures just for diagnostic purposes in benign reactive lymph nodal enlargement. Fine Needle Cytology (FNC) has gained a definitive role in the diagnosis of lymphadenopaties being an accurate, rapid, minimal invasive and cost-effective procedure useful for the clinical management and therapeutic decisions. This study reports the use of FNC in the diagnosis of reactive lymph nodes in infectious diseases, exploring possibilities and limitations of the technique in this specific clinical setting.

Fine needle cytology, infectious diseases and non-Hodgkin lymphoma.

The association between some infections and non-Hodgkin lymphomas (NHL) is well known. Human T-cell leukemia/lymphoma virus type 1 (HTLV-1) was the first oncogenic human retrovirus to be discovered and has been found to be associated with adult T-cell leukemia/lymphoma (ATLL). Epstein-Barr virus (EBV) has consistently been linked to both endemic and sporadic Burkitt lymphoma (BL), as well as to Hodgkin lymphoma (HL), post-transplantation proliferative disorders, extra-nodal NK-T-cell lymphoma (nasal type) and B-cell NHL arising in HIV patients; HCV infection is also associated to low-grade lymphoproliferative disorders that can progress to NHL. Bacterial infections have also been associated to NHL; chronic gastritis caused by Helicobacter pylori is responsible for mucosa-associated lymphoid tissue (MALT) NHL and high prevalence of Chlamydia psittaci infections has been reported in ocular adnexal lymphomas. In both these conditions, infection may contribute to the development of lymphomas, as proven by the clinical responses eradicating antibiotic therapies. Histological diagnosis coupled with immunohistochemical and molecular procedures are needed for a definitive diagnosis, but Fine Needle Cytology (FNC) combined with ancillary techniques can also produce correct diagnoses in most cases. In patients suffering from NHL, FNC also plays an important role in differential diagnosis between relapse of primary disease and reactive lymph nodes enlargement. This review explores the role of FNC in the diagnosis and classification of NHL trying to highlight possibilities and the limitations of the technique.