Histopathology of <i>B</i>. <i>pahangi</i> infected and uninfected limbs in a mouse model.

<p>BALB/c mice were injected with 50 L3 of <i>B</i>. <i>pahangi</i> into the RHS footpad and with HBSS into the LHS footpad and analysed on the days p.i. as indicated. <b>(A)</b> uninfected mouse, Day 21, HBSS-injected limb: there is no inflammation in the fascial plane <b>(B)</b> infected mouse, Day 7: a filarial nematode is present within the lumen of a dilated lymphatic vessel in the fascial plane. <b>(C)</b> infected mouse, Day 14 prominent lymphangiectasis and lymphangitis in the fascial plane. <b>(D)</b> infected mouse, Day 21: marked inflammatory infiltration obliterating the lumen of a lymphatic vessel. Inset: remnants of degenerate nematodes within the lumen surrounded by inflammatory cells. <b>(E)</b> infected mouse (same animal as section C), Day 14: the inflammatory infiltrate affecting the lymphatic vessel in the fascial plane is characterised by the presence of many eosinophils highlighted by the modified haematoxylin eosin method. <b>(F)</b> infected mouse, Day 21: nematodes surrounded by intense inflammatory infiltration comprising large numbers of eosinophils highlighted by the modified Congo Red protocol.</p

<i>In vivo</i> imaging of MPO-specific bioluminescence in mice infected with <i>B</i>. <i>pahangi</i> L3 larvae.

<p>BALB/c mice were injected with 50 L3 of <i>B</i>. <i>pahangi</i> into the RHS footpad (<i>B</i>.<i>p</i>) and with HBSS into the LHS footpad. On day 4 (<b>A</b>) and day 11 (<b>B</b>) post-infection mice were imaged using an IVIS spectrum 20 minutes after subcutaneous injection of 200 mg/kg luminol. Representative images of 3 mice per group are shown. The colour scale indicates bioluminescence radiance in photons/second/cm²/steradian. Graphs show the bioluminescence total flux (in photons/second) within the same footpad region of interest (ROI, red ovals in images). Each symbol shows the total flux for a single mouse, lines indicate the means (n = 7 mice) and error bars show SD (**<i>p</i> < 0.01 using a Mann-Whitney test).</p

<i>In vivo</i> imaging of MPO-specific bioluminescence in mice implanted with adult <i>B</i>. <i>pahangi</i>.

<p>BALB/c mice were infected intraperitoneally by transplantation with 10 female adult <i>B</i>. <i>pahangi (B</i>.<i>p)</i>. Sham-operated mice (Sham) received no worms and control mice (Ctl) received an intraperitoneal injection of HBSS but underwent no surgery. On day 17 post-infection mice were imaged using an IVIS spectrum 20 minutes after subcutaneous injection of 200 mg/kg luminol. Representative images of 2 mice per group are shown. The colour scale indicates bioluminescence radiance in photons/second/cm²/steradian. Graphs show the bioluminescence total flux (in photons/second) within the same abdominal region of interest. Each symbol shows the total flux for a single mouse, lines indicate the means (n = 4–6 mice) and error bars show SD (*<i>p</i> < 0.05 using a one-way ANOVA with Dunn’s post-test).</p

Additional file 1: of Serum miR-22 as potential non-invasive predictor of poor clinical outcome in newly diagnosed, uniformly treated patients with diffuse large B-cell lymphoma: an explorative pilot study

Fig. S1. Kaplan-Meier Overall Survival curves. Correlation between the indicated miRNAs and the Overall Survival of 47 DLBCL patients from TCGA data analysis. (PDF 204 kb

Image_2_Immunogenicity of SARS-CoV-2 vaccination in patients undergoing autologous stem cell transplantation. A multicentric experience.jpeg

COVID-19 disease has a strong impact on hematological patients; those receiving autologous hematopoietic stem cell transplantation (aHSCT) represent a particularly vulnerable group, in which the effectiveness of vaccination is very variable. Chiarucci et al. showed that patients affected by non-Hodgkin lymphoma (NHL) and treated with rituximab experienced a lower rate of immunization against SARS-CoV-2 (54%), as well as significantly lower IgG antibody titers. In our multicenter retrospective observational study, we included 82 patients who underwent aHSCT, divided into two groups: 58 patients vaccinated after aHSCT (group A) and 24 vaccinated before getting transplantation (group B). In group A, 39 (67%) patients had positive serology, and the rate of positivity increased with time after aHSCT. In the subgroup of patients with NHL, the administration of rituximab predicted negative serology, particularly when administered in the 6 months before vaccination (13% response rate). Patients affected by plasma cells had a higher rate of positivity (83% overall), independently of the time to aHSCT. In group B, no patient who initially showed positive serology became negative after transplantation, so the aHSCT did not affect the response to the vaccination. Our study confirmed the role of rituximab as a negative predictor of response to SARS-CoV-2 vaccination, whereas the conditioning and transplantation procedure itself seemed to be less important.</p

Image_1_Immunogenicity of SARS-CoV-2 vaccination in patients undergoing autologous stem cell transplantation. A multicentric experience.jpeg

COVID-19 disease has a strong impact on hematological patients; those receiving autologous hematopoietic stem cell transplantation (aHSCT) represent a particularly vulnerable group, in which the effectiveness of vaccination is very variable. Chiarucci et al. showed that patients affected by non-Hodgkin lymphoma (NHL) and treated with rituximab experienced a lower rate of immunization against SARS-CoV-2 (54%), as well as significantly lower IgG antibody titers. In our multicenter retrospective observational study, we included 82 patients who underwent aHSCT, divided into two groups: 58 patients vaccinated after aHSCT (group A) and 24 vaccinated before getting transplantation (group B). In group A, 39 (67%) patients had positive serology, and the rate of positivity increased with time after aHSCT. In the subgroup of patients with NHL, the administration of rituximab predicted negative serology, particularly when administered in the 6 months before vaccination (13% response rate). Patients affected by plasma cells had a higher rate of positivity (83% overall), independently of the time to aHSCT. In group B, no patient who initially showed positive serology became negative after transplantation, so the aHSCT did not affect the response to the vaccination. Our study confirmed the role of rituximab as a negative predictor of response to SARS-CoV-2 vaccination, whereas the conditioning and transplantation procedure itself seemed to be less important.</p

Table_1_Immunogenicity of SARS-CoV-2 vaccination in patients undergoing autologous stem cell transplantation. A multicentric experience.docx

COVID-19 disease has a strong impact on hematological patients; those receiving autologous hematopoietic stem cell transplantation (aHSCT) represent a particularly vulnerable group, in which the effectiveness of vaccination is very variable. Chiarucci et al. showed that patients affected by non-Hodgkin lymphoma (NHL) and treated with rituximab experienced a lower rate of immunization against SARS-CoV-2 (54%), as well as significantly lower IgG antibody titers. In our multicenter retrospective observational study, we included 82 patients who underwent aHSCT, divided into two groups: 58 patients vaccinated after aHSCT (group A) and 24 vaccinated before getting transplantation (group B). In group A, 39 (67%) patients had positive serology, and the rate of positivity increased with time after aHSCT. In the subgroup of patients with NHL, the administration of rituximab predicted negative serology, particularly when administered in the 6 months before vaccination (13% response rate). Patients affected by plasma cells had a higher rate of positivity (83% overall), independently of the time to aHSCT. In group B, no patient who initially showed positive serology became negative after transplantation, so the aHSCT did not affect the response to the vaccination. Our study confirmed the role of rituximab as a negative predictor of response to SARS-CoV-2 vaccination, whereas the conditioning and transplantation procedure itself seemed to be less important.</p

Table_2_Immunogenicity of SARS-CoV-2 vaccination in patients undergoing autologous stem cell transplantation. A multicentric experience.docx

COVID-19 disease has a strong impact on hematological patients; those receiving autologous hematopoietic stem cell transplantation (aHSCT) represent a particularly vulnerable group, in which the effectiveness of vaccination is very variable. Chiarucci et al. showed that patients affected by non-Hodgkin lymphoma (NHL) and treated with rituximab experienced a lower rate of immunization against SARS-CoV-2 (54%), as well as significantly lower IgG antibody titers. In our multicenter retrospective observational study, we included 82 patients who underwent aHSCT, divided into two groups: 58 patients vaccinated after aHSCT (group A) and 24 vaccinated before getting transplantation (group B). In group A, 39 (67%) patients had positive serology, and the rate of positivity increased with time after aHSCT. In the subgroup of patients with NHL, the administration of rituximab predicted negative serology, particularly when administered in the 6 months before vaccination (13% response rate). Patients affected by plasma cells had a higher rate of positivity (83% overall), independently of the time to aHSCT. In group B, no patient who initially showed positive serology became negative after transplantation, so the aHSCT did not affect the response to the vaccination. Our study confirmed the role of rituximab as a negative predictor of response to SARS-CoV-2 vaccination, whereas the conditioning and transplantation procedure itself seemed to be less important.</p

Histological and immunohistochemical analysis of different tissues.

<p>SCID mice were inoculated iv with 5×10⁶ AML-NS8 cells, monitored for clinical signs and sacrificed upon manifestations of terminal disease. Different tissues were collected and fixed for histology and immunohistochemistry. (<b>A</b>) Clinical signs and histopathological characterisation of AML-NS8 <i>in vivo</i> disseminated model. (<b>B, C</b>) Representative pictures (Axio Scope Zeiss, magnification×200) of the indicated tissues stained with H&E (left panels) or anti-human HLA-A,B,C (right panels). Black/white bar, 100 µm.</p

Karyotype and Genomic aberrations of AML-NS8 from different sources.

a<p>M: metaphases.</p>b<p>ND: not done.</p>c<p>x: presence of aberration.</p