Human Herpesvirus-8 Infection Leads to Expansion of the Preimmune/Natural Effector B Cell Compartment

BACKGROUND: Human herpesvirus-8 (HHV-8) is the etiological agent of Kaposi's sarcoma (KS) and of some lymphoproliferative disorders of B cells. Most malignancies develop after long-lasting viral dormancy, and a preventing role for both humoral and cellular immune control is suggested by the high frequency of these pathologies in immunosuppressed patients. B cells, macrophages and dendritic cells of peripheral lymphoid organs and blood represent the major reservoir of HHV-8. Due to the dual role of B cells in HHV-8 infection, both as virus reservoir and as agents of humoral immune control, we analyzed the subset distribution and the functional state of peripheral blood B cells in HHV-8-infected individuals with and without cKS. METHODOLOGY/PRINCIPAL FINDINGS: Circulating B cells and their subsets were analyzed by 6-color flow cytometry in the following groups: 1- patients HHV-8 positive with classic KS (cKS) (n = 47); 2- subjects HHV-8 positive and cKS negative (HSP) (n = 10); 3- healthy controls, HHV-8 negative and cKS negative (HC) (n = 43). The number of B cells belonging to the preimmune/natural effector compartment, including transitional, pre-naïve, naïve and MZ-like subsets, was significantly higher among HHV-8 positive subjects, with or without cKS, while was comparable to healthy controls in the antigen-experienced T-cell dependent compartment. The increased number of preimmune/natural effector B cells was associated with increased resistance to spontaneous apoptosis, while it did not correlate with HHV-8 viral load. CONCLUSIONS/SIGNIFICANCE: Our results indicate that long-lasting HHV-8 infection promotes an imbalance in peripheral B cell subsets, perturbing the equilibrium between earlier and later steps of maturation and activation processes. This observation may broaden our understanding of the complex interplay between viral and immune factors leading HHV-8-infected individuals to develop HHV-8-associated malignancies

Global patient outcomes after elective surgery: prospective cohort study in 27 low-, middle- and high-income countries.

BACKGROUND: As global initiatives increase patient access to surgical treatments, there remains a need to understand the adverse effects of surgery and define appropriate levels of perioperative care. METHODS: We designed a prospective international 7-day cohort study of outcomes following elective adult inpatient surgery in 27 countries. The primary outcome was in-hospital complications. Secondary outcomes were death following a complication (failure to rescue) and death in hospital. Process measures were admission to critical care immediately after surgery or to treat a complication and duration of hospital stay. A single definition of critical care was used for all countries. RESULTS: A total of 474 hospitals in 19 high-, 7 middle- and 1 low-income country were included in the primary analysis. Data included 44 814 patients with a median hospital stay of 4 (range 2-7) days. A total of 7508 patients (16.8%) developed one or more postoperative complication and 207 died (0.5%). The overall mortality among patients who developed complications was 2.8%. Mortality following complications ranged from 2.4% for pulmonary embolism to 43.9% for cardiac arrest. A total of 4360 (9.7%) patients were admitted to a critical care unit as routine immediately after surgery, of whom 2198 (50.4%) developed a complication, with 105 (2.4%) deaths. A total of 1233 patients (16.4%) were admitted to a critical care unit to treat complications, with 119 (9.7%) deaths. Despite lower baseline risk, outcomes were similar in low- and middle-income compared with high-income countries. CONCLUSIONS: Poor patient outcomes are common after inpatient surgery. Global initiatives to increase access to surgical treatments should also address the need for safe perioperative care. STUDY REGISTRATION: ISRCTN5181700

Clinical characteristics of cKS patients.

a<p>Mean ± standard error.</p>b<p>cKS patients were classified according to our classification that takes into account the prevalent type of lesions, localization, clinical behaviour, evolutive pattern and presence of complications ^(53,54).</p><p>A indicates slow evolution; B, rapid evolution; rapid denotes an increase in the total number of nodules/plaques or in the total area of plaques in three months following the last examination.</p

HHV-8 load in isolated PBMCs, B cells and non-B cells from cKS patients and healthy HHV-8-seropositive (HSP) controls according to their peripheral blood B cells count.

a<p>cKS patients were classified according to our classification that takes into account the prevalent type of lesions, localization, clinical behaviour, evolutive pattern and presence of complications.</p>b<p>gEq =  genome Equivalents.</p

B cells and their non-memory subsets are increased in patients with cKS.

<p>The number of total B cells and CD27⁻ B cells was significantly higher in cKS patients (grey bars) than HCs (white bars). All the subsets composing the preimmune/natural effector compartment, namely transitional, pre-naïve, naïve and MZ-like B lymphocytes, were increased in cKS patients, while the subsets composing the antigen-experienced T cell-dependent compartment, namely IgM-only, switched and CD27⁻ memory B cells, were unaffected. Data shown as mean ± SE. <i>P</i>-values calculated using the Student <i>t</i> test for independent samples. *<i>P</i><.05; **<i>P</i><.01; ***<i>P</i><.001.</p

B cells from cKS patients show a low state of activation.

<p>B cells from cKS patients (grey bars) expressed lower levels of the costimulatory molecules CD80 and CD86 and higher levels of CD20 than B cells from HCs (white bars). The expression of the indicated markers on total B cells and their CD27⁻ and CD27⁺ subsets is shown. Data presented as mean ± SE of mean fluorescence intensity (MFI) values. <i>P</i>-values calculated using the Student <i>t</i> test for independent samples. *<i>P</i><.05; **<i>P</i><.01; ***<i>P</i><.001.</p

Acronychiabaueri Analogue Derivative-Loaded Ultradeformable Vesicles: Physicochemical Characterization and Potential Applications

Elastic and ultradeformable liposomes were synthesized and physicochemically characterized to make suitable topical formulations for delivering the anti-inflammatory and anticancer compound 3-(4′-geranyloxy-3′-methoxyphenyl)-2-trans-propenoic acid. The average sizes of elastic and ultradeformable liposomes are below 300 nm, while the size distribution and Z-potential are below 0.3 and − 25 mV, respectively. The presence of 3-(4′-geranyloxy-3′-methoxyphenyl)-2-trans-propenoic acid does not affect the physicochemical parameters of nanovesicles. Elastic and ultradeformable liposomes show a zero order release kinetic and are stable at room temperature for a long time with or without 3-(4′-geranyloxy-3′-methoxyphenyl)-2-trans-propenoic acid. The ultradeformable liposomes are more deformable than elastic liposomes. These differences may depend on sodium cholate derivatives making nanoformulations. The 3-(4′-geranyloxy-3′-methoxyphenyl)-2-trans-propenoic acid-loaded elastic and ultradeformable liposomes can provide innovative nanotherapeutics-based natural compounds for the potential treatment of cutanous inflammation

B cells from cKS patients show increased resistance to spontaneous apoptosis and unaffected <i>in vivo</i> turnover.

<p>Apoptotic cells were detected by annexin V binding after 24 h culture in unstimulated conditions. <i>A</i>, representative flow cytometric analysis showing annexin V binding gated on total B cells, CD27⁻, MZ-like (CD27⁺IgD^lo) and switched memory (CD27⁺IgD⁻) B cells, as indicated; comparison between one HC (upper row) and one cKS patient (lower row). B, B cells from cKS patients (grey bars) showed significantly lower annexinV binding than B cells from HCs (white bars); this lower annexin V binding was evident on CD27⁻ and MZ-like (CD27⁺IgD^lo) B cells, roughly composing the preimmune/natural effector compartment, but not on antigen-experienced switched memory (CD27⁺IgD⁻) B cells. Data shown as mean ± SE. <i>C, In vivo</i> cell turnover was estimated by Ki67 staining of freshly isolated PBMCs. Representative flow cytofluorimetric analysis showing Ki67 binding gated on total B cells, CD27⁻ and CD27⁺ B cells, as indicated; comparison between one HC (upper row) and one cKS patient (lower row). <i>D</i>, The proportion of Ki67⁺ cells within total B cells, CD27⁻ and CD27⁺ B cells did not differ between cKS patients (grey bars) and HCs (white bars). Data shown as mean ± SE. <i>P</i>-values calculated using the Student <i>t</i> test for independent samples.*<i>P</i><.05; **<i>P</i><.01.</p

Analysis of B cells from healthy HHV-8-seropositive (HSP) compared with HHV-8-seronegative controls (HC) and cKS patients (cKS).

a<p>Mean ± standard error.</p>b<p>Mann-Whitney U test was used to detect significant differences.</p