New approach in vaginal prolapse repair: mini-invasive surgery associated with application of platelet-rich fibrin.

INTRODUCTION AND HYPOTHESIS: Platelet-rich fibrin (PRF) matrix is an autologous leukocyte and PRF biomaterial. PRF is a fibrin matrix polymerized in a tetramolecular structure with the incorporation of platelets, leukocytes, cytokines, and circulating stem cells. The three-dimensional structure of PRF is optimal for migration of endothelial cells and fibroblasts. It permits rapid angiogenesis and easier remodeling of fibrin in a more resistant connective matrix. In vaginal surgery, PRF may act as a graft material with better healing and better functional outcome. METHODS: We performed a prospective observational study on ten consecutive women requiring surgery for prolapse recurrence (stage II or higher). These women had high risks for recurrence, erosion with graft materials, and intraoperative and postoperative complications with traditional pelvic reconstructive surgical procedures. ICS score and P-QoL Questionnaire results were assessed preoperatively and postoperatively. Surgery consisted of anterior, posterior, or apical repair plus PRF. Follow-up was performed at 1, 6, 12, 18, and 24 months. RESULTS: Anatomically, the success rate was 80%. Prolapse symptoms improved by 100%. Sexual activity increased by 20% without dyspareunia. The surgical time was satisfactory (mean, 38.5 min). There were no intraoperative or postoperative complications. CONCLUSIONS: The use of PRF for site-specific prolapse repair is associated with a good functional outcome because of the healing and mechanical properties of PRF.

Down regulation of human natural killer cell-mediated cytolysis induced by blood transfusion: role of transforming growth factor-ß(1), soluble Fas ligand, and soluble Class I human leukocyte antigen.

BACKGROUND: Human natural killer (NK) cells are thought to play a role in antiviral response and tumor immune surveillance. The molecular mechanisms of down regulation of NK-cell activity observed after red blood cell (RBC) transfusion is still undefined. STUDY DESIGN AND METHODS: Both effects of blood transfusion (ex vivo) and supernatants (SNs) derived from RBC units unstored (RBC-0) or stored for 5 or 30 days (RBC-5 or -30, respectively) in vitro were analyzed on NK cell-mediated cytolytic activity. RESULTS: We have found that NK cells isolated from transfused patients on Day 3 lysed the NK-sensitive target cells K562 to a lesser extent than before transfusion. This down regulation of NK-cell activation was evident also for NK-cell killing mediated through the engagement of NK cell-activating receptors as NKG2D, NKp30, NKp46, and CD16. Transfused patients reacquired NK cell-mediated cytolytic activity from Day 5 to Day 7 after transfusion. SN from RBC-30, but not from RBC-0 or RBC-5, strongly inhibited the generation of lymphokine-activated killer (LAK) cells and lysis of the NK-resistant target cell Jurkat in a dose-dependent manner. Transforming growth factor-ß1 (TGF-ß1) blocking antibodies partially restored the generation of LAK activity. In addition, the depletion of both soluble Class I human leukocyte antigens (sHLA-I) and soluble Fas ligand (sFasL) from SN of RBC-30 completely restored the generation of LAK activity. CONCLUSIONS: Altogether, these findings would support the idea that blood transfusion-mediated down regulation of NK-cell activity is mediated by sHLA-I, sFasL, and TGF-ß1.

sHLA-I Contamination, a novel mechanism to explain ex vivo/in vitro modulation of IL-10 synthesis and release in CD8(+) T lymphocytes and in neutrophils following intravenous immunoglobulin infusion.

BACKGROUND: Numerous mechanisms have been proposed to explain the beneficial action of intravenous immune globulin (IVIG) in autoimmune and systemic inflammatory disorders; among others, they could decrease pro-inflammatory cytokine levels and also induce anti-inflammatory cytokines. MATERIALS AND METHODS: Ex vivo analysis of cells from ten IVIG recipients showed significant increase of IL-10 mRNA and intra-cellular IL-10 molecules in both leukotypes. RESULTS: In vitro comparable results were obtained incubating CD8(+) T lymphocytes and neutrophils from healthy donors with IVIG. sHLA-I and/or sFasL immunodepletion abolished IL-10 modulation. Co-culture with contaminant-free IgM or MabThera did not exert any mRNA modulation. Finally, IgM or MabThera plus purified sHLA-I molecules enhanced IL-10-mRNA in both leukotypes to levels comparable to those obtained with IVIG incubation. CONCLUSION: As IVIG infusion involves administration of soluble contaminants, these data consent to speculate that IVIG might modulate IL-10 via the immunomodulatory activities of sHLA-I contaminant molecules inducing transcriptional and post-transcriptional modulation of IL-10 in CD8(+) T lymphocytes and neutrophils.

Immunomodulatory effects of blood transfusions: the synergic role of soluble HLA Class I free heavy-chain molecules detectable in blood components.

BACKGROUND: Over the past decades, the weight of the published literature demonstrates that blood transfusions can induce clinically significant immunosuppression in recipients. Several studies showed significant improved clinical outcomes in the patients receiving leukoreduced transfusions, compared with control patients who received nonleukoreduced transfusions. Moreover, the immunosuppressive potential of blood products grows with the time of their storage and becomes highest in nonleukoreduced blood products stored for a long time. STUDY DESIGN AND METHODS: The interest was previously focused on the determination of immunomodulatory soluble molecules such as soluble HLA Class I (sHLA-I) and soluble Fas ligand (sFasL) in different blood components and on the evaluation of their immunomodulatory activities. On this basis, whether soluble beta2-microglobulin free HLA Class I heavy chains (sHLA-beta2fHC) could be detected and immunochemically characterized in different blood components was evaluated. Immunomodulatory activity of detectable sHLA-beta2fHC molecules was evaluated by apoptosis inducing capacity in interleukin-2-activated antigen-specific cytotoxic T lymphocytes (CTL). RESULTS: Double-determinant immunoenzymatic assay indicates that sHLA-beta2fHC levels in red blood cells stored for up to 30 days and in random-donor platelets are significantly (p < 0.001) higher than in other blood components, and the immunochemical characterization suggests that the major source of sHLA-beta2fHC molecules might be the residual white cells that undergo membrane damage during storage. Finally, allogeneic CD8+ CTL apoptosis induction confirmed biofunctionality of sHLA-beta2fHC molecules. CONCLUSION: These data are comparable with those previously reported dealing with contaminant soluble molecules in allogeneic and autologous blood components, suggesting that sHLA-beta2fHC molecules could contribute to the immunosuppressive effects of blood transfusions.

Nonleukoreduced red blood cell transfusion induces a sustained inhibition of neutrophil chemotaxis by stimulating in vivo production of transforming growth factor-beta1 by neutrophils: role of the immunoglobulinlike transcript 1, sFasL, and sHLA-I.

BACKGROUND: Red blood cell (RBC) transfusion has been linked to increased susceptibility to infections in critically ill patients and to augmented incidence of postoperative infections. The mechanisms by which transfusions can induce immunosuppression are only partially defined. Recently, it has been demonstrated that RBC supernatants inhibit neutrophil migration. Such inhibitory activity is due to transforming growth factor (TGF)-beta1 contained in the supernatants that desensitize neutrophils to subsequent chemotaxic stimulation. STUDY DESIGN AND METHODS: In ancillary experiments, it was observed that plasma from transfused patients maintained its capacity of inhibiting neutrophil chemotaxis several days after RBC transfusion. Thus, this study was planned to investigate the mechanism(s) responsible for the prolonged inhibition of neutrophil chemotaxis observed after RBC transfusion. RESULTS: Plasma samples obtained from subjects who underwent RBC transfusion display a capability of inhibiting neutrophil chemotaxis, which is detectable up to 15 days after the transfusion. The inhibition is related to the capacity of FasL and HLA-I molecules contained in RBC supernatants to induce in vivo TGF-beta1 synthesis by neutrophils. The induction of TGF-beta1 secretion in neutrophils by HLA-I molecules depends on immunoglobulinlike transcript 1/CD85 triggering. CONCLUSION: The property of RBC transfusion of inducing a sustained inhibition of neutrophil chemotaxis seems to be a potential mechanism that concurs to the susceptibility to infections in patients who receive transfusions. Furthermore, our findings, showing neutrophil production of TGF-beta1 in response to FasL and HLA-I molecules, confirm that neutrophils are endowed not only with effector functions but also with immunomodulatory properties possibly involved in the regulation of inflammatory processes.

Transforming growth factor-beta1 in supernatants from stored red blood cells inhibits neutrophil locomotion.

Studies comparing transfusion and nontransfusion patients suggest an increased risk of postoperative infections in transfusion groups. Supernatants of blood components have been shown to affect the function of T lymphocytes and natural killer cells. Here, we found that supernatants from stored red blood cells (RBCs) inhibit human neutrophil migration in response to formyl peptides and stimulate neutrophil locomotion. These effects can be observed with high dilutions of RBC supernatants, such as 1:5 x 106 (vol/vol), able to trigger locomotion as well as desensitization of the cells to alternative chemoattractants. The phenomenon might be mediated by chemoattractants present in the supernatants. As RBC supernatants failed to mobilize intracellular free calcium, the chemoattractants should belong to the group of pure chemoattractants, that is, soluble Fas ligand (sFasL) and transforming growth factor-beta1 (TGF-beta1), known to act without increasing calcium levels. Recombinant TGF-beta1, but not sFasL, was found to reproduce the ability of RBC supernatants to both inhibit neutrophil response to formyl peptides and stimulate neutrophil locomotion. Moreover, TGF-beta1-immunodepleted supernatants did not display neutrophil-directed activities. Finally, RBC supernatants from RBCs stored after depletion of leukocytes were incapable of affecting neutrophil function. With neutrophils acting as a first-line antimicrobial defense, the ability, shown here, of high dilutions of RBC supernatants to inhibit neutrophil chemotaxis through TGF-beta1 may be a relevant determinant of infections in the postoperative period for transfusion patients. Consistently, the neutrophil chemotactic response to formyl peptide was inhibited by the plasma obtained from 5 transfusion patients.