Leukocyte Depletion Research Articles

Treatment of children and adolescents with ulcerative colitis by adsorptive depletion of myeloid lineage leucocytes as monotherapy or in combination with low dose prednisolone after failure of first-line medications

BackgroundCurrently available drugs for the treatment of ulcerative colitis (UC) include salicylates, thiopurines, corticosteroids and new anti-tumour necrosis factor (TNF)-α biologics. Among these medications, corticosteroids in children and adolescents may adversely affect the patients’ growth and development. Further, UC patients have elevated and activated myeloid lineage leucocytes including the CD14 + CD16+ monocytes, which release TNF-α as a significant exacerbating factor. Accordingly, depletion of these cells by granulocyte/monocyte adsorption (GMA) should alleviate inflammation and promote UC remission. The objective of this study was to evaluate the efficacy of GMA in children and adolescents in whom conventional first-line medications had failed to induce remission.MethodsIn a single centre setting, between 2007 and 2012, a total of 24 consecutive children and adolescents, age 11–19 years were given mesalazine or sulphasalazine as a first-line medication. Seventeen patients relapsed or did not respond to the first-line medications, and received GMA with the Adacolumn, 2 sessions in the first week, and then weekly, up to 11 sessions. Patients who achieved a decrease of ≥5 in the clinical activity index (CAI) were to continue with GMA, while non-responders were to receive 0.5 to 1.0 mg/kg/day prednisolone (PSL) plus additional GMA sessions similar to GMA responder cases. At entry and week 12, patients were clinically and endoscopically evaluated, allowing each patient to serve as her/his own control.ResultsSeven patients achieved remission with the first-line medications and did not receive GMA. Five patients did not respond to the first 5 GMA sessions and received PSL plus GMA, while 12 patients responded to the first 5 GMA sessions and received additional sessions. At entry, the average CAI was 12.7 ± 2.5, range 8–17, and the average endoscopic index was 8.5 ± 1.5, range 7–11. The corresponding values at week 12 were 2.1 ± 0.2, range 1–4 (P < 0.001) and 2.4 ± 0.2, range 1–4 (P < 0.001). PSL was tapered to 0 mg within 3 months.ConclusionsWith the strategy we applied in this study, all 24 consecutive patients achieved remission. In growing patients with active UC refractory to first-line medications, GMA was associated with clinical remission and mucosal healing, while in non-responders to GMA monotherapy, addition of a low dose PSL enhanced the efficacy of GMA and tapering of the PSL dose soon after remission was not associated with UC relapse. Therefore, the majority of young corticosteroid naive UC patients in whom first-line salicylates have failed may respond to GMA and be spared from additional drug therapy. Avoiding corticosteroids at an early stage of UC should ensure better long-term clinical course.

UK approach to assessing assays and filters designed to reduce the risk of transfusion-transmitted vCJD

Three cases of vCJD transmission by blood transfusion have been reported in the UK, and a fourth case discovered at post-mortem. Modelling has been conducted to predict the number of cases that may occur in the future through transfusion, based on estimates of prevalence, infectivity and susceptibility, and a number of steps have been taken to reduce the risk of transmission. These include deferral of previously transfused donors, leucocyte depletion of all components, importation of plasma for certain patient groups and for fractionation, and the collection of the majority of platelets from single donors (by apheresis). However, even with these interventions, some future cases are still predicted. The UK-wide Advisory Committee on the Safety of Blood, Tissues and Organs (SaBTO) considers the evidence for clinical and cost-effectiveness of any proposed intervention, such as prion assays and filters, and makes recommendations to the governments of the UK. The development of prion assays is challenging as prions do not generate an immune response, do not have nucleic acid and are present in blood in very low concentrations against a high background of normal prion protein. It is critically important that prion assays show high levels of sensitivity and — especially —specificity for a healthy blood donor population. Assessment is impacted by the very short supply of positive human samples, necessitating the use of animal models. Filters that are capable of removing prions from blood components have been developed and CE marked, but it is again necessary to use animal models to study their efficacy. Guidelines have been produced for the assessment of the quality of red cells filtered through these devices, and a clinical safety study has recently been completed. In conclusion, the evaluation of screening assays and prion filters is challenging, time-consuming and costly, but these evaluations are critical to policy making.

Ultra-pure platelet isolation from canine whole blood

BackgroundSeveral research applications involving platelets, such as proteomic and transcriptomic analysis, require samples with very low numbers of contaminating leukocytes, which have considerably higher RNA and protein content than platelets. We sought to develop a platelet purification protocol that would minimize contamination, involve minimal centrifugation steps, and yield highly pure platelet samples derived from low volume whole blood samples from healthy dogs.ResultsUsing an optimized OptiPrep density gradient technique, platelet recovery was 51.56% with 99.99% platelet purity and leukocyte contamination of 100 leukocytes per 108 platelets, on average. Platelet samples were subjected to additional purification with CD45-labeled Dynabeads after density barrier centrifugation resulting in a 95-fold depletion of residual leukocytes. Platelets purified using these methods remained inactivated as assessed by Annexin V and P-selectin labeling with flow cytometry.ConclusionsThe use of OptiPrep density gradient is a quick method for obtaining highly purified platelet samples from low volumes of canine whole blood with minimal contamination. Additional depletion of residual leukocytes can be achieved using CD45-labeled beads. These platelet samples can then be used for many downstream applications that require ultra-pure platelet samples such as RNA and protein analysis.

Extracorporeal tumor cell filtration during extended liver surgery: first clinical use of leukocyte depletion filters – a case series

BackgroundDuring oncologic surgery, intraoperative manipulation of tumor tissue is almost unpreventable and causes a high risk of tumor cell dissemination into venous blood. A tumor cell-reducing effect of leukocyte adhesion filter systems has been shown under in vitro conditions.MethodsIn a first clinical attempt, leukocyte adhesion filters were integrated into veno-venous bypass systems in four patients undergoing extended liver surgery for secondary hepatic malignancies.Practicability, handling, and safety aspects as well as potency of cell removal and clinical side effects of the filter system were analyzed.ResultsAll patients tolerated the application of the system without problems during operative and postoperative follow-up. Immunohistochemical staining of perioperative blood samples detected cytokeratin positive (CK+) cells in three cases during the hepatic mobilization.ConclusionsEffectiveness of CK+ cell depletion and safety of the procedure was shown. The presented surgical technique represents a safe and innovative tool; however, clinical significance has to be examined in a larger patient cohort.

Impact of Pre-Storage and Bedside Filtered Leukocyte-Depleted Blood Transfusions on Infective Morbidity after Colorectal Resection: A Single-Center Analysis of 437 Patients

Leukocyte-depleted blood transfusions were introduced to reduce transfusion-associated immunomodulation, but the clinical effects of different types of leukocyte depletion have been analyzed rarely. The aim of this survey was to analyze the clinical impact of pre-storage leukocyte-depleted blood transfusions (considered as pre-storage or bedside-filtered) on post-operative complications in patients undergoing elective or urgent colorectal resection. Data were collected retrospectively from the medical records of 437 consecutive patients who underwent colorectal resection from 2005 to 2010. All patients requiring transfusion received pre-storage or bedside-filtered leukocyte-depleted red blood cell concentrates according to availability at the blood bank. The outcomes were measured by the analysis of post-operative morbidity in patients receiving the different types of transfusions or having other potentially predictive risk factors. The overall morbidity rate, infective morbidity rate, and non-infective morbidity rate were, respectively, 35.6%, 28.1%, and 21.0%. Two hundred five patients (46.9%) received peri-operative transfusions. On multivariable analysis, leukocyte-depleted transfusion (odds ratio [OR] 3.33; 95% confidence interval [CI] 2.14-5.20; p<0.001) and both pre-storage (OR 2.82; 95% CI 1.73-4.59; p<0.001) and bedside-filtered (OR 4.69; 95% CI 2.54-8.67; p<0.001) transfusions were independent factors for post-operative morbidity. Prolonged operation (p=0.035), American Society of Anesthesiologists score≥3 points (p=0.023), diagnosis of cancer rather than benign disease (p=0.022), and urgent operation (p=0.020) were other independent predictors of post-operative complications. Patients transfused with bedside-filtered blood showed significantly higher rates of infective complications (51.4% vs. 31.8%; p=0.006), but not non-infectious complications (35.7% vs. 32.6; p=0.654) than patients who received pre-storage transfusions. Leukocyte-depleted blood transfusions and, in particular, bedside-filtered blood have a significant negative effect on infectious complications after colorectal resection.

OP0079 Salivary Gland Epithelial Cells (SGECS) Promote the Differentiation of B Cells

BackgroundSGECs have been shown to possess a key regulatory role in the inflammatory responses of patients with Sjögren’s syndrome (SS). Emerging studies present evidence for their capacity to participate in...

Uncontrolled donors with controlled reperfusion after sixty minutes of asystole: a novel reliable resource for kidney transplantation.

BackgroundOrgan shortage leads to usage of kidneys from donors after sudden cardiac death, or uncontrolled donors (UDCD). Ischemic injury due to cessation of circulation remains a crucial problem that limits adoption of UDCD. Our clinical investigation was to determine the applicability of kidneys obtained from UDCD and resuscitated by extracorporeal perfusion in situ after 60 minutes of asystole.MethodsIn 2009–2011, organ procurement service of St. Petersburg, obtained kidneys from 22 UDCD with critically expanded warm ischemic time (WIT). No patients were considered as potential organ donors initially. All donors died after sudden irreversible cardiac arrest. Mean WIT was 61.4±4.5 minutes. For kidney resuscitation, the subnormothermic extracorporeal abdominal perfusion with thrombolytics and leukocyte depletion was employed. Grafts were transplanted into 44 recipients. The outcomes of transplantation of resuscitated kidneys were compared to outcomes of 87 KTx from 74 brain death donors (BDDs).ResultsImmediate functioning of kidney grafts was observed in 21 of the 44 recipients, with no cases of primary non function. By the end of the first post-transplant year there was an acute rejection rate of 9.1% (4 episodes of rejection) in the UDCD group versus 14.2% (13 episodes of rejection) in the BDD group. The actual 1-year graft survival rate was 95.5% (n = 42) in UDCD group, and 94.6% (n = 87) in BDD group. Creatinine levels at the end of the first year were 0.116±0.008 and 0.115±0.004 mmol/l in UDCD and BDD groups, respectively.ConclusionsUDCD kidneys with critically expanded WIT could be succefully used for transplantation if in situ organ “resuscitation” perfusion is included into procurement protocol. The results of 1-year follow-up meet the generally accepted criteria for graft survival and function. In situ reperfusion may exert a therapeutic effect on grafts before procurement. This approach could substantially expand the organ donors' pool.

Circulating Blood Endothelial Nitric Oxide Synthase Contributes to the Regulation of Systemic Blood Pressure and Nitrite Homeostasis

Mice genetically deficient in endothelial nitric oxide synthase (eNOS(-/-)) are hypertensive with lower circulating nitrite levels, indicating the importance of constitutively produced nitric oxide (NO•) to blood pressure regulation and vascular homeostasis. Although the current paradigm holds that this bioactivity derives specifically from the expression of eNOS in endothelium, circulating blood cells also express eNOS protein. A functional red cell eNOS that modulates vascular NO• signaling has been proposed. To test the hypothesis that blood cells contribute to mammalian blood pressure regulation via eNOS-dependent NO• generation, we cross-transplanted wild-type and eNOS(-/-) mice, producing chimeras competent or deficient for eNOS expression in circulating blood cells. Surprisingly, we observed a significant contribution of both endothelial and circulating blood cell eNOS to blood pressure and systemic nitrite levels, the latter being a major component of the circulating NO• reservoir. These effects were abolished by the NOS inhibitor L-NG-nitroarginine methyl ester and repristinated by the NOS substrate L-arginine and were independent of platelet or leukocyte depletion. Mouse erythrocytes were also found to carry an eNOS protein and convert (14)C-arginine into (14)C-citrulline in NOS-dependent fashion. These are the first studies to definitively establish a role for a blood-borne eNOS, using cross-transplant chimera models, that contributes to the regulation of blood pressure and nitrite homeostasis. This work provides evidence suggesting that erythrocyte eNOS may mediate this effect.

The Molecular Link Between C-C-Chemokine Ligand 2-Induced Leukocyte Recruitment and Hyperalgesia

The chemokine C-C-chemokine ligand 2 (CCL2) (formerly known as MCP, macrophage chemotactic protein) is one of the important genes upregulated in different types of pain both in animals and humans. CCL2 governs the recruitment of C-C chemokine receptor 2-expressing monocytes into inflamed tissue. In contrast to neutrophilic chemokines, intraplantar injection of CCL2 in Wistar rats recruited macrophages and neutrophils and simultaneously lowered nociceptive thresholds. CCL2-induced hyperalgesia was abolished by prior systemic leukocyte depletion by cyclophosphamide and was reconstituted by local adoptive transfer of donor macrophages but not of neutrophils. Antagonists against transient receptor potential vannilloid 1 inhibited thermal and against transient receptor potential ankyrin 1 blocked mechanical hyperalgesia. Peripheral but not central activation of cyclooxygenase-2 (Cox-2) were critical for CCL2-induced hyperalgesia. In vitro CCL2 did not directly stimulate Cox-2 expression or prostaglandin E2 formation but slightly enhanced the formation of reactive oxygen species in monocytes and macrophages. In vivo, increased immunoreactivity for 4-hydroxy-2-nonenal (4-HNE), a downstream product of reactive oxygen species and known inducer of Cox-2, was observed and colocalized with Cox-2 in ED1 (CD68) positive infiltrating cells. No hyperalgesia, 4-HNE, or Cox-2 immunoreactivity was seen in leukocyte-depleted rats that were reconstituted with macrophages in the absence of CCL2, supporting the important role of CCL2. PerspectiveCCL2 plays a dual role: 1) promoting monocyte/macrophage recruitment into tissue; and 2) potentially stimulating macrophages in the tissue to produce 4-HNE and subsequently Cox-2, all resulting in the induction of hyperalgesia via transient receptor potential vannilloid 1 and transient receptor potential ankyrin 1. This encourages pharmacological efforts targeting CCL2/C-C chemokine receptor 2 and macrophages for treatment of inflammatory pain.

Spdef Null Mice Lack Conjunctival Goblet Cells and Provide a Model of Dry Eye

Goblet cell numbers decrease within the conjunctival epithelium in drying and cicatrizing ocular surface diseases. Factors regulating goblet cell differentiation in conjunctival epithelium are unknown. Recent data indicate that the transcription factor SAM-pointed domain epithelial-specific transcription factor (Spdef) is essential for goblet cell differentiation in tracheobronchial and gastrointestinal epithelium of mice. Using Spdef(-/-) mice, we determined that Spdef is required for conjunctival goblet cell differentiation and that Spdef(-/-) mice, which lack conjunctival goblet cells, have significantly increased corneal surface fluorescein staining and tear volume, a phenotype consistent with dry eye. Microarray analysis of conjunctival epithelium in Spdef(-/-) mice revealed down-regulation of goblet cell-specific genes (Muc5ac, Tff1, Gcnt3). Up-regulated genes included epithelial cell differentiation/keratinization genes (Sprr2h, Tgm1) and proinflammatory genes (Il1-α, Il-1β, Tnf-α), all of which are up-regulated in dry eye. Interestingly, four Wnt pathway genes were down-regulated. SPDEF expression was significantly decreased in the conjunctival epithelium of Sjögren syndrome patients with dry eye and decreased goblet cell mucin expression. These data demonstrate that Spdef is required for conjunctival goblet cell differentiation and down-regulation of SPDEF may play a role in human dry eye with goblet cell loss. Spdef(-/-) mice have an ocular surface phenotype similar to that in moderate dry eye, providing a new, more convenient model for the disease.

In vivo depletion of leukocytes and platelets following injection of T cell-specific antibodies into mice

In vivo T cell depletion experiments are widely used to establish the role of these cells in a variety of immunological processes. Different clones of monoclonal antibody targeting the CD3 molecular complex (mainly 145-2C11 and 17A2) have been successfully used for T cell depletion. In the present work, we assessed the specificity of monoclonal antibody-mediated CD3 T cell depletion in mouse peripheral blood. We showed that treatment of BALB/C mice with monoclonal antibodies (clones 145-2C11 and 17A2) not only efficiently depletes T cells in vivo, but also leads to a substantial reduction in B cell, granulocyte and platelet counts. In contrast, T cell depletion using a combination of anti-CD4 and anti-CD8 antibodies was efficient and produced less deleterious effects on other blood cell populations. Therefore, the results obtained from T cell depletion experiments using anti-CD3 antibodies must be interpreted with caution prior to draw definitive conclusions on the role of T cells in a given immunological process.

Abstract 1454: Evaluation of negative selection strategy for enrichment of circulating melanoma cells (CMCs) from patients with metastatic melanoma.

Abstract Introduction. Circulating tumor cells (CTCs) have emerged as a predictive biomarker in the treatment of metastatic breast, colorectal and prostate cancers. Non-epithelial tumors such as melanoma also present evidence of circulating cells that have been difficult to detect due to technological limitations. Previous clinical studies have shown presence of melanoma specific marker mRNAs circulating in the blood, used as a surrogate of the circulating melanoma cell (CMC). This study was designed to use a negative cell selection platform to enrich CMCs from blood. The approach involved depletion of red blood cells by centrifugation and lysis, and white blood cells by immunomagnetic tagging with pan-leukocyte (CD45) antibody and magnetic separation. The enriched non-hematopoietic cell analysis was optimized for high sensitivity and specificity of CMC detection using a panel of antibodies against known melanoma markers. The secondary goal was to estimate the number of CMCs per milliliter of blood based on immunophenotype and morphology criteria. Methods. The study was reviewed and approved by the IRB. The leukocyte depletion was accomplished using a fluid dynamics-based quadrupole magnetic separating device (1) which, in principle, enriches CMCs without affecting them by immunomagnetic reagents. CMC cytospin smears were immunostained against known melanoma markers (Melan-A and S100). A high magnification microscopy imaging and a computer-aided image analysis were developed for efficient CMC enumeration. Results. Based on preliminary studies with SKMEL-28 melanoma cell line, normal adult blood controls and blood samples from advanced melanoma patients, the CMC was defined as an intact, nucleated cell, positive for Melan A or S100 and negative for CD45. Separations of SKMEL-28 cells spiked at approximately one SKMEL-28 cell per million leukocytes resulted in high efficiency separation with a mean recovery of (93.0 ±5.4 S.E.M.)% of SKMEL-28 cells. Samples obtained from apparently healthy donor blood (n =5) had a baseline of 0-3 positive cells per mL blood, whereas samples from stage IV melanoma patients (n =8) exhibit CMC counts ranging from 78 to 28,500 cells per mL of blood. Conclusions. Our study combines negative selection platform with immunophenotypic and morphological criteria for CMC enrichment, identification and enumeration. Interestingly, our preliminary results indicate the presence of high numbers of CMCs in malignant melanoma patients, providing basis for larger studies on correlations with clinical outcomes of the disease. Moreover, negative selection method lends itself to in vitro or in vivo culture bioassays, and molecular marker assays such as qRT-PCR, genomic PCRs, FISH or microarrays. Citation Format: Powrnima Joshi, Barbara Jacobs, Adeeb Derakhshan, Lee R. Moore, Pierre Triozzi, Ernest C. Borden, Maciej Zborowski. Evaluation of negative selection strategy for enrichment of circulating melanoma cells (CMCs) from patients with metastatic melanoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1454. doi:10.1158/1538-7445.AM2013-1454

Disease Amelioration With Tocilizumab in a Treatment-Resistant Patient With Neuromyelitis Optica

Neuromyelitis optica (NMO) is an autoimmune disease of the central nervous system in which aberrant antibody responses to the astrocytic water channel aquaporin 4 have been described. Experimental evidence is emerging that NMO is partly driven by the proinflammatory cytokine interleukin 6 (IL-6), which propagates the survival of disease-specific B cell subclasses, and deviates CD4+ T helper cell differentiation toward IL-17-producing T helper 17 cells. Tocilizumab is a recombinant humanized monoclonal antibody against the IL-6 receptor approved for treatment of rheumatoid arthritis. To study clinical and paraclinical effects of tocilizumab in a patient with NMO. Case report. Academic neurology department. A patient with highly active aquaporin 4–seropositive NMO who failed numerous immunosuppressive interventions, including high-dose corticosteroids, mitoxantrone, plasma exchange (PE), rituximab (anti-CD20), and alemtuzumab (anti-CD52), before receiving tocilizumab. Clinical disability, magnetic resonance imaging, cytokines and transcription factors levels in the cerebrospinal fluid, and peripheral blood mononuclear cells. A patient who continued to accumulate neurological disability and magnetic resonance imaging activity while receiving numerous immunoactive therapies stabilized, and eventually improved clinically and on magnetic resonance metrics after treatment initiation with tocilizumab. Treatment and clinical response correlated with a significant reduction of IL-6 levels in the CSF as well as a diminished expression of signal transducer and activator of transcription 3. Tocilizumab might be effective in NMO, here in a patient not responding to leukocyte depletion. Our findings further support data that implicate IL-6 as a critical molecule in the immunopathogenesis of NMO, and a critical role for T cells in the pathogenesis of this disorder.

Distinct functions of chemokine receptor axes in the atherogenic mobilization and recruitment of classical monocytes

We used a novel approach of cytostatically induced leucocyte depletion and subsequent reconstitution with leucocytes deprived of classical (inflammatory/Gr1hi) or non-classical (resident/Gr1lo) monocytes to dissect their differential role in atheroprogression under high-fat diet (HFD). Apolipoprotein E-deficient (Apoe−/−) mice lacking classical but not non-classical monocytes displayed reduced lesion size and macrophage and apoptotic cell content. Conversely, HFD induced a selective expansion of classical monocytes in blood and bone marrow. Increased CXCL1 levels accompanied by higher expression of its receptor CXCR2 on classical monocytes and inhibition of monocytosis by CXCL1-neutralization indicated a preferential role for the CXCL1/CXCR2 axis in mobilizing classical monocytes during hypercholesterolemia. Studies correlating circulating and lesional classical monocytes in gene-deficient Apoe−/− mice, adoptive transfer of gene-deficient cells and pharmacological modulation during intravital microscopy of the carotid artery revealed a crucial function of CCR1 and CCR5 but not CCR2 or CX3CR1 in classical monocyte recruitment to atherosclerotic vessels. Collectively, these data establish the impact of classical monocytes on atheroprogression, identify a sequential role of CXCL1 in their mobilization and CCR1/CCR5 in their recruitment.

A negative selection system PowerMag for effective leukocyte depletion and enhanced detection of EpCAM positive and negative circulating tumor cells

BackgroundCirculating tumor cells (CTCs) can be enriched from peripheral blood by leukocyte depletion. In this study, a PowerMag system was designed to enhance leukocyte depletion and CTC detection. MethodsThe efficiency of PowerMag in leukocyte depletion and recovery of CTCs was evaluated. The cell populations in leukocyte-depleted cell filtrate were characterized by immunofluorescence staining using anti-EpCAM and anti-CD45 antibodies. The suitability of PowerMag in monitoring cancer patient CTCs was assessed by analyzing blood samples collected at baseline and 2weeks after baseline from the patients with colorectal cancer (CRC, n=24) and head and neck squamous cell carcinoma (HNSCC, n=28). ResultsOnly few contaminated CD45+-leukocytes were retained in the leukocyte-depleted cell filtrate. The enriched CTCs were viable with a recovery rate of 46–62%. Immunofluorescence staining of the nucleated cells enriched by PowerMag revealed 2 major cell populations EpCAM+CD45− and EpCAM−CD45−. The number for both cell types was significantly increased in cancer patients when compared to healthy control. After chemotherapy, the number of EpCAM+CD45− and EpCAM−CD45− cells was decreased more prominently for CRC and HNSCC patients, respectively. ConclusionsPowerMag holds great promise as a platform for leukocyte depletion and CTC detection in a clinical setting.

P330 Treatment of children and adolescents with ulcerative colitis by adsorptive depletion of elevated myeloid lineage leucocytes as mono-therapy or in combination with low dose prednisolone

P330 Treatment of children and adolescents with ulcerative colitis by adsorptive depletion of elevated myeloid lineage leucocytes as mono-therapy or in combination with low dose prednisolone

P341 Therapeutic depletion of myeloid lineage leucocytes in ulcerative colitis patients instead of pharmacological intervention: clinical and endoscopic features of responders and non-responders

P341 Therapeutic depletion of myeloid lineage leucocytes in ulcerative colitis patients instead of pharmacological intervention: clinical and endoscopic features of responders and non-responders

Therapeutic depletion of myeloid lineage leukocytes in patients with generalized pustular psoriasis indicates a major role for neutrophils in the immunopathogenesis of psoriasis

Generalized pustular psoriasis (GPP) is a chronic autoimmune disease characterized by fever, erythema, and neutrophilic pustules over large areas of the skin. GPP does not respond well to pharmacologic intervention. We sought to assess efficacy of selectively depleting the myeloid lineage leukocytes in patients with GPP. Fifteen patients with persistent moderate to severe GPP despite conventional therapy were included. Eligible patients had more than 10% of their skin area covered by pustules. Treatment with oral etretinate, cyclosporine, methotrexate, prednisolone, and topical prednisolone/vitamin D3 was continued if had been initiated well in advance of study entry. Five sessions of adsorptive granulocyte and monocyte apheresis (GMA) with the Adacolumn (JIMRO Co Ltd, Takasaki, Japan) were administered (1 session/wk over 5 weeks) to selectively deplete Fcγ receptor and complement receptor bearing leukocytes. Efficacy was assessed by measuring the skin areas covered by pustules at baseline and 2 weeks after the last GMA session. One patient did not complete the first GMA session. Based on the GPP severity scores relative to entry, the overall scores improved (n = 14, P = .0027), and the area of erythroderma (P = .0042), pustules (P = .0031), and edema (P = .0014) decreased. Likewise, Dermatology Life Quality Index improved (P = .0016), reflecting better daily function and quality of life. Twelve patients were judged as responders (85.7%), and 10 patients maintained the clinical response for 10 weeks after the last GMA session without any change in medication. This study was unblinded and without a placebo arm. GMA in this clinical setting was safe and effective, suggested a major role for granulocytes/monocytes in the immunopathogenesis of GPP.

Risk of Cytomegalovirus Infection and Disease after Umbilical Cord Blood Transplantation in Children

Pediatric data regarding cytomegalovirus (CMV) infections in pediatric patients receiving umbilical cord blood (UCB) transplantation are sparse. To determine whether UCB transplantation increases the risk of CMV infection and disease compared with other graft sources. The medical files of patients who underwent allogeneic hematopoietic stem cell transplantation at CHU Ste-Justine (Montreal, Quebec) from April 2000 to December 2006 were retrospectively reviewed. A Cox proportional hazard model was used to assess the effect of potential predictors of outcomes. A total of 176 patients with a median age of nine years (range 0.1 to 18 years) underwent hematopoietic stem cell transplantation. The source of stem cells were UCB, bone marrow and peripheral blood stem cells in 86, 86 and four of the cases, respectively. CMV infection occurred in 29 patients (16%). At day 100 post-transplantation, the rate of CMV infection was 13% in UCB transplant recipients (11 of 86) versus 20% in those with other sources of graft (18 of 90) (P=0.19). Positive CMV serology of the recipient and leukocyte depletion were two independent variables associated with an increased risk of CMV infection. Among infected patients, six developed CMV disease (20.7%). The rate of CMV disease one year after infection was 49% in patients who received UCB (five of 11) and 6% in others (one of 18). This difference was significant by univariate (P=0.01) but not by multivariate analysis. In the setting of the current study, with a moderate CMV infection rate (16.5%), UCB transplantation did not appear to increase the risk of CMV infection and disease.

Role of Intraoperative Red Cell Salvage and Autologus Transfusion in Metastatic Spine Surgery: A Pilot Study and Review of Literature

Study DesignProspective cohort study.PurposeThere has been no research examining the use of intraoperative cell salvage during metastatic spinal surgery. The present work is a pilot study investigating the role of cell salvage during metastatic spine surgery.Overview of LiteratureThere is no spinal literature about role of cell salvage and autologus transfusion in metastatic spinal cancer.MethodsSixteen spinal metastases patients who received red cell salvage using a leucocyte depletion filter were enrolled. Of these, ten patients who received salvaged blood transfusion were included in the final analysis. Data collection involved looking at the case notes, operating room records and the prospectively updated metastatic spinal cancer database maintained in the spinal department. Cell salvage data was recovered from the central cell salvage database maintained in the anesthetic department.ResultsAmount of salvaged blood ranged from 120 to 600 mL (average, 318 mL). The average drop in hemoglobin was 1.65 units (range, 0.4-2.7 units). Three patients (30%) required postoperative allogenic blood transfusion. The average follow up was 9.5 months (range, 6-6 months). One patient developed new lung metastasis, at seven months. No patient developed new liver metastases. Preoperatively, six patients had diffuse skeletal metastases. Of this subgroup, three developed new skeletal metastases. No cases showed any wound related problems in the postoperative period.ConclusionsIn our study transfusion of intraoperatively salvaged blood did not result in disseminated metastatic cancer. We would suggest that red cell salvage might have a role during metastatic spine surgery.