Immunological Typing Research Articles

Pneumococcal infection in children with recurrent respiratory diseases

In frequently ill children with recurrent respiratory diseases (RRD), violations of local immunity and microbiocenosis of the mucous membranes can contribute to the development of acute and chronic bronchopulmonary diseases. Vaccination against major pathogens, including S. pneumoniae, can help prevent respiratory infections in children.Objective: to study the microbiocenosis and local immunological factors of the nasopharyngeal mucosa in children with RRD, the serotype composition of S. pneumoniae in children with community-acquired pneumococcal pneumonia (CAP) and pneumococcal carriers, as well as the correspondence of the serotype landscape to the composition of modern pneumococcal vaccines.Materials and methods: The study included 150 patients (104 children with RRD and 46 patients with CAP). To assess the condition of the nasopharyngeal mucosa cultural methods, morphofunctional, immunological and capsule PCR typing of S. pneumoniae isolates were performed. Statistical analysis was performed using the Statistica 10.0 package.Results: The age structure was dominated by children aged 3 to 6 years (36%). The majority of patients with RRD had pronounced nasopharyngeal mucosal dysbiosis (n=60), as well as destruction of epithelial cells (n=97). These factors may predispose to the development of bronchopulmonary diseases, including community-acquired pneumonia (CAP). Capsule PCR typing of S. pneumoniae isolates from the nasopharynx was performed in 46 patients. It has been established that in the structure of nasopharyngeal carrier S. pneumoniae in children with CAP, serotypes “3” and 19F prevailed, which were also the main pathogens in pneumococcal CAP. In nasopharyngeal pneumococcal carrier, the thirteen–valent pneumococcal conjugate vaccine (PCV13) overlapped 39.6% (95%CI 27.6–53.1%) of serotypes, in pneumococcal VP – 57.2% (95% CI 36.5-75.5%), while PCV20 overlapped 22.6% more S. pneumoniae serotypes in children with CAP.Conclusion: The strategy for the prevention of acute respiratory infections in children is immunization against pneumococcal infection, which should be carried out taking into account the regional prevalence of S. pneumoniae serotypes.

Limbic encephalitis – A case report of atypical dementia syndrome with potentially therapeutic consequence

IntroductionLimbic encephalitis (LE) is a subacute or chronic, non-infectious inflammation of the brain, usually occurring in adulthood, with predominant involvement of mesiotemporal structures and a clinical manifestation consisting mainly of new memory impairment, affective disorder, temporal lobe epilepsy, psychoses, etc.ObjectivesTo point out the importance of knowledge of potentially treatable dementia syndromes such as atypical manifestation of probably LE.MethodsWe present a clinical case of a 47-years-old woman with an atypical dementia syndrome and typical radiological findings corresponding to a LE, among others, but without the previously known immunological antibodies.ResultsAccording to the literature, the diverse subsyndromes of LE can be subsumed under the two main categories of “paraneoplastic” and “non-paraneoplastic”. In addition to the acute and subacute courses, there is increasing evidence for chronic, slowly progressive courses, which expand the spectrum of potentially treatable dementia syndromes. Understanding and knowledge of the broad, clinical syndrome of LE have increased dramatically in recent years. Both nosological classification through differentiated diagnosis and specific therapeutic protocols have become increasingly developed and established. Nevertheless, there are rare clinical cases with a clinical phenotype and radiological findings that correspond to LE, but are both non-paraneoplastic in origin and seronegative with respect to the previously known immunological typing by autoantibodies. This gray area of nosological entity represents a diagnostic and therapeutic challenge.ConclusionsThe authors would like to point out the importance of an adequate diagnosis of the forms of LE that have been nosologically classified so far and are partly well treatable. Limbic encephalitis is an important differential diagnosis in dementia, especially in young patients with atypical courses. There is a need for further research regarding better diagnosis and therapy of the so far immunologically unidentifiable forms of clinical LE.Literature:Bazir Ahmad et al., Practical Neurology 2011Guidelines of the German Neurological Society (DGN), 2008Leypoldt et al., Akt Neurol 2012Prüss et al., Neurology 2012Disclosure of InterestNone Declared

Identification of IRF-associated molecular subtypes in clear cell renal cell carcinoma to characterize immunological characteristics and guide therapy.

Recently studies have identified a critical role for interferon regulatory factor (IRF) in modulating tumour immune microenvironment (TME) infiltration and tumorigenesis. Based on IRF1-9 expression profiles, we classified all ccRCC samples into three molecular subtypes (clusters A-C) and characterized the prognosis and immune infiltration of these clusters. IRFscore constructed by principal component analysis was performed to quantify IRF-related subtypes in individual patients. We proved that IRFscore predicted multiple patient characteristics, with high IRFscore group having poorer prognosis, suppressed TME, increased T-cell exhaustion, increased TMB and greater sensitivity to anti- PD-1/CTLA-4 therapies. Furthermore, analysis of metastatic ccRCC (mccRCC) molecular subtypes and drug sensitivity proved that low IRFscore was more sensitive to targeted therapies. Moreover, IRFscore grouping can be well matched to the immunological and molecular typing of ccRCC. qRT-PCR showed differential expression of IRFs in different cell lines. Evaluating IRF-related molecular subtypes in individual ccRCC patients not only facilitates our understanding of tumour immune infiltration, but also provides more effective clinical ideas for personalised treatment.

Identification and characterization of neutrophil heterogeneity in sepsis

BackgroundAlthough the immune function of neutrophils in sepsis has been well described, the heterogeneity of neutrophils remains unclear during the process of sepsis.MethodsIn this study, we used a mouse CLP model to simulate the clinical scenario of patients with sepsis, neutrophil infiltration, abnormal distribution and dysfunction was analyzed. LPS was used to stimulate neutrophils in vitro to simulate sepsis; single-cell gene sequencing technology was used to explore the immunological typing. To explore the immunological function of immunosuppressive neutrophils, PD-L1 knockout neutrophils were cocultured with lymphocytes from wild-type mice.ResultsWe found that neutrophils presented variant dysfunction at the late stage of sepsis, including inhibition of apoptosis, seriously damaged chemotaxis and extensive infiltration into the tissues. Single-cell RNA sequencing revealed that multiple subclusters of neutrophils were differentiated after LPS stimulation. The two-dimensional spatial distribution analysis showed that Foxp3+ T cells were much closer to Ly-6G than the CD4+ and CD8+ cells, indicating that infiltrated neutrophils may play immunomodulatory effect on surrounding T-regs. Further observations showed that LPS mediates PD-L1 over expression through p38α-MSK1/-MK2 pathway in neutrophils. The subsets of highly expressed PD-L1 exert immunosuppressive effect under direct contact mode, including inhibition of T cell activation and induction of T cell apoptosis and trans-differentiation.ConclusionsTaken together, our data identify a previously unknown immunosuppressive subset of neutrophils as inhibitory neutrophil in order to more accurately describe the phenotype and characteristics of these cells in sepsis.

Diagnostic Value of Cytomorphology in Acute Leukemia

To investigate the diagnostic value of cytomorphology (including cytochemical staining) in newly diagnosed acute leukemia, so as to improve the importance of cytomorphology. The clinical data of 119 cases of acute leukemia diagnosed in our hospital from April 2016 to June 2018 were analyzed retrospectively. According to morphologic and immunological typing, accordance rate to final diagnosis was compared. The diagnostic accordance rate of simple morphological typing was 76.5%, and the diagnostic accordance rate of simple immunological typing was 79.8%, the difference of diagnostic coincidence rate was not significant between the two groups of acute leukemia. Cytomorphology is the cornerstone of the diagnosis of acute leukemia, it has similar value to immunological classification in the diagnosis of leukemia and should pay enough attention. MICM comprehensive diagnosis can improve the final diagnosis rate, showing a guidance significance for the treatment and prognosis of patients with acute leukemia.

Group B Streptococcal Infections.

1. Tara M. Randis, MD, MS* 2. Jacqueline A. Baker, MD† 3. Adam J. Ratner, MD, MPH*,‡ 1. Departments of *Pediatrics and 2. ‡Microbiology, New York University School of Medicine, New York, NY 3. †Department of Pediatrics, Columbia University College of Physicians & Surgeons, New York, NY * Abbreviations: CDC: : Centers for Disease Control and Prevention CFR: : case fatality rate CSF: : cerebrospinal fluid EO: : early-onset GBS: : group B Streptococcus HIV: : human immunodeficiency virus IAP: : intrapartum antibiotic prophylaxis LO: : late-onset PCR: : polymerase chain reaction Despite recommendations for universal screening and intrapartum antibiotic prophylaxis strategies for pregnant women, group B Streptococcus remains a major cause of neonatal disease. After completing this article, readers should be able to: ### Organism Streptococcus agalactiae , or group B Streptococcus (GBS), was first recognized as a distinct entity in the 1930s by Rebecca Lancefield, who used immunologic typing of carbohydrate antigens as a means to classify streptococci. (1) Early studies by her group and others indicated that GBS was an uncommon cause of human disease and was more frequently isolated as an etiologic agent of bovine mastitis. Sporadic case reports from that time demonstrated the potential for GBS to cause invasive infections, especially in peripartum women, although such infections were believed to be infrequent. However, over the period of the 1950s to the early 1970s, GBS became the major cause of neonatal sepsis in the United States and worldwide. ### Colonization and Pathogenesis GBS asymptomatically colonizes the lower gastrointestinal and female genital tracts. Colonization rates …

Arrival of rabbit haemorrhagic disease virus 2 to northern Europe: Emergence and outbreaks in wild and domestic rabbits (Oryctolagus cuniculus) in Sweden.

Incursion of rabbit haemorrhagic disease virus (RHDV) into Sweden was documented in 1990 and it is now considered endemic in wild rabbit (Oryctolagus cuniculus) populations. Rabbit haemorrhagic disease virus 2 (RHDV2), a new, related lagovirus was first detected in France in 2010, and has spread rapidly throughout Europe and beyond. However, knowledge of RHDV2 in northern Europe is sporadic and incomplete, and in Sweden, routinely available diagnostic methods to detect rabbit haemorrhagic disease (RHD) do not distinguish between types of virus causing disease. Using RHDV2-specific RT-qPCR, sequencing of the VP60 gene and immunological virus typing of archived and prospective case material from the National Veterinary Institute's (SVA) wildlife disease surveillance programme and diagnostic pathology service, we describe the emergence of RHDV2 in Sweden in both wild and domestic rabbits. The earliest documented outbreak occurred on 22 May 2013, and from May 2013 to May 2016, 10 separate incidents of RHDV2 were documented from six different municipalities in the southern half of Sweden. Phylogenetic analysis of the VP60 gene shows clear clustering of Swedish isolates into three separate clusters within two different clades according to geographic location and time, suggesting viral evolution, multiple introduction events or both. Almost all cases of RHD examined by SVA from May 2013 to May 2016 were caused by RHDV2, suggesting that RHDV2 may be replacing RHDV as the predominant cause of RHD in Sweden.

Immunological subtypes analysis of Uygur diffuse large B-cell lymphoma in Xinjiang and their prognostic significance.

This study aims to explore the application of Choi's typing method in the immunological typing of diffuse large B-cell lymphoma (DLBCL) in Xinjiang Autonomous Region and its prognostic significance. Seventy-eight cases of DLBCL tumor tissues from Xinjiang were collected to detect the expression of germinal center B (GCB) cell-expressed transcript-1, FOXP1, CD10, bcl-6, and MUM1 using an immunohistochemical method. Then, immunological typing was carried out using Choi's typing method, and the survival analysis was performed using Kaplan-Meier method. Cox proportional hazard model was used to analyze the prognostic factors. GCB-cell-like-DLBCL and non-GCB-DLBCL accounting for 29.5% (23/78) and 70.5% (55/78), respectively. The 3-year overall survival of GCB-DLBCL was 58%, significantly higher than that of non-GCB-DLBCL (39%, P < 0.05). Multivariate analysis showed that International Prognostic Index and immunological typing were two independent prognostic factors for Uygur patients with DLBCL. Non-GCB-DLBCL is the main type of DLBCL in Xinjiang and Choi's typing method can be a helpful indicator to determine the prognosis of the Uygur DLBCL in Xinjiang.

Cost-Effectiveness of the Management of Rh-Negative Pregnant Women

ObjectiveThe purpose of this study was to determine the most cost-effective option to prevent alloimmunization against the Rh factor. MethodsA virtual population of Rh-negative pregnant women in Quebec was built to simulate the cost-effectiveness of preventing alloimmunization. The model considered four options: (1) systematic use of anti-D immunoglobulin; (2) fetal Rh(D) genotyping; (3) immunological determination of the father’s Rh type; (4) mixed screening: immunological determination of the father’s Rh type, followed if positive by fetal Rh(D) genotyping. Two outcomes were considered, in addition to the estimated costs: (1) the number of babies without hemolytic disease, and (2) the number of surviving infants. ResultsIn a first pregnancy, two options emerged as the most cost-effective options: systematic prophylaxis and immunological Rh typing of the father, with overlapping confidence intervals between them. In a second pregnancy, the results were similar. In all cases (first or second pregnancy or a combination of the two) fetal genotyping was not found to be a cost-effective option. ConclusionRoutine prophylaxis and immunological Rh typing of the father are the most cost-effective options for the prevention of Rh alloimmunization. Considering that immunological typing of the father would probably not be carried out by the majority of clinicians, routine prophylaxis remains the preferred option. However, this could change if the cost of Rh(D) fetal genotyping fell below $140 per sample.

Immunosuppression and lung cancer of donor origin after bilateral lung transplantation

Analysis of databases from transplant recipients revealed a 3–5 fold higher risk to develop de novo malignancies under continued immunosuppression. The underlying mechanisms are poorly understood. Here we describe a patient who received a bilateral lung transplantation for end-stage ‘Usual Interstitial Pneumonia’ (UIP) resulting in idiopathic lung fibrosis. The recipient presented with a non-small cell lung carcinoma (NSCLC) in the donor lung 7 months later. Molecular and immunological typing of the tumor revealed a cancer of donor origin with a prominent intratumoral immune cell infiltrate without detectable effector function. This is a unique case of de novo outgrowth of a NSCLC of donor origin under continued immunosuppression, supporting the concept of tumor immunosurveillance in vivo.

Injury of the coronary endothelium at implantation increases endothelial dysfunction and intimal hyperplasia after heart transplantation

Background Coronary endothelial dysfunction occurs early after heart transplantation and predicts the development of intimal thickening characteristic of cardiac allograft vasculopathy. Objectives To assess the effects of removal of the endothelium by balloon injury of coronary arteries of allografts without rupture of the internal elastic lamina at the time of implantation and on coronary endothelial dysfunction, and to assess the development of accelerated atherosclerosis after heart transplantation. Methods A porcine model of heterotopic heart transplantation with preoperative immunologic typing, enabling progressive rejection without immunosuppression, was used to study the effect of endothelial removal on these 2 end points. Endothelium-dependent relaxations of epicardial coronary arteries from allografts submitted to endothelial denudation after harvest, arteries from allografts not undergoing denudation, and native coronary arteries were compared 30 days after graft implantation by using standard organ chamber experiments. Intimal thickening was measured by light microscopy with a semiquantitative scale (0 to 4+ grading). Results Relaxations to serotonin and to bradykinin were significantly decreased in denuded arteries compared with nondenuded allograft arteries. There was a significant increase in the incidence of severe intimal hyperplasia in denuded arteries compared with nondenuded arteries, which were both significantly increased compared to native coronary arteries. Conclusions Endothelial injury at implantation worsens the endothelial dysfunction as a result of rejection after heart transplantation and compounds the intimal thickening leading to cardic allograft vasculopathy. All efforts should be deployed to maintain a morphologically intact and functional endothelium at the time of graft implantation.

Autoimmune Diabetes mellitus, Hodgkin’s Disease and Graves’ Ophthalmopathy in a Patient with 8.1 Ancestral Haplotype

In this report, we describe the case of a 43-year-old woman affected by type 1 diabetes mellitus diagnosed 8 years before, who developed Graves' disease 2 years after chemotherapy and mantle radiotherapy treatment for Hodgkin's disease. Bilateral Graves' ophthalmopathy appeared four months before our observations. Intravenous methyl-prednisolone therapy was started, but was interrupted due to severe metabolic failure. Autoantibodies (anti-islet cells, anti-thyroid, thyroid-stimulating, non-organ-specific) were positive. Since the clinical picture suggested a genetic immunological ground predisposing to autoimmunity, we evaluated her HLA haplotype. Genomic typing of the patient permitted identification of the 8.1 ancestral haplotype, a Caucasoid haplotype unique in its association with many immunopathological diseases. Moreover, we also observed a haplotype unusual in Caucasians, trans DRB1*1101, DQA1*0103, DQB1*0603. To our knowledge, HLA-related genetic risk of developing thyroid autoimmunity after neck irradiation has never been studied. Although we cannot confirm a direct association between the 8.1 ancestral haplotype or DRB1*1101, DQA1*0103, DQB1*0603 and the diseases described, we suggest considering immunological parameters and HLA typing in candidate patients for mantle radiation therapy for Hodgkin's disease or other tumors. HLA haplotype determination could be useful in identifying the patients at raised risk of developing autoimmune diseases after irradiation, thus permitting a more appropriate follow-up schedule.

Inhibiting the NO pathway with intracoronary L-NAME infusion increases endothelial dysfunction and intimal hyperplasia after heart transplantation

Background: The endothelium protects the vascular wall through the nitric oxide (NO) release. Coronary endothelial dysfunction occurs early after heart transplantation and predicts the development of intimal thickening typical of graft coronary vasculopathy. Objective: We designed this study to examine the effect of endothelial NO synthase (eNOS) inhibition on the endothelial dysfunction caused by rejection and on the development of accelerated atherosclerosis after heart transplantation. Methods: To study the effect on these 2 end-points of inhibiting eNOS with intracoronary L-nitro arginine methyl ester (L-NAME; 1 mg/kg/day), infused with an osmotic pump for 30 days, we used a porcine model of heterotopic heart transplantation with pre-operative immunologic typing, to permit slow rejection without the need for immunosuppression. The endothelium-dependent relaxations of allografted coronary arteries, allografted arteries infused with L-NAME, allografted arteries mounted with the pump, and vehicle and native coronary arteries were compared 30 days after graft implantation using standard organ chamber experiments. We evaluated intimal thickening using a semi-quantitative scale (0–4+ grading). Results: A significant decrease in relaxations to serotonin (5-HT) occurred in allografted arteries infused directly with L-NAME compared with allografted arteries from swine receiving 5-HT, and relaxations in the latter were decreased compared with those of swine receiving the vehicle and native coronary arteries ( p < 0.05). We found no significant differences in endothelium-dependent relaxations to bradykinin among coronary rings from all groups. We observed a significant increase in the prevalence and severity of intimal thickening in allografted coronary arteries infused with L-NAME compared with allografts not infused ( p < 0.05), which had significantly more intimal thickening compared with native coronary arteries ( p < 0.05). Conclusion: These results demonstrate that inhibiting the NO pathway worsens the endothelial dysfunction caused by rejection after heart transplantation and accelerates the intimal thickening process, leading to graft coronary vasculopathy. Strategies designed to preserve endothelial integrity and function of the endothelial NO pathway should be used to prevent graft coronary vasculopathy.

Anaesthetic management and outcome in right-lobe living liver-donor surgery.

We reviewed retrospectively the anaesthetic management and perioperative course of eight right hepatectomies for living liver donation. After preoperative psychiatric evaluation, eight ASA I-II individuals donated the right lobe of their liver to a family member. A graft-recipient body weight ratio of 0.8-1.0% was required for patient selection. Indications for liver transplantation were: hepatitis C viral-related cirrhosis in six patients; combined hepatitis C and B viral cirrhosis in one patient; multifocal hepatocellular carcinoma--four lesions, involving both liver lobes--of hepatitis C viral-related cirrhosis in another patient. Indication for adult-to-adult living-donor liver transplantation was retained in the latter because of rapid deterioration of liver disease, rare recipient's blood group and extended, unresectable hepatocellular carcinoma. Hepatitis C viral-related cirrhosis was casually the primary indication for adult-to-adult living-donor liver transplantation in this group. The condition of the donated hepatic lobe was optimized by appropriate drug and perfusion management. Preoperative investigations included: blood tests (full cell count and film, thyroid function tests, pregnancy tests, full virological tests and bacteriological cultures, and immunological typing), chest radiograph, electrocardiogram plus Doppler cardiac ultrasound, spirometry, aminopyrine breath test, liver Doppler examination, magnetic resonance imaging, angiography and cholangiography and a volumetric study of the whole liver and the right lobe. Haemoglobin and lactate concentrations, liver function tests and international normalized ratio were measured before and after operation. The volume and weight of the resected right lobe was calculated. Anaesthesia was induced with propofol 300 mL h(-1) and sufentanil 0.3 microg kg(-1) intravenously; cisatracurium, 0.15 mg kg(-1), was given to facilitate tracheal intubation. Anaesthesia was maintained during normocapnic ventilation of the lungs with oxygen 40% in air, isoflurane 1-1.5 MAC and sufentanil. Routine anaesthetic monitoring included electrocardiography, pulse oximetry, invasive blood pressure, central venous pressure, urine output, state of neuromuscular blockade and core temperature. Periods of hypotension (<80% of the preoperative blood pressure) or haemodynamic instability (requiring inotropic or vasoactive support) were registered. Total blood loss and transfusion (homologous, autologous or cell-saver blood) requirements were measured; volume replacements were derived. Data are presented as mean (range). There was no morbidity or mortality and no periods of intraoperative hypotension or haemodynamic instability. The operation time averaged 619 (525-780)min. Four donors were extubated in the operating room immediately after surgery; the others were extubated in the intensive care unit, where the mean extubation time was 16.3 (5-25)h after arrival. The estimated blood loss was 967 (550-1,600)mL. No homologous blood was administered; five donors received autologous blood, intraoperatively; three donors received a cell-saver blood transfusion. Intraoperative fluid replacement was with crystalloids, colloids and 4% albumin. Total urine output was 1,472 (700-3100)mL. Although intraoperative hypothermia occurred all subjects were normothermic at the end of operation. The pre- and immediately postoperative haemoglobin concentration averaged 13.6 (9.8-15.6) and 10.5 (6.9-13.0)gdL(-1), respectively. On the first postoperative day, the haemoglobin was 11.7 (8.4-15.1)gdL(-1). The donors' liver function tests were transiently elevated in the initial postoperative period. The intensive care unit discharge time was 2 (1-3) days. The hospital stay was 13 (7-17) days. There was no morbidity or mortality. The study demonstrates that right-lobe living-donor surgery was well tolerated, without intraoperative hypotension or haemodynamic instability, without perioperative anaesthetic or surgical complications, and with an excellent general outcome.

Anaesthetic management and outcome in right-lobe living liver-donor surgery

Background and objective: We reviewed retrospectively the anaesthetic management and perioperative course of eight right hepatectomies for living liver donation. Methods: After preoperative psychiatric evaluation, eight ASA I-II individuals donated the right lobe of their liver to a family member. A graft-recipient body weight ratio of 0.8-1.0% was required for patient selection. Indications for liver transplantation were: hepatitis C viral-related cirrhosis in six patients; combined hepatitis C and B viral cirrhosis in one patient; multifocal hepatocellular carcinoma - four lesions, involving both liver lobes - of hepatitis C viral-related cirrhosis in another patient. Indication for adult-to-adult living-donor liver transplantation was retained in the latter because of rapid deterioration of liver disease, rare recipient's blood group and extended, unresectable hepatocellular carcinoma. Hepatitis C viral-related cirrhosis was casually the primary indication for adult-to-adult living-donor liver transplantation in this group. The condition of the donated hepatic lobe was optimized by appropriate drug and perfusion management. Preoperative investigations included: blood tests (full cell count and film, thyroid function tests, pregnancy tests, full virological tests and bacteriological cultures, and immunological typing), chest radiograph, electrocardiogram plus Doppler cardiac ultrasound, spirometry, aminopyrine breath test, liver Doppler examination, magnetic resonance imaging, angiography and cholangiography and a volumetric study of the whole liver and the right lobe. Haemoglobin and lactate concentrations, liver function tests and international normalized ratio were measured before and after operation. The volume and weight of the resected right lobe was calculated. Anaesthesia was induced with propofol 300 mL h−1 and sufentanil 0.3 μg kg−1 intravenously; cisatracurium, 0.15 mg kg−1, was given to facilitate tracheal intubation. Anaesthesia was maintained during normocapnic ventilation of the lungs with oxygen 40% in air, isoflurane 1-1.5 MAC and sufentanil. Routine anaesthetic monitoring included electrocardiography, pulse oximetry, invasive blood pressure, central venous pressure, urine output, state of neuromuscular blockade and core temperature. Periods of hypotension (<80% of the preoperative blood pressure) or haemodynamic instability (requiring inotropic or vasoactive support) were registered. Total blood loss and transfusion (homologous, autologous or cell-saver blood) requirements were measured; volume replacements were derived. Results: Data are presented as mean (range). There was no morbidity or mortality and no periods of intraoperative hypotension or haemodynamic instability. The operation time averaged 619 (525-780) min. Four donors were extubated in the operating room immediately after surgery; the others were extubated in the intensive care unit, where the mean extubation time was 16.3 (5-25) h after arrival. The estimated blood loss was 967 (550-1600) mL. No homologous blood was administered; five donors received autologous blood, intraoperatively; three donors received a cell-saver blood transfusion. Intraoperative fluid replacement was with crystalloids, colloids and 4% albumin. Total urine output was 1472 (700-3100) mL. Although intraoperative hypothermia occurred all subjects were normothermic at the end of operation. The pre- and immediately post-operative haemoglobin concentration averaged 13.6 (9.8-15.6) and 10.5 (6.9-13.0) g dL−1, respectively. On the first postoperative day, the haemoglobin was 11.7 (8.4-15.1) g dL−1. The donors' liver function tests were transiently elevated in the initial postoperative period. The intensive care unit discharge time was 2 (1-3) days. The hospital stay was 13 (7-17) days. There was no morbidity or mortality. Conclusions: The study demonstrates that right-lobe living-donor surgery was well tolerated, without intraoperative hypotension or haemodynamic instability, without perioperative anaesthetic or surgical complications, and with an excellent general outcome.

Surgical Experience With Retroperitoneal Heterotopic Heart Transplantation in the Large White Domestic Swine

Accelerated coronary atherosclerosis following heart transplantation is the main limiting factor for long-term survival, aside from graft failure and complications due to immunosuppression. Graft coronary vasculopathy is due to chronic rejection of the vascular wall leading to intimal hyperplasia in coronary arteries. Numerous heterotopic heart transplantation models have been used in different species to study the immunology and pathophysiology following graft implantation. This study reports our experience with the retroperitoneal heterotopic heart transplantation in Large White domestic swine using immunological typing. This approach mimics the kinetics of slow low-grade rejection in clinical human heart transplantation. One hundred and fifty-four retroperitoneal ( n = 154) heterotopic heart transplantations were performed using Large White swine sampled for the majorhistocompatibility class (MHC) class I antigen and blood type using the microlymphotoxicity technique. Acute rejection studies were performed by intentional mismatch of the swine lymphocyte alloantigen (SLA) and chronic rejection studies were done in allografts implanted in donor-recipients matched for blood type and class I antigen to assess the effects of rejection per se, hypercholesterolemia, intracoronary L-NAME infusion, and endothelial denudation on the development of graft coronary vasculopathy. Assessment of in vitro coronary arterial vascular reactivity in standard organ chamber experiments comprised the core of vascular biology studies in this large animal model. Eighty (52%) transplanted recipients survived until the elective date of sacrifice at 60 days, 14 (9.1%) died during the surgery, 21 (13.6%) died < 24 h after the transplant, and 8 (5.2%) died of late deaths. The retroperitoneal heterotopic heart transplantation model with blood typing and determination of the SLA class I antigen is a useful model for the study of immunological and vascular events due to graft rejection after heart transplantation.

Adhesion Molecule Expression, Clinical Features and Therapy Outcome in Childhood Acute Lymphoblastic Leukemia

In view of the relevance of adhesion molecule expression for the mechanisms of homing, trafficking and spreading of malignant cells, we have investigated the expression of surface adhesion molecules in lymphoblasts from 57 acute lymphoblastic leukemia (ALL) cases and tried to correlate the adhesive phenotype with immunological typing, prognostic factors at diagnosis and clinical follow-up. Blasts from all cases expressed adhesion molecules at high rates. β1 integrin chain (CD18) was consistently found on blasts from most ALL cases; among integrins of the β2 family, LFA-1 was detected in 58% of cases, in the virtual absence of other α chains. CD54 and CD58 were expressed in variable proportions by ALL blasts and CD44 was detected in the majority of the malignant cells, whereas the CD62L selectin was only present in 24% of cases. B-lineage ALL's displayed similar adhesion molecule phenotypes irrespective of maturational stages of the leukemic cells. We found a significantly reduced expression of β2 αL integrins in the hybrid ALL cases (CD13 and/or CD33 positive). However, these cases did not show differences in clinical presentation and behaviour in comparison with patients of other groups. We did not find a significant correlation between adhesion molecule expression and well established risk factors (age, white blood cell count, central nervous system involvement, chromosomal abnormalities), with the exception of splenomegaly, that was significantly associated with CD18 expression. In the follow-up, no evidence of significant correlation between adhesive phenotype and adverse events such as leukemic relapse and death was found. In conclusion, although expression of adhesion molecules on lymphoblasts confirms the phenotypic heterogeneity of ALL, it appears that this is not relevant for the clinical aspects of the disease and for prognosis.

Role of immunological lymphocyte subset typing as a screening method for lymphoid malignancies in daily routine practice

Role of immunological lymphocyte subset typing as a screening method for lymphoid malignancies in daily routine practice

Role of immunological lymphocyte subset typing as a screening method for lymphoid malignancies in daily routine practice

The major diagnostic role of peripheral lymphocyte subset typing is to distinguish between malignant and reactive conditions. The present study evaluates the screening efficacy of flow cytometric lymphocyte subset typing for the presence of a lymphoid malignancy. Four hundred samples were analyzed with a combination of anti-T-, B-, and natural killer (NK)-cell monoclonal antibodies. Two hundred and twenty (55%) samples showed a normal distribution of lymphocyte subsets, 73 (18%) samples exhibited unspecific alterations of lymphocyte subsets, 19 (5%) samples exhibited a reactive phenotype typical of Epstein-Barr virus/cytomegalovirus (EBV/CMV) infection, and 88 (22%) samples expressed a phenotype suggestive of lymphoma. The most predictive independent factor of a lymphoma-specific phenotype was the absolute lymphocyte count (P = 0.0001, odds ratio 73.225). Seventy-eight percent of samples containing >/=4 x 10(9)/l lymphocytes and 2% of samples with lymphocyte counts <4 x 10(9)/l exhibited a lymphoma-specific phenotype. The specificity of the referring clinical comment was the second best predictor of a lymphoma-specific typing outcome (P = 0.0001, odds ratio 19.589). The independent predictive values of lymphocyte morphology and of relative lymphocyte counts were of borderline significance. The use of flow cytometric lymphocyte subset typing as a diagnostic screening method for lymphoma should be restricted to cases of unexplained elevation of absolute lymphocyte counts with or without morphological atypias and to cases with definite clinical symptoms of lymphoma.

EBV-RELATED MONOCLONAL GAMMOPATHY (MGUS) IN LONG-TERM RENAL ALLOGRAFT RECIPIENTS IS ASSOCIATED WITH ACTIVATION OF CD8+ T CELLS WHICH MAY PREVENT DEVELOPMENT OF B-CELL LYMPHOMA

698 The EBV is associated with a spectrum of B-cell lymphoproliferative diseases (PTLD) that develop following organ transplantation. According to Hanto PTLDs can be classified on their clinical, histologic, immunologic cell typing, cytogenetic, Ig gene rearrangement, and virological characteristics into four disease groups: i) uncomplicated posttransplant infectious mononucleosis, ii) benign polyclonal polymorphic B-cell hyperplasia, iii) early malignant transformation in polyclonal polymorphic B-cell lymphoma, iv) monoclonal polymorphic B-cell lymphoma. In addition, a significant proportion of long-term transplant patients (in our center 10%) develop monoclonal gammopathy of undetermined significance (MGUS). We found an association between MGUS in long-term kidney transplant patients and high expression of EBV-latency related EBER-1 transcripts in peripheral blood B cells (at least five times elevated, p< 0.001). By contrast gene transcripts associated with the lytic virus cycle were rarely detectable. A similar latency-associated EBV expression pattern is found in B-cell lymphoma (PTLD type iv). The onset of disease after transplantation, age of patients, virological characteristics, appearance of monoclonal Ig is comparable in both groups. What may be the reason for the different clinical course in patients with EBV-related B-cell lymphoma or MGUS? Interestingly, in contrast to control long-term transplant patients (A) and patients with B-cell lymphoma(B) we found very low numbers of circulating CD19+ B cells in the peripheral blood of MGUS patients (C) (A: 5.3 +/- 2.5%; B: 15 +/- 4%; C: 2.8 +/- 1.3%[p<0.01 C vs A and B]). In addition, patients with MGUS showed significantly higher numbers of CD8+ T cells expressing memory/effector cell markers (LFA-1+++, CD57+) (p>0.05). These data demonstrate an association between EBV infection and both B-cell lymphoma and MGUS. Obviously, T cells control the expansion of EBV-infected B-cell clones in MGUS but not in lymphoma patients as suggested by the expansion of circulating CD8+ T-effector cells and the parallel decrease of CD19+ B cells in MGUS patients. The development of lymphoma in some MGUS patients suggest distruption of this sensitive balance. Therefore, CD19+ B-cell counts should be carefully monitored in MGUS patients for early detection of ongoing PTLD in these patients.