Generation and characterization of ex vivo propagated autologous CD8+ cells used for adoptive immunotherapy of patients infected with human immunodeficiency virus

Cytolytic T lymphocytes play an important role in host defense against viral infections, including human immunodeficiency virus (HIV). In a phase I clinical trial (protocol 080 of the AIDS Clinical Trials Group), generation of CD8+ effector cells from peripheral blood of patients with acquired immunodeficiency syndrome (AIDS)-related complex (ARC) or AIDS and safety of autologous adoptive transfer of these cells were evaluated. For therapeutic infusions, CD8+ T cells were purified by positive selection on anti-CD8 monoclonal antibody-coated flasks from leukapheresed peripheral blood of seven patients. These CD8+ T cells were cultured in the presence of interleukin-2 and phytohemagglutinin for up to 3 weeks to obtain cells sufficient for therapeutic infusions (10(8) to 10(10)). All 31 cell cultures established from the seven patients and used for therapy were highly enriched in CD8+ (mean, 97%), CD8+HLA-DR+ (50%), cytotoxic CD8+CD11b- (82%), and memory CD29+ (78%) T lymphocytes. In vitro expanded CD8+ cells had excellent cytotoxic function at the time they were used for therapy, including HIV-specific activity against autologous targets infected with vaccinia vectors expressing HIV-IIIb antigens, gag, pol, and env. Anti-HIV activity of cultured CD8+ cells was significantly higher than that of autologous fresh peripheral blood lymphocytes. Our results show that CD8+ T lymphocytes obtained from peripheral blood of symptomatic HIV-infected patients can be purified, cultured to obtain large numbers of cells with enhanced anti-HIV activity, and safely infused into patients with AIDS as a form of immunotherapy.     

Hepatitis G virus infection in patients with hepatitis C virus infection undergoing liver transplantation

BACKGROUND & AIMS: Hepatitis G virus (HGV) is transmissible by blood transfusion, but its role in chronic liver disease is unknown. The aim of this study was to determine the prevalence of HGV infection in patients infected with hepatitis C virus (HCV) undergoing transplantation and evaluate the effects of HGV coinfection on the course of posttransplantation HCV infection. METHODS: One hundred twenty-four patients infected with HCV undergoing liver transplantation were studied. Serum samples were tested for HCV and HGV RNA; HCV RNA was quantitated by branched DNA assay, and HCV genotype was determined. RESULTS: The prevalence of pretransplantation and posttransplantation HGV infection was 24% and 28%, respectively. Pre-transplantation HGV infection was positively correlated with posttransplantation HGV infection (P < 0.001). Pretransplantation clinical features were not different in patients infected with HCV with and without HGV infection. Posttransplantation HCV RNA levels were not significantly different in patients with and without HGV coinfection, but HCV genotype 1b was more frequent in patients with HGV coinfection. There were no differences in the histological severity of posttransplantation liver disease, graft, and patient survival between patients with and without HGV infection. CONCLUSIONS: Although HGV coinfection is frequent in patients with end- stage HCV disease undergoing liver transplantation, there is no association between the presence of HGV coinfection and the severity of liver disease post-transplantation, graft, or patient survival. (Gastroenterology 1996 Dec;111(6):1569-75)     

Decreased inducible expression of CD80 and CD86 in human monocytes after ultraviolet-B irradiation: its involvement in inactivation of allogenecity

Ultraviolet-B (UV-B) irradiation of antigen presenting cells (APCs) modifies their allogenecity, resulting in inhibition of the proliferative response of T cells in mixed lymphocyte reaction (MLR). Costimulation by the CD28 ligand CD80 (B7/B7-1) and CD86 (B70/B7-2) plays an important role during T-cell proliferation by augmenting synthesis of interleukin-2 (IL-2) and other cytokines. In this study, we demonstrated induced expression of both CD80 and CD86 during allogeneic MLR, though human freshly isolated monocytes express CD86 constitutively with a much lower level of CD80. A monoclonal antibody (MoAb) against CD86, but not CD80, efficiently inhibited allogeneic T- cell proliferative responses stimulated with highly purified monocytes. UV-B exposure (0 to 1,000 J/m2) of monocytes inhibited the proliferation of T lymphocytes in MLR in a dose-dependent manner. Flow cytometric analysis showed that UV-B exposure of monocytes impaired the constitutive expression of CD54 (intercellular adhesion molecule-1) by 24 hours after irradiation, but the effect on CD86 was relatively less. The surface expression of CD80, CD86, CD54, and HLA-DR on monocytes was further augmented by interferon (IFN)-gamma; this cytokine-induced expression was dose-dependently reduced by UV-B irradiation. Similarly, the upregulation of these molecules following allogeneic MLR was downregulated by UV-B irradiation. UV-B irradiation of monocytes inhibited the expression of IL-2 mRNA in monocyte-stimulated allogeneic MLR. In contrast, the addition of anti-CD28 MoAb at the onset of MLR prevented, at least partially, the reduction of IL-2 mRNA. These results strongly suggest that the impairment of inducible expression of CD86 and CD80 may contribute to the reduced MLR response following exposure of monocytes of UV-B.     

Identification of risk groups for development of central nervous system leukemia in adults with acute lymphocytic leukemia

The risk of development of CNS leukemia was investigated in 153 adults with acute lymphocytic leukemia (ALL) who received systemic combination chemotherapy without CNS prophylaxis. Overall, 31 patients (20%) developed CNS leukemia after a median of 6 months of therapy; the estimated 1-year incidence of CNS leukemia was 21% (SE, 3.9%). Characteristics significantly associated with CNS involvement included the presence of elevated hemoglobin creatinine, alkaline phosphatase, fibrinogen, and lactic dehydrogenase levels; B-cell leukemia; and high leukemic cell proliferative activity. Multivariate analysis identified lactic dehydrogenase levels of greater than or equal to 600 U/L and greater than or equal to 14% of cells in the S + G2M compartment to have independent additive poor prognostic significance. Patients were categorized into different risk groups for CNS leukemia with 1-year incidences ranging from 4% to 55%. While related to a high occurrence of CNS leukemia at diagnosis (33%) and subsequently (100%), the low incidence of B-cell disease excluded it from the multivariate analysis. The use of systemic chemotherapy containing multiple agents with good CNS penetration and in high doses (VAD regimen) in 90 patients was associated with a trend for lower CNS leukemia at 1 year (15% v 31%), especially in the low-risk category. We propose to develop future therapies for adults with ALL that include risk-oriented CNS prophylactic approaches.     

Tissue and corneal donation and transplantation in the UK

NHS Blood and Transplant (NHSBT) was established in 2005 as a Special Health Authority when the National Blood Authority and UK Transplant merged. This helped to bring tissue banking and organ transplantation services under one umbrella organization. This merger means that ~!95% of all deceased donors (whether tissue, organ or both) are now facilitated by one organization. NHSBT Tissue Services is the largest tissue establishment in the UK, and is a multi-tissue bank that specializes in the consent, retrieval, processing, storage, and dispatch of donated tissue coordinated from a purpose built, state-of-the-art tissue bank in Liverpool. Tissue donations can come from either tissue-only donors or solid organ donors who also donate tissue. Annually there are ~450 multi-tissue donors and 2500 eye donors in the UK, resulting in many thousands of transplants, including 3564 cornea transplants in 2010-2011. The separation of tissue- and organ-specific donors is largely artificial, and while organ transplantation can be life-saving, tissue transplantation can also have a dramatic effect on a patient's quality of life. It is hoped that all donors, both organ and tissue, will be recognized for the gift they make to society after their death.     

Serologic evidence that Scianna null (Sc:-1,-2) red cells lack multiple high-frequency antigens

Sera from three unrelated persons whose red cells (RBC) had the common Scianna phenotype (Sc:1,-2) contained IgG alloantibodies directed against high-frequency RBC antigens. In each case, sera or eluates or both failed to react only with Scianna null (Sc:-1,-2) cells, although an eluate from one person was compatible with a sibling's Sc:1,-2 cells. Cross-testing cells with sera or eluates, or both, from the three persons revealed no mutual compatibility. These studies show the existence of three additional RBC antigens phenotypically related to the Scianna blood group system. Sc:-1,-2 cells lack these antigens, which indicates that Scianna null cells lack multiple high-frequency antigens.     

肺泡膜弥散和肺毛细血管血量特征在不同级别慢性阻塞性肺病患者的表现

目的:了解不同级别慢性阻塞性肺病患者肺泡毛细血管膜弥散量和肺泡毛细血管血量的特点。方法:收集2006-07/2007-03就诊于四川大学华西医院呼吸科的慢性阻塞性肺病患者154例,依据慢性阻塞性肺病方案分级,0级64例,Ⅰ级38例,Ⅱ级26例,Ⅲ和Ⅳ级共26例。行肺通气功能、肺容量、一氧化碳弥散量、肺泡毛细血管膜弥散量和肺泡毛细血管血量测定。结果:154例患者均进入结果分析。①0级与I级慢性阻塞性肺病患者,无论一氧化碳弥散量、肺泡毛细血管膜弥散量和肺泡毛细血管血量都正常。随着慢性阻塞性肺病程度加重,一氧化碳弥散量、肺泡毛细血管膜弥散量和肺泡毛细血管血量均有不同程度降低。Ⅱ级慢性阻塞性肺病时一氧化碳弥散量和肺泡毛细血管膜弥散量下降明显,肺泡毛细血管血量改变不明显。其中肺泡毛细血管膜弥散量出现异常明显早于一氧化碳弥散量和肺泡毛细血管血量,而且较严重。Ⅲ和Ⅳ级慢性阻塞性肺病的肺泡毛细血管血量下降明显。②一氧化碳弥散量、肺泡毛细血管膜弥散量和肺泡毛细血管血量与慢性阻塞性肺病级别、残气量/肺总量成负相关,与第1秒用力呼气容积/用力肺活量和第1秒用力呼气容积占预计值百分比成正相关,而且,肺泡毛细血管膜弥散量与各指标的相关性最好。结论:随着慢性阻塞性肺病程度的加重,一氧化碳弥散量、肺泡毛细血管膜弥散量和肺泡毛细血管血量均有下降。肺泡毛细血管膜弥散量的异常比一氧化碳弥散量和肺泡毛细血管血量较早出现而且较严重。肺泡毛细血管膜弥散量的测定可以监测疾病发展并早期发现慢性阻塞性肺病气体交换的异常。     

α-促黑素细胞激素和白细胞介素8在病理性瘢痕组织中的表达及意义

目的:检测α-促黑素细胞激素、白细胞介素8在增生性瘢痕、瘢痕疙瘩、正常瘢痕和正常皮肤组织中的表达,探讨其在病理性瘢痕中的作用及意义。方法:①对象:收集2005-08/10中南大学湘雅医院烧伤整形科手术切下的瘢痕标本46例(其中增生性瘢痕18例、瘢痕疙瘩12例、正常瘢痕16例)和正常皮肤标本12例(患者和供皮者均知情同意)。②实验过程:标本用40g/L甲醛固定,行连续切片,厚4μm,应用链菌素生物素-过氧化物酶标记物免疫组织染色法染色。③实验评估:观察α-促黑素细胞激素和白细胞介素8在上述标本组织中的表达及相关性,采用着色强度和阳性细胞率二者评分相结合的方法判断结果。结果:①增生性瘢痕、瘢痕疙瘩中α-促黑素细胞激素的表达评分值和强阳性率高于正常瘢痕、正常皮肤(P<0.05);增生性瘢痕与瘢痕疙瘩比较、正常瘢痕与正常皮肤比较差异无显著性意义(P>0.05)。②增生性瘢痕、瘢痕疙瘩中白细胞介素8的表达评分值和强阳性率高于正常瘢痕、正常皮肤(P<0.05);增生性瘢痕与瘢痕疙瘩比较、正常瘢痕与正常皮肤比较差异无显著性意义(P>0.05)。③α-促黑素细胞激素、白细胞介素8在增生性瘢痕、瘢痕疙瘩和正常瘢痕中表达呈正相关(P<0.05),在正常皮肤中无相关性。结论:α-促黑素细胞激素、白细胞介素8在增生性瘢痕、瘢痕疙瘩中的表达显著高于正常瘢痕和正常皮肤,且二者在病理性瘢痕和正常瘢痕组织中表达呈正相关,提示α-促黑素细胞激素、白细胞介素8对促进病理性瘢痕的发生、发展可能起一定作用。