Immunomodulatory dendritic cells generated from nonfractionated bulk peripheral blood mononuclear cell cultures induce growth of cytotoxic T cells against renal cell carcinoma

Dendritic cells (DCs) loaded with tumor antigens have the potential to become a powerful tool for clinical cancer treatment. Recently, the authors showed that a tumor-specific immune response can be elicited in culture via stimulation with autologous renal tumor lysate (Tuly)-loaded DCs that were generated from cytokine-cultured adherent peripheral blood mononuclear cells (PBMCs). Here, the authors show that immunomodulatory DCs can be generated directly from nonfractionated bulk PBMC cultures. Kinetic studies of DC differentiation and maturation in PBMC cultures were performed by monitoring the acquisition of DC-associated molecules using fluorescence-activated cell sorting analysis to determine the percentage of positive immunostained cells and the mean relative linear fluorescence intensity (MRLFI). Compared with conventional adherent CD14+ cultures, which have mostly natural killer, T, and B cells removed before cytokine culture, bulk PBMC cultures exhibited an early loss of CD14+ cells (day 0 = 78.8%, day 2 = 29.6% versus day 0 = 74%, day 2 = 75%) with an increase in yield of mature DCs (DC19- CD83+) (day 5 = 17%, day 6 = 21%, day 7 = 22% versus day 5 = 11%, day 6 = 15%, day 7 = 23%). Although a comparable percentage of DCs expressing CD86+ (B7-2), CD40+, and HLA-DR+ were detected in both cultures, higher expression levels were detected in DCs derived from bulk culture (CD86 = MRLFI 3665.1 versus 2662.1 on day 6; CD40 = MRLFI 1786 versus 681.2 on day 6; HLA-DR = MRLFI 6018.2 versus 3444.9 on day 2). Cytokines involved in DC maturation were determined by polymerase chain reaction demonstrating interleukin-6 (IL-6), IL-12, interferon-gamma, granulocyte-macrophage colony-stimulating factor, and tumor necrosis factor-alpha mRNA expression by bulk culture cells during the entire 9-day culture period. This same cytokine mRNA profile was not found in the conventional adherent DC culture. Autologous renal Tuly (30 micrograms protein/10(7) PBMCs) enhanced human leukocyte antigen expression by DCs (class I = 7367.6 versus 4085.4 MRFLI; class II = 8277.2 versus 6175.7 MRFLI) and upregulated cytokine mRNAs levels. Concurrently, CD3+ CD56-, CD3+ CD25+, and CD3+ TCR+ cell populations increased and cytotoxicity against autologous renal cell carcinoma tumor target was induced. Specific cytotoxicity was augmented when cultures were boosted continuously with IL-2 (20 U/mL biological response modifier program) plus Tuly stimulation. These results suggest that nonadherent PBMCs may participate in enhancing DC maturation. Besides the simplicity of this culture technique, bulk DC cultures potentially may be used with the same efficiency as conventional purified DCs. Furthermore, bulk culture-derived DCs may be used directly in vivo as a tumor vaccine, or for further ex vivo expansion of co-cultured cytotoxic T cells to be used for adoptive immunotherapy.     

Processing of urushiol (poison ivy) hapten by both endogenous and exogenous pathways for presentation to T cells in vitro.

The antigen processing requirements for urushiol, the immunogen of poison ivy (Toxicodendron radicans), were tested by presentation of urushiol to cultured human urushiol-responsive T cells. Urushiol was added to antigen-presenting cells (APC) either before or after fixation with paraformaldehyde. Three distinct routes of antigen processing were detected. CD8+ and CD4+ T cells, which were dependent upon processing, proliferated if urushiol was added to APC before fixation, but did not proliferate when urushiol was added to APC after fixation. Processing of urushiol for presentation to CD8+ T cells was inhibited by azide, monensin, and brefeldin A. This suggests that urushiol was processed by the endogenous pathway. In contrast, presentation of urushiol to CD4+ T cells was inhibited by monensin but not by brefeldin A. This was compatible with antigen processing by the endosomal (exogenous) pathway. Finally, certain CD8+ T cells recognized urushiol in the absence of processing. These cells proliferated in response to APC incubated with urushiol after fixation. Classification of contact allergens by antigen processing pathway may predict the relative roles of CD4+ and CD8+ cells in the immunopathogensis of allergic contact dermatitis.     

Regulation of mouse immuno-responses by a natural suppressor cell clone from bone marrow

Natural suppressor (NS) activity is mediated by several kinds of cell lineages in bone marrow. We demonstrated that a NS cell, clone 7–31, was obtained after limiting dilution with bone marrow culture supplemented with WEHI supernatant. Clone 7–31, was capable of non-specific suppression of the generation of cytotoxic T lymphocytes without major histocompatibility complex restriction. Suppression of cytotoxic thymus-dependent lymphocyte (CTL) generation was also induced with a cell-free supernatant from the 7–31 cells. Interleukin-2 (IL-2) containing concanavallin-A-conditioned medium could not reverse the suppression. The supernatant did not inhibit the proliferation of IL-2-dependent CTLL-2 cells and rapidly growing tumour cells and fibroblasts. Thus, this bone marrow suppressor cell produces a soluble factor that inhibits the generation of CTL in vitro, and prolongs the acceptance of skin allografts in in vivo.     

肿瘤病人T细胞核仁嗜银蛋白和辅助性T细胞亚群变化

目的:探讨肿瘤病人外周血单个核细胞(PBMC)中T细胞核仁嗜银蛋白(AgNORs)变化和辅助性T(Th)细胞亚群变化的关系及其临床意义。方法PBMC经多克隆T细胞活化剂刺激,以图像分析法检测AgNORs区面积与细胞核仁面积的百分比(1.S％):PBMC被离子霉素和佛波醇酯活化5h后,以酶联免疫斑点法(ELISPOT)检测IFNY、和IL-4产生细胞的频度(％):以离子霉素加佛波醇酯刺激24h,用ELISA试刑盒检测培养上清液中的IFNY、和IL-4水平。结果 肿瘤病人(n=86)和正常人(n=44),PBMC中活化T细胞的AgNORs表达分别为4.9±1.21.S％和.7.82±1.28 1.S％,肿瘤组非常显著下降,P<0.05:肿瘤组PBMC中1112细胞额度显著上升,Th1／Th2比值显著下降:同时,肿瘤组PBMC产生IFN和IL-4水平发生相应变化,变化方式与Th亚群变化相似:在肺癌组中,转移的与未转移的,复发的与未复发的相比,Th1细胞数、Th1／Th2之比和IFNY、产生水平非常显著下降:在肿瘤组AgNORs表达与Th1细胞频度、Th1／Th2比值、IFNY产生和肿瘤病人生活状态的Kamofsky评分正相关。结论 现有病例观察结果表明,肿瘤病人PBMC小T细胞表达AgNORs下降,与患者Th1细胞功能下降正相关。     

ABO血型不合的同胞异基因外周血干细胞移植

目的探讨HLA配型相合、ABO血型不合的同胞异基因外周血干细胞移植(alloPBSCT)的疗效。方法对27名HLA配型相合、ABO血型不合的血液恶性肿瘤患者作同胞alloPBSCT(实验组,供、受者ABO血型主侧不合的有15例,次侧不合的有10例,主次侧均不合的有2例),其中急性髓细胞白血病(AML)6例、急性淋巴细胞白血病(ALL)8例、慢性粒细胞性白血病(CMLLP)10例、骨髓增生异常综合征(MDSRAEBT)2例、非霍奇金氏淋巴瘤(ⅣB)1例;并选用同期的35名ABO血型相合的移植患者作比较(对照组)。移植物抗宿主病(GVHD)的预防采用霉酚酸酯(MMF)、环孢菌素A(CSA)和短程甲氨喋呤(MTX)三联预防方案。结果62例全部造血重建。实验组:27名alloPBSCT患者均未出现急性溶血反应,主侧不合者红系造血明显延迟,供/受者血型为A/O的患者中有3例(3/7)发生纯红细胞再生障碍性贫血(PRCA),27名患者于移植后25～153d血型成功转变为供者型;实验组GVHD发生率、VOD发生率、CMV感染、HC发生率及疾病复发率、死亡率与对照组相比差异无统计学意义(P>0.05)。结论ABO血型不合可以进行alloPBSCT,并且不影响干细胞移植的植活、GVHD及其它移植相关并发症的发生和预后。供/受者血型为A/O是主侧ABO血型不合患者alloPBSCT后PRCA发生的高危因素。     

Regulation of cytolytic cell populations from human peripheral blood by B cell stimulatory factor 1 (interleukin 4)

The effects of B cell stimulatory factor 1 (BSF-1) on the generation of human CTL and lymphokine-activated killer (LAK) cells in vitro were investigated. Both L-2 and BSF-1 were potent helper factors for the generation of antigen-specific CTL in MLC; detection of optimal BSF-1-induced CTL activity in this system occurred when BSF-1 was added to cultures after an initial period of activation during which exogenous BSF-1 was not present. In contrast to IL-2, BSF-1 failed to induce an LAK cell population, as detected with Daudi tumor targets, in cultures that had not been allosensitized. Furthermore, when both lymphokines were added together at culture initiation, BSF-1 inhibited the IL-2-driven generation of cytolytic cells. The differential ability of BSF-1 to promote the generation of CTL but not LAK could have important implications for lymphokine-mediated immunotherapy.     

强直性脊柱炎患者外周血T细胞亚群表达的临床意义

目的 探讨强直性脊柱炎(AS)患者外周血中T细胞亚群分布以及在AS细胞免疫中所发挥的作用.方法 采用流式细胞技术检测30例未经治疗的活动期AS患者及30例健康对照人群外周血中的CD4+、CD8+T细胞亚群,并探讨其与Bath AS功能指数(BASFI)、Bath AS测量指数(BASMI)、病程、年龄及血沉(ESR)、超敏C反应蛋白(hs-CRP)的相关性.结果 AS患者CD4+水平和CD4+/CD8+比值分别为(29.24±9.22)%和0.96±0.49,较正常对照组的(40.09±6.86)%和1.70±0.67显著降低(P均＜0.01);CD8+水平为(32.91±6.86)%,较正常对照组的(25.60±5.97)%明显增高(P＜0.01);AS患者外周血CD4+水平与BASMI呈负相关(r=-0.479,P＜0.01).CD8+与血沉、hs-CRP、BASFI、BASMI呈正相关(r值分别为0.373、0.430、0.462、0.530,P均＜0.05).CD4+/CD8+比值与hs-CRP、BASFI、BASMI呈负相关(r值分别为-0.465、-0.473、-0.426,P均＜0.05).CD4+、CD8+、CD4+/CD8+比值与AS患者的年龄、病程均无明显相关性(P均＞0.05).结论 AS患者外周血存在T淋巴细胞亚群紊乱,其水平与AS的严重程度、活动性有关,提示T细胞亚群失衡在AS的发生发展中具有重要意义.     

血液透析患者血浆PGE2含量与T细胞亚群和NK细胞活性的关系

目的 探讨血液透析患者外周血前列腺素E2(PGE2)含量与T细胞亚群和NK细胞活性的关系。方法 应用流式细胞仪和酶联免疫分析法检测透析、未透析和长期维持性透析(透析时间大于7年)患者外周血PGE2、T细胞表面CD3、及其亚群CD4及CD8抗原表达。结果 尿毒症未透析组(NHD组)的外周血NK细胞和T细胞的CD3^ 、CD4^ 、CD8^ 表达及CD4^ /CD8^ 比值均显著低于正常对照组(P＜0．05)；PGE2含量明显增高，并与CD4^ ／CD8^ 细胞比值和NK细胞活性间呈负相关。透析后，尿毒症患者PGE2水平降低，CD3^ 、CD4^ 和CD8^ 值及CD4/^ CD8^ 比值均显著上升(P＜0．05)，NK细胞活性接近于正常对照组。然而透析时间大于7年的尿毒症患者血浆PGE2水平又显著增高，外周血淋巴细胞数值、CD3^ 细胞浓度、CD4^ /CD8^ 细胞比值及NK细胞活性均显著低于正常组(P均＜0．05)。结论 PGE2异常增高与细胞免疫功能失衡有关，血液透析能够降低PGE2浓度，改善患者的免疫功能。但长期维持性透析患者免疫功能又呈现恶化状态。     

肿瘤病人T细胞核仁嗜银蛋白和辅助性T细胞亚群变化

目的　探讨肿瘤病人外周血单个核细胞 (PBMC)中T细胞核仁嗜银蛋白 (AgNORs)变化和辅助性T(Th)细胞亚群变化的关系及其临床意义。方法　PBMC经多克隆T细胞活化剂刺激 ,以图像分析法检测AgNORs区面积与细胞核仁面积的百分比 (I.S % ) ;PBMC被离子霉素和佛波醇酯活化 5h后 ,以酶联免疫斑点法 (ELISPOT)检测γ干扰素 (IFNγ)和白细胞介素 4(IL 4)产生细胞的频度( % ) ;以离子霉素加佛波醇酯刺激 2 4h ,用ELISA试剂盒检测培养上清液中的IFNγ 和IL 4水平。结果86例肿瘤病人和 44名正常人 ,PBMC中活化T细胞AgNORs表达分别为 ( 4 90± 1 2 0 )I S %和 ( 7 82±1 2 8)I S % ,肿瘤组下降非常显著 (P <0 0 5 ) ;肿瘤组PBMC中Th2细胞频度显著上升 ,Th/Th2比值显著下降 ;同时 ,肿瘤组PBMC产生IFNγ 和IL 4水平发生相应变化 ,变化方式与Th亚群变化相似 ;肺癌转移组与未转移组、复发组与未复发组相比 ,Th1细胞数、Th1/Th2之比和IFNγ 的产生下降非常显著 ;在肿瘤组AgNORs表达与Th1细胞频度、Th1/Th2比值、IFNγ 产生和肿瘤病人生活质量评分呈正相关。结论　肿瘤病人PBMC中T细胞表达AgNORs下降 ,与患者Th1细胞功能下降呈正相关。     

Factors influencing the timing of peripheral blood stem cell collection (PBSC)

High-dose conditioning regimens followed by autologous peripheral blood stem cell rescue are frequently used for the treatment of solid tumors and hematological malignancies. In 24 patients up to four peripheral stem cell collections (PBSC) were performed after priming with various chemotherapies and G-CSF (300 micrograms s.c. per day). In 16 patients (group A) more than 2 x 10(6) CD 34 positive cells per kg bodyweight could be collected; fewer were harvested in the remaining eight patients (group B). The amount of collected CD 34 positive cells correlated with the median number of these cells in the peripheral blood at the start of PBSC. The two groups differed both in recovery time after priming-induced cytopenia (4 vs 6 days from nadir) and in the number of WBC (21 x 10(6) mL-1 vs 6.1 x 10(6) mL-1) and platelets (133 x 10(6) mL-1 vs 58 x 10(6) mL-1) reached at first day of PBSC. No difference between the two groups was seen according to age, duration of disease or disease status. However, the intensity of prior treatment was significantly greater in group B than in group A. These observations indicate that the toxicity of previous chemotherapy is the most important factor for the mobilization of sufficient CD 34 positive cells into the peripheral blood.     

"Helper" and "suppressor" T lymphocytes regulating blood cell formation in man

Human hemopoietic progenitor cells have been characterized as null lymphocytes. These primitive precursors are subject to interaction with mature cells which are derived from the thymus. Such T lymphocytes exist in several subpopulations which can exhibit "helper" and "suppressor" activity in various immune responses. Evidence for the involvement of these cells in hematopathological disorders has been presented and the possibility of a physiological role for helper and suppressor T cells in hemopoiesis has been discussed. It is proposed that "steady state" blood cell formation is maintained, at least in part, by the preponderance of suppressors in the bone marrow, while the dominant helper population in peripheral blood can recirculate under the influence of corticosteroids with consequent stimulation of hemopoiesis. The influence of histocompatibility antigens on this process of cellular regulation remains to be determined.     

T cell receptor V-segment frequencies in peripheral blood T cells correlate with human leukocyte antigen type

We compared T cell receptor (TCR) V-segment frequencies in human leukocyte antigen (HLA) identical siblings to sibling pairs who differ at one or both HLA haplotypes using four V beta-specific and one V alpha-specific monoclonal antibody. In every one of nine families HLA-identical sibs had the most similar patterns of V-segment frequencies in their peripheral blood, whereas totally mismatched sibs were, in general, the most dissimilar; HLA haploidentical sibs tended to be intermediate between the two groups. The degree of similarity among HLA-identical sibs was comparable to that observed among three pairs of identical twins suggesting that HLA is the major genetic component influencing TCR V-segment frequency. Consistent with this observation, it was found that the frequency of T cells expressing particular V beta segments was skewed towards either CD4+ or CD8+ cells indicating that T cells expressing some V beta genes may be positively selected primarily by class I or class II major histocompatibility complex proteins. Finally, it was observed that individuals who express the HLA class I specificity, B38, tend to express high levels of V alpha 2.3+ cells among their CD8+ T cells. These observations represent definitive proof that human V-segment frequencies are profoundly influenced by the HLA complex.     

The impact of peripheral blood stem cell transplants (PBSCT) on platelet usage

Peripheral blood stem cells (PBSC) are rapidly replacing bone marrow cells for autologous transplantation. This introduction, largely without randomized prospective trials, has occurred because of the ease of PBSC collection and the associated rapid haematological recovery with its lower costs and reduced blood product exposure. The administration of haematopoietic growth factors during recovery from high-dose chemotherapy increases the number of circulating haematopoietic progenitor cells to levels 1000-fold greater than levels normally found in blood. The CD34+ cell number, CFU-GM and CFU-Meg are commonly employed parameters used to assess the quality of PBSC harvests. This review examines the impact that PBSCT has had on haematological practice and patient care illustrated by our local practice in Cardiff.     

Human natural killer cells isolated from peripheral blood do not rearrange T cell antigen receptor beta chain genes

The lineage of NK cells and their relationship to T lymphocytes have been controversial issues. Since rearrangement of the T cell antigen receptor beta chain genes occurs early in the ontogeny and differentiation of all T cells, this can be used as an unequivocal marker to discriminate T from non-T lymphocytes. Recent studies (16-18) examining T cell antigen receptor gene rearrangement and expression in certain IL-2-dependent NK cell lines and leukemias have revealed that some lines rearrange C beta genes, whereas others do not. However, it is important to establish whether these cell lines are representative of the major population of NK cells freshly derived from the host. Herein, we have purified granulocytes, CD16+ NK cells and T lymphocytes from human peripheral blood, prepared genomic DNA from each cell type, and then examined the organization of their T cell antigen receptor genes by restriction enzyme analysis using a C beta cDNA as probe. The C beta genes were in germline configuration in NK cells and granulocytes. In contrast, peripheral blood T lymphocytes showed rearrangement of the C beta gene. These data support the hypothesis that the majority of human peripheral blood NK cells are fundamentally distinct from T lymphocytes in lineage and nonself recognition.     

A 180-kilodalton protein from Mycobacterium tuberculosis defined by a human T cell clone

A high molecular weight protein from Mycobacterium tuberculosis (M. tuberculosis) has been identified, that is recognized by peripheral blood mononuclear cells from several tuberculous patients and by a T cell clone derived from a patient with tuberculous pleurisy. Purification of this fraction demonstrated biological activity to reside in a 180-kDa protein component. This mycobacterial protein appears to exist in some, but not all mycobacteria as the clone reacts to M. tuberculosis, BCG M. kansasii, M. flavescens and M. fortuitum, but not to M. intracellulare, M. scrofulaceum or a variety of gram-positive or gram-negative bacteria, or to PPD. Specific anti-genic challenge of the T cell clone in the presence of irradiated antigen presenting cells results in proliferation and interleukin-2 (IL-2) production. Proliferation is restricted to HLA Class II antigens as antigenic recognition occurs only in the presence of either one of the two parental DR haplotypes. Peripheral blood mononuclear cells from several other patients with pulmonary tuberculosis also proliferate in response to this antigen, emphasizing the relevance of T cell cloning techniques in identifying important mycobacterial antigens.     

外周血辅助性T细胞17与调节性T细胞的平衡与白癜风临床特征的相关性

目的分析白癜风患者外周血辅助性T细胞17(Th17)与调节性T细胞(Treg)的平衡与临床特征的关系。方法选取白癜风患者109例,其中进展期患者56例,稳定期患者53例,另选取同期本院健康体检者59例为对照组。采用流式细胞术检测3组受试者外周血Th17、Treg细胞数量;采用酶联免疫吸附法(ELISA)检测3组受试者白介素-17(IL-17)、叉头盒蛋白P3(Foxp3)水平;采用Pearson法分析白癜风患者Th17、Treg细胞及IL-17、Foxp3的关系。结果对照组、稳定期组、进展期组Th17细胞含量、Th17/Treg、IL-17水平逐渐升高,Treg细胞含量、Foxp3水平逐渐降低,差异有统计学意义(P0.05)。随着白癜风皮损面积的增加,Th17细胞含量、Th17/Treg、IL-17水平逐渐升高,Treg细胞含量、Foxp3水平逐渐降低,差异有统计学意义(P0.05)。白癜风患者Th17细胞与Treg细胞、IL-17与Foxp3均呈显著负相关(r=-0.449、-0.497,P0.05)。结论 Th17/Treg细胞平衡可能与白癜风的发生及临床特征有关。     

Analysis of T helper and antigen-presenting cell functions in cord blood and peripheral blood leukocytes from healthy children of different ages.

The development of antigen-specific functional T lymphocyte immunity in infants and children is an area of immunology that needs elucidation. Leukocytes from cord blood (CBL) and from PBL of children of different ages who were in the hospital for minor surgical procedures were compared with PBL from healthy adults for their ability to generate T helper cell (Th) responses assessed by in vitro proliferation and IL-2 production after stimulation with: influenza A virus (FLU); tetanus toxoid (TET); adult allogeneic PBL that were either undepleted (ALLO) or depleted of adherent antigen presenting cells (ALLONW); and PHA. CBL generated Th responses to ALLONW, ALLO, and PHA, but not to FLU or TET. PBL from infants between 6 and 13 mo of age responded to ALLO and PHA; none responded to FLU or ALLONW, and two of four responded weakly to TET. PBL from children between 13 and 26 mo of age responded to all stimuli except FLU, to which only one child responded marginally. PBL from children older than 36 mo responded to all stimuli at levels comparable to those of PBL from adults. The use of undepleted and adherent cell-depleted CBL and PBL from children of different ages as allogeneic stimulators of responses generated by PBL from adults indicated that the antigen presenting function of CBL and PBL from children 13 mo or older are sufficiently developed to present alloantigen, whereas PBL from children younger than 13 mo are not. Therefore, our results indicate that age-dependent differences exist in both T helper and antigen-presenting functions of CBL and PBL from children of different ages. Surprisingly, CBL appear to be more efficient in antigen-presenting function than PBL from children younger than 13 mo. These findings are important for establishing developmental parameters of T helper cell immunity relevant for pediatric infection and transplantation in infants and children.     

Antigen-specific suppressor T cell interactions. II. Characterization of two different types of suppressor T cell factors specific for L-glutamic acid50-L-tyrosine50 (GT) and L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT)

We have previously reported that two types of suppressor T cell factors (TsF) specific for L-glutamic acid50-L-tyrosine50 (GT) or L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT) can be distinguished based upon differences in their ability to suppress responses by allogeneic mice. Injection of GAT or GT induces a suppressor T cell subset that produces an antigen-binding, I-J+, genetically unrestricted, specific suppressor factor (TsF1). Injection of this factor plus small amounts of antigen induces a second-order suppressor T cell that produces an antigen- binding, I-J+, genetically restricted, specific suppressor factor (TsF2). In this report, we demonstrate that these two factors are also biochemically distinct. Monoclonal TsF1 molecules are composed of a single polypeptide chain that bears both the antigen-binding site and I- J determinant, whereas TsF2 molecules are composed of two disulfide- linked polypeptide chains, one of which is antigen-binding and I-J-, and the other, nonantigen-binding, I-J+. The antigen-binding chain must be added at culture initiation to achieve suppression, but the I-J+ chain can be added as late as day 3 with complete suppression observed. However, isolated chains from TsF2-producing hybridomas derived from three different haplotypes were unable to suppress immune responses when chains from heterologous TsF2 were mixed. Indirect evidence is presented that suggests that this restriction is because the chains fail to interact rather than the inability of the target cells to recognize both chains.     

Use of a human plaque-forming cell assay to study peripheral blood bursa-equivalent cell activation and excessive suppressor cell activity in humoral immunodeficiency.

A plaque assay that detects human mononuclear blood cells producing immunoglobulin (Ig)M antibody to sheep erythrocytes was investigated for its usefulness in studying B-cell activation and regulation in 24 patients with humoral immunodeficiency. Cells from 3 of 15 patients with common variable agammaglobulinemia produced some plaques (range 40--160/10(6) cells; normal range 80--1240/10(6)), but those from the other 12, from all 7 with x-linked agammaglobulinemia and from the 2 with x-linked immunodeficiency with hyper-IgM failed to produce any detectable plaques. In co-cultures of patient and normal cells a very good correlation was seen between results of the plaque assay and an IgM biosynthesis assay in detecting excessive suppressor cell activity. Cells from 7 of 15 common variable agammaglobulinemics, from 3 of 7 x-linked agammaglobulinemics, and from both patients with hyper-IgM caused significant suppression of IgM biosynthesis and(or) plaque formation by normal cells. The observations in the last two groups and discordance for excess suppressor activity in identical twins with common variable agammaglobulinemia suggest that the activity develops secondarily to whatever their primary defects may be. Culturing non-T cells from common variable agammaglobulinemics exhibiting excessive suppressor cell activity with normal T cells resulted in plaque formation in four of five patients so studied; in all five the suppressor activity was found in the T-cell population. The availability of a plaque assay for the study of blood cells from immunodeficient patients provides a new probe to examine the cellular nature of such defects.     

落新妇甙对肺移植大鼠外周血T细胞活化的影响

背景:目前,对于排斥反应的研究主要是通过阻断T细胞活化来达到抑制排斥反应.而在众多体外试验中,落新妇甙及其类似物显示了典型的选择性免疫抑制特色.目的:观察落新妇甙对大鼠移植肺活化T细胞的影响及动态变化.探讨落新妇甙对大鼠移植肺活化T细胞的作用.方法:50只人鼠随机分为3组.其中10只不做移植,抽取外周血没为空白组.剩余40只大鼠随机均分供,受体,左侧单肺原位移植建模后再随机均分为2组:对照组肺移植后,用生理盐水灌胃;落新妇甙组移植后用落新妇甙灌胃.移植前、移植后第2,5天,分别用荧光标记抗体双染色结合流式细胞技术检测各阶段大鼠T细胞表达的活化抗原CD69、CD25和CD71的表达.结果与结论:移植后第2天,落新妇甙组检测的CD69、CD25和CD71抗原均显著高于空折组(P<0.01),但低于对照组,差异有显著性意义(P<0.05).移植后第5天,落新妇甙组CD69、CD25和CD71抗原基本降至正常水平,显著低于对照组(P<0.01).结果提示落新妇甙能显著抑制急性排斥反应诱导的T细胞活化,对肺移植排斥反应有较强的抑制作用.