Analysis of the immunoglobulin heavy chain gene variable region of 101 cases with peripheral B cell neoplasms and B cell chronic lymphocytic leukemia in the Japanese population

We have analyzed the immunoglobulin heavy chain (VH) gene variable regions (CDR2 and FW3) of 101 Japanese cases with peripheral B cell neoplasms. When all except one case with a deletion were graphed by frequency of replacement mutation, the 100 cases could be separated into two groups: 24 cases with zero, one and two mutations (germline or low frequency of somatic mutation); and 76 cases with three or more mutations (medium to high frequency of somatic mutation). While most mantle cell lymphoma cases (11/13) showed germline or low frequency of somatic mutation, all cases of mucosa-associated lymphoid tissue (MALT) lymphoma (11/11), follicular lymphoma (three of three cases), plasma cell myeloma (seven of seven cases) and most cases of diffuse large B cell lymphoma (DLBCL; 42/47) belonged to the latter group. These 76 cases, therefore, may be considered to show somatic hypermutation. More than half of chronic lymphocytic leukemia/small lymphocytic lymphoma cases (CLL/SLL; eight of 13) showed a hypermutated VH gene and the ratio of replacement mutation: silent mutation in CDR2 of CLL/SLL was considerably higher compared with DLBCL and MALT lymphoma, showing somatic hypermutation. When comparing VH gene type of B cell-CLL (B-CLL) among our series and those in the literature, more cases of CD5+ B-CLL in the Western literature have the VH5 and VH6 family types, while more cases in Japan are reported to have VH4 family. The occurrence of VH families in B-CLL between Japanese and Western people seems to be comparable.     

Increased dimeric IgA-producing B cells in tonsils in IgA nephropathy determined by in situ hybridization for J chain mRNA.

The origin of mesangial IgA deposits in IgA nephropathy (IgAN) remains obscure. A significant proportion of deposited immunoglobulin is dimeric (J chain-positive). Previous studies of J chain expression within lymphoid tissue in IgAN have utilized antibodies which other investigators have found to be non-specific. To address this problem, we have developed and in situ hybridization (ISH) method for the detection of J chain mRNA within IgA plasma cells. Tonsils from 12 patients with IgAN and 12 controls were studied using (i) non-isotopic ISH for J chain mRNA, and (ii) combined immunofluorescence (IF) and fluorescent ISH. J chain mRNA-positive cells were identified in germinal centres, and within the subepithelial and interfollicular zones. A greater number of J chain mRNA-positive cells were found in the germinal centres of patients (mean 57.7 +/- 4.6 cells/10(5) micron2) compared with controls (mean 36.9 +/- 3.5 cells/10(5) micron2) (P < 0.001). Combined IF and fluorescent ISH showed a greater proportion of J chain mRNA-positive interfollicular IgA cells in patient tonsils (32 +/- 3.4%) compared with controls (21 +/- 2.3%; P < 0.02). These results indicate a shift towards dimeric IgA production in the tonsils in IgAN. In addition, the finding of excess numbers of J chain-positive Iga-negative cells within germinal centres suggests that an abnormality may be present at the B cell differentiation stage before IgA switching. These results further highlight immune abnormalities within the tonsil as a central feature of abnormal polymeric IgA biology in this common form of glomerulonephritis.     

Antibody against the human J chain inhibits polymeric Ig receptor-mediated biliary and epithelial transport of human polymeric IgA

To emphasize the requirement for a J chain in native polymeric immunoglobulins for their selective transport into exocrine secretions, IgG, purified from two different antisera specific for the human J chain, was shown to: (i) bind in vitro to human polymeric IgA (pIgA) by density gradient ultracentrifugation; (ii) inhibit binding in vitro of rat secretory component to human pIgA; (iii) inhibit hepatic transport of human pIgA into rat bile in vivo; and (iv) inhibit apical transcytosis of pIgA in vitro by polarized human polymeric immunoglobulin receptor (pIgR)-expressing Madin-Darby canine kidney cells. Inhibition of biliary transport increased with the molar ratio of anti-J chain antibodies against pIgA and their incubation time. Anti-J chain F(ab ′ )₂ and Fab fragments also inhibited biliary transport, excluding a role for phagocytic clearance or excessive size of the immune complexes. Anti-human-Fcα Fab, bound to human pIgA in complexes of larger size than those with anti-J chain Fab, did not inhibit biliary transport of human pIgA. Propionic acid-denatured human pIgA, although containing J chains, was very poorly transported into rat bile. Altogether, our data strongly support, now also by in vivo experiments, the crucial role of the J chain of native pIgA in its selective pIgR-mediated transport into secretions, as suggested long ago by in vitro data only. Recent data on J chain-knockout mice, with low IgA levels in bile and feces, cannot explain the role of the J chain in contributing to the secretory component/pIgR-binding site of normal pIgA, but otherwise agree with our study.     

人胎中后期肠系膜淋巴结及相关T、B细胞的发育

目的从形态学的角度探讨人胎中后期肠系膜淋巴结的发育。方法采用常规组织学H-E染色和免疫组织化学法染色及图像分析进行研究,观察人胎中后期肠系膜淋巴结及相关T、B细胞的发育情况,相关数据做统计学处理。结果自第7个月龄起,在胚胎早期已形成的淋巴结周边的皮质继续增厚,可辨别浅层皮质与副皮质区,浅层皮质中的初级淋巴小结随胎龄的增加而增多,至出生时淋巴结内均未发现生发中心,仍为初级淋巴小结。早期的髓质随胎龄的增加被淋巴结周边增厚的皮质挤向淋巴结的中央。胚胎发育到第7～8个月龄时,随着淋巴结发育加速,淋巴结皮质逐渐增厚,可观察到分布在皮质内的CD3、IgM阳性细胞出现较强的阳性反应,第9～10个月龄时,CD3、IgM阳性细胞出现强阳性反应,灰度具有显著性差异P0.05。结论①出生时肠系膜淋巴结形态结构基本成熟,淋巴小结仍为初级淋巴小结。②人胎肠系膜淋巴结具有潜在的CD3、IgM合成和释放能力,对腹腔内免疫乃至整个免疫系统有重要作用。     

Subclass composition and J-chain expression of the 'compensatory' gastrointestinal IgG cell population in selective IgA deficiency.

The subclass distribution of IgG-producing immunocytes was examined by two-colour immunohistochemistry in gastrointestinal mucosa of 14 patients with selective serum IgA deficiency providing the following biopsy material: gastric (n = 1); jejunal (n = 12); colonic (n = 1); and rectal (n = 2). All except two patients suffered from various infections, and coeliac disease was observed in six of them. Control reference data were based on biopsies from immunologically intact subjects, including histologically normal jejunal (n = 10) and large bowel (n = 10) mucosa and stomach mucosa with slight chronic gastritis (n = 8). The total mucosal population of immunoglobulin-producing cells per 500 microns gut length unit was only slightly decreased in IgA deficiency because of an increased number of IgG (30%) and especially IgM (71%) immunocytes. The IgG1 immunocyte proportion in the proximal gut (median 87%) was higher than that in the comparable controls (gastric 69%, jejunal 66%). A similar trend was seen in the distal gut (69%) compared with controls from the large bowel mucosa (55%). Conversely, IgG2 and IgG3 cell proportions were significantly decreased compared with the respective controls from the proximal gut. The same was true for IgG4, which also was significantly reduced in jejunal mucosa. Paired staining for cytoplasmic J chain and immunoglobulin isotype showed 71% positivity for jejunal IgG-producing cells in IgA deficiency, which was somewhat reduced compared with comparable controls (89%). J chain appeared to be preferentially expressed by IgG1 cells (75%), but was also found in IgG2 (70%), IgG3 (32%) and IgG4 cells (33%). IgM-producing cells showed a J-chain positivity (99%) in IgA deficiency similar to normal (100%). Our results suggested that the block in mucosal B cell differentiation to IgA expression in the proximal gut is mainly located immediately upstream to the CH alpha 1 gene, giving excessive terminal maturation of J-chain-positive IgG1 immunocytes.     

Differential expression of chemokine receptors on human IgA+ and IgG+ B cells

Organ-specific lymphocyte homing is dependent on the expression of tissue-specific homing receptors and selected chemokine receptors. During the effector phase of an immune response, IgA and IgG antibody-secreting cells (ASC) are differently distributed in the body. Still, B cell expression of L-selectin and the mucosal homing receptor integrin alpha4beta7 is not related to the isotype produced, but only to the site of antigen encounter. In this study, we examined if differences in chemokine responsiveness between IgA+ and IgG+ B cells could explain their different tissue localization. Circulating CD19+ B cells were isolated and their expression of IgA, IgG, and selected chemokine receptors was determined by flow cytometry. Few Ig+ cells expressed CCR2, CCR3, or CCR9, and there was no difference in the expression of these receptors between IgA+ and IgG+ cells. In contrast, CCR4, CCR5, and CXCR3 was expressed on significantly more IgG+ than IgA+ cells. The function of chemokine receptors on memory B cells and ASC was then tested in the transwell system. IgG+ memory cells migrated to a higher extent than IgA+ cells towards the CXCR3 ligand CXCL11/I-TAC, while there was only a small migration towards the CCR4 ligand CCL17/TARC and the CCR9 ligand CCL25/TECK. ASC migrated poorly to all chemokines tested. In conclusion, this study shows that IgG+ and IgA+ memory B cells have a differential expression of the Th1 associated chemokine receptor CXCR3, as well as of CCR4 and CCR5. In contrast, none of the studied chemokine receptors was preferentially expressed by IgA+ cells.     

Immature B cells from human and mouse bone marrow can change their surface light chain expression

The capacity of bone marrow-derived surface immunoglobulin-positive (sIg⁺) human and mouse immature B cells, generated either in vitro or in vivo, to change their light (L) chain expression, has been assayed by the number of cells which change in vitro from one type of L chain to the other type, or to no sIg at all. Immature sIg⁺ B cells were generated in vitro from sIg^? precursor cells from human or mouse bone marrow. The immature sIg⁺ cells expressed RAG-1. Human sIg⁺ cells expressed xfr; and λ L chains in ratios between 1:1 and 3:1, whereas in mouse cells, this ratio ranged from 10:1 to 20:1. Upon reculture of the human and mouse xfr;⁺sIg⁺ cells, about half of them remained xfr;⁺, a quarter became λ⁺, and another quarter became sIg^?. Between 1 and 3% expressed both xfr; and λ chains. Of the human λ⁺ cells, about two-thirds remained λ⁺, only 1 to 2% became xfr;⁺, while the other third became sIg^?. Again, between 1 and 3% expressed both xfr; and λ L chains. These results indicate that expression of sIgM in the B cell membrane does not terminate L chain gene rearrangement, and that some order exists in xfr; versus λ gene rearrangements. Hence, human and mouse xfr;⁺ immature B cells can become λ⁺, but very few of the λ⁺ cells can become xfr;⁺, and both can become sIg^?. Further, human CD10⁺/sIg⁺ xfr;⁺ and λ⁺ cells and mouse B220^low/sIg^low xfr;⁺ cells enriched from bone marrow, i.e. immature B cells differentiated in vivo, changed their Ig phenotype upon in vitro culture, but in lower frequencies. By contrast, human and mouse mature B cells did not change their L chain or Ig phenotype. Hence, at least a part of the sIg⁺ immature B cells in bone marrow retain the capacity to change their L chain and Ig phenotype, and this capacity is lost when they become mature, peripheral B cells.     

Vasoactive intestinal peptide stimulates immunoglobulin production and growth of human B cells.

The effect of vasoactive intestinal peptide (VIP) on human lymphoblastoid B cell lines and tonsil B cells was studied. VIP increased immunoglobulin production and proliferation by lymphoblastoid B cell line, GM-1056, in a dose-dependent manner. As little as 10(-12) M of VIP was effective, and higher concentrations of VIP induced an approximately five-fold increase in IgA production. Moreover, this enhancement was blocked by VIP antagonist. Similarly, VIP enhanced IgM and IgG production by other lymphoblastoid B cell lines, CBL and IM-9, respectively. In contrast to VIP, another neuropeptide substance P (SP) or somatostatin failed to enhance immunoglobulin production and thymidine uptake. VIP also enhanced IgA production and thymidine uptake by purified tonsil B cells. However, in contrast to B cell lines, VIP failed to enhance IgM and IgG production by tonsil B cells. SP or somatostatin failed to enhance immunoglobulin production or thymidine uptake by tonsil B cells. These results indicate that VIP acts as B cell stimulatory factor and that VIP may also have preferential effect on IgA production on tonsil B cells.     

人胎肠系膜淋巴结组织发生及相关T、B细胞的发育

目的:从形态学的角度探讨人胎肠系膜淋巴结组织发生及相关T、B细胞的发育.方法:收集人胎33例,测量顶臀长,按Patten法确定胎龄.采用常规组织学H-E染色,免疫组织化学法,观察人胎肠系膜淋巴结组织发生及相关T、B细胞的发育.结果:9周,人胎肠系膜淋巴结原基形成,IgM阳性细胞出现,散在分布;11周,出现CD3阳性细胞,原基中有高内皮微静脉;15周,形成早期髓质;23周,IgM阳性细胞集聚形成小结状.连续切片显示,CD3阳性细胞分布在小结深面形成薄层副皮质区,可辨认皮质和髓质;至28周时,淋巴小结内均未发现生发中心.结论:9周淋巴结原基出现.15周早期髓质形成,髓质形成早于皮质.23周皮、髓质明显可辨,皮质内出现淋巴小结,但至28周时小结仍为初级淋巴小结;9周出现B细胞;11周出现T细胞.     

Delineation of producing ability of IgG and IgA subclasses by naive B cells in newborn infants and adult individuals

Neonatal B cells with the naive (sIgD⁺) phenotype are able to generate IgG- and IgA-producing cells as well as IgM production in the presence of memory CD4⁺ T cells expressing L-selectin (CD62L) in pokeweed mitogen-stimulated cultures. We used this system to examine comparatively the ability of naive B cells to produce IgG and IgA subclasses in newborn infants and adult individuals. Naive B cells were enriched from both donors on the basis of sIgD positivity, and memory (CD45RO⁺) CD4⁺ T cells with CD62L expression were isolated from adults. We here demonstrate some differences in profiles of IgG and IgA subclass production between neonatal and adult naive B cells. In neonatal B cells, IgG1 and IgG3 were predominantly produced, but IgG2 and IgG4 production was virtually absent. Similar to neonatal B cells, adult naive B cells produced mainly IgGq and IgG3, although memory (sIgD’) B cells from adults secreted all of the IgG subclasses. It should be noted that low but detectable levels of IgG2 and IgG4 were found in adults’naive B cell cultures. Although IgA produced by neonatal B cells was exclusively IgA1, IgA2-secreting cells were identifiable in adult naive B cells. The results suggest that further class switch of naive B cells to IgG2, IgG4 and IgA2 in addition to IgG1 and IgG3 may be controlled by their own age-dependent maturation process.     

Mast cells crosstalk with B cells in the gut and sustain IgA response in the inflamed intestine

B lymphocytes are among the cell types whose effector functions are modulated by mast cells (MCs). The B/MC crosstalk emerged in several pathological settings, notably the colon of inflammatory bowel disease (IBD) patients is a privileged site in which MCs and IgA⁺ cells physically interact. Herein, by inducing conditional depletion of MCs in red MC and basophil (RMB) mice, we show that MCs control B cell distribution in the gut and IgA serum levels. Moreover, in dextran sulfate sodium (DSS)-treated RMB mice, the presence of MCs is fundamental for the enlargement of the IgA⁺ population in the bowel and the increase of systemic IgA production. Since both conventional B-2 and peritoneal-derived B cells populate the intestine and communicate with MCs in physiological conditions and during inflammation, we further explored this interplay through the use of co-cultures. We show that MCs finely regulate different aspects of splenic B cell biology while peritoneal B cells are unresponsive to the supporting effects provided by MCs. Interestingly, peritoneal B cells induce a pro-inflammatory skewing in MCs, characterized by increased ST2 and TNF-α expression. Altogether, this study uncovers the versatility of the B/MC liaison and highlights key aspects for the resolution of intestinal inflammation.     

Dendritic cells from human mesenteric lymph nodes in inflammatory and non‐inflammatory bowel diseases: subsets and function of plasmacytoid dendritic cells

Plasmacytoid dendritic cells (pDC) in mesenteric lymph nodes (MLN) may be important regulators of both inflammatory and non‐inflammatory mucosal immune responses but human studies are rare. Here we compare pDC from human MLN and peripheral blood (PB) by phenotype and function. MLN from patients with or without inflammatory bowel disease (IBD) undergoing colon surgery and PB from patients with IBD and from controls were used to isolate mononuclear cells. The pDC were analysed by flow cytometry for the expression of CD40, CD80, CD83, CD86, CCR6, CCR7, CX3CR1, CD103 and HLA‐DR. Purified pDC from MLN and PB were stimulated with staphylococcus enterotoxin B (SEB), CpG‐A, interleukin‐3 (IL‐3), SEB + IL‐3, CpG‐A + IL‐3 or left unstimulated, and cultured alone or with purified allogeneic CD4⁺ CD45RA⁺ HLA‐DR‐ T cells. Subsequently, concentrations of IL‐1β, IL‐2, IL‐4, IL‐6, IL‐8, IL‐10, IL‐12, IL‐17, interferon‐α (IFN‐α), IFN‐γ and tumour necrosis factor‐α (TNF‐α) in culture supernatants were determined by multiplex bead array. The PB pDC from IBD patients exhibited an activated and matured phenotype whereas MLN pDC and control PB pDC were less activated. CpG‐A and CpG‐A + IL‐3‐stimulated MLN pDC secreted less IL‐6 and TNF‐α compared with PB pDC from controls. Compared with co‐cultures of naive CD4 T cells with PB pDC, co‐cultures with MLN pDC contained more IL‐2, IL‐10 and IFN‐γ when stimulated with SEB and SEB + IL‐3, and less IFN‐α when stimulated with CpG‐A. MLN pDC differ phenotypically from PB pDC and their pattern of cytokine secretion and may contribute to specific outcomes of mucosal immune reactions.     

人外周血NK细胞在细胞因子刺激下扩增后穿孔素、颗粒酶B基因表达的变化

目的 探讨经过纯化及在多种细胞因子的作用下扩增后的NK细胞,其穿孔素(per-furin)、颗粒酶B(granzyme B)表达水平的改变与细胞杀伤率的关系.方法 应用竞争性定量RT-PCR方法 检测了8例供者纯化、扩增后NK细胞的穿孔素、颗粒酶B基因表达水平,同时检测其对K562细胞的杀伤率.结果 经纯化、扩增后的NK细胞,在多种细胞因子作用下穿孔素和颗粒酶B的基因表达水平明显提高,且.IL-2+IL-15组、IL-2+IL-12+IL-15组基因的表达量均显著高于其他组,NK细胞对K562的杀伤率结果 与基因表达水平一致.结论 应用细胞因子后可使NK细胞的穿孔素、颗粒酶B基因表达水平明显提高,同时提高了NK细胞的杀伤功能.     

孟鲁司特对人鼻黏膜上皮细胞黏蛋白MUC2和MUC5B mRNA表达的影响

目的 探讨白三烯受体拮抗剂孟鲁司特对人鼻黏膜上皮细胞黏蛋白MUC2和MUC5BmRNA表达的影响。方法 应用下鼻甲黏膜组织进行鼻黏膜上皮细胞的原代培养,取第2代培养的鼻黏膜上皮细胞分成对照组、IL-1β组、孟鲁司特+IL-1β组和孟鲁司特组。IL-1β组加入含IL-1β(10 μg/L)的无血清培养基;孟鲁司特+IL-1β组先加入孟鲁司特(0.01 mol/L)孵育8h后,再加入含IL-1β（10μg/L）的无血清培养基;孟鲁司特组加入含孟鲁司特(0.01 mol/L)的无血清培养基;对照组仅加入无血清培养基。培养24h后通过荧光定量PCR检测各组上皮细胞黏蛋白MUC2和MUC5B mRNA的表达。结果对照组MUC2和MUC5B mRNA和孟鲁司特组水平相近[MUC2:(2.93±1.57)×106拷贝/μg、( 1.63±0.47)× 106拷贝/μg;M UC5B:(8.21±3.54)×105拷贝/μg(5.15±2.16)×105拷贝/μg]。而孟鲁司特+IL-1β组MUC2和MUC5B mRNA定量表达均低于IL-1β组[(3.48±1.41)× 106拷贝/μg比(6.63±1.73)× 10拷贝/μg,MUC5B：(1.75±0.69)×106拷贝/μg比(3.40±2.79)×107拷贝/μg,均P＜0.05],但高于对照组和孟鲁司特组（均P＜0.05）。结论 孟鲁司特可降低IL-1β诱导的鼻黏膜上皮细胞黏蛋白MUC2和MUC5BmRNA的表达,但不影响正常状态鼻黏膜上皮细胞黏蛋白mRNA的表达,可能对炎性细胞因子诱导的鼻黏膜上皮细胞黏蛋白mRNA的表达有抑制作用。     

Localization and identification of granzymes A and B-expressing cells in normal human lymphoid tissue and peripheral blood.

Cytoplasmic granules from activated natural killer (NK) and cytotoxic T lymphocytes (CTL) contain a pore-forming protein, perforin, and several homologous serine proteinases called granzymes. Expression of these proteins correlates with the cytolytic potential of cytotoxic lymphocytes. Using a panel of MoAbs specific for human granzyme A and B, respectively, expression of these proteinases in non-pathological lymphoid tissue and peripheral blood lymphocyte (PBL) subpopulations was investigated. Using immunohistochemistry and double stainings, the phenotype of granzyme-expressing cells in lymphoid tissue was investigated. Granzyme-positive cells were detected in all lymphoid tissues tested. No large differences in the number and distribution between granzyme A- and granzyme B-positive cells were observed. The highest number of positive cells was located in the red pulp of the spleen. Significant numbers were detected in tonsil, lymph nodes, liver and thymus. Low numbers were present in the lamina propria of non-inflamed stomach, small intestine and colon. Phenotypic analysis and cell sorting showed that most of the granzyme-positive cells in lymphoid tissue and PBL consisted of CD3-CD16+CD56+ lymphocytes. Hardly any granzyme-positive CD3+CD8+ CTL were present in peripheral blood. The synthesis of granzyme A as well as B by both CD3+CD16+CD56+ and CD3+CD8+ cells in peripheral blood was increased upon IL-2 stimulation. These results indicate that in normal lymphoid tissue the predominant cytolytic cell population is formed by the NK cells, and activated CTL are rare.     

Frequencies of HIV-reactive B cells in seropositive and seronegative individuals.

Peripheral blood mononuclear cells (PBMC) from HIV-infected seropositive (HIV+) but not from normal, seronegative (HIV-) individuals are known to produce anti-HIV antibodies in vitro, in the absence or presence of pokeweed mitogen (PWM). Previous studies showed that up to 20-40% of spontaneously immunoglobulin-secreting B cells from HIV+ individuals are HIV-specific. To analyse the frequency of anti-HIV B cells among 'total' peripheral blood B cells in the present study, we used a limiting dilution assay in which EL-4 thymoma cells induce clones of immunoglobulin-secreting cells in activated as well as resting B cells. Anti-HIV B cells were detected not only in 11/12 HIV+ individuals (with frequencies from 1/910 to 1/21,500 B cells cultured; one negative test was from a person undergoing seroconversion), but also in 4/9 HIV- normal blood donors (1/16,200 to 1/49,000 B cells cultured) and in 3/6 newborns from HIV- mothers (1/11,800 to 1/26,600 B cells cultured). The mean frequency was nine times higher in the HIV+ individuals than in the normal donors. As in previous studies, only the cells from HIV+ individuals generated anti-HIV antibodies in PBMC bulk cultures with or without PWM. The relative proportion of specific anti-HIV antibody/total immunoglobulin in PBMC bulk cultures was 800 times higher by the mean than in EL-4 B cell cultures from HIV+ individuals (whereby the total immunoglobulin secretion for equal numbers of B cells cultured was 500 times lower for PBMC). These different results obtained with different assays suggest that in seropositives most anti-HIV B cells belong to an activated B compartment which is quite small, even in a disease with B cell hyperactivity. Therefore, the specific B cells are strongly diluted among the EL-4 cell-responsive, total B cells. On the other hand, the EL-4 assay can detect HIV-reactive B cells in the B cell repertoire of normal, non-infected individuals.     

慢性乙型肝炎外周血单个核细胞内HBV-DNA与血清HBV-DNA及HBeAg表达关系的研究

目的研究慢性乙型肝炎患者外周血单个核细胞(PBMC)内HBV-DNA和血清中HBV-DNA表达量、e抗原表达的关系。方法采用聚合酶链反应(PCR)检测208例慢性乙型肝炎患者PBMC内HBV-DNA,应用荧光定量聚合酶链反应(FQ-PCR)检测血清中HBV DNA含量,应用酶联免疫吸附(ELISA)法检测乙肝血清标志物。结果208例慢性乙型肝炎患者PBMC内HBV-DNA阳性106例、阴性102例。HBV-DNA(PBMC)阳性组、阴性组血清HBV-DNA定量≥1.0E5患者比例分别为91.5%(97例)、45.1%(46例)(χ2=52.12,P<0.01);HBeAg阳性率分别为76.4%(81例)、50.9%(52例)(χ2=21.55,P<0.01)。结论PBMC内HBV-DNA的检测与血清中HBV-DNA定量检测及HBeAg阳性率存在明显的正相关。提示血清HBV-DNA高载量的HBeAg阳性患者外周血单个核细胞感染HBV-DNA明显增加。