介绍一种制备特异性TF和特异性iRNA的方法

以乙肝疫苗、人喉癌细胞膜抗原为抗原，猪脾细胞为效应细胞，经体外免疫后收集应答细胞，制备ＰＳＨＢＶ－ＴＦ ＰＳＡＣ－ｉＲＮＡ。通过抗原特异性细胞免疫功能试验证实，ＰＳＨＢＶ－ＴＦ和ＰＳＡＣ－ｉＲＮＡ都能转移特异性细胞免疫功能。采用体外免疫法制备ＰＳＨＢＶ－ＴＦ和ＰＳＡＣ－ｉＲＮＡ是可行的，并且具有诸多优点。     

APAAP免疫印迹技术对抗副流感病毒1，3型单克隆抗体识别抗原表位的研究

副流感病毒１，３型（ＰａｒａｉｎｆｌｕｅｎｚａＶｉｒｕｓｔｙｐｅ１，３：ＰＩＶ＿１．３）单克隆抗体（ＭｃＡｂ）应用免疫印迹技术（Ｗｅｓｔｅｒｎｂｌｏｔｔｉｎｇ）识别抗原表位特性。结果表明：ＰＩＶ＿１．３抗原经还原剂处理后，ＳＤＳ－ＰＡＧＥ５％～１５％梯度胶电泳，能分辨二十多条清晰的蛋白带。转印后采用碱性磷酸酶抗碱性磷酸酶复合物（ＡＰＡＡＰ）染色，本底浅，呈玫瑰红色带，优于ＨＲＰ染色结果。ＰＩＶ＿１的５株ＭｃＡｂ：ＩＣ＿５、ＩＨ＿６、ＩＨ＿２、ＩＣ＿１０、３Ｄ＿５分别与６８ｋＤ～５０ｋＤ、６８ｋＤ、５８ｋＤ～２７ｋＤ、５５ｋＤ～５０ｋＤ、５０ｋＤ对应ＰＩＶ＿１的蛋白质抗原表位起反应。ＰＩＶ＿３的６株ＭｃＡｂ：２Ａ＿１０、５Ｇ＿３、２Ｄ＿１１、２Ｅ＿１０、２Ｂ＿１２、４Ｆ＿１２与７０ｋＤ、６８ｋＤ、６０ｋＤ～５０ｋＤ、５５ｋｐ～４０ｋＤ、５５ｋＤ、４０ｋＤ对应的蛋白质抗原表位特异结合，说明ＰＩＶ＿１．３的１１株ＭｃＡｂ同对应抗原表位点的结合分布较广，有利于对ＰＩＶ＿１．３抗原的快速、敏感、特异检测。     

Endocarditis with ruptured cerebral aneurysm caused by Cardiobacterium valvarum sp. nov

A fastidious gram-negative bacterium was isolated from the blood of a 37-year-old man who had insidious endocarditis with a sudden rupture of a cerebral aneurysm. Characterization of the organism through phylogenetic and phenotypic analyses revealed a novel species of Cardiobacterium, for which the name Cardiobacterium valvarum sp. nov. is proposed. C. valvarum will supplement the current sole species Cardiobacterium hominis, a known cause of endocarditis. Surgeries and antibiotic treatment cured the patient's infection and associated complications. During cardiac surgery, a congenital bicuspid aortic valve was found to be the predisposing factor for his endocarditis.     

Stimulating capacity of fresh and cultured human leukaemic lymphoid and myeloid cells in `one-way'' mixed lymphocyte reaction

Fresh normal peripheral blood B lymphocytes possess a strong stimulating capacity while fresh thymus cells or fresh peripheral T lymphocytes possess a weak, but significant stimulating capacity on allogeneic lymphocytes in `one-way' mixed lymphocyte reaction. Fresh leukaemic T lymphoid cells from patients with T-cell ALL or T-cell CLL exert little or no stimulation on allogeneic lymphocytes. Fresh leukaemic B lymphoid cells from patients with B-cell CLL or B-cell HCL, on the other hand, exert a lesser stimulation on allogeneic lymphocytes, as compared to that of normal B lymphocytes. Leukaemic myeloblasts from patients with AML or Ph<sup>1</sup>(+) CML-BP exert significantly higher stimulation than leukaemic lymphoid cells in `one-way' mixed lymphocyte reaction (P<0.05). Cultured leukaemic T lymphoid cells (MOLT-4) possess no stimulating capacity, cultured leukaemic B lymphoid cells (BALM-2) possess a moderate degree of stimulating capacity and cultured leukaemic, possibly myeloid, cells (NALM-1 and K562) possess vigorous stimulation on allogeneic lymphocytes. The stimulating capacity of NALM-1 or K562 cells is significantly higher than that of BALM-2 cells (P<0.01 or P<0.05, respectively) and that of MOLT-4 cells (P<0.001). These observations suggest that the stimulating capacity of leukaemic T or B lymphoid cells may have been completely or partially lost during the process of leukaemogenesis. Since we do not have an opportunity to study the stimulating capacity of normal myeloblasts, it is not known whether the stimulating capacity of leukaemic myeloblasts, which is found to be very strong on allogeneic lymphocytes, may have been modified during the process of leukaemogenesis.     

Cloning, expression and characterization of a murine-human chimeric antibody with specificity for pre-S2 surface antigen of hepatitis B virus

The cloning, expression and characterization of a murine-human chimeric antibody with specificity for the pre-S2 surface antigen (Ag) of hepatitis B virus (HBV) is described. The heavy and light chain variable region (VH and VL) genes encoding the murine monoclonal antibody (mAb) were isolated and combined with human γ 1 and κ constant region genes, respectively. The expression vectors containing the chimeric heavy and light chain genes were sequentially electroporated into murine Sp2/0 hybridoma cells and transfectomas secreting chimeric antibody were isolated. The chimeric antibody was purified and characterized by ELISA, Western analysis and competition immunoassay, demonstrating that the transfectoma functionally express and secrete murine-human chimeric antibody which retained the specificity and affinity of the parental murine mAb.     

Assessment of HBV persistent infection in an adult population in Taiwan

In order to study the prevalence of hepatitis B virus (HBV) in the adult population of Taiwan, we screened for the presence of HBV DNA in 205 blood samples from adult (20-59-year-old) volunteers. According to the serological markers of HBV, samples were divided into three groups: group I (173 subjects) was negative for both HBsAg and HBeAg; group II (14 subjects) was positive for both HBsAg and HBeAg; and group III consisted of 18 subjects who were HBsAg-positive but HBeAg-negative. Plasma HBV DNA was not detected in group I, but it was found in 85.7% and 11.8% of samples in group II and group III, respectively. A free-form HBV DNA was found in 14.3% of the leukocyte samples in group II. Furthermore, an integrated form of HBV DNA was detected in the leukocytes of two cases of group I who remained healthy based on clinical data. HBV DNA was also detected in the spermatozoa and liver cells of one of the cases.     

Bam32 links the B cell receptor to ERK and JNK and mediates B cell proliferation but not survival

Bam32 is an adaptor protein recruited to the plasma membrane upon B cell receptor (BCR) crosslinking in a phosphoinositol 3-kinase (PI3K)-dependent manner; however, its physiologic function is unclear. To determine its physiologic function, we produced Bam32-deficient mice. Bam32(-/-) B cells develop normally but have impaired T-independent antibody responses in vivo and diminished responses to BCR crosslinking in vitro. Biochemical analysis revealed that Bam32 acts in a novel pathway leading from the BCR to MAPK/ERK Kinases (MEK1/2), MAPK/ERK Kinase Kinase-1 (MEKK1), extracellular signal-regulated kinase (ERK), and c-jun NH2-terminal kinase (JNK), but not p38 mitogen-activated protein kinase (p38). This pathway appears to be initiated by hematopoietic progenitor kinase-1 (HPK1), which interacts directly with Bam32, and differs from all previously characterized BCR signaling pathways in that it is required for normal BCR-mediated proliferation but not for B cell survival.     

Pulmonary granuloma caused by Pseudomonas andersonii sp nov

Pulmonary granuloma is a common lesion for which gram-negative bacteria are rarely implicated as a cause. Hence, most physicians are unaware of this etiology. We isolated a gram-negative bacterium from a surgically resected pulmonary granuloma in a 42-year-old, nonimmunocompromised woman. Within the necrotizing granuloma, numerous organisms also were demonstrated by Gram stain, suggesting a cause-disease relationship. Characterization of the bacterium by sequence analysis of the 16S ribosomal gene, cellular fatty acid profiling, and microbiologic studies revealed a novel bacterium with a close relationship to Pseudomonas. We propose a new species for the bacterium, Pseudomonas andersonii. These results suggest that the differential diagnosis of a lung granuloma also should include this gram-negative bacterium as a potential causative agent, in addition to the more common infections caused by acid-fast bacilli and fungi. This bacterium was shown to be susceptible to most antibiotics that are active against gram-negative bacteria.