Neutralization of the AIDS retrovirus by antibodies to a recombinant envelope glycoprotein

Mammalian cell lines have been engineered to produce a secreted form of the AIDS retrovirus envelope glycoprotein. The recombinant protein has been isolated from growth-conditioned culture media and used to immunize animals. Antibodies directed against the recombinant molecule were found to react with the envelope glycoprotein produced in virus-infected cells. Furthermore, these antibodies were able to directly inactivate the AIDS retrovirus in a neutralization assay in vitro. The expression system reported here should provide sufficient quantities of the AIDS retrovirus envelope protein for biological and vaccination studies.     

Isolation of a new human retrovirus from West African patients with AIDS

The etiological agent of AIDS, LAV/HTLV-III, is common in Central Africa but is not endemic in other areas of that continent. A novel human retrovirus, distinct from LAV/HTLV-III, has now been isolated from two AIDS patients from West Africa. Partial characterization of this virus revealed that it has biological and morphological properties very similar to LAV but that it differs in some of its antigenic components. Although the core antigens may share some common epitopes, the West African AIDS retrovirus and LAV differ substantially in their envelope glycoproteins. The envelope antigen of the West African virus can be recognized by serum from a macaque with simian AIDS infected by the simian retrovirus termed STLV-IIImac, suggesting that the West African AIDS virus may be more closely related to this simian virus than to LAV. Hybridization experiments with LAV subgenomic probes further established that this new retrovirus, here referred to as LAV-II, is distantly related to LAV and distinct from STLV-IIImac.     

Simian AIDS: isolation of a type D retrovirus and transmission of the disease

A type D retrovirus related to but distinct from Mason-Pfizer monkey virus was isolated in vitro from the blood of two rhesus monkeys (Macaca mulatta) with simian acquired immunodeficiency syndrome (SAIDS). Three juvenile rhesus monkeys that were injected intravenously with tissue culture fluids containing this virus developed SAIDS after 2 to 4 weeks.     

重组逆转录病毒载体系统的建立及其感染滴度的研究

选择新霉素磷酸转移酶II（Neo^R)做为标记基因,将重组逆转录病毒载体经包装细胞馐后得到的重组病毒浓缩后,应用该重组逆转录病毒载体系统体外感染靶细胞NIH3T3以测定其滴度,其滴度可达到10^6,具有理想的感染靶细胞的能力。     

Expression in Escherichia coli of open reading frame gene segments of HTLV-III

Human T-cell lymphotropic virus type III (HTLV-III), the causative agent of the acquired immune deficiency syndrome (AIDS), was recently isolated and its genomic structure analyzed by DNA cloning methods. In the studies reported here a combined cloning and expression system was used to identify HTLV-III encoded peptides that react immunologically with antibodies in sera from AIDS patients. Cloned HTLV-III DNA was sheared into approximately 500-base-pair fragments and inserted into an "open reading frame" expression vector, pMR100. The inserted DNA was expressed in Escherichia coli transformants as a polypeptide fused to the lambda CI protein at its amino terminus and to beta-galactosidase at its carboxyl terminus. Sera from AIDS patients containing antibodies to HTLV-III were then used to screen for immunoreactive fusion proteins. Twenty clones, each specifying a fusion protein strongly reactive with AIDS serum, were identified. DNA sequence analysis indicated that the HTLV-III fragments were derived from the open reading frame DNA segments corresponding to the gag and pol gene coding regions and also the large open reading frame region (env-lor) located near the 3' end of the viral genome.     

Proviral DNA of a retrovirus, human T-cell leukemia virus, in two patients with AIDS

The acquired immune deficiency syndrome (AIDS) is characterized by T-lymphocyte dysfunction and is frequently accompanied by opportunistic infections and Kaposi's sarcoma. Human T-cell leukemia virus (HTLV) is associated with T-cell malignancies and can transform T lymphocytes in vitro. In an attempt to find evidence of HTLV infection in patients with AIDS, DNA from samples of peripheral blood lymphocytes from 33 AIDS patients was analyzed by Southern blot-hybridization with a radiolabeled cloned HTLV DNA probe. Analysis of DNA from both the fresh (uncultured) lymphocytes and from T cells cultured with T-cell growth factor revealed the presence of integrated HTLV proviral sequences in lymphocytes from two of the patients, both of whom had antibody to HTLV. The proviral sequences could not be detected in blood samples obtained from these individuals at a later date, consistent with the possibility that the population of infected cells had become depleted.     

半滑舌鳎GAPDH基因开放读码框的克隆及序列分析

甘油醛-3-磷酸脱氢酶(GAPDH)广泛存在于真核生物中,对许多生理活动具有重要作用.参考金头鲷和斑马鱼的甘油醛-3-磷酸脱氢酶(GAPDH)基因序列设计一对简并引物,提取半滑舌鳎肌肉组织总RNA,采用RT-PCR技术对GAPDH基因进行克隆测序及序列分析.结果表明,从半滑舌鳎肌肉组织中成功克隆了GAPDH基因的开放读码框(ORF),序列全长1002 bp,编码333个氨基酸,分子量为36.0235 ku,等电点PI为7.75.序列同源性分析显示,该序列与其他鱼类的GAPDH氨基酸序列具有较高的同源性.     

Apobec3 encodes Rfv3, a gene influencing neutralizing antibody control of retrovirus infection

Recovery from Friend virus 3 (Rfv3) is a single autosomal gene encoding a resistance trait that influences retroviral neutralizing antibody responses and viremia. Despite extensive research for 30 years, the molecular identity of Rfv3 has remained elusive. Here, we demonstrate that Rfv3 is encoded by Apobec3. Apobec3 maps to the same chromosome region as Rfv3 and has broad inhibitory activity against retroviruses, including HIV. Not only did genetic inactivation of Apobec3 convert Rfv3-resistant mice to a susceptible phenotype, but Apobec3 was also found to be naturally disabled by aberrant messenger RNA splicing in Rfv3-susceptible strains. The link between Apobec3 and neutralizing antibody responses highlights an Apobec3-dependent mechanism of host protection that might extend to HIV and other human retroviral infections.     

Epidemiology of HIV infection and AIDS in the United States

By the end of 1987, nearly 50,000 cases of acquired immunodeficiency syndrome (AIDS) had been reported since 1981, 20,745 in the past year alone. Black and Hispanic adults and children have reported rates 3 to 12 times as high as whites. This can be largely attributed to higher reported rates in black and Hispanic intravenous (IV) drug abusers, their sex partners, and infants. In 1986, reported AIDS deaths increased adult male and female mortality in the United States by an estimated 0.7 and 0.07%, respectively, with much greater increases in selected age groups or areas of the country. The greatest variation in infection with the human immunodeficiency virus (HIV) (0 to 70%) has been found in surveys of IV drug abusers, while surveys of homosexual men reveal infection rates of 20 to 50%. Infection with HIV ranged from 0 to 2.6% in limited sexually transmitted disease clinic surveys of heterosexual men and women without a history of IV drug abuse or known sexual contact with persons at increased risk. The modes of HIV transmission are now well understood, but a large amount of biologic variability in efficiency of transmission remains to be explained. The period between initial infection with HIV and the development of AIDS is variable, but the risk for disease progression increases with duration of infection.     

HTLV-III infection in brains of children and adults with AIDS encephalopathy

Unexplained debilitating dementia or encephalopathy occurs frequently in adults and children with the acquired immune deficiency syndrome (AIDS). Brains from 15 individuals with AIDS and encephalopathy were examined by Southern analysis and in situ hybridization for the presence of human T-cell leukemia (lymphotropic) virus type III (HTLV-III), the virus believed to be the causative agent of AIDS. HTLV-III DNA was detected in the brains of five patients, and viral-specific RNA was detected in four of these. In view of these findings and the recent demonstration of morphologic and genetic relatedness between HTLV-III and visna virus, a lentivirus that causes a chronic degenerative neurologic disease in sheep, HTLV-III should be evaluated further as a possible cause of AIDS encephalopathy.     

Feedback of speech muscle activity during silent reading: rapid extinction

Surface electromyograms of the laryngeal muscles were made while subjects read silently. Those who showed an increase in electrical activity over that at relaxation were provided with auditory feedback of the muscle activity.This treatment resulted in immediate and long-lasting cessation of the subvocalazation. This method should prove valuable in treating some reading problems.     

Detection of human cytomegalovirus in peripheral blood lymphocytes in a natural infection

In situ hybridization was used to detect human cytomegalovirus (HCMV) in the peripheral blood mononuclear cells of some naturally infected (seropositive) individuals. A subpopulation of cells hybridized specifically to a portion of the HCMV genome that is heavily transcribed during the immediate-early period of infection. The hybridization signal was markedly reduced by base hydrolysis and ribonuclease, and therefore the probe appears to be detecting viral RNA. A fluorescence-activated cell sorter was used to select lymphocytes bearing the OKT4 and OKT8 markers. Hybridization with the HCMV probe revealed a higher proportion of positive cells in the OKT4 than in the OKT8 subset. This observation specifically identifies lymphocytes as a cell population involved in natural HCMV infection and suggests that lymphocytes may be a reservoir for maintaining infection and may also serve as a vehicle for its spread by blood transfusion.     

Initial function determination for the open reading frame (ORF) region of Pib gene via rice transformation

Three plant binary expression vectors—pNAR501, pNAR502 and pNAR503—were constructed, carrying fragments of exon2-exon3, 5′partial deletion exon1 and 5′partial deletion exon1-exon2-exon3 of Pib gene driven by 35S. These three vectors were transformed into the japonica rice variety Nipponbare through agrobacterium-mediated transformation. More than 30 transgenic rice plants were obtained and confirmed by polymerase chain reaction (PCR), Southern hybridization and the hygromycin resistance test in seed germination of their progeny. A rice blast resistance test for in vitro leaves of T_o transgenic plants in the tillering stage showed higher resistance to the races of E1, F1 and G1 of rice blast than that of the control Nipponbare. However, results of rice blast resistance test for seedlings of T₁ transgenic plants in the 3-to 4-leaf stage were different. All T₁ transgenic seedlings had a lower level of resistance to E1, F1 and G1 races than that of the control Nipponbare. Translated from Journal of Nanjing Agricultural University, 2006, 29(3): 1–5 [译自: 南京农业大学学报]     

Dual infection of the central nervous system by AIDS viruses with distinct cellular tropisms

Human immunodeficiency virus (HIV) is the causative agent of the acquired immune deficiency syndrome (AIDS). A large number of AIDS patients show evidence of neurologic involvement, known as AIDS-related subacute encephalopathy, which has been correlated with the presence of HIV in the brain. In this study, two genetically distinct but related viruses were isolated from one patient from two different sources in the central nervous system: brain tissue and cerebrospinal fluid. Both viruses were found to replicate in peripheral blood lymphocytes, but only virus from brain tissue will efficiently infect macrophage/monocytes. The viruses also differ in their ability to infect a brain glioma explant culture. This infection of the brain-derived cells in vitro is generally nonproductive, and appears to be some form of persistent or latent infection. These results indicate that genetic variation of HIV in vivo may result in altered cell tropisms and possibly implicate strains of HIV with glial cell tropism in the pathogenesis of some neurologic disorders of AIDS.     

Pyrimidine nucleotide synthesis: regulatory control during transformation of lymphocytes in vitro

The incorporation of uridine into RNA in lymphocytes undergoing transformation induced by phytohemagglutinin parallels its incorporation into the intratcellular pool of acid-soluble nucleotides and coincides with a 20-fold increase in the specific activity of uridine kinase. This increase is dependent upon synthesis of both RNA and protein and is subject to both repression and end-product inhibition by cytidine.