Immunomodulation during treatment of polymyositis with plasmapheresis and immunosuppressive drugs

Immunologic studies were carried out in a patient with polymyositis (PM), who showed increasing muscle strength and decreasing serum creatine phosphokinase levels during 20 weeks of treatment with plasmapheresis in conjunction with prednisone and cyclophosphamide. After an initial rise, serum IgG declined with treatment. Natural killer (NK) lymphocytes were reduced by 74%, B cells by 95%, and T cells by 38%. Spontaneous proliferation of peripheral blood mononuclear cells increased dramatically. Within the CD4+ T cell subset there was increasing maturation as shown by a rise in percent mature (CD29+) cells and reciprocal decline of immature (CD45RA+) cells. At the same time CD4+ T cells became increasingly activated as shown by HLA-DR expression. The percentage of CD8+ T cells increased strongly with treatment, and they showed increased activation and expression of the cytotoxic CD29+ and CD11b- phenotypes. CD8+ T cells exhibiting CD45RA or CD11b+ suppressor phenotypes were overall unchanged; however, on follow-up a proportion of CD8+ cells expressed the activated suppressor effector (CD11b—CD28—) phenotype. In addition to control of PM by the possible deletion of activated autoreactive B and T lymphocyte clones with cyclophosphamide, the activation and maturation of CD4+ T cells during treatment may have downregulated the autoreactive disease process, either through direct antiidiotypic suppression or by induction of the observed increase in cytotoxic and suppressor CD8+ T cells. © 1994 Wiley-Liss, Inc.     

Update on new antivirals under development for the treatment of double-stranded DNA virus infections

All the currently available antiviral agents used in the treatment of double-stranded (ds) DNA viruses, with the exception of interferon-α, inhibit the same target, the viral DNA polymerase. With increasing reports of the development of resistance of herpes simplex virus (HSV), cytomegalovirus (CMV), and hepatitis B virus (HBV) to some of these drugs, new antiviral agents are needed to treat these infections. Additionally, no drugs have been approved to treat several DNA virus infections, including those caused by adenovirus, smallpox, molluscum contagiosum, and BK virus. We report the status of 10 new antiviral drugs for the treatment of dsDNA viruses. CMX-001 has broad activity against dsDNA viruses; 3 helicase-primase inhibitors, maribavir, and FV-100 have activity against certain herpesviruses; ST-246 inhibits poxviruses; GS-9191 inhibits papillomaviruses; and clevudine and emtricitabine are active against HBV. Most of these drugs have completed at least phase I trials in humans, and many are in additional clinical trials.     

Antibodies in patients with recurrent respiratory papillomatosis treated with lymphoblastoid interferon

Serum specimens from 53 evaluable patients enrolled in a clinical trial of lymphoblastoid interferon in recurrent respiratory papillomatosis were screened for the presence of interferon-binding antibodies by an indirect enzyme immunoassay and evaluated for neutralizing antibody measured as the inhibition of antiviral activity. Immunoglobulin G antibodies that specifically bound lymphoblastoid interferon were detected in 66% (35 of 53) of patients; neutralizing antibody was detected in 11 of the 35 patients having binding antibody (and in none of the patients who were negative for binding antibody). The incidence of detectable neutralizing antibody in this study population was 20.8% (11 of 53), which is markedly higher than in previous reports of lymphoblastoid interferon in patients with other diseases (i.e., less than 1% incidence). The cumulative dose received at the time of detection of neutralizing antibody ranged from 163 to 385 MU per square meter of body surface. Neutralizing antibody was detectable at a median time of 120 days after initiation of interferon therapy, and binding antibody appeared earlier in those patients (median 59 days) than in patients in whom only binding antibody was produced (median 116 days). Despite the tendency of binding antibody to appear either in patients in whom neutralizing antibody was eventually formed, the detection of binding antibody was not necessarily predictive of the subsequent development of neutralizing antibodies. Binding antibody persisted after neutralizing antibodies had become undetectable.(ABSTRACT TRUNCATED AT 250 WORDS)     

The role of interferon in the protective effect of a synthetic double-stranded polyribonucleotide against intranasal vesicular stomatitis virus challenge in mice

Intravenous injection of polyinosinic acid/polycytidylic acid [(poly rI).(poly rC)] offered significant protection against intranasal challenge of young mice with vesicular stomatitis virus (VSV). Optimal protection was obtained when a single dose was administered 2 hr before virus challenge, but repeated doses were effective when started as late as 3 days after virus challenge. The therapeutic ratio or ratio of maximum tolerated dose to minimum effective dose for a single intravenous injection of (poly rI).(poly rC) 2 hr before virus inoculation was >/=8 mg/kg:0.004 mg/kg or >/=200.Dose-response curves for interferon production and antiviral protection by (poly rI).(poly rC) were closely parallel. Equivalent doses of poly rI or poly rC alone did not exert any interferon-inducing capacity or protective effect on intranasal VSV challenge. Several factors, which are known to potentiate or antagonize interferon production, increased or decreased the interferon-inducing capacity and antiviral protection of either (poly rI).(poly rC) or maleic acid/divinyl ether copolymer (MA/DVE) in parallel. Interferon production and antiviral protection by MA/DVE were enhanced by arginine but abolished by prior treatment with MA/DVE; DEAE-dextran (intraperitoneally), kinetin riboside and isopentenyladenosine, and prior injection of endotoxin reduced both interferon production and antiviral protection by (poly rI).(poly rC).Treatment with exogenous interferon in amounts which closely mimicked the levels of circulating interferon produced endogenously by an effective dose of (poly rI).(poly rC) gave protection against intranasal VSV which was identical with that dose of (poly rI).(poly rC). This strongly suggests that interferon production accounts for the whole protective effect of (poly rI).(poly rC) in the intranasal VSV assay.     

Antibodies against Yersinia enterocolitica in patients with Reiter's syndrome

Reiter's syndrome can be induced by several different bacteria. A frequent cause in Finland is Yersinia enterocolitica serotypes 03 and 09, but these strains are rarely found in the United States. Although this does not exclude the possibility that U.S. patients with Reiter's syndrome have been infected with Yersinia, it is more likely that they develop Reiter's syndrome as a consequence of infection by non-Yersinia arthritis-causing organisms that share certain determinants with Yersinia organisms. We used radioimmunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis to analyze the serum antibodies against iodine 125-labeled, detergent-solubilized serotype 03 Y. enterocolitica. Our results demonstrated that most serum samples of United States subjects precipitate three to five radioactively labeled Yersinia molecules. A Yersinia antigen of 88K appeared to be of possible discriminatory value. Protein A-reacting antibodies directed against this antigen were detected in only two of twenty-five patients with rheumatoid arthritis and only seven of 44 normal control subjects, compared with 18 of 27 patients with Reiter's syndrome (p less than 0.005) and eight of 16 patients with ankylosing spondylitis (p less than 0.01). Our results indicate that, despite the relatively rare occurrence of Y. enterocolitica serotypes 03 and 09 infection in the United States, examination of the immune response to the serotype 03 Yersinia strain is a promising approach to the study of Reiter's syndrome in the United States.     

Rapid detection of herpes simplex virus DNA by in situ hybridization with photobiotin-labelled double-stranded DNA probes

An assay for rapid detection of herpes simplex virus in infected cells is described. The assay utilizes in situ hybridization with photobiotin-labelled double-stranded DNA probes prepared from HSV-1 DNA cloned in plasmid vectors. The assay provided an alternative method for earlier detection of virus in cell cultures with the ease of preparation of photobiotin-labelled double-stranded DNA.     

Sequence-dependent quenching of fluorescein fluorescence on single-stranded and double-stranded DNA

Fluorescein is commonly used to label macromolecules, particularly proteins and nucleic acids, but its fluorescence is known to be strongly dependent on its direct chemical environment. In the case of fluorescein-labeled nucleic acids, nucleobase-specific quenching originating in photoinduced charge transfer interactions results in sequence-dependent chemical environments. The resulting sequence specificity of fluorescent intensities can be used as a proximity detection tool, but can also lead to biases when the abundance of labeled nucleic acids is quantified by fluorescence intensity. Here we comprehensively survey how DNA sequences affect fluorescence intensity by preparing permutational libraries containing all possible 5mer contexts of both single-stranded and double-stranded DNA 3′ or 5′ end labeled with fluorescein (6-carboxyfluorescein, FAM). We observe the expected large quenching of fluorescence with guanine proximity but also find more complex fluorescence intensity changes depending on sequence contexts involving proximity to all four nucleobases. A terminal T (T > A ≈ C ≫ G) in both 3′ and 5′ labeled single strands results in the strongest fluorescence signal and it changes to a terminal C (C ≫ T > A ≫ G) in double-stranded DNA. Therefore, in dsDNA, the terminal G·C base pair largely controls the intensity of fluorescence emission depending on which of these two nucleotides the dye is attached to. Our data confirms the importance of guanine in fluorescence quenching while pointing towards an additional mechanism beyond the redox potential of DNA bases in modulating fluorescein intensity in both single and double stranded DNA. This study should help in designing better nucleic acid probes that can take sequence-dependent quenching effects into account.

Fluorescein is commonly used to label macromolecules, particularly proteins and nucleic acids, but its fluorescence is known to be strongly dependent on its direct chemical environment.     

Renal cell carcinoma: treatment with interferon

All IFN types--alpha, beta, and gamma--appear to have some antitumor activity against RCC. IFNa has been extensively studied and has demonstrated objective response rates between 15% and 20% when administered in a variety of doses, routes, and schedules. Intermediate dose levels may be associated with greater response rates than low dose levels, and high dose levels are poorly tolerated and usually require dose reduction because of toxicity. Among the means of administration, intramuscular or subcutaneous routes are favored because of logistic advantages; in the low- and intermediate-dose ranges chronic sequential administration (daily, three times a week, or five days per week) is tolerable and may ameliorate toxicity; none of these therapeutic recommendations can be proven to be superior, with respect to response, to several other alternatives. No survival advantage can yet be proven to result from IFN therapy for patients with RCC. Studies evaluating combinations of IFNa and other IFNs or cytotoxic agents have demonstrated increased toxicity. Although responses have been seen in the limited number of studies performed to date, these studies do not appear to support in vivo suggestions of dramatic synergism between these agents. Knowledge of the therapeutic use of IFN is in its infancy. Although the response rates described in this review are unimpressive, they are commensurate with the best available conventional therapy for RCC. As clinical strategies for the use of IFN improve, so too, might the therapeutic efficacy in RCC improve.     

Lactoferrin interacts with deoxyribonucleic acid: a preferential reactivity with double-stranded DNA and dissociation of DNA-anti-DNA complexes

LF was found to bind to deoxyribonucleic acid as assessed by immunofluorescence studies on cell nuclei, affinity chromatography of DNA on immobilized LF, and gel chromatography of an LF-DNA reaction mixture. LF immobilized on Sepharose 4-B was reacted with 125I-labeled DNA in both its double-stranded and single-stranded configurations; dsDNA eluted with a 0.69M NaCl buffer, whereas ssDNA eluted with a 0.25M NaCl buffer. Additional evidence for a preferential reactivity with dsDNA was provided by the enzymatic treatment of preformed dsDNA-LF and ssDNA-LF complexes with S1 endonuclease, and DNAse 1--DNase digestion alone liberated free LF. The interaction of LF with DNA partially inhibited the binding of anti-DNA antibodies from patients with SLE, as assayed in a standard Farr assay. Furthermore, DNA-anti-DNA (labeled with 125I-IgG) complexes could be dispersed in vitro by the addition of LF. It is hypothesized that the release of LF by neutrophils chemotactically attracted to DNA-anti-DNA complexes may act as a feedback loop to modulate the inflammatory response in SLE.     

Antibodies against arylcyclohexylamines and their similarities in binding specificity with the phencyclidine receptor

Rabbit antibodies were generated against five unique epitopes of phencyclidine (PCP)-like molecules to determine the molecular requirements for arylcyclohexylamine binding to the PCP receptor. Three of the haptens contained the three ring structures of PCP. A fourth hapten was synthesized from a derivative of the highly potent PCP analog, 1-[1-(2-thienyl)cyclohexyl]piperidine. The fifth hapten, 5-[N-(1'-phenylcyclohexyl)amino]pentanoic acid, was used as a haptenic model for N-ethyl-1-phenylcyclohexylamine, one of the most potent arylcyclohexylamines. These haptens were bound covalently to bovine serum albumin and were then used as antigens to immunize rabbits. The affinities and cross-reactivity patterns of the resulting five antibodies were studied in a [3H]PCP radioimmunoassay using standard curves of various arylcyclohexylamines. The dissociation constants ranged from 1.9 to 51.6 nM. From the average IC50 values of the radioimmunoassay dose-response curves, the relative potency of each ligand to PCP was determined. Least-squares linear regression was used to correlate these data with relative potency data from two [3H]PCP receptor binding assays and a PCP drug discrimination assay in the rat. Only relative potency data from the anti-5[N-(1'-phenylcyclohexyl)amino]pentanoic acid antibody showed a significant correlation with data from the three pharmacological studies (r2 = 0.80, 0.57 and 0.78, respectively; p less than .05 in all cases). These data indicated the 5-[N-(1'-phenylcyclohexyl)amino]pentanoic acid hapten contained the pharmacologically active features needed for arylcyclohexylamine binding to the PCP receptor.     

血小板糖蛋白特异性抗体在骨髓增生异常综合征患者中的表达及其意义

本研究探讨血小板糖蛋白特异性抗体在骨髓增生异常综合征（MDS）患者发病中的作用。采用改良的MAIPA法检测血浆血小板糖蛋白特异性抗体（抗GPⅡb／Ⅲa抗体和抗GPIb／Ⅸ抗体）。若患者的OD值大于正常OD值的均数＋3倍标准差则为阳性。结果表明：MDS组总阳性率为16．67％（5／30），ITP组总阳性率为46．67％（14／30），两者之间差异有显著性（P〈0．05）。结论：部分MDS患者的糖蛋白特异性抗体为阳性，这表明部分MDS患者的血小板减少与免疫因素有关；血小板糖蛋白特异性抗体所致的血小板破坏在MDS患者的血小板减少中可能具有一定作用，这为MDS患者的免疫抑制剂治疗提供了新的依据。     

Mitochondrial DNA depletion in HIV-infected patients is more pronounced with chronic hepatitis C and enhanced following treatment with pegylated interferon plus ribavirin

BACKGROUND: Mitochondrial DNA (mtDNA) damage seems to be responsible for many of the toxicities associated with the long-term use of nucleoside analogues in HIV-infected patients. These adverse effects, mainly lipoatrophy, seem to be even more pronounced in subjects with hepatitis C virus (HCV) co-infection. However, there is no information about a possible additive effect of HCV on mtDNA depletion nor about the impact of ribavirin use in HIV/HCV-coinfected individuals. PATIENTS AND METHODS: mtDNA was measured in peripheral blood mononuclear cells (PBMC) collected from 192 individuals classified into 4 groups: HIV-neg/HCV-neg (control group, n = 11), HIV-pos/HCV-neg (56), HIV-neg/HCV-pos (18) and HIV-pos/HCV-pos (107). A duplex real-time NASBA assay was used to quantify mtDNA on maximal platelet-depleted specimens and all experiments were run in duplicate. The mtDNA copy number per cell was estimated taking as reference the nuclear DNA copy number. RESULTS: The mean mtDNA values in the control group was 757 copies/cell, while it was 428, 349 and 296 for HIV-pos, HCV-pos and HIV/HCV-coinfected individuals, respectively (P < 0.001 for all groups relative to the control group). No significant differences were observed when comparing patients with HIV or HCV infections alone, but coinfected individuals showed a lower mtDNA copy number than patients infected with HIV (P < 0.001) or with HCV (P = 0.089). In a subset of 18 patients with HIV/HCV-coinfection, treatment with pegylated interferon plus ribavirin produced a further reduction in mtDNA (mean value, 189 copies/cell; P = 0.009). CONCLUSIONS: HIV and HCV may independently cause mtDNA depletion in PBMC. Coinfection may result in more pronounced mtDNA depletion. The administration of interferon plus ribavirin may further enhance mtDNA depletion. These findings may explain the greater risk of lipoatrophy of antiretroviral therapy in HIV-infected patients with HCV coinfection and why anti-HCV therapy may aggravate this effect.     

慢性乙型肝炎病人干扰素治疗期间生存质量的调查研究

慢性乙型肝炎有自然加重的趋势,如不实施有效的治疗,会逐渐发展为肝硬化或肝癌.抗病毒治疗是治疗慢性乙型肝炎的根本方法,它通过清除或抑制病毒复制,可减轻肝脏炎症,延缓或阻止病情进展.随着科学的进步,治疗成功的概念界定为在保证病人生存质量的同时,改善症状,延长生命,因此有关病人生存质量的研究非常重要.     

多发性肌炎病人的观察与护理

目的探讨多发性肌炎病人的护理。方法对45例多发性肌炎住院病人进行药物护理、并发症护理、休息与肢体功能锻炼、饮食护理、心理护理、出院宣教等综合护理。结果45例病人肌力均恢复正常,临床痊愈3例,好转35例,无效7例。结论做好多发性肌炎病人的护理对提高病人生活质量至关重要。     

多发性肌炎病人的观察与护理

[目的]探讨多发性肌炎病人的护理。[方法]对45例多发性肌炎住院病人进行药物护理、并发症护理、休息与肢体功能锻炼、饮食护理、心理护理、出院宣教等综合护理。[结果]45例病人肌力均恢复正常，临床痊愈3例，好转35例，无效7例。[结论]做好多发性肌炎病人的护理对提高病人生活质量至关重要。