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1.
Zhonghua Yu Fang Yi Xue Za Zhi ; 55(5): 598-605, 2021 May 06.
Article in Chinese | MEDLINE | ID: mdl-34034399

ABSTRACT

Objective: To analyze the associations between prenatal and 1-year-old exposure to antibiotics and allergic symptoms in children aged 6-11 months and 18-23 months. Methods: In this study, a prospective birth cohort study was adopted. A total of 2 122 pregnant women were enrolled in Maternal and Child Health Care Center of Ma'anshan from June 2015 to June 2016, and they were followed up from the beginning of pregnancy to children's 24 months of age. Excluding 564 pairs of mothers and children who were lost to follow-up or with incomplete information on the use of antibiotics and children's allergic symptoms, a total of 1 558 pairs of mothers and children were included in the analysis of this study. The parents and children's general demographic information, early-life antibiotic exposure and other data were collected, the information about allergic symptoms in children aged 6-11 months and 18-23 months were investigated by reference to the "International Study of Asthma and Allergies in Childhood (ISAAC)". The univariate and multivariate binary unconditional logistic regression model was used to was used to estimate associations between the effects of early-life antibiotic exposure on allergic symptoms in 2-year-old children. Results: The antibiotic usage rate of pregnant women during pregnancy was 3.4% (53), and the antibiotic usage rates of children between 0 to 2 months, 3 to 5 months, and 6 to 11 months were separately 15.2%(237), 15.5%(242) and 17.3%(269). The total prevalence of allergic diseases in children aged 6 to 11 months was 24.1% (375 children), and the total prevalence of allergic diseases in children aged 18 to 23 months was 22.0% (342 children). After adjust parental (maternal) education level, family monthly income per capita, parental (maternal) allergy history, parental (maternal) age at pregnancy, mother's Body Mass Index (BMI) before pregnancy, exposure to second-hand smoke during pregnancy, delivery method, child gender, birth weight, preterm birth, the use of antibiotics when children were 3-5 months old (RR=1.61,95%CI:1.19-2.17) and 6-11 months old (RR=1.43,95%CI:1.06-1.93) were the risk factors for allergic symptoms at 6-11 months of age; and the use of antibiotics when children were 0-2 months old (RR=1.41, 95%CI: 1.03-1.95), 3-5 months old (RR=1.54, 95%CI: 1.12-2.11) and 6-11 months old (RR=1.58, 95%CI: 1.17-2.14) were the risk factors for allergic symptoms at 18-23 months of age. Conclusion: Children's exposure to antibiotics within 1 year of age was a risk factor for allergic symptoms in children aged 6-11 months and 18-23 months, children should avoid unnecessary antibiotic use in infancy.


Subject(s)
Premature Birth , Prenatal Exposure Delayed Effects , Anti-Bacterial Agents/therapeutic use , Child, Preschool , Cohort Studies , Female , Humans , Infant , Infant, Newborn , Mothers , Pregnancy , Prospective Studies
2.
Zhonghua Liu Xing Bing Xue Za Zhi ; 41(3): 320-325, 2020 Mar 10.
Article in Chinese | MEDLINE | ID: mdl-32294828

ABSTRACT

Objective: To describe the epidemiological characteristics of mobile phone use in early pregnancy, and to explore the relationship between pregnancy mobile use and infant sleep-wake behavior. Methods: During February 2015 to August 2016, 2 212 subjects who had their first antenatal examination at Maanshan Maternity and Child Health Hospital were recruited in this cohort study and followed until postpartum for 6 months. Information of phone use was collected through questionnaire in the third trimester. There were 1 779 pregnant reported hours of mobile phone use in the questionnaire. A total of 1 951 parent reported the night-wake times. Data on night-wake behavior in infants was collected during the 6 months study. Questionnaires were completed by parents when taking the physical examination. More than 3 times per night was defined as the night-wake frequency. Unconditional multivariate logistic regression was applied to analyze the association of pregnancy time of mobile phone use and the infant night-wake frequencies. Results: In this cohort study, the average age of 2 212 pregnant women was (26.95±3.82) years, with 1 983 of them were followed up to the time of delivery. The incidence of night-wake frequency was 28.3% (553/1 951) among these 6-month-old infants. After adjusted for feeding factors in the first trimester, frequencies of using the phone as "3 to 4 hour per day" and "5 hour and above per day" were both positively associated with the frequencies of night-wake behavior in infants. The adjusted OR (95%CI) were 1.49 (1.07-2.07) and 1.79 (1.31-2.46), respectively. Conclusions: The mobile phone use during pregnancy was associated with night-wake of infants. Mobile phone should be rationally used during pregnancy.


Subject(s)
Cell Phone Use/statistics & numerical data , Pregnancy Trimester, First , Sleep Initiation and Maintenance Disorders/epidemiology , Adult , Cohort Studies , Female , Humans , Infant , Pregnancy , Surveys and Questionnaires , Time Factors , Young Adult
3.
Zhonghua Liu Xing Bing Xue Za Zhi ; 39(4): 464-468, 2018 Apr 10.
Article in Chinese | MEDLINE | ID: mdl-29699038

ABSTRACT

Objective: To explore the interaction effect between mother's educational level and preschoolers' dietary pattern on attention-deficit/hyperactivity disorder (ADHD). Methods: In 2014, there were 16 439 children aged 3-6 years old from 91 kindergartens in Ma'anshan municipality of China. A semi-quantitative food frequency questionnaire and the 10-item Chinese version of the Conners' Abbreviated Symptom Questionnaire (C-ASQ) were administered to assess the usual dietary intake and symptoms on ADHD. Social-demographic information was collected through questionnaires. Unconditional logistic regression was used to analyze the multiplication interaction effect between mother's educational level and preschoolers' dietary pattern on ADHD. Excel software was used to analyze the additive interaction effect of mother's educational level and preschoolers'dietary pattern on ADHD. Results: Results showed that factors as: mother's low educational level[aOR=1.31 (1.13-1.52)], scores related to preschoolers in the top quintile of "food processing" [aOR=1.31 (1.16-1.48)] and "snack" [aOR=1.45 (1.29-1.63)]patterns showed greater odds while preschoolers in the top quintile of "vegetarian" [aOR=0.80 (0.71-0.90)]showed less odds for having ADHD symptoms. Both multiplication and additive interactions were observed between mothers with less education. The processed dietary patterns (OR=1.17, 95%CI: 1.11-1.25), relative excess risk of interaction (RERI), attributable proportion (AP) and the interaction index (SI) appeared as 0.21, 0.13 and 1.47, respectively. Multiplication interaction was observed between levels of mother's low education and the snack dietary pattern (OR=1.21, 95%CI: 1.14-1.29), with RERI, AP and SI as 0.49, 0.26 and 2.36, respectively. However, neither multiplication interaction or additive interaction was noticed between levels of mother's low education and the vegetarian dietary pattern (OR=0.97, 95%CI: 0.92-1.03), with RERI, AP and SI as 0.09, 0.05 and 1.15, respectively. Conclusions: Levels of mother's low education presented a risk factor for ADHD symptoms in preschool children. Both multiplication interaction and additive interaction were observed between mother's low education levels and the processed dietary pattern. Multiplication interaction was noticed between mother's education levels and the snack dietary pattern but not with the vegetarian dietary pattern.


Subject(s)
Attention Deficit Disorder with Hyperactivity/epidemiology , Diet , Educational Status , Feeding Behavior , Mothers , Attention Deficit Disorder with Hyperactivity/diagnosis , Child , Child Nutritional Physiological Phenomena , Child, Preschool , China , Female , Humans , Logistic Models , Risk Factors , Surveys and Questionnaires
4.
Eur J Clin Microbiol Infect Dis ; 33(9): 1525-32, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24756211

ABSTRACT

The aim of this study was to estimate the risk factors of bacterial vaginosis (BV) among rural married women of childbearing age in Anhui Province of China. A cross-sectional study was conducted and the method of stratified cluster sampling was used to identify a sample of 53,652 married women aged 18-49 years. All women were asked to complete an interviewer-administered standardized questionnaire, covering sociodemographic characteristics, history of menstruation, marriage and procreation, sexual life, personal hygienic behaviors, and reproductive tract infections (RTIs) knowledge, followed by the gynecological examination and laboratory inspection. A total of 53,286 married women aged 18-49 years were included in this analysis. The prevalence of BV was 11.99 % (6,391/53,286). Risk factors for BV included the minority nationality, women's lower education levels, husband's elder age, over 35 days of menstrual cycle, less than 3 days of menstruation, dysmenorrhea, usage of an intrauterine device (IUD), lack of RTIs knowledge, higher frequency of washing genitals before having sex with husband and changing underwear, lower frequency of sexual intercourse per month, and suffering from other RTIs. The results suggest that BV can be affected by many factors among rural married women of reproductive age, so comprehensive, scheduled programs at healthcare educations should be provided for women in order to prevent BV.


Subject(s)
Vaginosis, Bacterial/epidemiology , Adolescent , Adult , Bacteriological Techniques , China/epidemiology , Cross-Sectional Studies , Female , Gynecological Examination , Humans , Prevalence , Risk Factors , Rural Population , Surveys and Questionnaires , Young Adult
5.
Infect Immun ; 78(11): 4601-12, 2010 Nov.
Article in English | MEDLINE | ID: mdl-20713623

ABSTRACT

Although merozoite surface protein 1 (MSP-1) is a leading candidate vaccine antigen for blood-stage malaria, its efficacy in clinical trials has been limited in part by antigenic polymorphism and potentially by the inability of protein-in-adjuvant vaccines to induce strong cellular immunity. Here we report the design of novel vectored Plasmodium falciparum vaccines capable of overcoming such limitations. We optimized an antigenic insert comprising the four conserved blocks of MSP-1 fused to tandemly arranged sequences that represent both allelic forms of the dimorphic 42-kDa C-terminal region. Inserts were expressed by adenoviral and poxviral vectors and employed in heterologous prime-boost regimens. Simian adenoviral vectors were used in an effort to circumvent preexisting immunity to human adenoviruses. In preclinical studies these vaccines induced potent cellular immune responses and high-titer antibodies directed against MSP-1. The antibodies induced were found to have growth-inhibitory activity against dimorphic allelic families of P. falciparum. These vectored vaccines should allow assessment in humans of the safety and efficacy of inducing strong cellular as well as cross-strain humoral immunity to P. falciparum MSP-1.


Subject(s)
DNA Viruses/genetics , Erythrocytes/parasitology , Genetic Vectors , Malaria Vaccines , Malaria, Falciparum/prevention & control , Merozoite Surface Protein 1/metabolism , Adenoviruses, Human/genetics , Adenoviruses, Simian/genetics , Animals , Antibodies, Protozoan/blood , Chick Embryo , Drug Design , Female , Humans , Immunization , Immunization, Secondary , Malaria Vaccines/administration & dosage , Malaria Vaccines/genetics , Malaria Vaccines/immunology , Malaria, Falciparum/immunology , Malaria, Falciparum/parasitology , Merozoite Surface Protein 1/genetics , Merozoite Surface Protein 1/immunology , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Plasmodium falciparum/immunology , T-Lymphocytes/immunology , Vaccinia virus/genetics
6.
J Virol ; 82(8): 3822-33, 2008 Apr.
Article in English | MEDLINE | ID: mdl-18256155

ABSTRACT

Human adenovirus serotype 5 (AdH5) vector vaccines elicit strong immune responses to the encoded antigen and have been used in various disease models. We designed AdH5 vectors expressing antigen under the control of a human cytomegalovirus (HCMV) immediate-early promoter containing its intron A sequence. The transcriptional levels of antigen and immune responses to antigen for vectors with the HCMV promoter with the intron A sequence (LP) were greater than those for AdH5 vectors using the HCMV promoter sequence without intron A (SP). We compared an E1E3-deleted AdH5 adenoviral vector, which affords more space for insertion of foreign sequences, and showed it to be as immunogenic as an E1-deleted AdH5 vector. Neutralizing antibodies to AdH5 limit the efficacy of vaccines based on the AdH5 serotype, and simian adenoviral vectors offer an attractive option to overcome this problem. We constructed E1E3-deleted human and simian adenoviral vectors encoding the pre-erythrocytic-stage malarial antigen Plasmodium berghei circumsporozoite protein. We compared the immunogenicity and efficacy of AdC6, a recombinant simian adenovirus serotype 6 vector, in a murine malaria model to those of AdH5 and the poxviral vectors MVA and FP9. AdC6 induced sterile protection from a single dose in 90% of mice, in contrast to AdH5 (25%) and poxviral vectors MVA and FP9 (0%). Adenoviral vectors maintained potent CD8(+) T-cell responses for a longer period after immunization than did poxviral vectors and mainly induced an effector memory phenotype of cells. Significantly, AdC6 was able to maintain protection in the presence of preexisting immunity to AdH5.


Subject(s)
Adenoviruses, Simian/genetics , Cytomegalovirus/genetics , Malaria Vaccines/immunology , Malaria/prevention & control , Plasmodium berghei/immunology , Protozoan Proteins/immunology , Adenoviruses, Human/genetics , Adenoviruses, Human/immunology , Animals , CD8-Positive T-Lymphocytes/immunology , Cell Line , Female , Genetic Vectors , Immunologic Memory , Mice , Mice, Inbred BALB C , Plasmodium berghei/genetics , Promoter Regions, Genetic , Protozoan Proteins/genetics , T-Lymphocyte Subsets/immunology , Time Factors
7.
Gene Ther ; 13(5): 421-9, 2006 Mar.
Article in English | MEDLINE | ID: mdl-16319951

ABSTRACT

Vaccines based on replication-defective adenoviral vectors are being developed for infectious agents and tumor-associated antigens. Early work focused on vaccines derived from a common human serotype of adenovirus, that is, adenovirus of the serotype 5 (AdHu5). Neutralizing antibodies against AdHu5 virus, present in a large percentage of the human population, dampen the efficacy of vaccines based on this carrier. To circumvent this problem, we generated vectors derived from chimpanzee adenoviruses. Here we describe some basic parameters of vectors derived from chimpanzee adenoviruses C68 and C7, including growth characteristics, yields of infectious particles, effects of additional deletions in E3 and E4 and lengths of the inserted foreign sequence as they relate to the suitability for their eventual development as vaccine carriers for clinical use.


Subject(s)
Adenoviridae/genetics , Genetic Engineering , Genetic Vectors/genetics , Viral Vaccines/genetics , Adenoviridae Infections/immunology , Animals , Antigens/immunology , Bioreactors , CD8-Positive T-Lymphocytes/immunology , Cell Line , Female , Genetic Vectors/administration & dosage , Immunoblotting , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Pan troglodytes/virology , Transgenes , Viral Vaccines/administration & dosage
8.
J Virol ; 78(14): 7392-9, 2004 Jul.
Article in English | MEDLINE | ID: mdl-15220412

ABSTRACT

Two triple immunization vaccine regimens with adenoviral vectors with E1 deleted expressing Gag of human immunodeficiency virus type 1 were tested for induction of T- and B-cell-mediated-immune responses in mice and in nonhuman primates. The vaccine carriers were derived from distinct serotypes of human and simian adenoviruses that fail to elicit cross-neutralizing antibodies expected to dampen the effect of booster immunizations. Both triple immunization regimens induced unprecedented frequencies of gamma interferon-producing CD8(+) T cells to Gag in mice and monkeys that remained remarkably stable over time. In addition, monkeys developed Gag-specific interleukin-2-secreting T cells, presumably belonging to the CD4(+) T-cell subset, and antibodies to both Gag and the adenoviral vaccine carriers.


Subject(s)
AIDS Vaccines/immunology , Adenoviruses, Human/immunology , Gene Products, gag/immunology , Genetic Vectors , HIV Infections/prevention & control , HIV-1/immunology , AIDS Vaccines/administration & dosage , AIDS Vaccines/genetics , Adenoviruses, Human/genetics , Animals , B-Lymphocytes/immunology , Gene Products, gag/genetics , HIV Antibodies/blood , HIV Infections/immunology , HIV Infections/virology , Humans , Immunization , Immunization Schedule , Immunization, Secondary , Macaca mulatta , Mice , Mice, Inbred BALB C , Species Specificity , T-Lymphocytes/immunology , Transgenes
9.
Vaccine ; 22(5-6): 697-703, 2004 Jan 26.
Article in English | MEDLINE | ID: mdl-14741162

ABSTRACT

An E1-deleted adenoviral recombinant derived from the chimpanzee serotype 6 expressing a codon-optimized truncated form of gag of human immunodeficiency virus type 1 (HIV-1) was tested for induction of a transgene product-specific CD8+ T cell response upon oral immunization of mice. The vector was shown to induce gag-specific CD8+ T cells detectable at moderate frequencies of approximately 0.5-1.0% in the spleens and to provide partial protection in a surrogate challenge model based on intraperitoneal (i.p.) infection of mice with a vaccinia virus recombinant expressing gag (VVgag) of HIV-1. Frequencies of gag-specific CD8+ T cells could be augmented by using a different, i.e., heterologous, vaccine carrier based on a distinct recombinant virus or an alternative adenoviral serotype expressing the same form of gag for oral or systemic-booster immunization.


Subject(s)
Adenoviruses, Simian/genetics , CD8-Positive T-Lymphocytes/immunology , Genetic Vectors , HIV Antigens/immunology , HIV-1/immunology , Immunity, Cellular/immunology , Administration, Oral , Animals , Cytokines/biosynthesis , Female , Gene Products, gag/immunology , HeLa Cells , Humans , Immunization , Immunization, Secondary , Immunohistochemistry , Mice , Mice, Inbred BALB C , Peptides/immunology , Vaccines, Synthetic/immunology , Vaccinia virus/immunology
10.
J Virol ; 77(20): 10780-9, 2003 Oct.
Article in English | MEDLINE | ID: mdl-14512528

ABSTRACT

Adenovirus vectors with E1 deleted of the human serotype 5 (AdHu5) and the chimpanzee serotype 68 (AdC68) expressing the glycoprotein of the Evelyn Rokiniki Abelseth strain of rabies virus were tested upon oral application for induction of systemic and mucosal transgene product-specific antibody responses in mice. Both vectors induced systemic and mucosal antibodies to rabies virus, including virus-neutralizing antibodies and protection against a severe intracerebral challenge with a mouse-adapted strain of rabies virus. Pre-existing immunity of AdHu5 virus, which dampens induction of transgene product-specific immunity elicited by AdHu5 vectors given systemically did not impair the response induced by oral vaccination. Oral priming-boosting regimens with either heterologous or homologous adenoviral vectors used sequentially increased both mucosal and systemic antibody titers to rabies virus [corrected]


Subject(s)
Adenoviridae/genetics , Adenoviridae/immunology , Antibodies, Viral/biosynthesis , Genetic Vectors , Rabies Vaccines/immunology , Vaccines, Synthetic/immunology , Administration, Oral , Animals , Antibodies, Viral/immunology , Female , Mice , Mice, Inbred C57BL , Mice, Inbred ICR , Neutralization Tests , Rabies Vaccines/administration & dosage , Transgenes , Vaccination
11.
Virology ; 305(2): 397-405, 2003 Jan 20.
Article in English | MEDLINE | ID: mdl-12573585

ABSTRACT

Mice lacking CD4(+) T cells due to a knock-out mutation respond to vaccination with a replication-defective adenoviral recombinant expressing the glycoprotein of rabies virus with a long-lasting virus-neutralizing antibody response. The vaccine-induced B cells secrete antibodies that are mainly of IgG isotypes. The response can be enhanced upon booster immunization, indicating the induction of B cell memory in the absence of CD4(+) T cells. The antibody response is independent of CD8(+) T cells but requires the presence of CD3(+) cells carrying the NK1.1 markers.


Subject(s)
Antibodies, Viral/biosynthesis , Antigens, Viral , Antigens/analysis , CD4-Positive T-Lymphocytes/physiology , Glycoproteins/immunology , Proteins/analysis , Rabies Vaccines/immunology , Rabies virus/immunology , Vaccines, Synthetic/immunology , Viral Envelope Proteins/immunology , Adenoviridae/genetics , Animals , Antibody Formation , Antigens, Ly , Antigens, Surface , B-Lymphocytes/immunology , CD4 Antigens/physiology , Cell Line , Glycoproteins/genetics , Immunization , Immunophenotyping , Lectins, C-Type , Mice , Mice, Inbred C57BL , Mice, Knockout , NK Cell Lectin-Like Receptor Subfamily B , Transgenes , Viral Envelope Proteins/genetics
12.
Gene Ther ; 9(14): 963-71, 2002 Jul.
Article in English | MEDLINE | ID: mdl-12085245

ABSTRACT

To test whether hepatocytes engineered in vivo can serve as surrogate beta cells by similarly secreting mature insulin in a glucose-sensitive manner, we prepared adenoviral vectors encoding wild-type proinsulin (hIns-wt), a modified proinsulin cleavable by the ubiquitously expressed protease furin (hIns-M3), or each of the two beta cell specific pro-insulin convertases PC2 and PC3. Following a detailed in vitro characterization of the proteins produced by our vectors, we infected the liver and, for comparison, the muscle of a chemically induced murine model of type I diabetes. Insulin expression from the transduced tissues was extensively characterized and showed to be constitutive rather than regulated. To obtain regulated expression, we placed expression of hIns-M3 under the control of the dimerizer-inducible transcription system. Hormone secretion from mouse liver was negligible in the absence of the dimerizer drug rapamycin, was inducible in a dose-dependent manner upon its administration, and reversible following drug withdrawal. These data confirm liver as a promising target for in vivo expression of processed insulin. While suggesting that hepatocytes cannot provide authentic glucose-responsive regulation, these results demonstrate that pharmacological regulation is a promising alternative route to the controlled delivery of insulin following hepatic gene transfer.


Subject(s)
Diabetes Mellitus, Type 1/therapy , Genetic Therapy/methods , Hepatocytes/metabolism , Proinsulin/genetics , Sirolimus/therapeutic use , Adenoviridae , Animals , Cells, Cultured , Combined Modality Therapy , Diabetes Mellitus, Experimental , Dimerization , Female , Gene Expression , Genetic Engineering/methods , Genetic Vectors/pharmacology , Hepatocytes/drug effects , Insulin/metabolism , Insulin Secretion , Mice , Mice, Nude , Transfection/methods
13.
J Virol ; 75(23): 11603-13, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11689642

ABSTRACT

An adenovirus previously isolated from a mesenteric lymph node from a chimpanzee was fully sequenced and found to be similar in overall structure to human adenoviruses. The genome of this virus, called C68, is 36,521 bp in length and is most similar to subgroup E of human adenovirus, with 90% identity in most adenovirus type 4 open reading frames that have been sequenced. Substantial differences in the hexon hypervariable regions were noted between C68 and other known adenoviruses, including adenovirus type 4. Neutralizing antibodies to C68 were highly prevalent in sera from a population of chimpanzees, while sera from humans and rhesus monkeys failed to neutralize C68. Furthermore, infection with C68 was not neutralized from sera of mice immunized with human adenovirus serotypes 2, 4, 5, 7, and 12. A replication-defective version of C68 was created by replacing the E1a and E1b genes with a minigene cassette; this vector was efficiently transcomplemented by the E1 region of human adenovirus type 5. C68 vector transduced a number of human and murine cell lines. This nonhuman adenoviral vector is sufficiently similar to human serotypes to allow growth in 293 cells and transduction of cells expressing the coxsackievirus and adenovirus receptor. As it is dissimilar in regions such as the hexon hypervariable domains, C68 vector avoids significant cross-neutralization by sera directed against human serotypes.


Subject(s)
Adenoviridae/genetics , Capsid Proteins , Genetic Vectors , Amino Acid Sequence , Animals , Capsid/chemistry , Capsid/genetics , Cloning, Molecular , Genome, Viral , Humans , Mice , Models, Molecular , Molecular Sequence Data , Neutralization Tests , Open Reading Frames , Pan troglodytes , Protein Conformation , Sequence Homology, Amino Acid
14.
Cancer Res ; 61(16): 6201-12, 2001 Aug 15.
Article in English | MEDLINE | ID: mdl-11507073

ABSTRACT

Malignant mesothelioma remains an incurable disease for which immune-modulatory therapies, such as exogenous cytokines, have shown some promise. One such cytokine, IFN-beta, has potent antiproliferative and immunostimulatory activity in vitro, but its in vivo use has been limited by toxicity. We thus conducted studies evaluating intracavitary delivery of a replication-deficient adenoviral (Ad) vector encoding for the murine IFN-beta gene (Ad.muIFN-beta) in mouse models of malignant mesothelioma. In contrast to multiple injections of recombinant protein, a single i.p. injection of Ad.muIFN-beta into animals with established tumors elicited remarkable antitumor activity leading to long-term survival in >90% of animals bearing either AB12 or AC29 i.p. mesotheliomas. A control adenovirus vector had minimal antitumor effect in vivo. Significant therapeutic effects were also seen in animals treated with large tumor burdens. Importantly, treatment of i.p. tumor also led to reduction of growth in tumors established at a distant site (flank). A number of experiments suggested that these effects were attributable to an acquired CD8(+) T-cell-mediated response including: (a) the induction of long-lasting antitumor immunity; (b) loss of efficacy of Ad.muIFN-beta in tumor-bearing, immune-deficient (SCID, SCID/beige) mice; (c) detection of high levels of specific antitumor cytolytic activity from unstimulated splenocytes harvested from Ad.muIFN-beta-treated animals that was abolished by CD8(+) T-cell depletion; and (d) abrogation of antitumor effects of Ad.muIFN-beta in tumor-bearing CD8(+) T-cell-depleted animals. These data show that intracavitary IFN-beta gene therapy using an adenoviral vector provides strong CD8(+) T-cell-mediated antitumor effects in murine models of mesothelioma and suggest that this may be a promising strategy for the treatment of localized tumors such as mesothelioma or ovarian cancer in humans.


Subject(s)
Genetic Therapy/methods , Interferon-beta/genetics , Interferon-beta/immunology , Mesothelioma/therapy , Peritoneal Neoplasms/therapy , Adenoviridae/genetics , Animals , CD4-Positive T-Lymphocytes/immunology , Cell Division/immunology , Cytotoxicity, Immunologic , Dose-Response Relationship, Immunologic , Female , Genetic Vectors/genetics , Injections, Intraperitoneal , Interferon-beta/metabolism , Mesothelioma/genetics , Mesothelioma/immunology , Mice , Mice, Inbred BALB C , Mice, SCID , Peritoneal Neoplasms/genetics , Peritoneal Neoplasms/immunology , T-Lymphocytes, Cytotoxic/immunology , Transfection
15.
Mol Ther ; 3(5 Pt 1): 697-707, 2001 May.
Article in English | MEDLINE | ID: mdl-11356075

ABSTRACT

We investigated the cellular basis for secretion of inflammatory cytokines in mice following intravenous administration of adenoviral vectors (Ad). Serum inflammatory cytokines including interleukin-6 (IL-6), IL-12, and tumor necrosis factor-alpha (TNF-alpha) were detected as early as 6 h following intravenous injection of Ad-expressing Escherichia coli beta-galactosidase (Ad-lacZ). Ad-lacZ readily accumulated in the splenic marginal zone 1 h after intravenous infusion, where both dendritic cells (DCs) and macrophages were transduced and activated within 6 h. Flow cytometric analyses showed that the expression of Ia and CD86 antigens was markedly enhanced on splenic DCs indicating their activation in vivo by Ad-lacZ. Upon ex vivo culture, these early-activated splenic DCs spontaneously produced high levels of IL-6 and IL-12. By contrast, activated splenic macrophages spontaneously secreted only IL-6. Elimination of tissue macrophages and splenic DCs in vivo considerably reduced the early release of IL-12, IL-6, and TNF-alpha and significantly blocked the specific cellular immune response to Ad and the transgene product in vivo. Our findings indicate that preferential activation of DCs and macrophages may account for Ad-triggered acute inflammatory response in vivo in mice. Moreover, DCs and macrophages may play different roles in this process in terms of their abilities to produce distinct patterns of inflammatory cytokines.


Subject(s)
Adenoviridae/genetics , Cytokines/biosynthesis , Dendritic Cells/metabolism , Inflammation , Macrophages/metabolism , Animals , Antigens, CD/biosynthesis , B7-2 Antigen , Cells, Cultured , Dose-Response Relationship, Drug , Escherichia coli/enzymology , Flow Cytometry , Immunohistochemistry , Interleukin-12/biosynthesis , Interleukin-6/biosynthesis , Lac Operon , Liposomes/metabolism , Membrane Glycoproteins/biosynthesis , Mice , Mice, Inbred C57BL , Microscopy, Fluorescence , Spleen/cytology , Spleen/metabolism , T-Lymphocytes, Cytotoxic/metabolism , Time Factors , Tissue Distribution , Transgenes , Tumor Necrosis Factor-alpha/biosynthesis , beta-Galactosidase/genetics
16.
Mol Ther ; 3(5 Pt 1): 708-22, 2001 May.
Article in English | MEDLINE | ID: mdl-11356076

ABSTRACT

The innate immune response to intraportally infused adenoviral vector was evaluated in rhesus monkeys. A first-generation adenovirus-expressing lacZ (Ad-lacZ) was administered at a dose just below that which causes severe morbidity. The response to vector was evaluated for the initial 24 h following infusion. Clinical findings during this time were primarily limited to petechiae, consistent with the development of thrombocytopenia and biochemical evidence of disseminated intravascular coagulation. Serum transaminases were elevated and a lymphopenia developed. Tracking of fluorescent-labeled vector demonstrated distribution to macrophages and dendritic cells of the spleen and Kupffer cells of the liver. A systemic release of the cytokine IL-6 occurred soon after vector infusion. Analysis of splenic cells revealed acute activation of macrophages and dendritic cells followed by massive apoptosis. Bone marrow cultures demonstrated normal erythroid and primitive progenitors with a significant decrease in myeloid progenitors. Similar findings, except the abnormality in bone marrow cultures, were observed in monkeys who received an identical dose of Ad-lacZ in which vector genes were inactivated with psoralen and UV irradiation. These data suggest that inadvertent targeting of antigen-presenting cells following intraportal infusion of vector leads to a systemic cytokine syndrome which may be triggered by the viral capsid proteins.


Subject(s)
Adenoviridae/genetics , Genetic Vectors , Animals , Apoptosis , Bone Marrow Cells/cytology , Bone Marrow Cells/metabolism , Cells, Cultured , Dendritic Cells/metabolism , Ficusin/pharmacology , Flow Cytometry , Fluorescent Dyes/pharmacology , Interleukin-6/biosynthesis , Kupffer Cells/metabolism , Lac Operon , Liver/metabolism , Lymphopenia , Macaca mulatta , Macrophages/metabolism , Male , Methylcellulose/metabolism , Microscopy, Electron , Models, Biological , Spleen/cytology , Spleen/metabolism , Thrombocytopenia , Time Factors , Tissue Distribution , Transaminases/biosynthesis , Ultraviolet Rays , beta-Galactosidase/metabolism
17.
J Virol ; 75(11): 5222-9, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11333904

ABSTRACT

Adenovirus vectors have been studied as vehicles for gene transfer to skeletal muscle, an attractive target for gene therapies for inherited and acquired diseases. In this setting, immune responses to viral proteins and/or transgene products cause inflammation and lead to loss of transgene expression. A few studies in murine models have suggested that the destructive cell-mediated immune response to virally encoded proteins of E1-deleted adenovirus may not contribute to the elimination of transgene-expressing cells. However, the impact of immune responses following intramuscular administration of adenovirus vectors on transgene stability has not been elucidated in larger animal models such as nonhuman primates. Here we demonstrate that intramuscular administration of E1-deleted adenovirus vector expressing rhesus monkey erythropoietin or growth hormone to rhesus monkeys results in generation of a Th1-dependent cytotoxic T-cell response to adenovirus proteins. Transgene expression dropped significantly over time but was still detectable in some animals after 6 months. Systemic levels of adenovirus-specific neutralizing antibodies were generated, which blocked vector readministration. These studies indicate that the cellular and humoral immune response generated to adenovirus proteins, in the context of transgenes encoding self-proteins, hinders long-term transgene expression and readministration with first-generation vectors.


Subject(s)
Adenoviridae/genetics , Adenovirus E1 Proteins/genetics , Genetic Therapy/methods , Genetic Vectors/administration & dosage , Adenoviridae/immunology , Animals , Antibodies, Viral/blood , Cytokines/analysis , Erythropoietin/blood , Erythropoietin/genetics , Gene Deletion , Genetic Vectors/genetics , Genetic Vectors/immunology , Growth Hormone/blood , Growth Hormone/genetics , Injections, Intramuscular , Lymphocyte Activation , Macaca mulatta , Neutralization Tests , T-Lymphocytes, Cytotoxic/immunology , Transfection
18.
Gene Ther ; 8(6): 442-6, 2001 Mar.
Article in English | MEDLINE | ID: mdl-11313822

ABSTRACT

The availability of inducible expression systems makes regulatable control of therapeutic proteins an attainable goal in gene therapy. We delivered tetracycline-inducible transgenes to the subretinal space using recombinant adenoviruses. Upon administration of doxycycline, we demonstrated reversible expression of green fluorescent protein in the retinal pigment epithelium as well as modulation of human growth hormone produced in the retina and secreted in the blood stream. This mode of delivery and regulation offers a unique way to evaluate gene function in the eye and represents a novel method for introducing therapeutic proteins into the retina.


Subject(s)
Anti-Bacterial Agents/pharmacology , Gene Expression Regulation/drug effects , Genetic Therapy/methods , Genetic Vectors/administration & dosage , Pigment Epithelium of Eye/metabolism , Transduction, Genetic/methods , Adenoviridae/genetics , Animals , Doxycycline/pharmacology , Female , Green Fluorescent Proteins , Human Growth Hormone/genetics , Humans , Luminescent Proteins/genetics , Mice , Mice, Nude , Tetracycline/pharmacology
19.
Hum Gene Ther ; 12(1): 71-6, 2001 Jan 01.
Article in English | MEDLINE | ID: mdl-11177544

ABSTRACT

One of the most promising gene transfer vectors in human clinical trials is AAV2. The quality of the vector preparations is a key element in obtaining reliable and reproducible data in preclinical studies. However, established protocols either result in impure, low infectious virus (CsCl2 gradient centrifugation) or demand a high level of manual and technical skills (CsCl2 gradient centrifugation, iodixanol/heparin or HPLC purification). In this study, we present an easy-to-do single-step column purification (SSCP) of AAV2 by gravity flow based on its affinity to heparin, without ultracentrifugation. Various vector preparations generated by our method reproducibly showed high titers, infectivity, and purity. In vivo, our single-step column-purified AAV2 vectors mediate significantly higher transduction efficiency compared with conventional protocols. Investigators still unsatisfied with previously published techniques or new to the field of AAV production may find in our method an interesting alternative.


Subject(s)
Dependovirus/isolation & purification , Genetic Therapy , Genetic Vectors/isolation & purification , Animals , Blotting, Western , Cell Line , Chromatography, Ion Exchange/methods , Electrophoresis, Polyacrylamide Gel , Enzyme-Linked Immunosorbent Assay , Gene Transfer Techniques , Heparin/chemistry , Humans , In Vitro Techniques , Lac Operon/physiology , Mice , Mice, Inbred C57BL , Muscles/virology , Transduction, Genetic , Transfection , beta-Galactosidase/metabolism
20.
Hum Gene Ther ; 12(2): 205-15, 2001 Jan 20.
Article in English | MEDLINE | ID: mdl-11177557

ABSTRACT

Recombinant adeno-associated virus (rAAV) vectors allow efficient gene transfer and expression in the muscle; therefore, rAAVs represent a potential gene therapy vector for muscular dystrophies. For further investigations, we used a mouse muscular dystrophy model (gsg(-/-) mice) gamma-sarcoglycan, a subunit of the dystrophin-glycoprotein complex, is missing. gsg(-/-) mice develop progressive dystrophy representative of a severe human phenotype disease. We previously showed high levels and stable expression of gamma-sarcoglycan in myofibers after direct muscle injection into gsg(-/-) mice of a recombinant AAV vector (AAV.dMCK.gSG) carrying the gamma-sarcoglycan cDNA driven by a muscle-specific promoter (truncated version of muscle creatine kinase). Here, we show that when gamma-sarcoglycan expression is driven by the ubiquitous cytomegalovirus (CMV) promoter (AAV.CMV.gSG), lower levels of transgene expression are observed and are associated with a humoral response to gamma-sarcoglycan. When using an rAAV vector, expressing the highly immunogenic product gamma-galactosidase under the CMV promoter (AAV.CMV.LacZ), we measured a strong cellular and humoral immune response to the transgene after intramuscular injection into gsg(-/-) mice. This study suggests that restriction of transgene expression to the muscle is an important criterion for the treatment of muscular dystrophies and will aid in the design of protocols for gene therapy.


Subject(s)
Cytoskeletal Proteins/biosynthesis , Dependovirus/genetics , Gene Transfer Techniques , Membrane Glycoproteins/biosynthesis , Muscle, Skeletal/metabolism , Muscular Dystrophies/therapy , Promoter Regions, Genetic , Animals , Antigen-Presenting Cells/immunology , Blotting, Western , Creatine Kinase/genetics , Dendritic Cells/immunology , Dystrophin/biosynthesis , Genetic Vectors , Humans , Immunoenzyme Techniques , Macrophages/physiology , Mice , Mice, Inbred C57BL , Mice, Mutant Strains , Muscular Dystrophies/genetics , Muscular Dystrophies/immunology , Muscular Dystrophies/metabolism , Sarcoglycans , T-Lymphocytes, Cytotoxic , Transduction, Genetic , beta-Galactosidase/metabolism
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