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J Castilla

Publications and source records attributed to J Castilla.

At least 55 records · Page 3Linked to original sources
PubMed1998

Interference of coronavirus infection by expression of IgG or IgA virus neutralizing antibodies.

Mouse immunoglobulin gene fragments encoding the variable modules of the heavy (VH) and light (VL) chains of a transmissible gastroenteritis coronavirus (TGEV) neutralizing monoclonal antibody (MAb) have been cloned and sequenced. The selected MAb recognizes a highly conserved viral epitope and does not lead to the selection of neutralization escape mutants. Chimeric immunoglobulin genes with the variable modules from the murine MAb and constant modules of human gamma 1 and kappa chains were constructed using RT-PCR. These chimeric immunoglobulins were stably or transiently expressed in murine myelomas and COS cells, respectively. The secreted recombinant antibodies had radioimmunoassay (RIA) titers higher than 10(3) and reduced the infectious virus more than 10(4)-fold. Recombinant dimeric IgA showed a 50-fold enhanced neutralization of TGEV relative to a recombinant monomeric IgG1 which contained the identical antigen binding site. Epithelial cell lines stably-transformed with these constructs and expressing either recombinant IgG or IgA TGEV neutralizing antibodies reduced virus production by > 10(5)-fold after infection with homologous virus, although a residual level of virus production (< 10(2) PFU/ml) remained in less than 0.1% of the cells.

Animals↗
PubMed1998

Lactogenic immunity in transgenic mice producing recombinant antibodies neutralizing coronavirus.

Protection against coronavirus infections can be provided by the oral administration of virus neutralizing antibodies. To provide lactogenic immunity, eighteen lines of transgenic mice secreting a recombinant IgG1 monoclonal antibody (rIgG1) and ten lines of transgenic mice secreting recombinant IgA monoclonal antibodies (rIgA) neutralizing transmissible gastroenteritis coronavirus (TGEV) into the milk were generated. Genes encoding the light and heavy chains of monoclonal antibody (MAb) 6A.C3 were expressed under the control of regulatory sequences derived from the mouse genomic DNA encoding the whey acidic protein (WAP) and beta-lactoglobulin (BLG), which are highly abundant milk proteins. The MAb 6A.C3 binds to a highly conserved epitope present in coronaviruses of several species. This MAb does not allow the selection of neutralization escaping virus mutants. The antibody was expressed in the milk of transgenic mice with titers of one million as determined by RIA, and neutralized TGEV infectivity by one million fold corresponding to immunoglobulin concentrations of 5 to 6 mg per ml. Matrix attachment regions (MAR) sequences were not essential for rIgG1 transgene expression, but co-microinjection of MAR and antibody genes led to a twenty to ten thousand-fold increase in the antibody titer in 50% of the rIgG1 transgenic animals generated. Co-microinjection of the genomic BLG gene with rIgA light and heavy chain genes led to the generation of transgenic mice carrying the three transgenes. The highest antibody titers were produced by transgenic mice that had integrated the antibody and BLG genes, although the number of transgenic animals generated does not allow a definitive conclusion on the enhancing effect of BLG co-integration. Antibody expression levels were transgene copy number independent and integration site dependent. The generation of transgenic animals producing virus neutralizing antibodies in the milk could be a general approach to provide protection against neonatal infections of the enteric tract.

Animals↗
PubMed1998

The decline in CD4+ T lymphocytes as a function of the duration of HIV infection, age at seroconversion, and viral load.

BACKGROUND: The rate of CD4+ T-lymphocyte decline seen in HIV-infected patients is very variable. Although older patients, a longer duration of HIV infection, and a high level of plasma viraemia, have been associated with a faster fall in CD4+ T-cells, the relationship between these variables is still not well known. PATIENTS AND METHODS: In a cross-sectional study that included a total of 107 patients of known age and date at HIV seroconversion, the current CD4+ T-cell count and plasma viraemia were examined. Patients were not taking antiretroviral drugs, nor had received immunizations nor were suffering any intercurrent infections at the time of the study. RESULTS: The mean duration of HIV infection was 8.6+/-2.9 years. The mean CD4+ T-lymphocyte count was 367+/-264 x 10(6)/l. Mean plasma viraemia was 4.3+/-0.9 logs. In a linear regression model, the current CD4+ T-cell count was explained in 21.7% by the duration of HIV infection, while the level of plasma viraemia justified separately up to 37.0%. When both parameters were combined, they explained 58.8%. of the CD4+ lymphocyte values. In this model, a variation of one logarithm in the plasma viraemia had six times greater effect on the number of CD4+ lymphocytes than each year of HIV infection. When the age at seroconversion was added to the model, the CD4+ cell count allowed the explanation of up to 62.2% of cases. CONCLUSIONS: The age at seroconversion, the duration of HIV infection, and the level of plasma viraemia independently and substantially influence the current CD4+ lymphocyte count in HIV-infected subjects. However, other variables should exist (e.g. virus syncytium-inducing phenotype, host immunogenetic repertoire, etc.), contributing to explaining the different rate of CD4+ T-cell decline seen in HIV-infected subjects.

Adult↗
PubMed1998

Engineering passive immunity in transgenic mice secreting virus-neutralizing antibodies in milk.

Protection against enteric infections can be provided by the oral administration of pathogen-neutralizing antibodies. To provide passive immunity, 18 lines of transgenic mice secreting a recombinant monoclonal antibody (Mab) neutralizing transmissible gastroenteritis coronavirus (TGEV) into the milk were generated. The genes encoding a chimeric Mab with the variable modules of the murine TGEV-specific Mab 6A.C3 and the constant modules of a human IgG, isotype Mab were expressed under the control of regulatory sequences derived from the whey acidic protein, which is an abundant milk protein. The Mab 6A.C3 binds to a highly conserved epitope present in coronaviruses of several species, which does not allow the selection of neutralization escape mutants. Antibody expression titers of 10(6) were obtained in the milk of transgenic mice that reduced TGEV infectivity 10(6)-fold. The antibody was synthesized at high levels throughout lactation. Integration of matrix attachment region sequences with the antibody genes led to a 20- to 10,000-fold increase in the antibody titer in 50% of the transgenic animals. Antibody expression levels were transgene copy number independent and related to the site of integration. The generation of transgenic animals producing virus neutralizing antibodies in milk could provide an approach to protection against neonatal infections of the enteric tract.

Animals↗
PubMed1998

Sexual behaviour and condom use in the general population of Spain, 1996.

A national household survey of a representative sample of 9,984 individuals aged 15 years or over, carried out in 1996 using a combination of face-to-face interviews and self-completion questionnaires, was analyzed in order to describe the frequency of HIV sexual risk behaviours and condom use in Spain. Of a total of 8,101 persons (81%) who completed the questionnaire, 37% reported no sexual partner during the previous 12 months, 57% reported one partner and 6% reported more than one partner. Three per cent reported engaging in HIV sexual risk behaviour (i.e. more than one partner and failure to use a condom systematically), a pattern of behaviour which showed independent positive association with the male sex, an age of 20-59 years and being unmarried. Among those who had casual sexual partners during the preceding 12 months, 38% had always used condoms. In the multivariate analysis, failure to use a condom systematically with casual partners was associated with a higher age and being married. Of those who had regular partners during the previous 12 months, 26% had always used condoms, a finding associated with a lower age, higher educational level, unmarried status and non-cohabitation with the sexual partner. This survey has furnished some useful indicators for prevention purposes. Periodic repetition of this survey would enable possible changes in sexual risk behaviours to be detected.

Adolescent↗
PubMed1998

Transgenic mice secreting coronavirus neutralizing antibodies into the milk.

Ten lines of transgenic mice secreting transmissible gastroenteritis coronavirus (TGEV) neutralizing recombinant monoclonal antibodies (rMAbs) into the milk were generated. The rMAb light- and heavy-chain genes were assembled by fusing the genes encoding the variable modules of the murine MAb 6A.C3, which binds an interspecies conserved coronavirus epitope essential for virus infectivity, and a constant module from a porcine myeloma with the immunoglobulin A (IgA) isotype. The chimeric antibody led to dimer formation in the presence of J chain. The neutralization specific activity of the recombinant antibody produced in transiently or stably transformed cells was 50-fold higher than that of a monomeric rMAb with the IgG1 isotype and an identical binding site. This rMAb had titers of up to 10(4) by radioimmunoassay (RIA) and neutralized virus infectivity up to 10(4)-fold. Of 23 transgenic mice, 17 integrated both light and heavy chains, and at least 10 of them transmitted both genes to the progeny, leading to 100% of animals secreting functional TGEV neutralizing antibody during lactation. Selected mice produced milk with TGEV-specific antibody titers higher than 10(6) as determined by RIA, neutralized virus infectivity by 10(6)-fold, and produced up to 6 mg of antibody per ml. Antibody expression levels were transgene copy number independent and integration site dependent. Comicroinjection of the genomic beta-lactoglobulin gene with rMAb light- and heavy-chain genes led to the generation of transgenic mice carrying the three transgenes. The highest antibody titers were produced by transgenic mice that had integrated the antibody and beta-lactoglobulin genes, although the number of transgenic animals generated does not allow a definitive conclusion on the enhancing effect of beta-lactoglobulin cointegration. This approach may lead to the generation of transgenic animals providing lactogenic immunity to their progeny against enteric pathogens.

Amino Acid Sequence↗
PubMed1998

[Gonadal, hormonal and spermatic function in patients treated for cryptorchidism].

This paper studies the gonadal-hormonal and spermatic function of 48 adults (over 18 years old) who were treated in childhood for cryptorchidism by medical, surgical or both treatments. We measure plasmatic levels of FSH, LH, testosterone and prolactin of 34 patients and sperm samples of 31 patients. The testicular hormonal function was not disturbed except in four patients with high levels of prolactin (> 20 ng/ml) without symptoms. The spermatic function was disturbed in bilateral cryptorchidism mainly. The better sperm quality and testicular volume were at patients treated with hormonal treatment before surgical treatment.

Adult↗
PubMed1997

[AIDS and women].

Explore the source record for details and available documents.

Acquired Immunodeficiency Syndrome↗
PubMed1997

A pilot study on the efficacy of escalating dosage of alpha-interferon for chronic hepatitis C in HIV-infected patients. The Hepatitis/HIV Spanish Study Group.

Chronic liver disease caused by hepatitis C virus (HCV) seems to present a more accelerated course in HIV-infected patients, leading to cirrhosis and liver failure in a shorter period of time than in HIV-negative individuals. As efforts to increase life expectancy in HIV-infected people progress, substantial morbidity and mortality from HCV infection is likely to arise amongst subjects infected parenterally, such as injecting drug users, haemophiliacs and transfusion recipients. Preliminary results have suggested that alpha-interferon (IFN) treatment allows higher rates of response in HIV-infected patients with higher CD4+ lymphocyte counts, suggesting a primary dependence of IFN on a preserved immune system in order to act appropriately. In an open, multicentre, prospective trial we analysed whether the use of larger doses of IFN, through an escalating schedule, might overcome the limits imposed by immune dysfunction. An interim analysis performed in 29 patients concluded that escalating the dosage did not improve the rate of response to IFN. In fact, only one (8.3%) out of 12 patients without response after 3 months of being on IFN therapy achieved response after the dosage was increased from 5 MU to 8 MU s.c. three times a week. Moreover, he relapsed 3 months after completion of treatment for 1 year.

Adult↗
PubMed1997

Impact of human immunodeficiency virus infection on mortality among young men and women in Spain.

BACKGROUND: This paper describes the impact of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) mortality among young adults in Spain with specific reference to other causes of death. METHODS: Based on death registration data for the period 1980-1993, HIV/AIDS was compared against all other causes of death by gender, using specific rates in the 25-44 age group and standardized rates for potential years of life lost (PYLL). RESULTS: In 1993, HIV/AIDS was the leading cause of death among men aged 25-44 years (21.8% of all deaths) and the second leading cause of death among women (14.9%), exceeded only by cancer. Since 1982, the trend in the overall standardized mortality rate for men in the 25-44 age group has been reversed, showing a progressive increase. Similarly, since 1984 there has been a halt in the decline in female mortality. For both sexes, maintenance of these trends in mortality was largely ascribable to the effect of HIV/AIDS deaths which registered a marked rise, a rise far sharper than that witnessed for variations in all other causes studied. In 1993, the adjusted PYLL rate for HIV/AIDS for ages 1-70 rose to 615 per 100,000 population in men and 156 in women. These values accounted for 9.2% and 5.8% of PYLL for all causes, thereby ranking HIV/AIDS behind motor vehicle accidents as the second leading cause of premature death in men, and behind motor vehicle accidents and breast cancer as the third leading cause in women. For both sexes, the rise in the PYLL rate for HIV/AIDS from 1992 to 1993 proved far greater than that for all other causes of death. CONCLUSION: In Spain, HIV/AIDS has become the leading cause of death among young adults and is counteracting improvements in mortality due to other causes. It should therefore be regarded as a priority public health problem.

Adolescent↗
PubMed1997

Pulmonary and extrapulmonary tuberculosis at AIDS diagnosis in Spain: epidemiological differences and implications for control.

OBJECTIVE: To ascertain the differential factors associated with pulmonary versus extrapulmonary tuberculosis (TB) at AIDS diagnosis in Spain. DESIGN: Analysis of AIDS surveillance data. METHODS: Data about AIDS patients, aged 12 years and over, diagnosed in 1995 were taken from the Spanish AIDS Register. The respective proportions of cases presenting with pulmonary and extrapulmonary TB at AIDS diagnosis were analysed by gender, age, HIV transmission category, prison record, province, country of origin and CD4+ lymphocyte count. A multivariate analysis was carried out using logistic regression analysis. RESULTS: Of 6161 AIDS cases analysed, 20.1% presented with pulmonary TB and 20.4% with extrapulmonary TB. Overall, TB showed association with men, age under 30 years, injecting drug users (IDU), cases of heterosexual HIV transmission, and concurrent or past stay in prison. Frequency of TB proved no different among foreign-born patients. Pulmonary and extrapulmonary TB showed a similar distribution for most of the variables. Current prison inmates registered a high risk of pulmonary TB [adjusted odds ratio (OR), 4.2; 99% confidence interval (CI), 3.1-5.8] compared with patients without prison record, and ex-prison inmates registered an intermediate risk (OR, 2.3; 99% CI, 1.8-3.0). Among patients with TB at AIDS diagnosis, pulmonary TB was associated with subjects currently in prison (OR, 2.1; 99% CI, 1.5-3.0) and injecting drug use (OR, 1.5; 99% CI, 1.0-2.4). Pulmonary TB presented with higher CD4+ lymphocyte counts than extrapulmonary TB (P < 0.001). CONCLUSIONS: The results suggest the importance that recent transmission of TB may be having among young adults, IDU and prison inmates in particular, and calls for a review of control strategies.

AIDS-Related Opportunistic Infections↗
PubMed1997

Interference of coronavirus infection by expression of immunoglobulin G (IgG) or IgA virus-neutralizing antibodies.

Immunoglobulin gene fragments encoding the variable modules of the heavy and light chains of a transmissible gastroenteritis coronavirus (TGEV)-neutralizing monoclonal antibody (MAb) have been cloned and sequenced. The selected MAb recognizes a highly conserved viral epitope and does not lead to the selection of neutralization escape mutants. The sequences of MAb 6A.C3 kappa and gamma 1 modules were identified as subgroup V and subgroup IIIC, respectively. The chimeric immunoglobulin genes encoding the variable modules from the murine MAb and constant modules of human gamma 1 and kappa chains were constructed by reverse transcriptase PCR. Chimeric immunoglobulins were stably or transiently expressed in murine myelomas or COS cells, respectively. The secreted recombinant antibodies had radioimmunoassay titers (i.e., the highest dilution giving a threefold increase over the background) higher than 10(3) and reduced the infectious virus more than 10(4)-fold. Recombinant dimeric immunoglobulin A (IgA) showed a 50-fold enhanced neutralization of TGEV relative to a recombinant monomeric IgG1 which contained the identical antigen binding site. Stably transformed epithelial cell lines which expressed either recombinant IgG or IgA TGEV-neutralizing antibodies reduced virus production by > 10(5)-fold after infection with homologous virus, although a residual level of virus production (< 10(2) PFU/ml) remained in less than 0.1% of the cells. This low-level persistent infection was shown not to be due to the selection of neutralization escape mutants. The implications of these findings for somatic gene therapy with recombinant antibodies are discussed.

Animals↗
PubMed1997

The AIDS epidemic among Spanish drug users: a birth cohort-associated phenomenon.

OBJECTIVES: In Spain the number of new acquired immunodeficiency syndrome (AIDS) cases among injection drug users continues to rise. The time trend up to 1994 has been analyzed, with special attention paid to the different generations. METHODS: The source for injection drug use-related cases was the Spanish AIDS Register. Independent analyses of annual specific rates were run for each sex with the use of an age-period-cohort log-linear model. RESULTS: After adjustment for age and year of diagnosis, AIDS incidence related to injection drug use is associated with specific birth cohorts. Rising values are observed in the successive generations born during the 1950s, peaking in men born in 1962 and women born in 1964. In subsequent cohorts, there is a marked falloff in incidence for both sexes, but this decline is seen to halt in men from the 1972 birth cohort onwards. The overall period effect is upward, yet the trend flattens in the last years. There is a pronounced age effect with maximum values in men and women at ages 29 and 27, respectively. CONCLUSIONS: It is essential to urge avoidance of risk behaviors in new generations.

Acquired Immunodeficiency Syndrome↗