I. Castillo

Hospital Universitario Puerta de Hierro-Majadahonda, Махадаонда, Madrid, Spain

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Publications (179)

  • [Show abstract] [Hide abstract] ABSTRACT: Background: Blood transfusion safety is based on reliable donor screening for transmissible infections such as the hepatitis C virus (HCV) infection. Study design and methods: A novel HCV core-specific antibody was assayed on random single donations from 2007 first-time blood donors who tested negative for anti-HCV and HCV RNA on routine screening. Sample collection broke the code between donations and donors for ethical reasons. Results: Forty-two donations (2.1%) displayed reactivity in the novel test. The specificity of the reactivity was evaluated by a peptide inhibition assay, and testing against additional nonoverlapping HCV core peptide epitopes and other HCV antigens was performed on these samples. Six donations (14.3%; 0.30% from the total) were considered to contain anti-HCV after such supplemental testing. HCV RNA detection was also performed in peripheral blood mononuclear cells (PBMNCs) and serum or plasma samples from reactive donors after virus concentration by ultracentrifugation. HCV RNA tested negative in all PBMNCs samples, and a very low amount of viral genome was detected in serum or plasma concentrates from three anti-HCV core-reactive donors (7.1%) but not among concentrates from 100 randomly selected nonreactive donors. Sequencing of these polymerase chain reaction products revealed differences between the isolates that excluded partially sample contamination from a common source. Conclusion: These findings argue in favor of an ongoing occult HCV infection among these blood donors and account for some rather low, but perhaps not negligible, infection risk for such donations. Future studies involving larger samples of donations from traceable donors would enlighten the significance of these findings for the viral safety of the blood supply.
    Article · May 2016 · Transfusion
  • [Show abstract] [Hide abstract] ABSTRACT: Polymorphisms upstream interleukin (IL)-28B gene and serum levels of interferon gamma inducible protein-10 (IP-10) are associated with spontaneous and treatment-induced hepatitis C virus (HCV) clearance. Patients with seronegative occult HCV infection are anti-HCV and serum HCV-RNA negative but have viral RNA in liver and abnormal values of liver enzymes. We examined if the rs12979860 polymorphism of IL-28B and serum IP-10 levels differ between chronic and seronegative occult CV infection. IL-28B polymorphism was determined with allele specific TaqMan probes in total DNA isolated from peripheral blood mononuclear cells and IP-10 by an enzyme-linked immunosorbent assay in serum from 99 patients with seronegative occult HCV infection and 130 untreated patients with chronic hepatitis C. IL-28B genotypes were also determined in 54 healthy volunteers. Prevalence of the IL-28B CC genotype was significantly higher in seronegative occult HCV infection (52/99; 52.5%) than in chronic hepatitis C (32/130; 24.6%, p<0.0001) or healthy controls (19/54: 32.5%, p=0.039). Among patients with seronegative occult HCV infection, HCV-RNA load in liver was significantly lower in those with the IL-28B CC genotype than in those with CT+TT genotypes (2.8 × 10(5 ) ± 5.8 × 10(4) vs 4.1 × 10(5 ) ± 5.9 × 10(4) copies/µg of total RNA respectively; p = 0.023). Mean serum IP-10 levels were significantly lower in patients with seronegative occult HCV infection than in patients with chronic hepatitis C (160.8 ± 17.9 vs 288.7 ± 13.3 pg/ml respectively; p < 0.0001). These findings suggest that the host immune response plays an important role in seronegative occult HCV infection in comparison with chronic hepatitis C. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    Article · Jul 2015 · Journal of Medical Virology
  • Article · Nov 2014 · Journal of Hepatology
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    [Show abstract] [Hide abstract] ABSTRACT: Amplification of hepatitis C virus (HCV) RNA from blood detected occult HCV infections in 30.9% of 210 HCV-seronegative dialysis patients with abnormal liver enzyme levels that had evaded standard HCV testing practices. Isolated HCV core-specific antibody detection identified three additional anti-HCV screening-negative patients lacking HCV RNA amplification in blood who were considered potentially infectious. Together, these findings may affect management of the dialysis setting.
    Full-text available · Article · May 2014 · Journal of Clinical Microbiology
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    Inmaculada Castillo · Jorge Martinez-Ara · Teresa Olea · [...] · Rafael Selgas
    [Show abstract] [Hide abstract] ABSTRACT: The association of hepatitis C virus (HCV) infection and glomerulonephritis is well known. However, the relationship between immune-mediated glomerulonephritis and occult HCV, characterized by the presence of HCV-RNA in liver or in peripheral blood mononuclear cells in the absence of serological markers, is unknown. We tested this in 113 anti-HCV-negative patients; 87 with immune-mediated glomerulonephritis and 26 controls with hereditary glomerular nephropathies. All patients were serum HCV-RNA negative by conventional real-time PCR. Significantly, occult HCV-RNA (detectable viral RNA in peripheral blood mononuclear cells or in serum after ultracentrifugation) was found in 34 of 87 patients with immune-mediated glomerulonephritis versus 1 of 26 control patients. The serum creatinine levels were significantly higher in patients with immune-mediated glomerulonephritis with than in those without occult HCV (1.5 versus 1.1 mg/dl, respectively). A multivariate analysis adjusted for gender showed a significantly increased risk of occult HCV in patients with immune-mediated glomerulonephritis versus the controls (odds ratio of 13.29). Progression to end-stage renal disease tended to be faster in patients with immune-mediated glomerulonephritis and occult HCV than in the negative cases. Thus, occult HCV is strongly associated with immune-mediated glomerulonephritis and may have a role in the progression of the disease.Kidney International advance online publication, 19 March 2014; doi:10.1038/ki.2014.68.
    Full-text available · Article · Mar 2014 · Kidney International
  • [Show abstract] [Hide abstract] ABSTRACT: Hepatitis C virus (HCV) infection in the absence of detectable antibodies against HCV and of viral RNA in serum is called occult HCV infection. Its prevalence and clinical significance in chronic hepatitis B virus (HBV) infection is unknown. HCV-RNA was tested in the liver samples of 52 patients with chronic HBV infection and 21 (40%) of them were positive for viral RNA (occult HCV infection). Liver fibrosis was found more frequently and the fibrosis score was significantly higher in patients with occult HCV than in negative ones, suggesting that occult HCV infection may have an impact on the clinical course of HBV infection.
    Article · May 2013 · Journal of Medical Microbiology
  • [Show abstract] [Hide abstract] ABSTRACT: Occult hepatitis C virus (HCV) infection, defined as the presence of HCV RNA in liver and in peripheral blood mononuclear cells (PBMCs) in the absence of detectable viral RNA in serum by standard assays, can be found in anti-HCV positive patients with normal serum levels of liver enzymes and in anti-HCV negative patients with persistently elevated liver enzymes of unknown etiology. Occult HCV infection is distributed worldwide and all HCV genotypes seem to be involved in this infection. Occult hepatitis C has been found not only in anti-HCV positive subjects with normal values of liver enzymes or in chronic hepatitis of unknown origin but also in several groups at risk for HCV infection such as hemodialysis patients or family members of patients with occult HCV. This occult infection has been reported also in healthy populations without evidence of liver disease. Occult HCV infection seems to be less aggressive than chronic hepatitis C although patients affected by occult HCV may develop liver cirrhosis and even hepatocellular carcinoma. Thus, anti-HCV negative patients with occult HCV may benefit from antiviral therapy with pegylated-interferon plus ribavirin. The persistence of very low levels of HCV RNA in serum and in PBMCs, along with the maintenance of specific T-cell responses against HCV-antigens observed during a long-term follow-up of patients with occult hepatitis C, indicate that occult HCV is a persistent infection that is not spontaneously eradicated. This is an updated report on diagnosis, epidemiology and clinical implications of occult HCV with special emphasis on anti-HCV negative cases.
    Article · Jun 2012 · World Journal of Gastroenterology
  • Inmaculada Castillo · Javier Bartolomé · Juan A Quiroga · [...] · Vicente Carreño
    [Show abstract] [Hide abstract] ABSTRACT: Patients with occult hepatitis C virus (HCV) infection (HCV-RNA in liver without detectable anti-HCV and serum HCV-RNA) may have viral RNA in peripheral blood mononuclear cells (PBMCs) and in serum after ultracentrifugation, and may present HCV-specific T-cell responses, but it is unknown whether these markers persist to be detectable over time. To perform a prospective virological long-term follow up of patients with occult HCV. Viral markers were tested every 3-4 months during 55.7 ± 20.3 months in 37 patients with occult HCV who were under ursodeoxycholic acid treatment. Viral RNA was detectable in PBMCs of 31 patients during the follow up. In 23 of them, viral RNA in PBMCs was detected intermittently and in the other eight patients HCV-RNA was positive in a single sample. After ultracentrifugation, serum HCV-RNA was detected in 33 patients, being the viraemia intermittently detectable in 28, whereas in the remaining five patients, serum HCV-RNA was positive only once. Only one patient tested always HCV-RNA negative in PBMCs and in ultracentrifuged serum during follow up. Specific Core, NS3, and/or NS4 T-cell responses were found in 31 of the patients. The patient who was always HCV-RNA negative in PBMCs and in ultracentrifuged serum had specific HCV-T-cell responses. Occult HCV infection persists over time with fluctuating viraemia levels that induce and maintain specific T-cell responses against viral proteins.
    Article · Nov 2011 · Liver international: official journal of the International Association for the Study of the Liver
  • [Show abstract] [Hide abstract] ABSTRACT: INTRODUCTION AND AIMS: Anti-HBs formation is impaired in HD patients naturally infected with HBV (14 - 31% of cases) or undergoing a planned immunization (20 – 40% of cases). Our aim was to determine the potential association between development of anti-HBs and demographic, clinical and laboratory data of HD patients. Special attention was paid to IL-18 -1297C>T polymorphism, because IL-18 promotes interferon-γ production, which is heplful in immune responsiveness. METHODS: Studies were carried out in 347 Caucasian HD patients: 219 persons developed an anti-HBs titre >10 IU/L as a result of vaccination (patients with negative total antibodies to core antigen of HBV, anti-HBc, n = 125) or as a result of HBV transmission (patients with total anti-HBc positive, n = 94), whereas 128 patients did not develop an anti-HBs titre >10 IU/L in response to at least one full series of vaccination (patients with total anti-HBc negative, n = 106) or HBV transmission (patients with total anti-HBc positive, n = 22). The frequency of IL18 -1297C>T alleles and genotypes was identified by polymerase chain reaction-restriction fragment length polymorphism. Blood donors (n = 240) served as controls for a genetic study. The logistic regression analysis (the hierarchical model) was used to show variables associated with development of anti-HBs. This analysis included gender, age, 4 the most frequently observed causes of end stage renal disease (diabetic nephropathy, chronic glomerulonephritis, hypertensive nephropathy, chronic tubulointerstitial nephritis), vintage of renal replacement therapy (RRT), liver enzymes activities and the -1297C>T (rs360719) polymorphism of IL-18 gene. RESULTS: IL-18 CC genotype (OR 0.208, CI 0.046 – 0.927) and chronic glomerulonephritis as a cause of chronic renal failure (OR 0.535, CI 0.295 – 0.969) were positively associated with anti-HBs development, whereas in the other model TT (OR 4.572, CI 1.009 – 20.710) and CT (OR 5.146, CI 1.128 – 23.475) genotypes showed negative association with anti-HBs formation. Only patients, who developed anti-HBs, had the -1297C>T (rs360719) polymorphism of IL-18 gene comparable to that shown in blood donors. In dialyzed non-responders the frequency of -1297CC (rs360719) was lower than that in controls in both CC vs CT + TT model (p = 0.006, OR 0.166, 95% CI 0.038 – 0.718, sample power 82.6%) and CC vs TT model (p = 0.009, OR 0.170, 95% CI 0.039 – 0.745, sample power 82.0%). Prevalence of patients with chronic glomerulonephritis among groups bearing different IL-18 genotypes was not significantly different (27.8%, 18.6% and 20.7% in patients with CC, CT and TT genotypes, respectively). Independent variables which were not associated with anti-HBs titre >10 IU/L were age, gender, liver enzymes activities, vintage of RRT and remaining 3 main causes of end stage renal disease. CONCLUSIONS: In HD patients, IL-18 -1297CC genotype as well as causes of end-stage renal disease may play a role in anti-HBs development in response to HBV surface antigen.
    Article · Jun 2011 · CKJ: Clinical Kidney Journal
  • Inmaculada Castillo · Javier Bartolomé · Juan Antonio Quiroga · [...] · Vicente Carreño
    [Show abstract] [Hide abstract] ABSTRACT: The diagnosis of occult hepatitis C virus (HCV) infection is based on the presence of HCV-RNA in the liver. This study aimed to evaluate the use of combining non-invasive assays to diagnose occult HCV. A total of 122 patients with occult HCV (HCV-RNA in the liver without detectable anti-HCV and serum HCV-RNA) and 45 patients with cryptogenic chronic hepatitis (without HCV-RNA in the liver and negative for anti-HCV and serum HCV-RNA) were included. HCV-RNA was tested in peripheral blood mononuclear cells (PBMCs) and in 2 ml of ultracentrifuged serum. Anti-core HCV was examined by a non-commercial enzyme-linked immunosorbent assay. All controls were negative for the three HCV markers studied. Among patients with occult HCV, 36% were anti-core HCV positive, 57% had serum HCV-RNA after ultracentrifugation, and 61% had HCV-RNA in PBMCs. Combining the results of the assays, 91% of the patients were positive for at least one marker. Intrahepatic HCV-RNA load was significantly higher in patients who were positive simultaneously for the three HCV markers than in patients who were negative for all markers (P = 0.006) and than in those with one or two HCV markers (P = 0.039). Replication of HCV in liver was detected more frequently in patients with three (93%, P = 0.002), two (82%, P = 0.001), and one HCV marker (73%, P = 0.011) than in those without markers (27%). In conclusion, testing for all these markers allows diagnosis of occult HCV without the need for a liver biopsy and these assays may help to elucidate the clinical significance of occult HCV infection.
    Article · Sep 2010 · Journal of Medical Virology
  • [Show abstract] [Hide abstract] ABSTRACT: Hemodialysis induces production of the hepatocyte growth factor (HGF) and decrease of serum hepatitis C virus (HCV) RNA in patients with HCV infection, but it is not known if the hemodialysis schedule or type of membrane affect both the HGF production and HCV viremia. The effects on both parameters of alternate-day intermittent hemodialysis and short-daily hemodialysis and high and low flux membranes were investigated in 41 patients treated by hemodialysis. Sixteen (39%) patients were anti-HCV positive and 11 (69%) had HCV RNA. Twenty-six patients were on alternate-day intermittent and 15 on short-daily hemodialysis. High flux membranes were used for 29 patients and low flux membranes for 12 patients. A decrease in HCV RNA was observed at the end of hemodialysis (8.6 x 10(5) +/- 1.1 x 10(6) IU/ml vs. 4.4 x 10(5) +/- 7.3 x 10(5) IU/ml, P = 0.003). The proportion of HCV RNA decrease was similar in patients dialyzed with both schedules and with both types of membranes. The HGF levels increased from 2,605.9 +/- 1,428.7 to >8,000 pg/ml at 15 min. At the end of the session, the HGF levels decreased to 5,106.7 +/- 2,533.9 pg/ml. The HGF levels at the start of the next session were similar to those at baseline (2,680.0 +/- 1,209.3 pg/ml). The increase and dynamics of the HGF levels were similar in patient's hemodialyzed with both schedules and with both types of membranes. These results suggest that changes in HCV RNA and HGF levels during hemodialysis are not influenced by the schedule or type of membrane used.
    Article · May 2010 · Journal of Medical Virology
  • J. A. Quiroga · I. Castillo · J. Bartolome · [...] · V. Carreñno
    Article · Apr 2010 · Journal of Hepatology
  • Guillermina Barril · Inmaculada Castillo · Vicente Carreño
    Article · Nov 2009 · American Journal of Kidney Diseases
  • Article · Sep 2009 · Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology
  • Inmaculada Castillo · Javier Bartolomé · Juan Antonio Quiroga · [...] · Vicente Carreño
    [Show abstract] [Hide abstract] ABSTRACT: Family members of patients with chronic hepatitis C virus (HCV) infection are at increased risk of HCV infection but the prevalence of HCV among family members of patients with occult HCV infection is not known. Anti-HCV, serum HCV RNA and levels of liver enzymes were determined in 102 family members of 50 index patients with occult HCV infection and in 118 family members of 59 chronic hepatitis C index patients. HCV RNA and/or anti-HCV were detected in 10/102 (9.8%) relatives of patients with occult HCV infection and in 4/118 (3.4%) of patients with chronic hepatitis C. Fourteen additional family members (seven were relatives of index patients with occult HCV infection) had abnormal values of liver enzymes without serological markers of HCV infection. Two of these patients (who were relatives of two index patients with occult HCV infection) underwent a liver biopsy and were diagnosed with an occult HCV infection because HCV RNA was detected in the liver cells in the absence of serological HCV markers. In conclusion, the prevalence of HCV infection among family members of patients with occult HCV infection was similar to that found among family members of patients with chronic hepatitis C. This stresses the need to adopt strategies to prevent the transmission of HCV in the family setting of patients with occult HCV infection.
    Article · Jul 2009 · Journal of Medical Virology
  • I Castillo · J Bartolomé · J A Quiroga · [...] · V Carreño
    [Show abstract] [Hide abstract] ABSTRACT: Concentration of viral particles by ultracentrifugation of serum prior to PCR allows detection of hepatitis C virus (HCV) RNA in patients with undetectable viral RNA by conventional PCR assays. To analyse if HCV-RNA is detected after serum ultracentrifugation in chronic hepatitis C patients with a sustained virological response to antiviral therapy (defined as serum HCV-RNA negativity by conventional assays 6 months after the end of therapy). HCV-RNA was tested using real-time PCR in ultracentrifuged sera collected during the post-treatment follow-up (mean: 42 +/- 27 months) in 57 sustained virological responders (SVR). After serum ultracentrifugation, HCV-RNA was detected on at least one occasion during the follow-up in 29/57 (51%) SVR. Thirteen (23%) of these 57 SVR suffered a reactivation 18 +/- 8 months after the end of therapy (reappearance of serum HCV-RNA detectable by conventional assays). Among reactivated patients, 11/13 (85%) had HCV-RNA in ultracentrifuged serum samples (detectable 10 +/- 5 months before reactivation), while HCV-RNA was positive after ultracentrifugation in 18/44 (41%) long-term SVR (P = 0.01). Persistence of detectable HCV-RNA after serum ultracentrifugation was associated with reactivation (P = 0.001). Serum ultracentrifugation prior to PCR allows detection of HCV-RNA in SVR and its persistence may predict late reactivation.
    Article · Jul 2009 · Alimentary Pharmacology & Therapeutics
  • Article · Jun 2009 · Hepatology
  • J. A. Quiroga · S. Llorente · I. Castillo · [...] · V. Carreño
    Article · Apr 2009 · Journal of Hepatology
  • J. A. Quiroga · I. Castillo · S. Llorente · [...] · V. Carreno
    Article · Dec 2008 · Journal of Hepatology
  • Article · Nov 2008 · Journal of clinical microbiology