Sabine Chapuy-Regaud

Fibrin Deimination in Synovial Tissue Is Not Specific for Rheumatoid Arthritis but Commonly Occurs during Synovitides

Autoantibodies to deiminated (citrullinated) proteins are the most specific serological markers of rheumatoid arthritis (RA). Deimination is critical in generating the peptidic epitopes they recognize. In the synovial tissue (ST), deiminated forms of the α- and β-chains of fibrin are their major autoantigenic targets (anti-human fibrin(ogen) autoantibodies (AhFibA)). We investigated whether the presence of deiminated fibrin in the ST was specific for RA, because this could explain why AhFibA are RA specific. In 13 patients with RA and 19 patients with various other rheumatological disorders, knee ST biopsies were collected in macroscopically inflamed areas identified under arthroscopy. Synovitis was histopathologically confirmed in all of the biopsies. By immunoblotting, using antisera to fibrin, Abs to citrullyl residues, and AhFibA purified from RA sera, deiminated fibrin was evidenced in ST extracts from all of the patients. Moreover, variations in the degree of fibrin deimination were observed that were not related to the disease. Immunohistochemical analysis, using Abs to citrullyl residues and an antiserum to fibrin on adjacent serial sections of ST, confirmed the results because deiminated proteins colocalized with fibrin in RA as well as in control patients. Therefore, fibrin deimination in the ST is a general phenomenon associated to any synovitis, which does not necessarily induce a B autoimmune response with production of AhFibA.

New Natural Intergenotypic (2/5) Recombinant of Hepatitis C Virus

ABSTRACT A 9.2-kb sequence from a hepatitis C virus (HCV) strain found in southwest France was compared to sequences from reference strains in HCV sequence databases. We found a recombinant virus with genotype 2 at the 5′ end and genotype 5 at the 3′ end. The crossover point was located between genes NS2 and NS3. Recombination between HCV genotypes must now be considered in studies on HCV epidemiology and evolution and in predictions of the virus response to antiviral therapy. Knowing the location of the recombination point may also be useful for constructing infectious chimeric viruses.

Hepatitis E Virus Reinfections in Solid-Organ-Transplant Recipients Can Evolve Into Chronic Infections

BACKGROUND Hepatitis E virus (HEV) infections are a major cause of acute hepatitis in developing and industrialized countries. Little is known about anti-HEV immunity in solid-organ recipients. METHODS We screened 263 solid-organ recipients for anti-HEV immunoglobulin G (IgG) at transplantation. They were followed up for 1 year and tested for HEV RNA and anti-HEV antibodies 1 year after transplantation and if their liver enzyme activities increased. RESULTS A total of 38.4% had anti-HEV IgG at transplantation. The mean concentrations (±SD) of anti-HEV IgG at transplantation (8 ± 17.5 U/mL) and 1 year later (6.4 ± 12.0 U/mL, P = .4) were similar. There were 3 de novo HEV infections during the 1-year follow-up among patients who were HEV seronegative before transplantation, giving an annual incidence of 2.1%. We also identified 3 HEV reinfections among patients who were seropositive before transplantation through detection of HEV RNA, for an annual incidence of 3.3%. Their anti-HEV IgG concentrations were 0.3, 2.1, and 6.2 World Health Organization (WHO) units/mL before transplantation. Reinfection of the patient with the lowest IgG concentration at transplantation had evolved to a chronic infection. CONCLUSIONS Low anti-HEV antibodies (<7 WHO units/mL) seemed not to protect solid-organ recipients. HEV reinfection in immunocompromised patients can lead to chronic infection, as in primary infections.

Zika virus in semen and spermatozoa

The current Zika virus epidemic is a major challenge for the medical and scientific communities for at least two reasons: the severe clinical situation associated with Zika virus infection (neurological complications and adverse fetal outcomes) and the unexpected sexual viral transmission from men to women. These recent findings have shifted the paradigm of arbovirus–host interactions, modifying standard epidemiology and clinical patterns. High infectious Zika virus loads have been detected in semen, but data for viral persistence after symptomatic infections are scarce and even nonexistent for asympto matic ones, with the remaining key issue: how long does semen contain infectious Zika virus? As long as this question is unanswered, the adaptation of preventive measures such as the use of condoms and the abstinence of semen donation will be hampered. Here, we report the longitudinal follow up of Zika virus RNA in the semen of a 32-year-old man returning from French Guyana. At admission, the patient had moderate fever, maculopapular rash, myalgia, and arthralgia and was diagnosed for Zika virus infection upon detection of viral RNA in plasma and urine 2 days after onset of symptoms. The molecular diagnosis for dengue and chikungunya viruses was negative (in-house RT-PCR). The patient was seronegative for HIV, had a normal immunological status, and recovered in few days. Semen (11 samples), blood (ten), and urine (five) were prospectively collected for 141 days after symptom onset. Zika virus RNA was detected on each semen sample and was still positive after 141 days, with viral load decreasing from 8·6 log copies per mL to 3·5 log copies per mL. It was detected until day 37 in both blood (2·5 log copies per mL) and urine (3·7 log copies per mL; fi gure). Such a prolonged RNA Zika virus excretion is quite different from other fl aviviruses, which are rapidly cleared by the immune response and for which the detection of viral nucleic acids is typically limited to a short window after symptom onset. In addition to the present case, we investigated five other symptomatic men for the presence of Zika virus RNA in semen; RNA was still detected 69 days and 115 days after the symptom onset in two patients, but not detected at day 20 in the other three individuals (data not shown). These data suggest that the length of Zika virus excretion varies, probably depending on viral and host characteristics, but long-lasting excretion might be frequent among adults who experienced a symptomatic infection. Such long-lasting excretion of genomic material was recently described in semen from Ebola survivors. The viral persistence in semen is of major concern and could be related to a viral tropism for male sexual cells. Accordingly, we report the immunohistochemical detection of Zika virus in the head of spermatozoa obtained from the fi rst patient (fi gure; appendix). The proportion of infected spermatozoa was estimated at 3·52% (SD 0·71), as assessed by counting Zikavirus-positive sperma tozoa on three smears by fl uorescence micro scopy. The use of both confocal and stimulated emission depletion microscopy (appendix), allowed the unambiguous demonstration that Zika virus antigens are indeed present inside spermatozoa. Additional immuno stainings of sperm from a healthy non-infected control donor for Zika virus antigens or with a mouse IgG2a isotype control confi rmed the specifi city of the staining (data not shown). Spermatozoa do not express the candidate Zika virus entry receptor Axl. Because Sertoli cells highly express Axl and have essential roles in spermatogenesis, we hypothesise that they might be involved in Zika virus transmission to spermatozoa. Appropriately designed studies are needed to discover the modes of spermatozoa infection and length of risk of sexual transmission. Answers to these questions will allow public health authorities to recom mend urgently needed universal safe practices.

Performance of anti-HEV assays for diagnosing acute hepatitis E in immunocompromised patients

Hepatitis E virus is an emerging concern in immunocompromised patients, who may become chronically infected. This prompted us to assess the performance of two anti-HEV IgG and IgM assays for diagnosing acute HEV infections. The specificities of the assays were estimated by testing samples from 2 to 3 year-old French children and blood donors and their sensitivities by testing 40 immunocompromised patients acutely infected. Both anti-HEV IgM assays were highly specific (99.6% and 100%). The sensitivity of the Adaltis was 87.5%, and that of Wantai was 85%. The specificities of anti-HEV IgG Wantai (97.8%) and Adaltis tests (89.5%, p=0.1) were similar but the Wantai test was more sensitive (45%) than the Adaltis test (15%, p<0.001). None of the samples was anti-HEV IgM negative and IgG positive. We conclude that these anti-HEV IgM assays performed well in immunosuppressed subjects with acute hepatitis E and can be used as first line virological tools. Testing for anti-HEV IgG and IgM simultaneously at the acute phase did not improve the diagnostic performance. In contrast, molecular detection of HEV RNA appears essential to exclude an HEV infection in patients who are negative for anti-HEV IgM and to assess the evolution of hepatitis E 3 months thereafter.

Zika virus in semen of a patient returning from a non-epidemic area.

894 www.thelancet.com/infection Vol 16 August 2016 one or more other microorganisms (Staphylococcus aureus, Enterococcus faecalis, Staphylococcus hominis, Corynebacterium pseudodiphtericum, and Aeromonas spp). We treated one of our cases with only antituberculosis therapy without surgery, and treated four with surgery followed by antituberculosis therapy, including debridement, antibiotics, and implant retention in one case, and two-stage prosthesis exchange in the three other cases. The mean length of antituberculosis therapy was 11 months. Remission was observed in four cases with an average follow-up of 24 months. One patient died of systemic tuberculosis after 2 months of treatment. Despite the high volume of arthroplasty, tuberculosis prosthetic joint infection is rare. Early prosthetic joint infection might be explained by pre-existing tuberculous coxitis. In other cases, tuberculosis prosthetic joint infection is likely to be a haematogenous infection following systemic tuberculosis. Diagnosis of tuberculosis prosthetic joint infection is mainly reached through positive cultures of deep samples. Nevertheless, percutaneous biopsy has been shown to be an alternative diagnostic method, as used in two of our cases. On the basis of our data, we believe that tuberculosis prosthetic joint infection is certainly under-reported and should be considered in culture-negative prosthetic joint infections, and not only in patients with a past medical history of osteoarticular tuberculosis. Previous or concomitant infection with another pathogen should not exclude diagnosis. Careful histological analysis, systematic mycobacterial cultures, and the contribution of specific PCR detection could help physicians to diagnose tuberculosis prosthetic joint infection.

Zika Virus Infection and Prolonged Viremia in Whole-Blood Specimens

We tested whole-blood and plasma samples from immunocompetent patients who had had benign Zika virus infections and found that Zika virus RNA persisted in whole blood substantially longer than in plasma. This finding may have implications for diagnosis of acute symptomatic and asymptomatic infections and for testing of blood donations.

Performance of Two Commercial Assays for Detecting Hepatitis E Virus RNA in Acute or Chronic Infections

ABSTRACT We assessed the performance of the Ceeram and Altona assays, the first two commercially available hepatitis E virus (HEV) RNA assays, using serial dilutions of 4 HEV-positive reference samples (genotypes 3a, 3c, 3e, and 3f). Both assays provided good analytical sensitivity and high reproducibility for detecting genotype 3 HEV RNA.

Hepatitis E Pathogenesis

Although most hepatitis E virus (HEV) infections are asymptomatic, some can be severe, causing fulminant hepatitis and extra-hepatic manifestations, including neurological and kidney injuries. Chronic HEV infections may also occur in immunocompromised patients. This review describes how our understanding of the pathogenesis of HEV infection has progressed in recent years.

Performance of an antigen assay for diagnosing acute hepatitis E virus genotype 3 infection

BACKGROUND Hepatitis E virus (HEV) is a major cause of hepatitis worldwide. Its diagnosis is based on the detection of anti-HEV IgM and/or HEV-RNA. OBJECTIVE To evaluate the performance of the Wantaï HEV-antigen (Ag) ELISA(Plus) assay for diagnosing acute HEV infections. STUDY DESIGN Specificity was assessed using 100 blood samples containing no anti-HEV IgM, anti-HEV IgG, or HEV-RNA. Cross reactivity was assessed using samples positive for hepatitis C virus RNA (n=10), Epstein-Barr virus DNA (n=10) and cytomegalovirus DNA (n=10). Serial dilutions of 4 HEV RNA positive samples were used to estimate the corresponding viremia detected with the Ag assay. Blood samples from 33 immunocompetent and 31 immunocompromised patients with an acute HEV genotype 3 infection, HEV-RNA positive, were tested to assess diagnostic sensitivity. RESULTS The HEV-Ag assay was 100% specific, with no cross-reactivity. The lower viremias detected ranged from 10(3)copies/ml to 10(5)copies/ml (800-80,000UI/ml). Diagnostic sensitivity for an acute HEV infection was 91%, with no significant difference between immunocompetent (88%) and immunocompromised (94%) patients. The HEV-Ag assay was more frequently positive in immunocompromised patients at the acute phase (94%) than was the anti-HEV IgM test (71%; p=0.04). The HEV-Ag assay ratio was correlated with HEV-RNA viral load (ρ=0.54; p<0.0001). CONCLUSION The HEV-Ag assay performed well and could be suitable for laboratories with no molecular diagnosic facilities.