Biomedical subjects
Publications and source records attributed to F Morinet.
A dot-blot hybridization assay for serum B19 DNA is described using a non-radioactive (digoxigenin) M13 single-stranded DNA probe. The assay is sensitive (0.3 pg of B19 DNA) and has several advantages over similar assays which use radioactive or non-radioactive labelled plasmid probe for the routine detection of B19 DNA in sera.
A new anti-B19 IgM ELISA was developed taking advantage of antibody-capture with biotinylated fusion protein as antigen. Specificity was examined using serum IgM antibody positive for rubella, hepatitis B core antigen, cytomegalovirus and Epstein-Barr virus as well as with sera positive for rheumatoid factors or antinuclear antibodies. The specificity was found to be 96%. Of one hundred serum samples compared using the new ELISA or the standard MACRIA tests for the presence of B19 IgM, 88 gave the same results. Fifty-three were negative and 35 were positive. Six sera were ELISA-negative MACRIA-positive, and six MACRIA-negative ELISA-positive. Thus, the ELISA gave 90% agreement with MACRIA. In a clinical study with 725 sera from suspected B19 infections, 161 (22%) were found positive by ELISA. The positive sera were from patients suffering from arthritis (35%), rash (35%), acute or chronic erythroblastopenia (21%), pancytopenia (5%), vascular purpura (2%) and lymphadenopathy (2%). A series of serum specimens obtained from two-B19 infected individuals were also studied. The IgM antibody became undetectable after four months.
We report three cases of systemic lupus erythematosus (SLE) associated with necrotizing histiocytic lymphadenitis (Kikuchi's disease) and immunologically proven human parvovirus B19 infection. Simultaneous occurrence of SLE and Kikuchi's disease was a characteristic of the three cases. Kikuchi's disease is an uncommon disease that usually affects young women and is characterized by painless unilateral cervical lymph-node enlargement. T-cell regions of affected lymph nodes are exclusively involved with patchy paracortical necrosis surrounded by a polymorphous cell population of histiocytes and macrophages. However, lymphadenopathy in patients with SLE may be histologically indistinguishable from Kikuchi's necrotizing lymphadenitis. The cause of Kikuchi's disease remains uncertain, although infectious agents have been proposed. A positive IgM-specific anti-human parvovirus B19 antibody test in our three cases suggests that B19 can induce a necrotizing histiocytic lymphadenitis and possibly a clinical SLE flare. High-dose (1 mg/kg/day) and medium-dose (0.5 mg/kg/day) oral prednisone was an effective treatment for constitutional and visceral symptoms of Kikuchi's and SLE diseases.
Florid necrotizing lymphadenitis, characterized by segmental infarction and lymphoid hyperplasia, is an uncommon feature of systemic lupus erythematosus (SLE). Kikuchi's disease is a well-defined clinicopathological entity, with a strong preference for the cervical lymph nodes of young women. The etiology of histiocytic necrotizing lymphadenitis (HNL) remains unknown, although viral agents have been proposed. HNL may reflect a self-limited SLE-like autoimmune disease but full-blown SLE associated with this condition has not, to the best of our knowledge, been reported. Thus, ours is the first description of the coexistence of SLE and HNL in 3 patients with immunologically proven parvovirus B19 infections. SLE and HNL were diagnosed simultaneously in 2 patients, but was retrospective in the third, in whom anti-tuberculous therapy was ineffective. Patients 1 and 2 were treated with prednisone (1 mg/kg/d) and responded rapidly. These data suggest that both HNL and SLE flares can be caused by parvovirus B19 infection.
We report a new complication related to parvovirus B 19: acute lethal myocarditis in the context of a familial epidemic. Serological studies confirmed the existence of a recent infection and the B 19 structural proteins were detected by immunocytochemical techniques on myocardial sections.
Two cases of maternal-fetal human parvovirus B19 infection are reported. The first case involved a feto-placental anasarca occurring during the third trimester and complicated by in-utero death with expulsion of a fetus with multiple malformations. The second case involved meconial peritonitis during the second trimester with favourable outcome after cesarean section and resolution of the digestive syndrome. Proof of PV B19 infection was obtained by identifying M specific immunoglobulins and viral DNA. After a review of the clinical situations most often encountered and the particular features of these two cases, therapeutic attitudes are proposed for this infection of major gravity during pregnancy.
PURPOSE: To determine whether Epstein-Barr virus (EBV) could be involved in the pathogenesis of Sjögren's syndrome (SS). PATIENTS AND METHODS: In situ hybridization using the BamH1-W fragment of EBV DNA was performed using labial salivary gland biopsy specimens from 14 patients with SS (eight with primary SS and six with secondary SS) and 39 control subjects. Furthermore, labial salivary gland biopsy specimens from 12 patients with SS (seven with primary SS and five with secondary SS) and 24 control subjects were submitted to the polymerase chain reaction to detect EBV DNA. RESULTS: In situ hybridization detected EBV DNA in epithelial cells of labial salivary gland biopsy specimens from four of eight (50%) patients with primary SS, zero of six patients with secondary SS, and three of 39 (8%) control subjects. The difference between patients with primary SS and control subjects was statistically significant (p less than 0.02). The polymerase chain reaction detected EBV DNA in six of seven (86%) patients with primary SS, three of five (60%) patients with secondary SS, and seven of 24 (29%) control subjects. The difference between patients with primary SS and control subjects was statistically significant (p less than 0.01). CONCLUSION: Both newly developed techniques showed that the presence of EBV DNA was significantly increased in patients with primary SS in comparison with control subjects. In all the positive SS patients who underwent in situ hybridization, epithelial cells of the labial salivary gland were the target of EBV infection. Our results suggest that this virus may play a role in the pathogenesis of SS. We cannot yet determine whether EBV is directly responsible for the destruction of the gland, or if its presence is a secondary event following gland injury.
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Using in-situ hybridizaiton, we showed the presence of the Epstein-Barr (EB) virus genome in epidermal cells from a patient with chronic lymphocytic leukemia and unusual cutaneous lesions characterized clinically by a maculopapular eruption and histologically by epidermal cell degeneration and lymphoid cell infiltration. Such histologic changes are similar to those seen in graft-versus-host disease. The EB virus genome was mainly detected in the basal, germinative cells of the abnormal epithelium. Specimens of our patient's healthy skin were negative. The presence of EB virus DNA in skin lesions was confirmed by polymerase chain reaction adapted for analysis of paraffin-embedded tissue. These findings indicate that EB virus can infect the human epidermis and that the viral infection may produce a distinctive cutaneous disease.
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A permanent cell line, LEF1, has been established from the cells of an adult suffering from a Philadelphia positive acute lymphoblastic leukemia. The LEF1 cell line was obtained by maintaining peripheral blood cells from the patient in culture on a fibroblast feeder; subsequently, an autonomously growing cell population, independent of that feeder layer, developed. The karyotype of the cell line, 46, t(9;22)(q34;q11), was different from the karyotype at diagnosis which had 53 chromosomes and two Philadelphia chromosomes. Furthermore, compared with the initial leukemic blasts, the immortalized cell had three differences in surface phenotype (CD23+, CD11b-, CD10-). However, molecular studies indicated that the breakpoint in the 3' part of the first intron of the BCR gene was unchanged, confirming the leukemic origin of LEF1. The cell line was shown to be Epstein-Barr virus negative.
The human parvovirus B19 may produce persistent infections with chronic bone marrow failure in immunocompromised hosts. Despite poor data on the pathophysiologic basis of such infections, it is likely that immune humoral response plays an important part. Indeed, administration of immunoglobulin preparation may help to control the course of B19 chronic infections.
A new anti-B19 IgM ELISA was developed using the antibody-capture principle. Biotinylated fusion protein was used as antigen. The specificity of the test was analysed using sera IgM positive to rubella, hepatitis B core antigen, cytomegalovirus and Epstein Barr virus as well with sera positive for rheumatoid factors, antinuclear antibodies and with sera from normal blood donors. The specificity of the test was 96.18%. One hundred serum samples were tested by the new ELISA and the standard MACRIA tests for the presence of B19 IgM. Eighty-eight sera gave the same results with both tests. Fifty-three were negative and 35 were positive. Six sera were ELISA negative MACRIA positive and six MACRIA negative ELISA positive. In a clinical study with 725 sera from suspected B19 infections, 161 (22%) were recorded as positive by the ELISA test. The positive sera were from patients suffering from arthritis (35%), rash (35%), acute or chronic erythroblastopenia (21%), pancytopenia (5%), vascular purpura (2%) and lymphadenopathy (2%).
A 1.4 kb fragment (nucleotides 2430 to 3901) encoding portions of the human parvovirus B19 structural proteins was inserted into the pRIT2 plasmid expression vector containing the gene encoding staphylococcal Protein A under the control of the phage lambda promoter PR. The fusion protein was used to raise antibodies in rabbits. The sera were shown by immune electron microscopy to agglutinate B19 particles and were also shown to recognize the VP2 B19 capsid protein, by Western blot analysis. The B19 antigenicity of the fusion protein was confirmed by immunoblot and enzyme immunoassay with IgG and IgM anti-B19-positive reference human sera.
The occurrence of Epstein-Barr virus (EBV) specific DNase antibodies was measured in the sera of 400 patients. Three hundred sera were positive (greater than or equal to 20) for early antigen diffuse component (EA-D) antibodies, as measured by indirect immunofluorescence (IF). One hundred sera were EA-D antibody-negative but had IF IgG titers (greater than 40) to the viral capsid antigen (VCA). Twenty-nine of the anti-EA-D-positive sera, and 14 of the anti-EA-D-negative sera showed specific inhibition of EBV DNase. The mean age of positive male patients (39 years) for DNase antibodies was significantly higher (p less than 0.05) than the mean age of positive females (27 years). Thirteen of the 40 patients (29.5%) with acquired immunodeficiency syndrome (AIDS) had EBV DNase-neutralizing antibodies. No correlation between the level of anti-DNase antibodies and the antibody titer to EA-D or VCA was observed. These results support the notion that the EBV DNase antibody assay measures an independent index reflecting factual EBV disease.
Hybridization technology allows the detection of EBV (Epstein-Barr Virus) DNA in lymphoproliferative diseases in immunosuppressed patients. Moreover, in HIV infected patients, LIP (Lymphocytic Interstitial Pneumonitis), hairy leucoplasia and lymphoma are associated with EBV by DNA hybridization. These procedures are also useful for understanding interactions between EBV and epithelial cells.