Discrepancy of Serological and Molecular Patterns of Circulating Epstein-Barr Virus Reactivation in Primary Sjögren's Syndrome.

Primary Sjögren's syndrome (pSS) is characterized by B cell hyperactivation, production of autoantibodies and increased risk of B cell lymphomas. Serological profile of Epstein-Barr virus (EBV) reactivation and increase EBV DNA levels in exocrine glands are observed in pSS, but whether these abnormalities are accompanied with disturbed systemic EBV control or have any association with pSS activity remains to be investigated. In this observational study, we initially explored anti-EBV antibodies and cell-free DNA in 395 samples from a cross-sectional plasma collection of pSS patients included in ASSESS French national cohort. Results were assessed in relation with disease activity. Further, to assess cell-associated EBV DNA we organized a case-control study including 20 blood samples from pSS patients followed in University Hospital Center of Montpellier. Results were compared with matched controls. Robust response against EBV early antigen (EA) was observed in pSS patients with anti-SSA/B (Sjögren's syndrome A and B) and anti-SSA autoantibodies compared to anti-SSA/B negatives (P < 0.01 and P = 0.01, respectively). Increased beta-2 microglobulin, kappa and lambda light chains, and immunoglobulin G levels were more frequently observed in anti-EA seropositive pSS subjects compared to anti-EA negative subjects (P < 0.001; P = 0.001; P = 0.003, respectively). Beta-2 microglobulin was independently associated with anti-EA positivity in multivariate analysis (P < 0.001). Plasma cell-free EBV DNA and EBV cellular reservoir was not different between pSS patients and controls. We conclude that serological evidence of EBV reactivation was more frequently observed and more strongly associated with anti-SSA/B status and B cell activation markers in pSS. However, serological profile of EBV reactivation was not accompanied by molecular evidence of systemic EBV reactivation. Our data indicated that EBV infection remains efficiently controlled in the blood of pSS patients.

Transgenic protein production in silkworm silk glands requires cathepsin and chitinase of Autographa californica multicapsid nucleopolyhedrovirus.

The silkworm Bombyx mori represents an established in vivo system for the production of recombinant proteins. Baculoviruses have been extensively investigated and optimised for the expression of high protein levels inside the haemolymph of larvae and pupae of this lepidopteran insect. Current technology includes deletion of genes responsible for the activity of virus-borne proteases, which in wild-type viruses, cause liquefaction of the host insect and enhance horizontal transmission of newly synthesised virus particles. Besides the haemolymph, the silk gland of B. mori provides an additional expression system for recombinant proteins. In this paper, we investigated how silk gland can be efficiently infected by a Autographa californica multicapsid nuclear polyhedrosis virus (AcMNPV). We demonstrated that the viral chitinase and the cysteine protease cathepsin are necessary to permit viral entry into the silk gland cells of intrahaemocoelically infected B. mori larvae. Moreover, for the first time, we showed AcMNPV crossing the basal lamina of silk glands in B. mori larvae, and we assessed a new path of infection of silk gland cells that can be exploited for protein production.

Experimental respiratory syncytial virus infection of human peribronchial gland cells in vitro.

Respiratory syncytial virus (RSV) is a relevant agent of respiratory tract infections, especially in exacerbations of chronic lung diseases. Peribronchial submucosal glands are the main source of tracheobronchial mucus and therefore of major interest. The authors isolated and cultured human peribronchial gland cells and infected them with RSV. The course of infection was monitored by transmission electron, immuno-, and lectin fluorescence microscopy. Morphology shows virus factories with budding particles within cytoplasmatic vacuoles and virus release after 44 h of infection. Experimental infection of human peribronchial gland cells in primary culture appears to be a suitable model in pulmonary research.

Characterization of B cell lymphoma in patients with Sjögren's syndrome and hepatitis C virus infection.

OBJECTIVE: To characterize the clinical and immunologic patterns of expression, response to therapy, and outcome of patients with Sjögren's syndrome (SS) and associated hepatitis C virus (HCV) infection who developed B cell lymphoma. METHODS: Various international reference centers constituted a multicenter study group with the purpose of creating a registry of patients with SS-HCV who developed B cell lymphoma. A protocol form was used to record the main characteristics of SS, chronic HCV infection, and B cell lymphoma. RESULTS: Twenty-five patients with SS-HCV with B cell lymphoma were included in the registry. There were 22 (88%) women and 3 (12%) men (mean age 55, 58, and 61 years at SS, HCV infection, and lymphoma diagnosis, respectively). The main extraglandular SS manifestations were cutaneous vasculitis in 15 (60%) patients and peripheral neuropathy in 12 (48%); the main immunologic features were positive rheumatoid factor (RF) in 24 (96%) and type II cryoglobulins in 20 (80%). The main histologic subtypes were mucosa-associated lymphoid tissue (MALT) lymphoma in 11 (44%) patients, diffuse large B cell lymphoma in 6 (24%), and follicular center cell lymphoma in 6 (24%). Fifteen (60%) patients had an extranodal primary location, most frequently in the parotid gland (5 patients), liver (4 patients), and stomach (4 patients). Twelve (52%) of 23 patients died after a median followup from the time of lymphoma diagnosis of 4 years, with lymphoma progression being the most frequent cause of death. Survival differed significantly between the main types of B cell lymphoma. CONCLUSION: Patients with SS-HCV and B cell lymphoma are clinically characterized by a high frequency of parotid enlargement and vasculitis, an immunologic pattern overwhelmingly dominated by the presence of RF and mixed type II cryoglobulins, a predominance of MALT lymphomas, and an elevated frequency of primary extranodal involvement in organs in which HCV replicates (exocrine glands, liver, and stomach).

Venom apparatus of the endoparasitoid wasp Opius caricivorae Fischer (Hymenoptera: Braconidae): morphology and ultrastructure.

The morphology and ultrastructure of the venom apparatus of the endoparasitoid wasp, Opius caricivorae Fischer (Hymenoptera: Braconidae), were observed using light and electron microscopes. The venom apparatus consists of one venom reservoir and several gland filaments. The gland filaments join together at the end of the reservoir and consist of an outer single layer of secretory cells, a layer of degenerated epidermal cells, and an inner intima that encloses the lumen. The secretory cells are organelle rich, with abundant rough endoplasmic reticulum, mitochondria, and vacuole, in which vesicular organelles secrete the components of venom. The reservoir consists of a muscular sheath, secretory cells, and squamous cells. The intima has an unevenly thickened chitinous coat. The vesicular organelles of the reservoir secretory cells differ from those of the gland filament: the microvilli being much longer and radiating in all directions. The venom reservoir not only serves to store but also secretes the venom. Virus-like particles were discovered in the secretory cells of the gland filaments. The structural features of venom apparatus of this species are discussed in a biological context.

Hepatitis C virus, Sjögren's syndrome and B-cell lymphoma: linking infection, autoimmunity and cancer.

An increased prevalence of hematologic malignancies is often described in patients with Sjögren's syndrome (SS). Viruses have been proposed as possible etiologic or triggering agents of systemic autoimmune diseases (SADs), with hepatitis C virus (HCV) being one of the viruses most frequently associated with autoimmune features and with systemic autoimmune diseases such as mixed cryoglobulinemia or SS. Moreover, the association between HCV infection and hematologic malignancies, mainly non-Hodgkin's lymphoma (NHL), is supported by several studies. For these reasons, the recognized association of specific systemic autoimmune diseases (mainly SS and mixed cryoglobulinemia) with HCV infection, added to the possible evolution of any one of these entities into a B-cell NHL, suggests the possibility of a close relationship among SS, HCV and B-cell lymphoproliferative disorders, especially in patients with type II mixed cryoglobulinemia.

Human papillomaviruses, expression of p16, and early endocervical glandular neoplasia.

Adenocarcinoma in situ (ACIS) is the precursor of cervical adenocarcinoma (ACs), and its distinction from benign but morphologically atypical glandular epithelium may be difficult. The cyclin-dependent kinase inhibitor p16(ink4) is expressed in cervical squamous cell carcinomas, their precursors, and cervical ACs, and there is a strong relationship between p16 expression and the presence of human papillomavirus (HPV)-encoded E6/E7 transcription. This study analyzed 95 cases of benign and premalignant cervical glandular ACIS lesions for p16 antigen and the proliferative marker Ki-67; HPV E6/E7 transcripts were detected by RNA/RNA in situ hybridization. HPV 16 or 18 E6/E7 transcription and strong, diffuse p16 positivity were detected only in ACIS lesions. A high and moderate Ki-67 index was observed in 76% and 22% of ACIS, respectively. Thirty-three of 36 microglandular change, tubal, atypical tubal, and endometrial-type epithelia scored negative or weakly positive for p16. Distribution of staining in 3 strongly positive cases was heterogeneous. The diffuse distribution of p16 immunostaining in HPV16/18-positive glandular neoplasms supports a strong association with HPV infection and indicates that this biomarker may discriminate ACIS from its benign mimics. However, this distinction requires attention to staining distribution because p16 is focally expressed in tubal-endometrial epithelia and diffusely expressed in endometrium, indicating that in some cases the use of other biomarkers, such as Ki-67, may be necessary. Because endometrial glandular epithelia may also express p16, the diagnostic application of p16 immunohistochemistry to cytological samples is uncertain.

Cytomegalovirus infection in a pig in South Africa.

An 8-week-old piglet with dyspnoea, bilateral mucopurulent nasal discharge and mouth breathing was euthanased and a necropsy was performed. Apart from histological evidence of diffuse rhinitis, large intranuclear inclusion bodies, pathognomonic forporcine cytomegalovirus infection, were detected within mucous glands on the nasal turbinates. This is the first such case to be diagnosed in South Africa.

[A viral infection in the neck glandula of larvae of Dione junio (Lepidoptera: Nymphalidae)]. / Una infección viral en la glándula cervical de larvas de Dione junio (Lepidoptera: Nymphalidae).

Lepidopteran larvae may be attacked by different viruses, many of which belong to the Baculoviridae family. Whilst studying the ultrastructure of the neck gland in Dione junio larvae we found that in later instars the larvae showed symptoms of attack by two types of virus. The glands were prepared for optical and electron microscopy using sodium cacodylate buffer and standard procedures (0.1M, pH 7.2). The neck gland is composed of two oval internal sacks which communicate with the exterior via an extracellular channel. Each sack contains, in its external region, cells with large, irregular nuclei and a dense cytoplasm containing numerous small mitochondria. In infected larvae, the tissues are damaged and the nuclear polyhedrosis virus can be observed in several of the nuclei. In the cytoplasm another "rickettsia type" virus, may be observed. The pathogenic viruses present in D. junio larvae could be studied as potential biological controls of this pest.

Una infección viral en la glándula cervical de larvas de Dione junio (Lepidoptera: Nymphalidae) / A viral infection in the neck glandula of larvae of Dione junio (Lepidoptera: Nymphalidae)

Lepidopteran larvae may be attacked by different viruses, many of which belong to the Baculoviridae family. Whilst studying the ultrastructure of the neck gland in Dione junio larvae we found that in later instars the larvae showed symptoms of attack by two types of virus. The glands were prepared for optical and electron microscopy using sodium cacodylate buffer and standard procedures (0.1M, pH 7.2). The neck gland is composed of two oval internal sacks which communicate with the exterior via an extracellular channel. Each sack contains, in its external region, cells with large, irregular nuclei and a dense cytoplasm containing numerous small mitochondria. In infected larvae, the tissues are damaged and the nuclear polyhedrosis virus can be observed in several of the nuclei. In the cytoplasm another "rickettsia type" virus, may be observed. The pathogenic viruses present in D. junio larvae could be studied as potential biological controls of this pest.

Pathology of interdigital glands in a wild Japanese serow (Capricornis crispus) infected with parapoxvirus.

The interdigital glands of a Japanese serow infected with parapoxvirus had severe papular and nodular lesions that completely occupied the sac and duct of the gland. The lesions were characterized by acanthosis with hyperkeratosis. Intracytoplasmic inclusion bodies were detected in the vacuolated prickle cells. By electron microscopy, mature and immature viral particles were present in the cytoplasm. These glands act as scent glands, and lesions in this organ probably affected the ecological adaptation of this individual.

Morphologic changes in the nasal cavity associated with sialodacryoadenitis virus infection in the Wistar rat.

A sequential study of lesions of the nasal cavity associated with sialodacryoadenitis virus (SDAV) infection was made in the laboratory rat. Wistar rats were intranasally inoculated with approximately 10(3) TCID50 of the coronavirus SDAV. Transverse sections of four regions of the nasal cavity from inoculated and control animals were examined by light microscopy and immunohistochemistry at 2, 4, 6, 8, 10, and 14 days postinoculation (PI). Lesions were observed in the following regions of the upper respiratory tract: respiratory epithelium, transitional epithelium, olfactory epithelium, nasolacrimal duct, vomeronasal organ, and the submucosal glands of the nasal passages. In general, in structures lined by ciliated epithelial cells, there was focal to segmental necrosis with exfoliation of affected cells and polymorphonuclear cell infiltration during the acute stages, progressing to squamous metaplasia during the reparative stages. Repair in these regions was essentially complete by 14 days PI. In the olfactory epithelium and the vomeronasal organ, there was interstitial edema with necrosis and exfoliation of epithelial cells and minimal to moderate inflammatory cell response during the acute stages. Residual reparative lesions were still evident in the olfactory epithelium, the columnar epithelium and neuroepithelium of the vomeronasal organ, and the nasolacrimal duct at 14 days PI. Viral antigen was demonstrated by immunohistochemistry in all regions during the acute stages of the disease, with the exception of the vomeronasal organ. In view of these findings, infections of the respiratory tract with viruses such as SDAV could have significant effects on functions such as olfaction and chemoreception for > or = 2 weeks postexposure in this species.