Immune interferon secretion as an expression of immunological memory to transplantation antigens: in vivo generation of long-lived,recirculating memory cells

Leukocytes from C57BL/6 mice immunized against DBA/2 strain antigens by intraperitoneal injection of mastocytoma P 815 cells produced, when stimulated in the mixed leukocyte reaction assay with DBAI2 spleen cells, an earlier and more intense secretion of immune interferon than leukocytes from untreated mice. This secondary-type interferon response was independent of cell proliferation. The memory phenomenon was induced by long-lived, recirculating lymphocytes found in spleen, lymph nodes and thoracic duct, but not in the thymus. Memory cells could be recruited into inflammatory sites. They were shown to be specific for H-2 alloantigens, although some cross-reactivity with stimulating cells bearing unrelated H-2 antigens was observed. The possible anti-tumor, antiviral and immunoregulatory roles of this memory phenomenon, and its significance in transplantation immunity are discussed.     

Neuropathology of Early HIV-1 Infection

Early HIV-1 invasion of the central nervous system has been demonstrated by many cerebrospinal fluid studies; however, most HIV-1 carriers remain neurologically unimpaired during the so called “asymptomatic” period lasting from seroconversion to symptomatic AIDS. Therefore, neuropathological studies in the early pre-AIDS stages are very few, and the natural history of central nervous system changes in HIV-1 infection remains poorly understood. Examination of brains of asymptomatic HIV-1 positive individuals who died accidentally and of rare cases with acute fatal encephalopathy revealing HIV infection, and comparison with experimental simian immunodeficiency virus and feline immunodeficiency virus infections suggest that, invasion of the CNS by HIV-1 occurs at the time of primary infection and induces an immunological process in the central nervous system. This includes an inflammatory T-cell reaction with vasculitis and leptomeningitis, and immune activation of brain parenchyma with increased number of microglial cells, upregulation of major histocompatibility complex class II antigens and local production of cytokines. Myelin pallor and gliosis of the white matter are usually found and are likely to be the consequence of opening of the blood brain barrier due to vasculitis; direct damage to oligodendrocytes by cytokines may also interfere. These white matter changes may explain, at least partly, the early cerebral atrophy observed, by magnetic resonance imaging, in asymptomatic HIV-1 carriers. In contrast, cortical damage seems to be a late event in the course of HIV-1 infection. There is no significant neuronal loss at the early stages of the disease, no accompanying increase in glial fibrillary acid protein staining in the cortex, and only exceptional neuronal apoptosis. Although HIV-1 proviral DNA may be demonstrated in a number of brains, viral replication remains very low during the asymptomatic stage of HIV-1 infection. This makes it likely that, although opening of the blood brain barrier may facilitate viral entry into the brain, specific immune responses including both neutralising antibodies and cytotoxic T-lymphocytes, continuously inhibits viral replication at that stage.     

Identification of human herpesvirus 6 variants A and B by primer-specific real-time PCR may help to revisit their respective role in pathology.

BACKGROUND: Human herpesvirus 6 (HHV-6) isolates are classified into two variants, termed HHV-6A and HHV-6B, on the basis of distinct genetic, antigenic and biological characteristics, but the specific pathogenicity of each variant remains poorly understood. OBJECTIVES: To design a rapid, sensitive and specific real-time variant-specific PCR (VS-PCR) method to differentiate both variants in biological specimens. STUDY DESIGN: The VS-PCR was adapted from a real-time PCR assay, based on TaqMan technology, previously developed for the genome quantitation of both HHV-6 variants [Gautheret-Dejean A, Manichanh C, Thien-Ah-Koon F, Fillet AM, Mangeney N, Vidaud M, et al. Development of a real-time polymerase chain reaction assay for the diagnosis of human herpesvirus-6 infection and application to bone marrow transplant patients. J Virol Meth 2002;100:27-35], a consensual reverse primer (Taq2) being changed into two variant-specific primers named H6A and H6B. This method was applied to a large set of biological specimens obtained in different pathological contexts. RESULTS: The sensitivity threshold was about 10 copies/well for HHV-6A-specific PCR (PCR-A) and 1 copy/well for HHV-6B-specific PCR (PCR-B). Both assays showed a linear dynamic range from 10 to 100,000 copies of HHV-6 DNA. Regarding the specificity and the capacity of discrimination of each assay, one variant could be detected and identified in the presence of more than 1000 times higher concentrations of the other variant in virus mixtures. The comparison of the results obtained with this VS-PCR with those previously obtained with a classic PCR method allowed us to validate our new technique on a wide panel of biological samples, including numerous patients with severe HHV-6-related symptoms. The high prevalence of HHV-6B was confirmed in healthy individuals and immunocompromised patients. HHV-6A was identified in distinct samples from several patients exhibiting neurological disorders. CONCLUSIONS: We developed a new VS-PCR assay, able to differentiate HHV-6A and HHV-6B in biological samples, even in the case of mixed infections. Our study confirms the wide prevalence of HHV-6B and highlights the potential greater neuropathogenic role of HHV-6A in immunocompromised patients and young infants.     

Heparin and non-heparin-like dextrans differentially modulate endothelial cell proliferation: in vitro evaluation with soluble and crosslinked polysaccharide matrices

Proliferation of endothelial cells (ECs) is a cellular step of particular importance for implanted cardiovascular biomaterials. Heparin and some synthetic water-soluble non-anticoagulant polysaccharides derived from dextran and bearing anionic carboxymethyl and hydrophobic benzylamine groups were first investigated for their effects on EC proliferation in vitro. The results assessed by cell counting, 3H-thymidine uptake, and flow cytometry analysis, showed that the derivatized dextran-bearing hydrophobic groups stimulated the EC growth in the presence of serum, whereas native dextran or dextran-bearing anionic carboxymethyl groups were inactive and heparin was slightly inhibitory. Then, we showed that the derivatized dextran enhanced EC proliferation by potentiation of the mitogenic activities of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (FGF-2), two potent EC growth factors. In the presence of 2 nM of derivatized dextran, a 3-fold and 13-fold increase of 3H-thymidine uptake was obtained with VEGF and FGF-2, respectively. Finally, proliferation of ECs was investigated on crosslinked gels made of polysaccharides. It is of interest that EC proliferation was higher on gels containing the derivatized dextran than on plain hydrogels, and heparinized gels inhibited cell proliferation. From the obtained results, we propose that the synthetic non-heparin-like dextran may be of interest as a coating for the endothelialization of cardiovascular biomaterials.     

Papillary renal cell carcinoma

A series of 43 papillary renal cell carcinomas (PRCCs) were analyzed to investigate the prognostic value of the morphological subtyping (type 1/type 2) proposed by Delahunt and Eble. Twenty-six cases were type 1 (small cuboid cells arranged in single or double layers), 13 cases were type 2 (voluminous eosinophilic cells with irregular pseudostratification pattern), and four cases with oncocytic cells (large eosinophilic cells with round regular nuclei) were distinct from type 2 and grouped apart. All type-1 and oncocytoid-type PRCCs were staged pT1 or pT2, whereas 8/13 type-2 PRCCs were staged pT3 or pT4. Follow-up information (range, 3-113 months; median, 43 months) showed 12 deaths from disease: 2 in the type-1 group,10 in the type-2 group, 0 in the oncocytoid-type group. The Kaplan-Meier analysis showed that pejorative outcome was associated (P<0.001) with high stage (pT3/pT4), high nuclear grade (3/4), morphological type 2, absence of foam cells, and abundant fibrous stroma. The multivariate analysis showed that stage and morphological type were independently associated with survival (P<0.05). These results support the clinical interest of morphological subtyping of PRCCs in the prognosis evaluation of the patients. The four oncocytoid-type PRCCs had a favorable outcome, but additional data are required to evaluate this type of neoplasm.     

The migration of lymphocytes in the fetal lamb.

The migration of 51Cr-labeled autologous lymphocytes from intestinal or prescapular lymph was compared in fetal lambs and adult sheep. A subpopulation of lymphocytes present in intestinal lymph of adults which migrated to the small intestine was not found in fetal intestinal lymph. There were marked differences in the migration of fetal and adult lymphocytes to the lungs and liver. In spite of the absence of circulating antibodies or immunoglobulins and of extrinsic antigen in the immunologically virgin sheep fetus, the circulation of lymphocytes through the spleen and lymph nodes of fetal lambs was more intense than in the adult.     

The relationship between Caesarean section and subfertility in a population-based sample of 14 541 pregnancies

BACKGROUND: There has been a threefold increase in the rate of Caesarean section over the past 25 years. The long-term consequences of Caesarean section may include subsequent subfertility. METHODS: We investigated the relationship between Caesarean section and subfertility within a cohort of 14 541 pregnant women. RESULTS: A history of previous Caesarean section was associated with an increased risk of taking >1 year to conceive from the time of planning a pregnancy, adjusted odds ratio (OR) 1.53 [95% confidence interval (CI) 1.09, 2.14]. This association was stronger for women of parity > or =2, adjusted OR 2.97 (95% CI 1.72, 5.10). Nulliparous women with a history of subfertility were at increased risk of delivery by Caesarean section, adjusted OR 1.56 (1.22, 2.00) and OR 2.33 (1.64, 3.30) for durations of >1 and >3 years respectively. CONCLUSIONS: These findings suggest a complex relationship between Caesarean section and subfertility where subfertility may both precede and be a consequence of Caesarean section.     

ARX, a novel Prd-class-homeobox gene highly expressed in the telencephalon, is mutated in X-linked mental retardation

Investigation of a critical region for an X-linked mental retardation (XLMR) locus led us to identify a novel Aristaless related homeobox gene (ARX ). Inherited and de novo ARX mutations, including missense mutations and in frame duplications/insertions leading to expansions of polyalanine tracts in ARX, were found in nine familial and one sporadic case of MR. In contrast to other genes involved in XLMR, ARX expression is specific to the telencephalon and ventral thalamus. Notably there is an absence of expression in the cerebellum throughout development and also in adult. The absence of detectable brain malformations in patients suggests that ARX may have an essential role, in mature neurons, required for the development of cognitive abilities.