Kristina Broliden

Mucosal and Plasma IgA from HIV-1-Exposed Uninfected Individuals Inhibit HIV-1 Transcytosis Across Human Epithelial Cells

HIV-1-specific IgA has been described in the genital tract and plasma of HIV-1 highly exposed, persistently seronegative (HEPS) individuals, and IgA from these sites has been shown to neutralize HIV-1. This study examines the ability of IgA isolated from HEPS individuals to inhibit transcytosis across a tight epithelial cell layer. A Transwell system was established to model HIV-1 infection across the human mucosal epithelium. The apical-basolateral transcytosis of primary HIV-1 isolates across this mucosal model was examined in the presence and the absence of IgA isolated from the genital tract, saliva, and plasma of HEPS individuals enrolled in both a sex worker cohort in Nairobi, Kenya, and a discordant couple cohort in Italy. In the absence of IgA, HIV-1 primary isolates were actively transported across the epithelial membrane and were released on the opposite side of the barrier. These transcytosed HIV-1 particles retained their ability to infect human mononuclear cells. However, IgA purified from the mucosa and plasma of HEPS individuals was able to inhibit HIV-1 transcytosis. Inhibition was seen in three of six cervicovaginal fluid samples, five of 10 saliva samples, and three of six plasma samples against at least one of the two primary HIV-1 isolates tested. IgA from low risk, healthy control subjects had no inhibitory effect on HIV-1 transcytosis. The ability of mucosal and plasma IgA to inhibit HIV-1 transcytosis across the mucosal epithelium may represent an important mechanism for protection against the sexual acquisition of HIV-1 infection in HEPS individuals.

Frequency of human parvovirus B19 infection in intrauterine fetal death

BACKGROUND Parvovirus B19 is known to cause fetal death in the second trimester, mainly in combination with hydrops fetalis. However, the frequency of parvovirus-B19-associated non-hydropic fetal loss in the late second and third trimester has not been thoroughly investigated. We aimed to investigate the frequency of parvovirus B19 infection in unselected cases of intrauterine fetal death and to assess the sensitivity of different diagnostic procedures. METHODS Of 14147 deliveries in three hospitals in the major Stockholm area of Sweden, all cases of intrauterine fetal death (>22 gestational weeks) that occurred between January, 1998, and May, 1999 (n=47), referred cases of miscarriage (<22 gestational weeks, n=37), and induced abortions (n=29), were included in the study. Placental and fetal tissues were examined by means of parvovirus-B19-specific PCR, histopathology, and immunohistochemistry. Placental tissues from 53 normal pregnancies at term were also examined. FINDINGS Significantly more cases of intrauterine fetal death were positive for parvovirus B19 DNA (seven [15%]) than were normal pregnancies at term (zero, p=0.049). Furthermore, parvovirus B19 DNA was found in two (5%) of the miscarriages but not in any of the cases of induced abortion. Only three of nine DNA-positive cases had parvovirus-B19-associated inclusions and stained positive for viral proteins. All but one of the DNA-positive cases of intrauterine fetal death were non-hydropic. INTERPRETATION The presence of parvovirus B19 DNA in cases of late second-trimester and third-trimester fetal death is common, and most are non-hydropic. The sensitivity of conventional diagnostic procedures for intrauterine fetal death could be greatly improved by addition of parvovirus B19 PCR.

Functional HIV-1 specific IgA antibodies in HIV-1 exposed, persistently IgG seronegative female sex workers

Although HIV-specific cellular immune responses are found in a number of HIV highly-exposed, persistently seronegative (HEPS) cohorts, late seroconversion can occur despite pre-existing cytotoxic T lymphocytes (CTL), suggesting that a protective HIV vaccine may need to induce a broader range of HIV-specific immune responses. Low levels of HIV-specific IgA have been found in the genital tract and plasma of the majority of Nairobi HEPS sex workers and appeared to be independent of HIV-specific cellular responses. IgA purified from genital tract, saliva and plasma of most HEPS sex workers were able to neutralize infection of PBMC by a primary (NSI) clade B HIV isolate, as well as viral isolates from clades A and D, which predominate in Kenya. In addition, these IgA were able to inhibit transcytosis of infective HIV virions across a transwell model of the human mucosal epithelium in an HIV-specific manner. Preliminary work in other HEPS cohorts has suggested the recognition of different gp41 epitopes in HEPS and HIV-infected subjects. Although present at low levels, these IgA demonstrated cross-clade neutralizing activity and were able to inhibit HIV mucosal transcytosis, suggesting an important functional role in protection against HIV infection.

Parvovirus B19 infection: association with third‐trimester intrauterine fetal death

Objective To identify the presence of parvovirus B19 infection as a possible cause of fetal loss in the third trimester.

Mucosal immune responses in the genital tract of HIV-1-exposed uninfected women

The development of HIV‐1 vaccines and microbicides remains hindered by our limited understanding of correlates of immune protection to infection. Evidence indicating that resistance to HIV‐1 infection is indeed possible comes from HIV‐1‐exposed yet uninfected individuals, including cohorts of commercial sex workers and discordant couples. Despite their uninfected status some of these individuals have mucosal and systemic HIV‐1‐specific humoral and cellular immune responses in addition to their innate immune response. The combined contribution of innate and adaptive immunity as well as genetic factors is most likely of great importance for this protection against infection. Here we review data on the antibody responses and secreted immune molecules of the innate immune system in the female genital tract with emphasis on individuals who seem to resist HIV‐1‐infection despite repeated exposure to the virus.

Isolate-specific neutralizing antibodies in patients with progressive HIV-1-related disease.

Four individuals with increasing severity of HIV-1 infection were studied for serum neutralizing activity against their own virus isolates collected during 1-3 years. Sequential serum samples and HIV-1 isolates were available from these patients from the stage of lymphadenopathy to severe immunodeficiency. The replicative capacity of isolates changed from slow/low to rapid/high in each patient during the study period. Sequential virus isolates showed differences in sensitivity to neutralization by autologous as well heterologous area. Taken together with our previous results demonstrating that variant viruses resistant to neutralization by autologous sera emerge during the year following primary infection, neutralization-resistant variants seem to emerge during the entire course of HIV-1 infection. The ability to produce neutralizing antibodies to autologous virus appears to correlate with replicative capacity of the virus and the degree of immunodeficiency in the patient.

Abundant expression of HIV target cells and C-type lectin receptors in the foreskin tissue of young Kenyan men

A biological explanation for the reduction in HIV-1 (HIV) acquisition after male circumcision may be that removal of the foreskin reduces the number of target cells for HIV. The expression of potential HIV target cells and C-type lectin receptors in foreskin tissue of men at risk of HIV infection were thus analyzed. Thirty-three foreskin tissue samples, stratified by Herpes simplex virus type 2 status, were obtained from a randomized, controlled trial conducted in Kenya. The samples were analyzed by confocal in situ imaging microscopy and mRNA quantification by quantitative RT-qPCR. The presence and location of T cells (CD3(+)CD4(+)), Langerhans cells (CD1a(+)Langerin/CD207(+)), macrophages (CD68(+) or CD14(+)), and submucosal dendritic cells (CD123(+)BDCA-2(+) or CD11c(+)DC-SIGN(+)) were defined. C-type lectin receptor expressing cells were detected in both the epithelium and submucosa, and distinct lymphoid aggregates densely populated with CD3(+)CD4(+) T cells were identified in the submucosa. Although the presence of lymphoid aggregates and mRNA expression of selected markers varied between study subjects, Herpes simplex virus type 2 serostatus was not the major determinant for the detected differences. The detection of abundant and superficially present potential HIV target cells and submucosal lymphoid aggregates in foreskin mucosa from a highly relevant HIV risk group demonstrate a possible anatomical explanation that may contribute to the protective effect of male circumcision on HIV transmission.

Parvovirus B19 infection in pediatric solid‐organ and bone marrow transplantation

The clinical significance of parvovirus B19 infection in pediatric solid-organ and bone marrow transplanted patients is unclear. The overall prevalence of parvovirus B19 infection in these patients is about 1-2% during the first year after transplantation. The most common symptom is anemia, but leukopenia and thrombocytopenia have also been observed. Rare cases of hepatic dysfunction, myocarditis, vasculitis and respiratory failure have also been reported. Whereas serology is of limited value around the time of transplantation, it is recommended that a search for B19 DNA is included in first-line investigations in any transplanted patient with unexplained anemia. Specific antiviral therapy is not available, however, intravenous immunoglobulin produces rapid improvement in most cases. Although relatively rare, the severe complications following parvovirus B19 infection in the transplant setting can be avoided by early diagnosis and treatment.

A Human Monoclonal Antibody to Hiv-1 gp41 with Neutralizing Activity Against Diverse Laboratory Isolates

A potential component that may be useful for passive immunotherapy for HIV-1 is human monoclonal antibodies (HumAbs) possessing potent anti-HIV-1 activity that is directed against conserved regions of the envelope glycoprotein. Such antibodies would, in principle, have the ability to neutralize diverse isolates of HIV-1. To develop such reagents, hybridomas were derived by initial Epstein Barr virus transformation of peripheral blood mononuclear cells (PBMCs) from an asymptomatic HIV-1 seropositive donor followed by fusion with heteromyelomas, and secreted anti-HIV-1 antibodies were further characterized. The specificity of one HumAb, designated as clone 3, was determined by enzyme-linked immunosorbent assay (ELISA) and Western blotting analyses that indicated reactivity to the transmembrane envelope glyco-protein gp41. Synthetic pentadecapeptides overlapping by 10 amino acids were utilized for epitope mapping of clone 3; a decapeptide GCSGKLICTT in the transmembrane gp41 was identified as the epitope. Clone 3 bound to SupT1 cells infected with HTLV-IIIB in fluorescent activated cell sorting analysis. In addition, in vitro biological assays demonstrated that clone 3 possessed neutralization reactivity against diverse laboratory isolates as well as an AZT-resistant isolate. Therefore, clone 3 reactivity defines a conserved neutralizable site on the HIV-1 transmembrane glycoprotein. Clone 3 and the conserved immunogenic epitope on gp41 could be useful in passive and active immunotherapy for the acquired immunodeficiency syndrome (AIDS).

Upregulation of interferon-α and RANTES in the cervix of HIV-1-seronegative women with high-risk behavior

Objective:The expression of innate immune molecules associated with potential blocking activity of HIV-1 propagation was analyzed in the cervical tissue of a group of African HIV-1 IgG-negative commercial sex workers (CSWs) with an HIV-1-encountering risk behavior. Methods:Cervical biopsies from the superior portion of the ectocervix were assessed for innate immune molecules and evaluated in situ by computerized image analysis at the single-cell level. Results:A higher expression of interferon-α (IFNα) and RANTES was detected in CSWs and HIV-1-infected individuals as compared to low-risk HIV-1-uninfected controls (Neg Ctrls). Most (>90%) of RANTES-expressing cells were CD8+ cells as determined by confocal microscopy. In contrast, the expression of leukemia inhibitory factor (LIF) and secretory leukocyte protease inhibitor (SLPI) was comparable between the groups. The expression of β-defensin 2 was highest in HIV-1-infected individuals. Conclusions:Induction of IFNα and RANTES expression in cervical mucosa may contribute to protection of sexual HIV-1 transmission in subjects with a higher risk behavior.