Marianne Antonius Jakobsen

Gene therapy of X-linked severe combined immunodeficiency by use of a pseudotyped gammaretroviral vector

BACKGROUND X-linked severe combined immunodeficiency (SCID-X1) is caused by mutations in the common cytokine-receptor gamma chain (gamma(c)), resulting in disruption of development of T lymphocytes and natural-killer cells. B-lymphocyte function is also intrinsically compromised. Allogeneic bone-marrow transplantation is successful if HLA-matched family donors are available, but HLA-mismatched procedures are associated with substantial morbidity and mortality. We investigated the application of somatic gene therapy by use of a gibbon-ape-leukaemia-virus pseudotyped gammaretroviral vector. METHODS Four children with SCID-X1 were enrolled. Autologous CD34-positive haemopoietic bone-marrow stem cells were transduced ex vivo and returned to the patients without preceding cytoreductive chemotherapy. The patients were monitored for integration and expression of the gamma(c) vector and for functional immunological recovery. FINDINGS All patients have shown substantial improvements in clinical and immunological features, and prophylactic medication could be withdrawn in two. No serious adverse events have been recorded. T cells responded normally to mitogenic and antigenic stimuli, and the T-cell-receptor (TCR) repertoire was highly diverse. Where assessable, humoral immunity, in terms of antibody production, was also restored and associated with increasing rates of somatic mutation in immunoglobulin genes. INTERPRETATION Gene therapy for SCID-X1 is a highly effective strategy for restoration of functional cellular and humoral immunity.

Differences in female-male mortality after high-titre measles vaccine and association with subsequent vaccination with diphtheria-tetanus-pertussis and inactivated poliovirus: reanalysis of West African studies

BACKGROUND Females given high-titre measles vaccine (HTMV) have high mortality; diphtheria-tetanus-pertussis (DTP) vaccination might be associated with increased female mortality. We aimed to assess whether DTP or inactivated poliovirus (IPV) administered after HTMV was associated with increased female-male mortality ratio. METHODS In three trials from West Africa, 2000 children were randomised to HTMV or control vaccine at 4-5 months of age; a second vaccination was given at age 9-10 months (standard measles vaccine). Children in high-titre groups were given IPV or DTP-IPV. Another 944 children received HTMV as routine vaccination in Senegal. FINDINGS When we compared high-titre and control groups, no difference in mortality between the first and the second vaccination was noted. After the second vaccination, the female-male mortality ratio was 1.84 (95% CI 1.19-2.84) in children in the high-titre groups who received DTP-IPV or IPV, and 0.59 (0.34-1.04) in controls who received standard measles vaccine (p=0.007). Children who received HTMV but no additional DTP-IPV or IPV had a female-male mortality ratio of 0.83 (0.41-1.67). This ratio was 2.22 (1.04-4.71) for children who received DTP-IPV after routine HTMV and 1.00 (0.68-1.47) for those who did not. When we combined the results from all trials, the female-male mortality ratio was 1.93 (1.33-2.81) for those who received DTP or IPV after HTMV, and 0.96 (0.69-1.34) for those who did not (p=0.006). INTERPRETATION A change in sequence of vaccinations, rather than HTMV itself, may have been the cause of increased female mortality in these trials.

Report of the first nationally implemented clinical routine screening for fetal RHD in D- pregnant women to ascertain the requirement for antenatal RhD prophylaxis

BACKGROUND: A combination of antenatal and postnatal RhD prophylaxis is more effective in reducing D immunization in pregnancy than postnatal RhD prophylaxis alone. Based on the result from antenatal screening for the fetal RHD gene, antenatal RhD prophylaxis in Denmark is given only to those D− women who carry a D+ fetus. We present an evaluation of the first national clinical application of antenatal RHD screening.

Tuberculin Reaction, BCG Scar, and Lower Female Mortality.

Background: Recent studies have suggested that bacille Calmette-Guérin (BCG) immunization may have a nonspecific beneficial effect on infant survival and that the effect may be more pronounced among girls. In a prospective birth cohort, we examine whether a positive tuberculin skin test and BCG scar in response to BCG immunization were related to better overall survival in Guinea-Bissau and, if so, whether the effect was sex-specific. Methods: Skin tests and BCG scarring were monitored at ages 2 months (n = 2332) and 6 months (n = 1817) in children born from March 2000 to July 2002. A tuberculosis (TB) surveillance system allowed us to exclude from the analysis children with likely TB exposure. The children were followed for survival until 18 months of age. Results: Among children with a tuberculin skin test at 2 and 6 months of age, the mortality rate ratio for skin test reactors (>1 mm) versus nonreactors (0–1 mm) was 0.54 (95% confidence interval = 0.30–0.99). Comparing children with and without a BCG scar, the ratio was 0.55 (0.31–0.96). The effect of a skin test reaction or a BCG scar seemed stronger among girls; for those with positive reaction, the mortality ratio was 0.31 (0.11–0.88) among girls and 0.84 (0.39–1.82) among boys; and for BCG scar, the results were 0.41 (0.21–0.82) and 0.88 (0.34–2.30), respectively. Conclusions: A good response to BCG vaccination is related to lower child mortality. The effect seems most pronounced among girls. The findings may have implications for future vaccine trials and policy.

Peroxisome Proliferator‐Activated Receptor α, δ, γ1 and γ2 Expressions are Present in Human Monocyte‐Derived Dendritic Cells and Modulate Dendritic Cell Maturation by Addition of Subtype‐Specific Ligands

It has recently been shown by Chang et al. (J Immunol 2000;165:3584–91) that the maturation of dendritic cells (DC) in the presence of long‐chain fatty acids redirects DC into Th0/Th2‐inducing cells suggesting the involvement of a receptor for long‐chain fatty acids like members of the peroxisome proliferator‐activated receptors (PPAR) superfamily. Here, we show that immature and mature monocyte‐derived DC (Mo‐DC) express PPARα, PPARδ, PPARγ1 and PPARγ2 mRNA with the highest level of PPARγ1 mRNA. We were only able to observe the expression of PPARγ1 protein by Western blotting probably because the protein level of the other subtypes is below the detection limit. Synthetic ligands specific for PPARα, PPARδ or PPARγ added at day 0–6 have similar effect on the maturation of Mo‐DC driving the maturation of Mo‐DC with atypical phenotype, reduced expression of IL‐10, IL‐12 p35 and IL‐12 p40 mRNA and with reduced stimulatory effects in mixed leucocyte reaction (MLR). Our data suggest that naturally occurring PPAR ligands like fatty acids and fatty acid derivates have anti‐inflammatory effects by redirecting DC into a less stimulatory mode.

Routine noninvasive prenatal screening for fetal RHD in plasma of RhD-negative pregnant women—2 years of screening experience from Denmark

Prenatal and postnatal RhD prophylaxis reduces the risk of RhD immunization in pregnancies of RhD‐negative women. Based on the result from prenatal screening for the fetal RHD gene, prenatal RhD prophylaxis in Denmark is targeted to RhD‐negative women who carry an RhD‐positive fetus. Here, we present a 2‐year evaluation of a nationwide prenatal RHD screening.

Serum Concentration of the Growth Medium Markedly Affects Monocyte‐Derived Dendritic Cells' Phenotype, Cytokine Production Profile and Capacities to Stimulate in MLR

We have investigated how the maturation of monocyte‐derived dendritic cells (Mo‐DC) is affected by the serum concentration of the culture medium. Day 6 DC cultured in 1% human serum were a heterogeneous population of CD1a– and CD1a+ DC that were separated by flow sorting. In contrast, Mo‐DC generated in 10% human serum formed a homogenous population of CD1a– cells. Other phenotypically immature characteristics also varied, and three subsets were still distinguishable upon maturation in LPS. Furthermore, CD1a– DC and CD1a+ DC from 1% culture conditions were excellent stimulators in MLR, while DC cultured in 10% serum were poor stimulators. Similarly, different cytokine profiles of the three subsets were identified. DC cultured in 1% serum had low expression of interleukin‐12 (IL‐12) p40 and IL‐10 mRNA at day 6. Upon maturation, expression of IL‐12 p40 mRNA was upregulated in CD1a+ DC, whereas the level remained relatively low in CD1a– DC. In contrast, DC cultured in 10% had high levels of IL‐10 mRNA at day 6 that was downregulated upon maturation. We conclude that the differentiation of monocytes into DC is significantly influenced by the serum concentration of the growth medium with effects on phenotype, cytokine profile and stimulatory activity.

Testosterone treatment increases androgen receptor and aromatase gene expression in myotubes from patients with PCOS and controls, but does not induce insulin resistance.

Polycystic ovary syndrome (PCOS) is associated with insulin resistance and increased risk of type 2 diabetes. Skeletal muscle is the major site of insulin mediated glucose disposal and the skeletal muscle tissue is capable to synthesize, convert and degrade androgens. Insulin sensitivity is conserved in cultured myotubes (in vitro) from patients with PCOS, but the effect of testosterone on this insulin sensitivity is unknown. We investigated the effect of 7days testosterone treatment (100nmol/l) on glucose transport and gene expression levels of hormone receptors and enzymes involved in the synthesis and conversion of testosterone (HSD17B1, HSD17B2, CYP19A1, SRD5A1-2, AR, ER-α, HSD17B6 and AKR1-3) in myotubes from ten patients with PCOS and ten matched controls. Testosterone treatment significantly increased aromatase and androgen receptor gene expression levels in patients and controls. Glucose transport in myotubes was comparable in patients with PCOS vs. controls and was unchanged by testosterone treatment (p=0.21 PCOS vs. controls). These results suggest that testosterone treatment of myotubes increases the aromatase and androgen receptor gene expression without affecting insulin sensitivity and if testosterone is implicated in muscular insulin resistance in PCOS, this is by and indirect mechanism.

Identification of a novel STAT3 mutation in a patient with hyper-IgE syndrome

Abstract Here we describe a patient with hyper-IgE syndrome presenting with recurrent staphylococcal abscesses, pneumonia, and chronic mucocutaneous candidiasis, and report the identification of a novel STAT3 mutation at amino acid position 621, which has not previously been described. In addition, we review the immunological, infectious, and genetic features of hyper-IgE syndrome.

Postmortem Detection of Hepatitis B, C, and Human Immunodeficiency Virus Genomes in Blood Samples from Drug-Related Deaths in Denmark*

Abstract:  Blood‐borne viral infections are widespread among injecting drug users; however, it is difficult to include these patients in serological surveys. Therefore, we developed a national surveillance program based on postmortem testing of persons whose deaths were drug related. Blood collected at autopsy was tested for anti‐HBc, anti‐HBs, anti‐hepatits C virus (HCV), or anti‐human immunodeficiency virus (HIV) antibodies using commercial kits. Subsets of seropositive samples were screened for viral genomes using sensitive in‐house and commercial polymerase chain reaction (PCR) assays. Hepatitis B virus (HBV) DNA was detected in 20% (3/15) of anti‐HBc‐positive/anti‐HBs‐negative samples, HCV RNA was found in 64% (16/25) of anti‐HCV‐positive samples, and HIV RNA was detected in 40% (6/15) of anti‐HIV‐positive samples. The postmortem and antemortem prevalences of HBV DNA and HCV RNA were similar. Postmortem HIV RNA testing was less sensitive than antemortem testing. Thus, postmortem PCR analysis for HBV and HBC infection is feasible and relevant for demonstrating ongoing infections at death or for transmission analysis during outbreaks.