Spontaneous and natural cytotoxicity receptor-mediated cytotoxicity are effector functions of distinct natural killer subsets in hepatitis C virus-infected chimpanzees.

In humans, CD16 and CD56 are used to identify functionally distinct natural killer (NK) subsets. Due to ubiquitous CD56 expression, this marker cannot be used to distinguish between NK cell subsets in chimpanzees. Therefore, functional analysis of distinct NK subsets during hepatitis C virus (HCV) infection has never been performed in these animals. In the present study an alternative strategy was used to identify four distinct NK subsets on the basis of the expression of CD16 and CD94. The expression of activating and inhibiting surface receptors showed that these subsets resemble human NK subsets. CD107 expression was used to determine degranulation of the different subsets in naive and HCV-infected chimpanzees. In HCV-infected chimpanzees increased spontaneous cytotoxicity was observed in CD94(high/dim) CD16(pos) and CD94(low) CD16(pos) subsets. By contrast, increased natural cytotoxicity receptor (NCR)- mediated degranulation after NKp30 and NKp44 triggering was demonstrated in the CD94(dim) CD16(neg) subset. Our findings suggest that spontaneous and NCR-mediated cytotoxicity are effector functions of distinct NK subsets in HCV-infected chimpanzees.

Whole blood stimulation with Toll-like receptor (TLR)-7/8 and TLR-9 agonists induces interleukin-12p40 expression in plasmacytoid dendritic cells in rhesus macaques but not in humans.

Macaques provide important animal models in biomedical research into infectious and chronic inflammatory disease. Therefore, a proper understanding of the similarities and differences in immune function between macaques and humans is needed for adequate interpretation of the data and translation to the human situation. Dendritic cells are important as key regulators of innate and adaptive immune responses. Using a new whole blood assay we investigated functional characteristics of blood plasmacytoid dendritic cells (pDC), myeloid dendritic cells (mDC) and monocytes in rhesus macaques by studying induction of activation markers and cytokine expression upon Toll-like receptor (TLR) stimulation. In a head-to-head comparison we observed that rhesus macaque venous blood contained relatively lower numbers of pDC than human venous blood, while mDC and monocytes were present at similar percentages. In contrast to humans, pDC in rhesus macaques expressed the interleukin (IL)-12p40 subunit in response to TLR-7/8 as well as TLR-9 stimulation. Expression of IL-12p40 was confirmed by using different monoclonal antibodies and by reverse transcription-polymerase chain reaction (RT-PCR). Both in humans and rhesus macaques, TLR-4 stimulation induced IL-12p40 expression in mDC and monocytes, but not in pDC. The data show that, in contrast to humans, pDC in macaques are able to express IL-12p40, which could have consequences for evaluation of human vaccine candidates and viral infection.

A clinically relevant HIV-1 subunit vaccine protects rhesus macaques from in vivo passaged simian-human immunodeficiency virus infection.

OBJECTIVES: To investigate whether immunization with recombinant HIV-1 envelope protein derived from a clinical isolate could protect macaques from infection with an in vivo passaged chimeric simian-human immunodeficiency virus (SHIV). DESIGN AND METHODS: A total of 16 animals were studied from which three groups of four animals were immunized with vaccine formulations of the CC-chemokine receptor-5-binding recombinant gp120 of HIV-1W6.1D. Four weeks after the last immunization, all 16 animals were intravenously challenged with in vivo passaged SHIV derived from the same HIV-1 group B clinical isolate (W6.1D) as the vaccines. RESULTS: Vaccine protection from infection was demonstrated in 10 out of 12 macaques immunized with recombinant gp120. Complete protection from infection was achieved with all of the animals that received the SBAS2-W6.1D formulation, a potent inducer of both T-cell and humoral immune responses. Partial protection was achieved with SBAS1-W6.1D, a formulation based on immunomodulators known to induce T-cell responses in humans. In vaccinated animals that were infected, virus load was reduced and infection was delayed. CONCLUSIONS: In a relatively large number of primates, vaccine efficacy was demonstrated with a clinically relevant HIV-1 vaccine. These results reveal that it is possible to induce sterilizing immunity sufficient to protect from infection with SHIV which was passaged multiple times in vivo. Our findings have implications for current HIV-1 clinical vaccine trials and ongoing efforts to develop safe prophylactic AIDS vaccines.

Iodine deficiency induces thyroid autoimmune reactivity in Wistar rats.

The last 2 decades it has become clear that iodine deficiency has a modulating effect on the thyroid autoimmune response in humans. Also, in animals that spontaneously develop autoimmune thyroid disease, evidence is accumulating that a low iodine intake can modulate thyroid autoimmune reactivity. However, it is still not clear what the effect of a low iodine intake on thyroid autoimmune reactivity is in normal nonautoimmune animals. To study the relationship of a dietary low iodine intake on the thyroid autoimmune reactivity in nonautoimmune animals, normal Wistar rats (female) were kept on an enriched iodine diet (daily iodine intake of 100 micrograms iodine), a "for our area normal" (conventional) diet (COD; daily iodine intake of 7 micrograms iodine), a low iodine diet (LID; 2 days of 1% KCLO4, followed by iodine-deficient drinking water/pellets), or an extremely low iodine diet (LID+; 1% KCLO4 continuously in the drinking water and iodine-deficient pellets). The enriched iodine diet rats were euthyroid (T3, approximately 8 nM/liter: T4, approximately 50 nM/liter; TSH, approximately 2 ng/ml), had a normal thyroid weight (approximately 12.5 mg), and showed only minimal signs of local thyroid immune reactivity; low numbers of intrathyroidal dendritic cells (DC; approximately 35 DC/mm2), CD4+ cells (approximately 2 cells/mm2), and CD8+ cells (approximately 2.5 cells/mm2) were found in combination with low anticolloid antibody production (incidence of positive animals, 12.5%). The COD resulted in a normal thyroid function. The rats were euthyroid (range of T3, 1.6-1.2 nM/liter; T4, approximately 50 nM/liter; TSH, approximately 2 ng/ml) and had a normal thyroid weight (approximately 12.5 mg). However, some signs of thyroid autoimmune reactivity were found [number of intrathyroidal DC, approximately 40/mm2; approximately 3 CD4-positive (CD+) cells/mm2; approximately 3 CD8+ cells/mm2; together with a 30% incidence of anticolloid antibodies]. The LID and LID+ not only induced goiter formation [thyroid weight, 27.3 +/- 4.2 mg (mean +/- SD) after 12 weeks of LID and 38.4 +/- 5.3 mg after 4 weeks of LID+] and low production of T4 by the thyroid [28 +/- 3 nM/liter (mean +/- SD)] after 12 weeks of LID and 14 +/- 3 nM/liter after 2 weeks of LID+], but also induced various signs of thyroid autoimmune reactivity.(ABSTRACT TRUNCATED AT 400 WORDS)

Differential cytotoxic T-lymphocyte (CTL) responses in HIV-1 immunised sibling chimpanzees with shared MHC haplotypes.

Cell mediated immune responses to HIV-1 and CTL responses in particular differ dramatically in infected individuals. This may largely be influenced by the immunogenetic differences of different individuals such as those encoded by the MHC. These differences may be difficult to dissect due to the immunosuppressive nature of HIV-1 infection itself. In order to reduce the variables associated with effects of the virus, one recombinant viral antigen was chosen from a particular HIV-1 variant (rgp120 of the clinical isolate HIV-1w6.1D). To minimise differences between outbred hosts, we chose two sibling chimpanzees from which the family pedigree and genetic segregation with respect to polymorphic MHC molecules was known. Immunisation induced strong antigen specific antibody and T-helper immune responses. The magnitude and persistence of the humoral and T-helper immune responses were comparable in both chimpanzees. However, CTL responses were only observed in one sibling. These responses were subsequently mapped to several distinct epitopes. The CTL response to the immunodominant epitope was found to be presented in the context of a MHC molecule which was shared by both siblings. The absence of a CTL response in the other sibling is not yet understood, but could not be attributed to MHC alleles that were not shared by these two chimpanzees. These findings suggest that other polymorphic immunoregulatory mechanisms such as those involved in antigen processing and presentation influence host CTL responses to HIV-1.

HIV-1 vaccine-induced immune responses which correlate with protection from SHIV infection: compiled preclinical efficacy data from trials with ten different HIV-1 vaccine candidates.

The specific immune mechanisms necessary and/or sufficient to elicit HIV-vaccine protection remain undefined. Utilising the SHIV rhesus macaque model the immunogenicity as well as the efficacy of ten different HIV-1 vaccine candidates was evaluated. Comparison of the immune responses induced, with the ability of the vaccine to protect from SHIV infection provided a means to determine which type of immune responses were necessary for protection. Vaccine candidates included VLPs, DNA, subunit protein with novel adjuvant formulations, ISCOMs and pox-virus vectors. Protection from SHIV infection was achieved in approximately half of the animals which received a primary intravenous cell-free challenge. The presence of CTL in the absence of other effector responses did not correlate with protection from this route and type of challenge. Virus neutralising antibodies (Nab) appeared to be necessary but alone were insufficient for protection. If Ag-specific IFN-gamma and/or IL-4 as well as lymphoproliferative (LP) responses were found with the lack of a detectable IL-2 response, then protection was not observed. Immunity correlated with the magnitude of Nab responses, beta-chemokines and as well as balanced, qualitative T-helper responses.

Effect of thyroid hormones and other iodinated compounds on the transition of monocytes into veiled/dendritic cells: role of granulocyte-macrophage colony-stimulating factor, tumour-necrosis factor-alpha and interleukin-6.

Stimulation of human peripheral blood monocytes with the thyroid hormones tri-iodothyronine (T3) and thyroxine (T4) enhanced their ability to mature into cytologically and functionally characteristic veiled/dendritic cells. Veiled/dendritic cell transition induced by T3 and T4 was dependent on the production of granulocyte-macrophage colony-stimulating factor (GM-CSF), tumour necrosis factor-alpha (TNF alpha) and interleukin-6 (IL-6) in the culture, since the addition of antibodies specific for GM-CSF, TNF alpha and IL-6 to the culture system had blocking effects. The addition of antibodies to macrophage colony-stimulating factor and IL-1 had no effects. Contaminating T cells and B cells did not contribute to the transition of monocytes to veiled/dendritic cells, and it is therefore likely that the GM-CSF, TNF alpha and IL-6 produced in the culture system were derived from the monocytes themselves. Stimulation of the blood monocytes with an optimal concentration of metrizamide (14.5%), reverse T3 (rT3; 2 x 10(-10) M) or highly iodinated thyroglobulin (Tg; 2 x 10(-11) M) also resulted in an increased transition of monocytes to veiled/dendritic cells, but to a lesser extent in comparison with the thyroid hormones (T3, 31 +/- 6% and T4, 25 +/- 5% vs rT3, 22 +/- 8% and Tg with an iodination grade of 0.37%: 20 +/- 4% veiled/dendritic cells). Administration of anti-GM-CSF, anti-TNF alpha and anti-IL-6 to the culture system also had blocking effects on the transition from monocytes to veiled/dendritic cells induced by the iodinated compounds. The mechanisms by which such iodinated compounds act on the monocyte to veiled/dendritic cell transition can only be speculated on (interference H2O2-generating system?).

Two additional cases of the Ohdo blepharophimosis syndrome.

Two additional cases of the Ohdo blepharophimosis syndrome are described and compared to the 5 patients previously reported. Blepharophimosis, ptosis, dental hypoplasia, mental retardation, and deafness can be considered as common manifestations of the syndrome. Male patients show cryptorchidism and scrotal hypoplasia.

Intermittent thrombocytopenia and absent radii: report of a patient with additional unusual manifestations.

We report on a boy with absent radii and intermittent thrombocytopenia. He has many other manifestations of the thrombocytopenia absent radius (TAR) syndrome but in addition has manifestations not previously described: palatoschisis of the soft palate, subcricoid stenosis, duodenal atresia and extreme sensitivity of chromosomes to X-rays. Our patient could either represent a unique condition or unusual variability of TAR syndrome.

Autoimmune thyroid disease.

A considerable proportion of the thyroid diseases is due to aberrant immune reactions toward thyroid antigens. Autoreactivity is considered to be a normal process controlled by several suppressor mechanisms. Malfunction of these suppressor mechanisms may result in autoimmune disease. Mechanisms of self-tolerance, defects in autoimmune disease, thyroid autoantibodies, thyroid autoreactive T cells, several thyroid autoimmune diseases, and the role of iodine in thyroid autoimmune reactivity are addressed.

Multivitamin supplementation in oral contraceptive users.

The effects of oral contraceptives (OC) containing 30 micrograms of ethinyl oestradiol and of subsequent multivitamin and folic acid supplementation on vitamin A, total B2 [including its three individual constituents, i.e. riboflavine, RB; flavine-mono-nucleotide, FMN; and flavine-adenine-dinucleotide, FAD], B12, C and folate concentration in serum and red blood cells have been studied in a group of 59 non-pregnant female volunteers. The group taking OC comprised 28 women while 31 women were included in the group of non-OC users serving as the controls. The women were studied for four cycles. Blood samples were taken on days 3 and 23 of the first cycle to obtain baseline values of each analyte. Multivitamin and folic acid supplementation started on day 1 of the second cycle and this was continued daily throughout three consecutive cycles until the end of the study. Vitamin A levels were significantly higher and vitamin B12 levels were significantly lower in the group using OC. Comparison of the baseline values of vitamin total B2, FAD, C, serum and red blood cell folate as determined on days 3 and 23 of the first cycle of the two groups compared revealed no significant differences. Multivitamin and folic acid supplementation did not affect the concentrations of vitamin A and vitamin B12 with either group, whereas all other vitamins increased significantly in both groups. The consistency of each effect of multivitamin supplementation between the two groups was also tested. The degree of these effects was not statistically different between both groups. The results suggest that the vitamin status is indeed affected by OC treatment, but the effects of multivitamin supplementation are not different in OC and non-OC users. Supplementation during OC use or just after discontinuing treatment cannot be justified for healthy young women. However, in the case of women with a critical vitamin balance or higher folate needs, multivitamin supplementation may be considered.

Protein/creatinine ratio in the assessment of proteinuria during pregnancy.

Since the nature and the level of urinary protein excretion have considerable clinical implications in the course of pregnancy, the early detection of even minor degrees of hyperproteinuria is important. Quantitation of 24-h urinary protein excretion is the only definite method of establishing the presence of hyperproteinuria, but this method shows practical failures particularly in an outpatient setting. Dipstick analysis as a screening for hyperproteinuria also lacks reliability, since the normal rate of protein excretion in primigravid pregnancy has recently been shown to be less that 150 mg per day. In this study, therefore, we propose the determination of the protein/creatinine ratio in a single voided urine sample, obtained during normal daylight activity, as a screening test for hyperproteinuria during pregnancy which can replace the quantitation of 24-h urinary protein excretion.

Intracranial pressure and fetal heart rate in a hydrocephalic fetus during labor.

Intra-uterine pressure (IUP), intracranial pressure (ICP) and fetal heart rate (FHR) were simultaneously recorded during labor in a severely hydrocephalic fetus. After cephalocentesis, 600 ml of liquor cerebrospinalis was drained. ICP exceeded IUP, but the increase in ICP was less than the increase in IUP during most of the contractions. The FHR showed marked decelerations during uterine contractions and changed gradually into a persistent bradycardia. The pathophysiology of fetal heart rate patterns during labor is discussed and the literature has been reviewed.

Disturbed reproductive performance in extreme folic acid deficient golden hamsters.

We studied the effects of folic acid-deficient diets on maternal red blood cell folate level and reproductive performance in golden hamsters (Mesocricetus auratus, Waterhouse). Animals on the same day of oestrous cycle were put together and finally divided into three groups. Each group comprised at least 12 mature female hamsters. The animals were given three different diets, which were commenced before mating. The feeding regimens comprised a standard diet (group 1), or a folic acid-deficient diet which started either 2 weeks (group 2) or 16 weeks (group 3) prior to mating. The regimens were continued until sacrifice of the animals on day 9 of pregnancy. Pregnancy was realized by caging a male with a selected female during the night of ovulation. Just prior to sacrifice, blood samples were drawn to measure maternal folic acid levels in red blood cells. The pregnant uteri were removed and the embryonic sacs were carefully dissected away from the uterus. The number of sacs per pregnant animal was noted. Extreme folic acid deficiencies were confirmed in the pregnant golden hamsters on a folic acid-deficient diet which started 16 weeks prior to mating as compared to those on a standard diet and on a folic acid-deficient diet which started 2 weeks prior to mating. This deficiency appears to interfere with normal reproductive performance. It caused early embryonic loss several days after mating and resulted, finally, in restoration of the oestrous cycle.

Aortic valve replacement in the second trimester of pregnancy: a case report.

A 36 year old woman with left heart failure and right-sided hemiparesis due to endocarditis of the aortic valve underwent urgent aortic valve replacement during the 24th week of gestation. High-flow high-pressure normothermic perfusion during cardiopulmonary bypass was performed. Peri-operative fetal heart rate and uterine contractions were monitored. Severe fetal heart rate decelerations and loss of variability as well as uterine contractions were observed during surgery. Uterine contractions were treated medically. Pregnancy was carried to term and a healthy baby was delivered vaginally. The effect of the nonpulsating cardiac pump during extracorporeal circulation on the fetus will be discussed. Recommendations for the peri-operative management of the fetal unit are made.

The effects of periconceptional folic acid and vitamin supplementation on maternal folate levels and on neurulating hamster embryos in vivo.

We studied the effects of periconceptional oral supplementation of folic acid and vitamins on the maternal red blood cell (RBC) folate level of golden hamsters (Mesocricetus auratus, Waterhouse). The effects of folate deficiency and supplementation on the process of neurulation and the incidence of resorptions were evaluated. Groups of at least eleven mature virgin female hamsters were placed on one of six specific regimens which started two weeks prior to mating and continued until sacrifice on day nine of pregnancy. Just prior to sacrifice, blood samples were drawn by cardiac puncture to measure maternal folic acid levels in red blood cells. The staging of the embryos was based on O'Rahill's modification of Streeter's developmental horizons in human embryos. Considerable variation was observed in the stage of embryonic development of 9-day-old hamster embryos, both between littermates and between litters of the same gestational age. A high overall incidence of open neural tubes was observed, which did not reflect neural tube closure failures. A folate-free diet caused retardation of the embryonic development, although maternal folate levels were unaffected (p < 0.01). The RBC folate levels in the animals which received oral supplementation with folic acid was significantly higher than that in the unsupplemented controls (p < 0.001). Folic acid supplementation alone increased the RBC folate level significantly more than combined supplementation with multivitamins (p < 0.001). The data indicate that oral supplementation with folic acid and/or multivitamins produce adequate maternal RBC folate levels. An inadequate maternal folate intake can affect the growth of neurulating embryos even if the maternal RBC folate levels are still sufficient.(ABSTRACT TRUNCATED AT 250 WORDS)

Emergency aortic valve replacement in the 24th-week of pregnancy.

The incidence of heart disease during pregnancy has declined from 3.6% to approximately 1.5% over the last 25 years as a result of better medical and surgical care. Rheumatic heart disease still accounts for 75% of cases and the incidence of bacterial endocarditis during pregnancy is about 1 in 8000. Seaworth and Durack reviewed 124 cases found in the literature of a 40 year period. The experience of any anesthetist with such cases will thus be limited. In this case report a patient is described who required emergency aortic valve replacement in the 24th week of pregnancy. The effect of the non-pulsatile flow during the extracorporeal circulation on the foetal unit will be discussed. Recommendations for the perioperative management include a normothermic high flow, high pressure perfusion technique, monitoring of foetal heart rate and uterine activity and vigorous tocolysis using beta sympathomimetics and progesterone.

[Allogeneic bone marrow transplantation in the treatment of (lysosomal) storage diseases]. / Allogene beenmergtransplantatie bij de behandeling van (lysosomale) stapelingsziekten.

The first report of a positive effect of allogeneic bone marrow transplantation (BMT) on the clinical course in a patient with a lysosomal storage disease was described in 1981. Since then, over 200 patients have been treated in this way but data are scarce and fragmentary. Allogeneic BMT involves replacement of the patient's haemopoietic system by that of a donor. The new cells that repopulate the body can correct the metabolic disturbance. Most experience with allogeneic BMT was gained in patients with mucopolysaccharidosis type I, metachromatic leukodystrophy and adrenoleukodystrophy. Allogeneic BMT reduces the amount of storage material in internal organs: skeletal abnormalities and neurological symptoms are at best stabilized. Transplantation-related mortality and morbidity are high. The applicability of allogeneic BMT is limited.

[Phenylketonuria in spite of screening]. / Fenylketonurie ondanks screening.

A girl with psychomotor retardation is described in whom the diagnosis phenylketonuria (PKU) was made at the age of 6.5 years. Previous investigations were not carried out as she was screened for PKU when she was a baby. Since the nationwide neonatal screening for PKU was started in 1974, 4 children have been detected with a false negative test result.

[Early diagnosis of fragile X mental retardation syndrome]. / Vroegtijdige herkenning van het fragiele-X-mentale-retardatie-syndroom.

Conducting an inquiry among 32 parents of 46 children with fragile-X mental retardation, we investigated the problems on early recognition of the syndrome. From the first call for medical help until establishment of the diagnosis, on average 2 years elapsed. The family history is very important, since in 82.5% of the cases mental retardation also occurred in other family members. We recommend that investigations should be performed in all children who do not repeat words and/or do not walk without help, at the age of 18 months. A high priority must then be given to specific investigations on the X-chromosomal fragile site.