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Die Hypothese, daβ durch die Gabe von hochdosierten intravenös verabreichten Immunglobulinen (HDivIg) die Hyperbilirubinämie bei Rhesusinkompatibilität positiv beeinfluβt und die Zahl von Austauschtransfusionen verringert werden kann, wurde in einer kontrollierten randomisierten multizentrischen Studie untersucht. Neugeborene mit Rhesuskonstellation und einem positiven direkten Coombs-Test erhielten randomisiert nur konventionelle Phototherapie oder zusätzlich einmalig Gammaglobuline in einer Dosierung von 500 mg/kg i. v. über 2 h. Austauschtransfusionen wurden durchgeführt, wenn die Bilirubinwerte 2 mg/dl oberhalb der modifizierten Kurven nach Polácek lagen. Der Verlauf bei 32 Neugeborenen konnte ausgewertet werden. Von 16 mit HDivIg behandelten Kindern benötigten 2 (12,5%) eine Austauschtransfusion gegenüber 11 von 16 Kindern (68%) in der Kontrollgruppe (p < 0,005). Durch Gabe von HDivIg kann die Frequenz von Austauschtransfusionen bei Kindern mit Rhesusinkompatibilität signifikant gesenkt werden. HDivIg beeinfluβt dabei wahrscheinlich nicht die Elimination des Bilirubins, sondern die Bilirubin-produktion, möglicherweise über Interaktion mit neonatalen Fc-Rezeptoren im retikuloendothelialen System.

Zur Therapie des schweren Asthma bronchiale müssen in der Regel Steroide eingesetzt werden. Angesichts der bekannten Nebenwirkungen wird nach potentiellen the-rapeutischen Alternativen gesucht; eine davon stellen intravenös zu verabreichende Immunglobuline (ivlg) dar. In einer Studie, an der 5 Kinder mit Asthma bronchiale und IgG-Subklassenmangel teilnahmen, konnte bei 4 der 5 Patienten eine Besserung des Asthmas nach mehrmonatiger Behandlung mit ivlg demonstriert werden. In einer anderen Studie, in der 8 immunkompetente Kinder mit steroidabhängigem Asthma bronchiale einer Therapie mit ivlg zugeführt wurden, konnte ebenfalls eine deutliche Besserung verzeichnet und eine Reduktion der Steroidmedikation erreicht werden; auβerdem kam es zu einer Verminderung der Hautreaktivität im Prick-Test. Auch wenn die derzeit vorliegenden Studien nur kleine Fallzahlen umfassen, scheint der Einsatz von ivlg insbesondere beim steroidabhängigen Asthma eine potentielle therapeutische Alternative darzustellen. Gerechtfertigt und notwendig ist nun die Durchführung gröβerer randomisierter doppelblinder Studien zur Untersuchung von optimaler Dosierung, klinischer Wirksamkeit und von Wirkmechanismen.

Seit der Arbeit der Gruppe um Imbach aus dem Jahre 1981 über die Wirkung hochdosierter intravenöser Immunglobuline bei der idiopathischen thrombozytopenischen Purpura sind viele Untersuchungen mit dem Ziel durchgeführt worden, den Wirkmechanismus besser zu verstehen. In dieser Übersicht versucht der Autor, die derzeit diskutierten und durch experimentelle Untersuchungen belegten Mechanismen zu illustrieren.

Zwei 9 bzw. 10 Jahre alte Kinder mit einer schweren systemischen Verlaufsform der juvenilen rheumatoiden Arthritis (Still-Syndrom) erhielten hochdosiert intravenös humanes Immunglobulin alle 4 Wochen über 7 bzw. 18 Monate. Eine deutliche Verminderung der arthritischen Entzündungsaktivität zeigte sich in der Abnahme des Ritchie-Indexes und der Anzahl geschwollener Gelenke sowie im Verschwinden der Gelenküberwärmung beim einen Patienten und im völligen Verschwinden aller artikulären Symptome beim anderen. Extraartikuläre Symptome wurden gleichfalls gebessert. Während der gesamten Therapiezeit waren keine erneuten Exazerbationen der Grundkrankheit aufgetreten. Untersuchungen der Akutphasereaktion zeigten eine deutliche therapiebedingte Abnahme von C-reaktivem Protein, α1-Glykoprotein und Thrombozytenzahl. Hämoglobinwerte und Eisenkonzentration stiegen an. Beim einen Patienten konnte eine vollständige stabile Remission erreicht werden, beim anderen Patienten zeigte sich ein regelmäßiger Wiederanstieg der Entzündungsaktivität kurz vor dem nächsten Therapiezyklus. Begleitende immunologische Untersuchungen ergaben eine Verminderung der Zytokinwerte im Serum für Interleukin-1β und Interleukin-6 sowie eine verminderte In-vitro-Produktion der Zytokine Interleukin-1β und Tumornekrosefaktor-α und machen eine verminderte Monozyten-Makrophagen-Aktivierung als möglichen Wirkmechanismus wahrscheinlich.

Phosphatidylserine molecules are translocated to the outer plasma membrane of lymphocytes undergoing apoptosis and can be detected by the binding of fluorochrome-conjugated annexin V. Using the annexin V assay, we examined CD4 and CD8 T cells from human immunodeficiency virus (HIV)-infected children for apoptosis upon isolation or following in vitro culture. Immediate ex vivo analysis or overnight culture showed significantly higher levels of apoptosis in CD8 cells than in CD4 cells. Following culture with the activating stimulus phytohemagglutinin or anti-CD3 monoclonal antibody, we observed an increase in the percentage of apoptotic CD4 cells, whereas there was no change in the rate of CD8 cell death. These results demonstrate that in HIV-infected children, CD8 apoptosis may occur at a greater rate than CD4 apoptosis in vivo; greater CD4 depletion may be observed due to more efficient mechanisms for peripheral lymphocyte replacement in the CD8 compartment. Furthermore, our data suggest that CD8 lymphocytes may be maximally activated in vivo, a condition which may lead to the exhaustion of CD8-mediated immunity. These findings clarify the differences between the CD4 and CD8 apoptotic responses to HIV.

Summary: The aim of this study was to obtain information on the feasibility (tolerance, safety) of antiretroviral combination therapy, including ritonavir, in children. In eight children (median age 8.9 years; range 3 to 13 years) with advanced HIV disease (median CD4+ lymphocyte count at baseline, 80 cells/μl;range 0 to 280 cells/μl), drug combinations including ritonavir (approximately 300 mg/m2 b.i.d.) were administered. In seven children, previous therapy using a combination of at least two nucleoside reverse transcriptase inhibitors (NRTI) had failed. Four patients had ritonavir added to an already existing regimen of two NRTI; two patients had one NRTI replaced by a new one; and in two patients two new NRTI were initiated. The number of CD4 T cells, plasma HIV RNA concentration, CBC and blood chemistry profile were monitored. Medication had to be discontinued in two children because of severe nausea and vomiting. In the remaining six children, ritonavir was tolerated and treatment was maintained for at least 6 months. The number of CD4 cells increased in five of six patients. The median number of CD4 cells increased from 66+/−110 cells/μl at baseline to 92+/−99 cells/μl, 161+/−88 cells/μl, and 252+/−25 cells/μl after 1,3 and 6 months of therapy, respectively. The plasma HIV RNA concentration decreased below the detection limit of 500 copies/ml in three children. In the remaining children a maximum reduction of 0.8, 1.0 and 1.8 log10 was observed. In one child the HIV RNA concentration reincreased after 6 months by 0.7 log10 above the nadir. Antiretroviral combinations including ritonavir were tolerated by six of eight children and produced substantial benefits with respect to increased numbers of CD4 cells and a decline in plasma viral RNA concentration. It can be concluded that the administration of ritonavir is possible in a significant proportion of HIV-infected children, and leads to improvement of the CD4 cell count and viral load.

Objective: Total serum IgE percentiles were derived for a population-based sample of 4082 white children from Germany by weighted analysis of measurements from the Multicenter Allergy Study cohort. Methods: The children of a prospective birth cohort were selected from a complete 1-year sample of newborns in 6 obstetric departments in 1990. Total IgE was determined at 1, 2, 3, 5, and 6 years of age in 1160 newborns of the cohort. By weighting these measurements for sex, atopic family history, and elevated cord blood IgE, total serum IgE percentiles were estimated for the original population-based sample of 4082 children. Results: IgE levels increased by age (P < .0001). We found statistically significant higher total IgE values in boys than in girls at each age (P < .05). Within the group of atopic children, this sex difference was not statistically significant. Conclusion: Our estimates of total serum IgE levels for a large population-based sample were lower than most values previously reported. We suggest that for both clinical and epidemiologic and genetic studies, IgE values should be expressed with percentiles. (J Pediatr 1999;134:453-8)

Three consecutive patients with the severe phenotype of leukocyte adhesion deficiency characterized by a defective expression of LFA-1, Mac-1 (CR3), and p150.95 on leukocytes have received HLA partially incompatible bone marrow transplantation (BMT). The degree of HLA incompatibility between related donors and recipients was 2 HLA antigens in one and one full haplotype in the two others. Graft-v-host disease (GVHD) prophylaxis consisted in T-cell depletion of the bone marrow inoculum and a 60-day course of cyclosporin A. A first attempt led to autologous recovery in one patient. The second transplant in this patient and the first transplant in the two others led to stable partial engraftment of lymphocytes and phagocytic cells, as shown by expression of adhesion molecules (LFA-1, Mac-1) on leukocytes and by HLA typing and restriction fragment-length polymorphism studies using minisatellite probes. Although the level of mixed chimerism was lower in one patient (7% to 30% donor cells) and greater than 50% in the two others, recovery of lymphocyte and phagocytic cell functions was sufficient enough to allow the patient to lead a normal life, infection free in the three cases. These patients, now 57, 32, and 19 months post- transplant, are in good condition without any therapy. These results lead us to propose that the LFA-1 molecule plays a role in HLA- incompatible graft rejection, probably by mediating adhesion of cytotoxic T and non-T lymphocytes to their targets.

ß-adrenoceptor binding in lymphocytes of asthmatic and non-asthmatic children and healthy adult volunteers was investigated with the radioligand 125-iodocyanopindolol (ICYP). Binding studies were performed with 4 to 5 different concentrations of ICYP. Receptor density and affinity were calculated by Scatchard plots. Resolution of ß-adrenoceptors into those of high and low affinity state was obtained from inhibition curves with salbutamol using Hofstee plots. Receptor density in B-cell enriched fractions was two to three-fold higher than in T-cells for all patients and volunteers studied (P less than 0.025). No difference in ß-adrenoceptor density on B and T-cells occurred neither in age-matched asthmatic and non-asthmatic children nor in adult volunteers. The affinity of ß-adrenoceptors did not differ for B and T-cells nor for the patients or volunteers studied. However, when two distinct binding states for ß-adrenoceptor agonists were obtained using salbutamol displacement curves it appeared that ß-adrenoceptors on T-cells were at a higher affinity state compared to those on B-cells in asthmatic and non-asthmatic children, as well as in adults. Since the ability of an agonist to activate adenylate cyclase correlates closely with the amount of high affinity receptor state formed in the presence of the agonist, increased intrinsic activity of the ß-adrenoceptor agonist on T-cells may be postulated. In conclusion, age-related control groups and determination of the B/T ratio are neccessary for interpretation of ß-adrenoceptor changes in bronchial asthma.

Infection with the human immunodeficiency virus (HIV) induces a polyclonal B-cell activation. Despite elevated serum immunoglobulin levels, a significant deterioration of the antigen specific humoral immune response exists in most cases. We studied the influence of HIV infection on the serum levels of IgG subclasses in children. We investigated 76 children (aged 15 months to 18 years) with HIV-1-infection. Most children (88%) showed elevated serum immunoglobulin levels. IgA (87%) and IgM (74%) were more often above normal levels for age than IgG (60%). IgG subclass serum levels were significantly altered. The increase in total IgG was mainly due to a marked augmentation of the IgG1 fraction. In most cases IgG3 was simultaneously elevated. Ten children (13%) had very low IgG4 levels (<0.03 g/l). Out of 61 patients older than 2 years 8 (13%) had a profound IgG2 deficiency with normal or elevated total IgG. Four of them also had low IgG4 levels (<0.03 g/l). A correlation between IgG2 deficiency and HIV infection according to the Centres for Disease Control classification for acquired immunodeficiency syndrome could not be demonstrated (three patients with symptomatic and five with asymptomatic infection).

L-Asparaginase l-Asp is widely used as an effective drug against childhood and adult acute lymphoblastic leukemia ALL. However, it is immunogenic in humans and may lead to hypersensitivity reactions. The immunological basis of these reactions is not clear. Since the presence of 1-Asp specific IgG-antibodies seems to correlate better with clinical reactions than IgE-antibodies and IgG-antibodies are known to be able to fix and activate the complement system, we speculated that the mechanism of anaphylaxis may be complement- rather than IgE-mediated. We analyzed 24 children with ALL age 2-15 yr for changes in the complement system during l-Asp infusions. Chemotherapy was administered according to the CoALL 82 protocol which is derived from the CoALL 8o protocol recently published. The formation of specific antibodies of IgM and IgG classes against l-Asp was monitored by a solid phase ELISA. The immunological responsiveness of individual patients varied over a wide range but both types of antibodies were induced. Anaphylactic reactions were observed on eight occasions in eight children. The infusions in the remaining 16 patients were tolerated without clinical reactions. Significant activation of complement was demonstrated in seven of eight reaction occasions and in none of the occasions without reactions. The most important complement activation parameter monitored was the C3 split product C3d measured in EDTA-plasma. We conclude that anaphylaxis to l-Asp in patients with ALL can be explained in most instances on the basis of complement activation induced by the formation of immune complexes of l-Asp and specific antibodies of IgM and IgG classes. Pediatr Res19 400-408, 1985

Sera from obligate heterozygotes for deficiency of the C8β subunit of the eighth component of human complement (C8) were analyzed for the molecular composition of C8. The C8α-γ and C8β subunits were separated by SDS-PAGE, visualized by immunoblotting, and the resulting bands were quantitated by laser densitom-etry. The laser densitometric absorption data were set to 100 arbitrary units (AU) for both subunits of pooled normal human sera. The AU values of individual normal sera ranged from 45 to 150 AU for C8α-γ (median 99 AU) and from 45 to 140 AU for C8β (median 101), whereas the C8α-γ/C8β-ratio varied from 0.7 to 1.4. Sera from C8β-deficient heterozygotes differed, as expected, from the normal sera for the markedly reduced levels of C8β (20 to 90 AU, median 55 AU) and for the higher C8α-γ/C8β-ratio (1.3 to 3.5). High voltage agarose gel electrophoresis was used to separate free and C8β-bound C8α-γ. The migration of free and C8β-bound C8α-γ subunit was checked by hemolytic overlay gels and by second dimension SDS-PAGE and immunoblotting. Immunochemical evaluation of C8α-γ using this system revealed about 5–14 free C8α-γ in sera with normal C8 and higher levels, from 33–71, in the C8βD heterozygous sera. Functional analysis confirmed the substantial increase of free C8α-β in the heterozygous group. We conclude that the C8 in C8βD heterozygous sera is characterized by increased amounts of free C8α-γ due to reduced concentrations of the C8β subunit. This finding may help to identify individuals heterozygous for C8β deficiency.

ABSTRACT. Hemolytic complement was found to be absent in the serum of an 11-yr-old girl (R.N.) with meningococcal septicemia. C1, C4, an C2 were slightly decreased, C3 was absent, C5-C9 within the normal range. B levels immunochemically and electrophoretic mobility of B were normal. C3d was >1000% of a pooled EDTA-plasma standard indicating hypercatabolism of C3. On incubation of the patient's serum with normal human serum activation of C3 occurred even in the presence of 0.04 M EDTA. The amount of C3b generated was, however, greater without any chelating agent or in Mg-EGTA. On gel filtration of the serum two protein containing peaks were found to be responsible for activation of C3: the IgG containing peak was able to activate C3 in normal human serum without chelating agents and in Mg-EGTA but not in the presence of EDTA. The IgM-containing peak activated the third component of complement even in the presence of EDTA. The factor responsible for this phenomenon was termed C3 converting factor (C3 CoF). The IgG fraction of the patients serum caused activation of C3 in Mg-EGTA. However, in the presence of EDTA no activation of C3 could be induced even if physiological concentrations of the patients IgG were added to normal human EDTA-plasma. Thus the activity of the patient's IgG did not differ from typical C3 nephritic factor. The decay of C2 in EAC42 intermediates in the presence of the patient's IgG was uninfluenced indicating that it did not carry autoantibody activity against the classical pathway convertase C4b,2a, an activity recently termed NFc. As the ability of the patient's serum to activate C3 in the presence of excess EDTA was still unexplained further analyses were performed. C3 CoF was found to be a β 2-euglobulin and could be separated from IgG by anion exchange chromatography following euglobulin precipitation. C3 conversion with this material was independent from Ca2+and Mg2+. It was partly heat resistant at 56° C and could not be inhibited by soy bean trypsin inhibitor at 5 mg/ml. In contrast, DFP at 5 mM concentration lead to approximately 50% inhibition indicating that C3 CoF might be a serine protease. Treatment of patient's EDTA plasma with anti-C3 or anti-B totally eliminated C3 CoF activity while antialbumin, anti-C2, anti-C4, or nonimmune rabbit-IgG did not. Precipitation with anti-IgG partly eliminated C3 CoF but absorption of serum with an anti-IgG affinity column totally eliminated C3 CoF; these observations suggest that C3 CoF may be a C3 NeF-stabilized alternative pathway C3 convertase.

Summary The pharmacokinetics of oral zidovudine in HIV-infected children and adults are reported. Fourty-six patients were investigated. For data analysis three groups of similar size were formed: young children 4 months -4 years, n=15 (group 1), older children up to 13 years, n=16 (group 2) and young adults, n=15 (group 3). After a single oral dose repeated blood samples were taken 1/2 hourly during a period of 4 hours and zidovudine concentrations in plasma were determined by high performance liquid chromatography. For better comparison of dose dependent parameters peak concentrations (Cmax) and the area under the time-concentration curves (AUC) were normalized either to the dose/body weight (bw) or the dose/body surface area (bs), respectively. Time to reach peak concentrations and mean terminal elimination half-life times (t1/2ß=63.4±47.6, 74.9±54.9 and 56.9±16.4 min in group 1, 2 and 3, respectively, mean ± SD) were not significantly different between the three groups. With normalization to dose/bw young children in comparison to adults had significantly lower Cmax (2.7±1.3 vs. 4.6±2.4 µmol/l, p=0.016) and AUC (226±108 vs. 373±224 µmol·min/l, p=0.038). Group 2 gave intermediate values. However, with normalization to dose/bs differences in Cmax (6.5±3.3, 7.3±4.2 and 6.8±3.6 µmol/l, in group 1, 2, and 3, respectively) and AUC (563±313, 691±351 and 555±342 µmol·min/l, in group 1, 2 and 3) were not significant between the three groups. It is likely that changes in body water content with age may account for most of these differences observed. In conclusion, a similar pharmacokinetic profile was found in children older than 3 months as compared to older children or adults.

We describe a patient with delayed umbilical cord detachment, recurrent bacterial infections, and inability to form pus, despite persistent leucocytosis. Immunofluorescence studies with specific monoclonal antibodies showed a severe deficiency in the expression of a-chains of the receptor for the C3bi fragment of C3, complement receptor type 3, and the lymphocyte function antigen 1 molecule, found on neutrophil, monocyte and lymphocyte membranes. These membrane antigen defects were responsible for abnormalities in adhesive cell functions. Polymorphonuclear leucoytes demonstrated a markedly reduced chemiluminescence response as well as an impaired nitroblue tetrazolium test and superoxide generation to a particulate stimulus (zymosan), while the responses to a soluble stimulus (phorbol myristate acetate) were normal. In addition, random migration und chemotactic response to zymosan-activated serum were impaired. The lymphocytes demonstrated abolished natural killer cell cytotoxicity as well as abnormal humoral immunity and a lack of antibody response to pertussis and tetanus antigens.

ABSTRACT: Sera from obligate heterozygotes for deficiency of the C8ß subunit of the eighth component of human complement (C8) were analyzed for the molecular composition of C8. The C8a-? and C8ß subunits were separated by SDS-PAGE, visualized by immunoblotting, and the resulting bands were quantitated by laser densitom-etry. The laser densitometric absorption data were set to 100 arbitrary units (AU) for both subunits of pooled normal human sera. The AU values of individual normal sera ranged from 45 to 150 AU for C8a-? (median 99 AU) and from 45 to 140 AU for C8ß (median 101), whereas the C8a-?/C8ß-ratio varied from 0.7 to 1.4. Sera from C8ß-deficient heterozygotes differed, as expected, from the normal sera for the markedly reduced levels of C8ß (20 to 90 AU, median 55 AU) and for the higher C8a-?/C8ß-ratio (1.3 to 3.5). High voltage agarose gel electrophoresis was used to separate free and C8ß-bound C8a-?. The migration of free and C8ß-bound C8a-? subunit was checked by hemolytic overlay gels and by second dimension SDS-PAGE and immunoblotting. Immunochemical evaluation of C8a-? using this system revealed about 5–14% free C8a-? in sera with normal C8 and higher levels, from 33–71%, in the C8ßD heterozygous sera. Functional analysis confirmed the substantial increase of free C8a-ß in the heterozygous group. We conclude that the C8 in C8ßD heterozygous sera is characterized by increased amounts of free C8a-? due to reduced concentrations of the C8ß subunit. This finding may help to identify individuals heterozygous for C8ß deficiency.

The nutritional status of an unselected group of 111 children from the village of Bouansa, People's Republic of the Congo, was studied. Comprehensive clinical examinations, anthropometrical measurements and analysis of albumin, prealbumin, ferritin, C-reactive protein (CRP), IgA, IgG, IgM, IgE, IgG- and IgM-circulating immune complexes (CIC) were carried out. The results show, by anthropometrical classification, a high prevalence of moderate malnutrition. Low levels of plasma proteins and high levels of immunoglobulins and CIC were found. No correlation between anthropometrical classification and plasma proteins was established. Children with increased levels of CRP showed low prealbumin values and increased levels of ferritin. Patterns of immunoglobulins and CIC were close to those found in other studies in tropical countries. To evaluate the anthropometrical and biochemical findings it is necessary to take into consideration the apparently healthy appearance of the children, which shows the degree of adaptation to the limited availability of food and the high rate of acute and chronic infections.

Selective homozygous deficiency of the second component of complement, C2, with increased susceptibility to infection was detected in five children of two unrelated families. Because the haemolytic activity of the alternative complement pathway (AP) was in the low normal range, we evaluated the AP activation pattern. Serum levels of factor B measured immunochemically and the haemolytic function of factor B were low normal. Levels of C3d were not increased. Activation products of factor B were undetectable indicating the absence of in vivo activation of AP. Activation of C3 in vitro by activators of the AP (zymosan A and lipopolysaccaride) was profoundly deficient in homozygous C2 deficiency while heterozygous carriers exhibited intermediate values. There was no correlation between serum levels of factor B and in vitro C3 activation. We conclude that defective AP activation may contribute to increased susceptibility to bacterial infections in some patients with homozygous C2 deficiency.

Hemolytic complement was found to be absent in the serum of an 11-yr-old girl (R.N.) with meningococcal septicemia. C1, C4, an C2 were slightly decreased, C3 was absent, C5-C9 within the normal range. B levels immunochemically and electrophoretic mobility of B were normal. C3d was >1000 of a pooled EDTA-plasma standard indicating hypercatabolism of C3. On incubation of the patient's serum with normal human serum activation of C3 occurred even in the presence of 0.04 M EDTA. The amount of C3b generated was, however, greater without any chelating agent or in Mg-EGTA. On gel filtration of the serum two protein containing peaks were found to be responsible for activation of C3 the IgG containing peak was able to activate C3 in normal human serum without chelating agents and in Mg-EGTA but not in the presence of EDTA. The IgM-containing peak activated the third component of complement even in the presence of EDTA. The factor responsible for this phenomenon was termed C3 converting factor (C3 CoF). The IgG fraction of the patients serum caused activation of C3 in Mg-EGTA. However, in the presence of EDTA no activation of C3 could be induced even if physiological concentrations of the patients IgG were added to normal human EDTA-plasma. Thus the activity of the patient's IgG did not differ from typical C3 nephritic factor. The decay of C2 in EAC42 intermediates in the presence of the patient's IgG was uninfluenced indicating that it did not carry autoantibody activity against the classical pathway convertase C4b,2a, an activity recently termed NFc. As the ability of the patient's serum to activate C3 in the presence of excess EDTA was still unexplained further analyses were performed. C3 CoF was found to be a 2-euglobulin and could be separated from IgG by anion exchange chromatography following euglobulin precipitation. C3 conversion with this material was independent from Ca2and Mg2. It was partly heat resistant at 56° C and could not be inhibited by soy bean trypsin inhibitor at 5 mg/ml. In contrast, DFP at 5 mM concentration lead to approximately 50 inhibition indicating that C3 CoF might be a serine protease. Treatment of patient's EDTA plasma with anti-C3 or anti-B totally eliminated C3 CoF activity while antialbumin, anti-C2, anti-C4, or nonimmune rabbit-IgG did not. Precipitation with anti-IgG partly eliminated C3 CoF but absorption of serum with an anti-IgG affinity column totally eliminated C3 CoF; these observations suggest that C3 CoF may be a C3 NeF-stabilized alternative pathway C3 convertase.

The Vß chain repertoire in peripheral blood T-cells was analyzed in 23 patients with juvenile rheumatoid arthritis (JRA). Of these, 15 patients had active disease as defined by tender and swollen joints. The ESR was elevated in all but three patients, C-reactive protein (CRP) was elevated in eight. Three patients were investigated during active and inactive phases of the disease. In the active phase of the disease T-cell composition was characterized by an increased number of CD4+ helper cells due to a marked increase in the CD4+CD45RA+ subgroup (34.0±10.9%,P<0.001) and a decrease in CD8+CD29+ T-cells (10.3±5.6%,P<0.05) compared to controls (15.4±10.0% and 17.8±12.3%, respectively). Using the monoclonal antibodies available to determine T-cell receptor (TCR) expression, patients with active disease demonstrated a significant predominance of T-cells bearing TCRs of the Vß5 family as determined by flow cytometry (7.6±6.7 vs 3.4±1.3 in controls,P=0.01). In active polyarthritis, up to 24% of peripheral blood T-cells expressed TCRs of the same family. In the majority of JRA patients and especially in non-active disease, no preferential TCRs were found compared to controls. However, even defining only a part of TCRs by immunofluorescence, in certain patients a preferential or dominant expression of a single Vß gene family in the T-cells was found, supporting the concept of an involvement of T-cells in the autoimmune pathogenesis of JRA.

Complete absence of Factor I (C3b inactivator) was found in the serum of a 3-year-old boy with recurrent polytope bacterial infections. Analysis of the complement (C) components of the patient's serum showed that while serum levels of the earlier C components (C1q, C1r, C1s, C4, and C2) were within the normal range, levels of C3, C5, Factor B, and properdin were decreased significantly. Factor H (ß1H) was also decreased and, by crossed immunoelectrophoresis, migrated to the alpha region (Ha). C-dependent biological functions such as chemotactic activity, opsonic factors, immune adherence inhibition, and intracellular killing of bacteria were defective. The patient has been treated with fresh frozen plasma (10 ml/kg body weight) every 4 weeks for 2 years. Most of the C-derived biological functions improved, and Factor H regained beta electrophoretic mobility (Hß) following plasma infusion. During the period of treatment, the child has been free of bacterial infections. Mild anaphylactic reactions to the plasma occurrred on two occasions; the other 22 infusions were well tolerated.

We investigated the ability of meconium, feces from human milk-fed (HMF) newborns, and feces from formula-fed (FF) newborns to inhibit adhesion of S-fimbriated E. coli to human buccal epithelial cells. S-fimbriae are a common property of E. colistrains causing sepsis and meningitis in neonates. Meconium had the highest content of neuraminic acid and the strongest inhibitory effect on bacterial adhesion. HMF also exerted high inhibitory activity while FF was markedly less active: To achieve inhibitory effects comparable to HMF a sixfold amount of FF was required. Glycoproteins from excretions were separated by gel chromatography. Fractions obtained were analyzed for adhesion-inhibiting activity. In all excretions analyzed, the mucin-containing fraction could be identified as the major inhibitory component. Inhibition was probably mediated by specific interaction of this fraction with S-fimbriae, as shown by binding of isolated fimbriae on Western blots after electrophoretic separation of glycoproteins. In conclusion, our data support the view that the mucin-containing fraction from meconium and human milk exerts antibacterial functions by preventing adhesin-mediated binding of pathogenic bacteria to mucosal epithelia.

We investigated the presence of factors in human milk that inhibit invasion of pathogenic bacteria. The effect of human milk fat globule membrane (HMFGM) components on adhesion of cloned S-fimbriated Escherichia coli to human buccal epithelial cells was analyzed. S fimbriae are a common feature of E. coli strains causing sepsis and meningitis in newborns and are bound to epithelia via sialyl-(alpha-2-3)galactoside structures. Human milk fat globules (HMFG) could be agglutinated by the above-mentioned bacteria. Agglutination could be inhibited by fetuin, human glycophorin, and alpha 1-acid glycoprotein. In addition, pretreatment of HMFG with Vibrio cholerae neuraminidase markedly reduced bacterium-induced agglutinations, indicating the involvement of neuraminic acid-containing glycoproteins. In contrast, lipid droplets of infant formula or artificial lipid emulsions (Intralipid) could not be agglutinated. HMFG were present in stools of breast-fed neonates as shown by indirect immunofluorescence staining with a monoclonal antibody directed against carbohydrate residues present on HMFGM. These HMFG could be agglutinated by bacteria. HMFG inhibited E. coli adhesion to buccal epithelial cells. To further characterize relevant E. coli binding structures, HMFGM components were separated by gel chromatography. The mucin fraction showed the most pronounced inhibitory effect on adhesion of S-fimbriated E. coli to human buccal epithelial cells. Our data suggest that HMFG inhibit bacterial adhesion in the entire intestine and thereby may provide protection against bacterial infection.

Two 4- and 5-year-old children suffering from refractory atopic dermatitis were treated with recombinant interferon-? (rIFN-?). rIFN-? was injected at 50 µg subcutaneously three times a week in the first child for 3 weeks, followed by three times 25 µg in week 4. In the other child two treatment courses of 4 weeks were given after a break of 2 weeks. Therapy was well tolerated. In child one reductions in eczematous body surface and severity of lesions were observed, while no beneficial effect was seen in the other. Clinical chemistry data remained unchanged. Immunological studies performed in parallel showed a decrease in total serum IgE of 50% in child 1, a decrease in spontaneous in vitro IgE production, an increase in in vitro production of interleukin-6, and a normalization of previously decreased in vitro lymphocyte responses to several mitogens. While marked immunological changes were noted during IFN-? treatment, clinical benefits were not encouraging. Diminished IFN-? production has been claimed to be a major pathogenic factor in atopic eczema. Our results indicate that the pathogenesis is more complex. Clinically, we were unable to confirm previous observations in adults. Further studies are needed before IFN-? can be recommended for therapy of pediatric atopic eczema.

The case of a 13-year-old girl with CVI is presented who required steroid treatment for myositis. After three weeks of treatment, the serum IgM level increased about ten-fold. Specific antibodies to vaccination antigens, which despite adequate vaccination were absent prior to any kind of treatment, could be synthesized following steroid treatment. The effects observed were only transient. Steroid influences on immunoregulatory T cells may have contributed to the improvement of humoral immunity in this patient.

Three consecutive patients with the severe phenotype of leukocyte adhesion deficiency characterized by a defective expression of LFA-1, Mac-1 (CR3), and p150.95 on leukocytes have received HLA partially incompatible bone marrow transplantation (BMT). The degree of HLA incompatibility between related donors and recipients was 2 HLA antigens in one and one full haplotype in the two others. Graft-v-host disease (GVHD) prophylaxis consisted in T- cell depletion of the bone marrow inoculum and a 60-day course of cyclosporin A. A first attempt led to autologous recovery in one patient. The second transplant in this patient and the first transplant in the two others led to stable partial engraftment of lymphocytes and phagocytic cells, as shown by expression of adhesion molecules (LFA-1, Mac-1) on leukocytes and by HLA typing and restriction fragment-length polymorphism studies using minisatellite probes. Although the level of mixed chimerism was lower in one patient (7% to 30% donor cells) and >50% in the two others, recovery of lymphocyte and phagocytic cell functions was sufficient enough to allow the patient to lead a normal life, infection free in the three cases. These patients, now 57,32, and 19 months post-transplant, are in good condition without any therapy. These results lead us to propose that the LFA-1 molecule plays a role in HLA- incompatible graft rejection, probably by mediating adhesion of cytotoxic T and non-T lymphocytes to their targets.

We investigated the ability of meconium, feces from human milk-fed (HMF) newborns, and feces from formula-fed (FF) newborns to inhibit adhesion of S-fimbriated E. coli to human buccal epithelial cells. S-fimbriae are a common property of E. colistrains causing sepsis and meningitis in neonates. Meconium had the highest content of neuraminic acid and the strongest inhibitory effect on bacterial adhesion. HMF also exerted high inhibitory activity while FF was markedly less active: To achieve inhibitory effects comparable to HMF a sixfold amount of FF was required. Glycoproteins from excretions were separated by gel chromatography. Fractions obtained were analyzed for adhesion-inhibiting activity. In all excretions analyzed, the mucin-containing fraction could be identified as the major inhibitory component. Inhibition was probably mediated by specific interaction of this fraction with S-fimbriae, as shown by binding of isolated fimbriae on Western blots after electrophoretic separation of glycoproteins. In conclusion, our data support the view that the mucin-containing fraction from meconium and human milk exerts antibacterial functions by preventing adhesin-mediated binding of pathogenic bacteria to mucosal epithelia.

To study the efficacy of intravenous immunoglobulin in symptomatic infection with human immunodeficiency virus (HIV) we enrolled 35 patients with CD4 lymphocyte counts below 300/µl in a randomized three-arm study. In addition to standard HIV treatment (e.g., zidovudine, aerosolized pentamidine), 13 patients were treated with 7.5 g and 11 with 40 g of a 7 S intravenous IgG preparation every 4 weeks over a period of 1 year. A control group of 11 patients remained on standard treatment. Clinical and laboratory parameters, including lymphocyte proliferation and in vitro immunoglobulin synthesis were evaluated prior to intravenous IgG administration. HIV-specific immunological abnormalities such as increased B-cell activation and B-cell immaturity were observed in all three study groups at the beginning of the study. Mitogen-induced lymphocyte proliferation was diminished. These disturbances were not influenced by intravenous IgG treatment. Further laboratory data and the course of the HIV infection (fever, antibiotic treatment, hospitalization, Candida and herpes simplex or cytomegalovirus infection) remained unchanged. Thus, our data with an observation period of 12 months do not support the use of intravenous IgG treatment in adult symptomatic HIV-infected patients with CD4 counts lower than 300/µl.

We describe an 11-year-old girl suffering from recurrent meningitis with a complete absence of the seventh component of complement (C7). Diagnosis was established by haemolytic titration and western blotting. The patient's serum lacked the 85 kDa C7 chain. Haemolytic activity of serum was reconstituted with either pooled normal human serum or with purified C7. The relatives (parents and one sister) had halfnormal levels of both immunochemically and functionally determined C7, indicating a heterozygous state for C7 deficiency.

Of 44 children born to human immunodeficiency virus (type 1) (HIV)-infected mothers, 11 have become seronegative. After the loss of maternal antibodies all children were analysed for several immunological functions and virological parameters in order to determine their HIV status. All children to date are clinically healthy and have normal immune functions. HIV-1 was detected by p24 antigen in one child, by in situ hybridization in nine children while viral cultures were all negative. These data suggest that the rate of vertical transmission of HIV-1 may be underestimated if seronegative children are considered to be not infected. They also suggest that molecular biological techniques are more sensitive than HIV antigen assay or viral cultures.

An 11-year-old boy with chronic granulomatous disease caused by cytochrome b deficiency developed right upper lung lobe aspergillosis. Intracerebral lesions developed on maximum doses of flucytosine and amphotericin B. Treatment with 16 mg/kg oral itraconazole was followed by a dramatic clinical improvement and almost complete disappearance of the intracerebral lesions. Plasma itraconazole levels were between 40 and 3440 ng/ml depending on concomitant medication. Toxicity was restricted to transient elevation of alkaline phosphatase and gamma glutamyl transferase. We conclude that further trials with itraconazole are justified in high risk patients in whom conventional therapy has failed.

A patient with glycogen storage disease (GSD) type Ib, neutrophenia, chronic inflammatory bowel disease and recurrent abscesses was treated with recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF). GM-CSF (and also granulocyte colony stimulating factor) therapy markedly increased the neutrophil counts and reduced the frequency of infections and inflammation. We conclude that myeloid growth factors are effective for the treatment and prevention of acute infections and chronic inflammatory complications in patients with GSD Ib.

Summary S-fimbriatedEscherichia coli, which cause sepsis and meningitis in the newborn, bind to sialic acid-containing glycoprotein structures on the surface of human buccal epithelial cells. The dependence of this binding on host age was examined. S-fimbriatedE. coli adhered in comparable numbers to cells in newborns, infants, children and adults (23.0 ± 8.6; 23.1 ± 11.5; 24.7 ± 7.9; 28.9 ± 8.8). Thus, the increased susceptibility of neonates to infections caused by S-fimbriatedE. coli cannot be explained by enhanced adhesion to epithelial cells.

The efficiency of human fibroblast interferon (IFN-ß) and human leukocyte interferon (IFN-a) was evaluated in two children with recurrent laryngeal papillomatosis. Both children failed to respond to fibroblast interferon, however the laryngeal papillomatosis disappeared when leukocyte interferon was applied. Possible explanations for these findings are discussed.

Abstract: Bronchopulmonary disease in patients with cystic fibrosis (CF) is a paradigm of neutrophil-dominated airway inflammation. We hypothesized that proinflammatory cytokines contribute to a localized neutrophil-dominated inflammatory state as present in CF airways. In a cross-sectional study, we analyzed 63 sputum samples from 33 CF patients for concentrations of the cytokines interleukin-lα (IL-lα), IL-lβ, IL-8, tumor necrosis factor-α (TNF-α, and granulocyte-colony stimulating factor (G-CSF) by means of enzyme-linked immunosorbent assay. Furthermore, the activity of neutrophil elastase (NE) in the sputum samples was determined using a specific chromogenic substrate. Compared to sputum samples from 10 healthy controls, there were significantly increased concentrations of IL-1β, IL-8 and TNF-α in the CF sputum samples. The concentration of IL-8 correlated significantly with NE activity in the CF sputum samples. In CF patients with airways chronically colonized withPseudomonas aeruginosa, IL-8 concentrations in sputum were significantly enhanced. In glucocorticoid-treated patients, IL-lα and G-CSF sputum concentrations were significantly lower when compared to levels in the other patients. These results show that there are high concentrations of proinflammatory cytokines in CF airways which may contribute to the localized neutrophil-dominated inflammatory state found clinically.

Introduction: Gastric carcinoma is a rare tumor in childhood with mostly fatal prognosis. It is well recognized that children with severe combined immunodeficiency have an increased risk for atrophic gastritis and gastric lymphoma, but this association has never been described in children with X-linked agammaglobulinemia (XLA or Bruton's disease). We report the first case of a boy with XLA (diagnosis was confirmed by mutation analysis of the Btk gene) who developed gastric carcinoma following atrophic gastritis.Case report: At the age of 15 years, the boy developed megaloblastic anemia, paresthesias, and ataxia due to vitamin B12-deficiency. A pathologic Schilling test confirmed a lack of intrinsic factor with reduced absorption of vitamin B12. At this time, gastroduodenoscopy revealed complete intestinal metaplasia with glandular epithelial dysplasia of the antral mucosa. Parenteral administration of vitamin B12 led to remission of all complaints. A second gastroduodenoscopy 9 months later showed a polypoid tumor near the pylorus. Histologic examination of biopsy material confirmed diagnosis of a highly differentiated gastric adenocarcinoma from the intestinal type. Complete gastrectomy with jejunal interposition was performed. The resected lymph nodes showed normal tissue without any hint for metastasis. As early as six months after operation, the boy was in good health without any abdominal discomfort. He showed sufficient weight gain. Today, ten years after diagnosis, the young man is still in good health, and repeated follow-up investigations gave no clue for relapse.Conclusion: XLA may be associated with atrophic gastritis and gastric carcinoma. Children with XLA should be regularly screened for vitamin B12-deficiency. In case of atrophic gastritis, only repeated gastroduodenoscopies may allow early diagnosis of gastric malignancies.