Comparison of RNA and DNA synthesis,spontaneous and PHA induced,between blood lymphoid cells of newborn infants,older infants,and adults

Spontaneous and PHA induced RNA and DNA synthesis was measured qualitatively by autoradiography and quantitatively by scintillation counting in blood lymphoid cells of newborn infants, older infants, and adults. Spontaneous RNA synthesis was found in transitional cells, large phagocytic lymphoid cells and lymphocytes. Transitional cells and phagocytic lymphoid cells also synthesized DNA spontaneously. Quantitatively, spontaneous RNA synthesis was active in both newborn infants and in adults, but significantly less so in older infants. PHA stimulation for 18 h increased RNA synthesis significantly in blood lymphoid cells of newborn infants and adults, and to a much lesser but still significant degree in older infants. Spontaneous DNA synthesis was significantly greater in newborn infants than in older infants and adults. PHA stimulation for 18 h had no effect on thymidine incorporation in any of the groups studied.Supported by grant Pr 75/7 from the Deutsche Forschungsgemeinschaft     

Method of birth alters interferon-gamma and interleukin-12 production by cord blood mononuclear cells

Many uncertainties exist regarding the capability of cord blood mononuclear cells (CBMC) to produce cytokines. A number of conflicting reports led us to examine the effects of method of birth on CBMC production of interferon-gamma (IFN-γ), interleukin-4 (IL-4) and interleukin-12 (IL-12). While constitutive production of IL-4 was found in both vaginally and cesarean-delivered infants, constitutive IFN-γ or IL-12 production was found in neither. CBMC from vaginally delivered infants responded to stimulation with concanavalin A/phorbol 12-myristate 13-acetate (Con A/PMA), phytohemagglutinin (PHA), and lipopolysaccharide (LPS) with significantly higher levels of IFN-γ than CBMC from unlabored cesarean section (CS) infants. Production of IL-12 was increased in the vaginally delivered group in response to LPS and PHA but not to ConA/PMA. In contrast, mode of delivery was not associated with differences in IL-4 production. These results indicate that mode of delivery significantly alters the capability of CBMC to produce some cytokines and therefore should be taken into account in interpreting fetal/neonatal mononuclear cell function studies.     

Mitogen-induced multiplication of lymphocytes from newborn infants

Marked lymphocyte responses to PHA, ConA and PWM were demonstrated in the first week of life. Prematurity up to 30 weeks gestational age did not influence the results. There were only small variations in the responses from day to day, no sex differences, and no differences between infants who were small for gestational age or of appropriate weight. Newborn infants with high serum IgM-concentrations had reduced lymphocyte responses to ConA, possibly through the influence of ConA on B-lymphocytes. Infants of mothers who had received corticosteroids prior to delivery had lower mean responses to all 3 mitogens, but the differences between the steroid- and comparable non-steroid groups were not significant. The results suggest that newborn infants have functioning cellular immune mechanisms.This study was supported, in part, by L. Meltzers Høyskolefond and Åndssvakesakens Forskningsfond     

Analysis of proliferating lymphocyte subpopulations in newborns and adults

Surface markers of peripheral lymphocyte subpopulations were determined before and after in vitro stimulation of neonatal and adult mononuclear cells with phytohaemagglutinin (PHA) and pokeweed mitogen (PWM). The type of cell responding to each mitogen was identified by a method combining autoradiography and the peroxidase-antiperoxidase (PAP) staining method. In comparison with adults, there is a lower proportion of OKT3-positive lymphocytes in neonates (43% vs. 72%). Analysis of lymphocyte subpopulations after stimulating mononuclear cells with PHA and PWM showed that in neonates nearly the same percentages of OKT4-and OKT8-positive proliferating cells (43% and 40%, respectively) could be observed, whereas in the proliferating cells from the adults, the OKT8 surface marker predominated (41% vs. 18%).Abbreviations PHA phytohaemagglutinin - PWM pokeweed mitogen - PAP peroxidase-antiperoxidase - MEM minimal essential medium - FCS fetal calf serum - MIF migration inhibition factor - LIF leukocyte migration inhibition factor - ConA concanavalin A     

Immunogenicity and immunomodulatory activity of bovine surfactant (SF-RI 1)

Respiratory distress syndrome in preterm infants can be treated successfully by endotracheal administration of a bovine surfactant preparation (SF-RI 1). Before the routine use of xenogenic surfactant preparations can be recommended, their immunogenicity as well as their in-vivo and in-vitro immunomodulatory activity have to be investigated. High titers of anti-surfactant antibodies were detected by a sensitive ELISA after immunizing rats, rabbits and mice with SF-RI 1. Repeated endotracheal administration of SF-RI 1 resulted in a humoral antibody response in three out of eight rabbits. After treatment of 34 preterm infants with SF-RI 1 (50-200 mg/kg), a humoral immune response to SF-RI 1 could not be detected. In-vitro restimulation of peripheral blood lymphocytes with SF-RI 1 after primary in-vivo administration did not result in cell proliferation as measured by 3H-thymidine incorporation. SF-RI 1 did not stimulate peripheral blood lymphocytes of neonates in vitro. The mitogenic response of these cells to stimulation with PHA, ConA or PWM was heavily impaired in the presence of SF-RI 1 concentrations increasing from 0.04 to 4 mg/ml. These data indicate that SF-RI 1 is immunogenic and that it may have an influence on lymphocyte proliferation in vivo.     

HLA specificities, lymphocyte subsets, and mitogenic response in Henoch-Sch?nlein purpura nephritis

The present study failed to show a significant association of Henoch-Sch?nlein Purpura Nephritis (HSPN) with any of the HLA specificities tested. No abnormalities pertaining to in vitro lymphocyte blastogenesis to various nonspecific mitogens (PHA, PWM, ConA) were detected during the clinical remission of HSPN. During the active phase, HSPN patients developed significant lymphocytosis. The absolute numbers of OKT4+ cells (helper cells) were significantly increased but absolute numbers of OKT8+ cells (suppressor cells) were not altered.     

Congenital malformation at Gunung Wenang Hospital Manado: a five-year spectrum.

A five-year evaluation of congenital malformation among newborn infants born at Gunung Wenang Hospital has been evaluated in an attempt to get the picture of the congenital malformation spectrum and the magnitude of its problems in Manado, Indonesia. The total incidence of congenital malformation in this study was 0.9%, of which 0.5% were major types. The most common major malformation were: cleft lip and palate, talipes, multiple malformation, anal atresia, omphalocele and congenital heart diseases. The minor types were: abnormal formation of the ears, incomplete descensus of the testis, hydrocele and finger defects. The risk of having a newborn with birth defects was highest among mother's first pregnancy and among grande multiparity.     

细菌内毒素对缺氧缺血性脑损伤新生大鼠远期智能的影响

目的：探讨细菌内毒素脂多糖（LPS）对缺氧缺血性脑损伤（HIBD）新生大鼠远期智能的影响，方法，建立新生大鼠LPS＋HIBD模型，于新生大鼠缺氧前4h给予腹腔注射LPS0．3mg/kg,HIBD组和对照组仅给予腹腔注射生理盐水，采用迷宫实验观察远期学习记忆能力的改变。结果：LPS＋HIBD组学习记忆能力明显低于HIBD组，表现在达标所需反映次数明显高于HIBD组（P＜0．05），而正确率支明显低于HIBD组（P＜0．05），结论：LPS对缺氧缺血引起的脑损伤有增敏作用，可进一步降低HIBD后的学习和记忆能力。     

Immune function in growth hormone-deficient children treated with biosynthetic growth hormone

Conflicting data regarding the immune function in growth hormone (GH) -deficient children or changes in immune parameters during substitutive GH therapy have been reported. We have studied the immune function in 13 patients with GH deficiency before and during treatment with biosynthetic GH (12 IU/m2 body surface/week) after 6 and 12 months of therapy. We found that the absolute number of total T lymphocytes and T-cell subsets (using monoclonal Ab as markers), Natural Killer cell activity (target K562) and response of lymphocytes to polyclonal mitogens (PHA, ConA, PWM) were all in the normal range and remained so after 6 and 12 months of therapy. The absolute number of B lymphocytes was in the normal range before treatment and after 6 months of therapy but dropped significantly after 12 months of treatment. Serum immunoglobulins (IgG, IgA, IgM) did not show a parallel drop and remained normal throughout the whole study. Our GH-deficient patients did not show any undue susceptibility to infections and our data thus seem to confirm that the immune function is basically intact in these children and that it is not suppressed by GH treatment. Although a drop in B lymphocytes was observed, the normal level of immunoglobulins and the normal functional response to PWM seem to demonstrate the maintenance of a normal humoral immune response.     

急性肺损伤新生大鼠血清及支气管肺泡灌洗液CC16的变化及意义

目的 探讨急性肺损伤(ALI)早期Clara细胞蛋白CC16的变化及意义.方法 70只健康新生SD大鼠随机分为无菌生理盐水对照组(NS组)及内毒素急性肺损伤组(LPS组,又按注射LPS后处死时间分为7个亚组).LPS组新生大鼠经腹腔注射5 ms/kg LPS,注射后按时间点收集肺组织标本行病理学观察及测定肺湿/干重比值,并收集血清.另取70只健康新生SD大鼠按上述方法分组(NS组及LPS组),收集支气管肺泡灌洗液(BALF),以双抗体夹心ELISA法检测血清及BALF中的CC16含量变化.结果 LPS注射0.5 h后新生大鼠即出现针尖样肺出血,随着时间延长,从局灶性向弥漫性发展;注射2、4 h新生大鼠肺湿/干重比明显增高,差异有非常显著性(P＜0.01),之后回落;BALF中CC16水平在注射LPS后迅速下降,各时点较NS组差异有非常显著性(P＜0.01);血清中CC16水平较NS组上升.以LPS 4 h达高峰,8、16 h组较前下降,但较NS组差异仍有非常显著性(P＜0.01).结论 ALI早期BALF与血清CC16浓度即发生改变,BALF中CC16下降,血清中CC16上升,血清CC16浓度与肺泡上皮及血管内皮完整性相关,血清CC16可作为早期诊断ALI的外周性生物学标志物.     

Deficient lymphokine production of newborn lymphocytes

Lymphokine production by newborn lymphocytes was assessed by measuring migration inhibition factor (MIF) and leukocyte inhibition factor (LIF) of isolated mononuclear cells from cord blood, 1-7-days-old newborns, and adult controls. Ficoll-Hypaque separated mononuclear cells were stimulated with phytohemagglutinin (PHA) or allogeneic lymphocytes in a mixed leukocyte culture (MLC), and the supernatants were harvested at optimal times for lymphokine assays. Thymidine incorporation into DNA was also assayed to calculate a proliferative index. MIF was assessed by the inhibition of adult mononuclear phagocyte cell migration under agarose; LIF was assessed by polymorphonuclear cell migration under agarose. Although the proliferative responses of cord and newborn cells are equivalent or greater than those of adult controls, the PHA-induced MIF production in cord blood and newborn lymphocytes was only 46% and 12.5% respectively of mean adult levels; MLC-induced MIF production was 44% and 7%, respectively of mean adult levels. PHA-induced LIF production in cord blood was 27% of adult levels. These differences are only appreciated if dilutions of the supernatants are assayed. Simultaneous assay of MIF and LIF production in dilution of supernatants from adult lymphocytes showed higher LIF activity, whereas in cord lymphocytes MIF activity was greater than LIF activity. This further emphasizes the non-identity of MIF and LIF. These results indicate another abnormality of T cellular immunity in newborns not detected by T-cell enumeration or proliferative responses and parallels other defects in specialized T cell function such as cytotoxicity and immune interferon production.     

Prophylactic treatment of endotoxic shock with monophosphoryl lipid A in newborn rats

Mortality due to gram-negative septic shock remains high despite advances in medical care. Induction of endotoxin tolerance might be a new treatment strategy to prevent septic shock in the newborn. The present study was performed to show that an injection in pregnant rats of monophosphoryl lipid A (MPL), a nontoxic derivative of lipopolysaccharide (LPS), induces tolerance to Salmonella enteritidis LPS and tumor necrosis factor alpha (TNF-alpha) in their offspring. MPL at a dose of 2 mg/kg was injected into pregnant rats on the 19th day of gestation. Their 0-day-old offspring later received an intraperitoneal injection of S. enteritidis LPS or TNF-alpha. Newborn rats of MPL-treated dams exhibited a higher survival rate, absence of lactacidemia and lower plasma TNF-alpha concentration in response to S. enteritidis LPS when compared to the newborn rats of saline-treated dams. Newborn rats of MPL-treated dams were more tolerant to TNF-alpha than those of saline-treated dams. MPL injection into pregnant rats did not increase plasma endotoxin concentration in the fetuses, suggesting no placental passage took place, but it did increase plasma TNF-alpha concentration. We concluded that an injection of MPL into pregnant rats induced tolerance to LPS in their offspring, which might be due to TNF-alpha-induced TNF-alpha tolerance.     

Immune Function in Growth Hormone-Deficient Children Treated with Biosynthetic Growth Hormone

ABSTRACT. Conflicting data regarding the immune function in growth hormone (GH) -deficient children or changes in immune parameters during substitutive GH therapy have been reported. We have studied the immune function in 13 patients with GH deficiency before and during treatment with biosynthetic GH (12 IU/m² body surface/week) after 6 and 12 months of therapy. We found that the absolute number of total T lymphocytes and T-cell subsets (using monoclonal Ab as markers), Natural Killer cell activity (target K562) and response of lymphocytes to polyclonal mitogens (PHA, ConA, PWM) were all in the normal range and remained so after 6 and 12 months of therapy. The absolute number of B lymphocytes was in the normal range before treatment and after 6 months of therapy but dropped significantly after 12 months of treatment. Serum immunoglobulins (IgG, IgA, IgM) did not show a parallel drop and remained normal throughout the whole study. Our GH-deficient patients did not show any undue susceptibility to infections and our data thus seem to confirm that the immune function is basically intact in these children and that it is not suppressed by GH treatment. Although a drop in B lymphocytes was observed, the normal level of immunoglobulins and the normal functional response to PWM seem to demonstrate the maintenance of a normal humoral immune response.     

内毒素血症新生大鼠肾组织核因子κB和转化生长因子β1表达及超微结构的动态变化

目的 探讨新生大鼠内毒素血症时肾脏损伤与修复的可能机制.方法 将80只健康7日龄Wistar大鼠随机分为2组,每组40只,对照组腹腔内注射0.9%氯化钠溶液0.1 ml;内毒素组(LPS组)腹腔注射等容积LPS 5 mg/kg.每组大鼠分别于腹腔注射后1、4、8和12 h处死.采用免疫组织化学方法检测肾组织中核因子(NF)-κB及转化生长因子(TGF)-β1表达的动态变化,用透射电镜观察.肾组织超微结构变化.结果 NF-κB在对照组基本无表达,LPS组主要在肾小管上皮细胞表达,于实验后1 h即有升高,8 h达高峰,12 h略有下降.TGF-β1在对照组肾小管中少量表达,LPS组1、4及8 hTCF-β1表达较对照组差异无显著性,而12 h显著升高.电镜下观察到,LPS组于实验后4 h可见肾小球上皮细胞足突融合减少,肾小管上皮细胞线粒体空泡形成,刷状缘无明显改变;12 h肾小球上皮细胞足突明显融合,系膜细胞线粒体嵴断裂,肾小管上皮细胞线粒体扩张.结论 NF-κB参与了新生大鼠内毒素血症时肾损害的病理过程,而TGF-β1可能促进肾组织修复,同时可能抑制NF-κB产生.     

Early diagnosis of congenital heart disease

Routine examination of apparently healthy newborn babies detects less than half of those with congenital cardiac malformations because they are asymptomatic and without signs. More severe cardiac malformations are not detected more easily. A normal clinical examination does not exclude serious congenital cardiac malformation. Left heart obstruction is easily overlooked but often causes serious deterioration in less than 3 weeks. It is important to arrange early echocardiography of babies with signs and to consider cardiac malformation in a sick baby even if a previous routine examination was normal. All babies with Down syndrome should have early expert cardiological assessment.     

Ileocolic intussusception: an unusual complication in a newborn with intestinal neuronal dysplasia

Intestinal neuronal dysplasia type B (IND B) represents a congenital malformation of the enteric nervous system causing disorders of intestinal motility, e. g., chronic constipation. We report a newborn who primarily suffered from intussusception and peritonitis. He required a subtotal colectomy for gangrene, but since IND B had not been expected at this time, no specific immunhistochemical workup for IND was initiated. Following recurrent episodes of ileus and subileus within the next years, colonic biopsies were taken and histotopochemical staining revealed IND B. The remaining colon required resection; an ileorectostomy was performed and the patient is now asymptomatic. This case report discusses the causality of IND B for intussusception and stresses that in newborn patients the clinical presentation may be misleading, and adequate histochemical evaluation is essential for early detection.     

Role of tumor necrosis factor-alpha and interferon-gamma in newborn host defense against Listeria monocytogenes infection.

The fetus and newborn are particularly susceptible to Listeria monocytogenes infection. We used a newborn rat animal model to investigate neonatal host defense against Listeria. In this animal model, newborn (3-d-old) rats are more susceptible to L. monocytogenes than older animals. Juvenile (23-d-old) L. monocytogenes-infected rats pretreated with lipopolysaccharide (LPS) had a lower bacterial load in blood than control animals, whereas LPS pretreated newborn rats had a higher bacterial load. Because LPS is a potent inducer of tumor necrosis factor (TNF) and TNF enhances host defense against this organism in adult animals, we assessed TNF content in splenic homogenates for animals of different ages. The age at which TNF was detectable in L. monocytogenes-Infected rats corresponded to the age at which LPS became active in preventing severe bacteremia. TNF was less than 1 unit/mL in splenic homogenates taken from rats less than 8 d of age, whereas 16-d-old rats infected with L. monocytogenes 1 d earlier had greater than 80 units/mL (p less than 0.0001 for 3-d-old versus 16-d-old rats). We also assessed the responsiveness of rats to exogenous TNF-alpha. Juvenile rats pretreated with TNF-alpha before L. monocytogenes infection had decreased bacterial load in spleen (p less than 0.02 versus controls) and better survival at 7 d (p less than 0.05 versus controls), whereas newborn rats did not improve with TNF-alpha pretreatment (p greater than 0.05 treated versus controls for splenic bacterial load and 7-d survival).(ABSTRACT TRUNCATED AT 250 WORDS)     

DiGeorge syndrome with hypogammaglobulinaemia: a patient with excess suppressor T cell activity treated with fetal thymus transplantation

A male infant with DiGeorge syndrome had hypogammaglobulinaemia with a normal number of B cells. CD3(+) T cells were reduced and the CD4(+)/CD8(+) ratio was reversed. Proliferative responses of T cells to mitogens and to allogeneic cells were low. The pokeweed mitogen (PWM)-induced B cell differentiation assay revealed a higher than normal suppressor T cell activity. This suggests that some T cells had differentiated into functionally mature cells resulting in an imbalance of regulatory T cell functions and that excess suppressor activity might play a role in hypogammaglobulinaemia. Fetal thymus transplantation improved both cellular and humoral immunity. The patient's susceptibility to viral and bacterial infections, proliferative response of T cells and serum Ig concentration returned to normal. The excess suppressor activity seen before transplantation disappeared. Hypocalcaemia did not improve. These results show that fetal thymus transplantation was effective not only in reconstituting cellular immunity but also in normalizing the imbalance of regulatory T cell functions in this patient with DiGeorge syndrome.Abbreviations MNC mononuclear cells - ConA concanavalin A - PHA phytohaemagglutinin P - PWM pokeweed mitogen - ³H-TdR tritiated thymidine - PBL peripheral blood lymphocytes     

静脉免疫球蛋白对新生儿脐带血淋巴细胞免疫抑制机制的研究

目的从脐带血单个核细胞(CBMC)和CD3^ T淋巴细胞膜表面CD25、CD45RA、CD45RO分子表达的角度,探讨静脉免疫球蛋白(IVIG)对新生儿免疫功能的抑制机制。方法利用IVIG和植物血凝素(PHA)不同组合对CBMC或CD3^ T淋巴细胞进行刺激培养,再利用四色免疫荧光抗体标记-流式细胞技术检测细胞表面CD25、CD45RA、CD45RO分子的表达情况。结果IVIG可以抑制PHA诱导的CBMC的活化,表现为CD25分子表达的明显抑制;并且随着CD25分子表达的抑制,CD4^ 细胞表面的CD45RO分子的表达也被抑制,阻止了CBMC中的CD4^ CD45RA^ 细胞向CD4^ CD45RO^ 细胞转换。IVIG也可以抑制PHA诱导的脐带血CD3^ T淋巴细胞CD25分子和CD45RO分子的表达,但这种抑制程度远远不如对CBMC作用明显。结论IVIG可以抑制脐带血T淋巴细胞的活化过程,这种抑制作用除了与IVIG对T淋巴细胞的直接作用外,还可能通过了其他免疫细胞或免疫分子的间接介导。IVIG对CD4^ CD45RO^ T淋巴细胞的抑制作用可能是IVIG抑制B淋巴细胞免疫球蛋白释放的重要机制之一。新生儿期应用IVIG有可能使免疫功能低下加重。     

Immune evaluation of 50 children with neuroblastoma at onset

Several studies document the importance of immunity in the host-tumor relationship in neuroblastoma patients, evidentiating a correlation between clinical compromission and immune impairment. In this study we evaluated some aspects of cellular and humoral immune capacity in 50 neuroblastoma patients at onset. The aim of this investigation was to define, if any, the common immune pattern of these patients, and to evidentiate a possible correlation with prognosis. Immune tests performed were serum immunoglobulin quantitation, absolute value of total T and B lymphocytes, and lymphocyte reactivity to phytohemagglutinin (PHA), Concanavalin A (ConA), and Pokeweed mitogen (PWM). In patients with localized or regional disease, diminished values of lymphocytes were observed in 4/16 cases, a datum highly correlated to a poor final outcome (P <0.01). Mitogen response and serum immunoglobulin levels were frequently altered, but no prognostic value was evidentiated. Thirty-one children with disseminated disease presented more frequent and extended abnormalities: No parameter was found to correlate with prognosis, except for an impaired PHA response that paradoxically assumed a favourable prognostic meaning. Our results suggest that only the total lymphocyte number can contribute to predict the survival ratio in patients with regional disease.