IMMUNOFLUORESCENT AND MORPHOLOGICAL STUDIES IN CONGENITAL NEPHROTIC SYNDROME1

Two kidneys from patients with congenital nephrotic syndrome obtained at nephrectomy at the age of 3 and 23 months, were investigated for possible immunopathological mechanisms for the kidney injury. Immuno-fluorescent staining with anti IgG, IgM, IgA and /SIC-globulins did not show attachment of immunolglobulins or complement to the glomeruli. Staining was found in some tubular casts and occasionally in tubular epithelium, as well. Electron microscopic studies of the glomeruli corroborated the idea that immunocomplexes are not conglomerated in the glomeruli because of the absence of typical deposits, which are usually found in association with positive immunofluorescent staining in various glomerulonephritides. Kidneys were also eluted with acidic buffer in order to remove antibodies from immunocomplexes. The eluates were analyzed for their immunoglobulin content and used for indirect immunofluorescent staining. The content of IgG of the kidney eluates was low and no difference of eluted IgG between the nephrotic kidneys and the control was apparent. No binding of immunoglobulin with structures of normal kidney could be seen in the indirect immunofluorescent study. Our results corroborate the previous results (10) that immunopathogenesis is not essential in the congenital nephrotic syndrome.     

肾活检免疫病理呈“满堂亮”现象临床意义的探讨(附50例病例分析）

目的 探讨肾活检免疫病理“满堂亮”（指肾组织免疫荧光染色IgG、IgA、IgM、C3、C1q同时阳性）的现象与临床肾脏疾病关系。方法 对1984－1999年有完整临床资料及肾活检免疫荧光呈“满堂亮”的50例闰例进行分析。结果 50例“满堂亮”的诊断为：（1）狼疮性肾炎21例,其病理改变Ⅱ型（系膜增生型）4例,Ⅲ型（局灶节段型）5段,Ⅳ型（弥漫增殖型）9例,Ⅴ型（膜型）3例。（2）乙型肝炎病毒相关肾炎13例,其病理为轻系膜增生型4例;膜性肾病7例;膜增生型肾小球肾炎2例。（3）过敏性紫瘢性肾炎（均为肾病型）9例。（4）IgA肾病6例。（5）抗中性粒细胞胞浆抗体阳性的急进性肾小球肾炎1例。结论 （1）“满堂亮”现象可见于多种肾小球疾病,最多见于狼疮性肾炎,是狼疮性肾炎最有特片性的免疫病理改变。（2）“满堂亮”现象也可出现于乙型肝炎病毒相关肾炎。（3）对呈‘满堂亮‘而临床诊断为非狼疮性肾炎的病例应追踪观察,尤应注意其与狼疮性肾炎的密切关系。以便发现些延迟出现狼疮血清学阳性反应的和（或）狼疮临床表现的狼疮患者。     

Pathologic and immune factors in thyroid disease.

Thyroid glands from 33 children with hyperthyroidism and nine with juvenile lymphocytic thyroiditis were examined histologically and for IgG, IgA, IgM, and C3 by immunofluorescent staining. There was no significant difference between glands with JLT and those with hyperthyroidism in the degree of lymphoid infiltration or lymphoid follicle formation. In thyroiditis there was no correlation between the degree of histologic abnormalities and the presence of immunofluorescent staining for IgG, IgM, or IgA. In hyperthyroidism there was a correlation between the degree of histologic abnormalities and the presence of IgG. In both groups of patients LI and LFF were distinctly more severe in glands positive for C3. Postsurgical hypothyroidism correlated with LI but not with LFF, IgG, or C3.     

The use of the kidney biopsy to predict allograft loss in a pediatric transplant population

Forty-seven biopsy specimens of renal allografts from pediatric transplant recipients were evaluated by light microscopy using a 117 item survey of histopathologic changes. The presence and distribution of immunofluorescent staining were also evaluated. Histopathologic findings associated with irreversible allograft rejection were determined using statistical methods and then correlated with one another. The effect on allograft survival when several of these features were present on the same specimen was assessed. Glomerular, arteriolar and small artery changes were associated with irreversible rejection. When several of these histologic findings occurred on the same biopsy specimen the allograft was always lost. Interstitial and tubular changes generally not predictive of rejection and immunofluorescent staining was not associated with graft loss.     

Lymphocyte subpopulations in the peripheral blood of children with coeliac disease

Circulating lymphocytes were enumerated in 28 children with coeliac disease and in 13 healthy controls by immunofluorescent staining using monoclonal antibodies for T surface phenotypic markers and polyvalent antisera for surface immunoglobulins (B cells). Proportion of peripheral T and B cells and the ratio of helper and suppressor T cells were not significantly different in coeliac children from those in controls. Authors conclude on the basis of the results that a significant impairment of circulating pool of lymphocytes in coeliac disease is improbable.     

Nosocomial respiratory syncytial virus infections in an intensive care nursery: rapid diagnosis by direct immunofluorescence.

A nosocomial outbreak of respiratory syncytial virus infections involved 8 of 17 infants in an Intensive Care Nursery and one additional infant in the adjoining Newborn Nursery. Immunofluorescent staining of nasopharyngeal specimens was positive in six of seven virologically confirmed cases (86%). One additional case with negative viral cultures was also identified by this technique. Viral isolation in tissue cultures required an average of 4.9 days, whereas results of immunofluorescent studies were available in two to four hours. Rapid identification of infected infants by immunofluorescence permitted prompt institution of infection control measures.     

The use of polymerase chain reaction assay versus conventional methods in detecting neonatal chlamydial conjunctivitis

PURPOSE: To compare the performance of polymerase chain reaction versus conventional methods (cell culture and direct immunofluorescent assay) in diagnosing neonatal chlamydial conjunctivitis and their correlations to the severity of conjunctivitis. METHODS: Consecutive cases of neonatal conjunctivitis were recruited over a year. Both eyes were clinically graded according to the severity of conjunctivitis and investigated using the three aforementioned chlamydial tests. Neonatal chlamydial conjunctivitis was assumed if one of these three tests was positive and there was clinical improvement after treatment. Sensitivity and specificity of each of the tests were analyzed. RESULTS: Three hundred sixty-eight sets of chlamydial tests were done for 184 neonates. The percentage of positive results was 93.8% and 71.9% for polymerase chain reaction and conventional methods, respectively. Using positive results in either cell culture or direct immunofluorescent assay as a standard to diagnose neonatal chlamydial conjunctivitis, the sensitivity and specificity of polymerase chain reaction were 92.0% and 97.7%, respectively. If we used polymerase chain reaction as a standard, the sensitivity and specificity of cell culture were 73.3% and 99.7%, respectively. A discrepancy was noted in the number of positive results between polymerase chain reaction and conventional methods in milder disease. CONCLUSIONS: Polymerase chain reaction might have a higher sensitivity and similar specificity in diagnosing neonatal chlamydial conjunctivitis compared to conventional methods, and it has an additional advantage as a diagnostic tool in mild disease.     

Morphological differences between glomerular epithelial cells (GEC) excreted during chemotherapy with antineoplastic drugs and GEC excreted in renal diseases

Abstract Background : To better understand the mechanisms of glomerular epithelial cell (GEC) injuries in various diseases, we compared GEC excreted during chemotherapy (antineoplastic drugs) and GEC excreted in renal diseases. Methods : For 19 patients undergoing chemotherapy (85 courses), 69 patients with IgA nephropathy and 16 patients with Henoch‐Schölein purpura nephritis, the number of excreted GEC and GEC casts were counted by an immunofluorescent study. The morphological features of GEC were also studied in an immunofluorescent study combined with Hoechst stain. Results : Glomerular epithelial cells were detected in 78% of the chemotherapy courses and in 94% of the patients with renal diseases. The GEC casts were observed in 2% of chemotherapy courses, while in renal diseases GEC casts were observed in 60% of the patients. Proteinuria (>30 mg/dL) and hematuria were not identified in any of the chemotherapy courses. The morphology and size of GEC were more variable than that in patients with nephropathy. Furthermore, GEC in patients undergoing chemotherapy often showed small nuclei and fragmented nuclei, which were rarely observed in patients with nephropathy. Conclusions : These results showed that the detachment of podocytes was not directly associated with proteinuria or hematuria. The findings also suggest that GEC are damaged via an apoptotic process by chemotherapy. On the contrary, GEC may be detached through a non‐apoptotic process in renal diseases.     

Diffuse mesangial sclerosis--light, immunofluorescent and electronmicroscopy findings

A 22-month-old child with familial infantile nephrotic syndrome due to diffuse mesangial sclerosis is presented. The case history of this patient is exceptional because of evidence of an immunologic pathogenesis of the renal disease, documented by characteristic immunofluorescent and electron-microscopy findings.     

Neonatal pemphigus vulgaris

The case history of a baby with neonatal pemphigus vulgaris is presented. This is the 13th case of pemphigus vulgaris during pregnancy reported in the literature. The correlations between the clinical, histologic, and immunofluorescent findings are discussed and a review of all previously reported cases is presented.     

Human herpesvirus-6 infection (exanthem subitum) without rash

Two patients with a three-day febrile episode in whom exanthem subitum was expected showed no appearance of skin eruption after subsidence of the fever. Virus was isolated from peripheral blood mononuclear cells in the acute stage, which was identified by the cytopathic effects of infected cells and the specific immunofluorescent staining of the cells with convalescent sera from exanthem subitum, but the virus could not be isolated from those in the convalescent stage. The morphology of the virus was similar to herpes group virus. There were seroconversions against representative strain of the causative agent of exanthem subitum in the two patients. The results indicated the presence of atypical clinical course of exanthem subitum.     

从北京地区急性呼吸道感染患儿的标本中分离到人偏肺病毒

目的 从北京患急性呼吸道感染患儿的临床标本中分离人偏肺病毒(HMPV),推动对该病毒的深入研究.方法 2008年5月至2009年4月收集门诊和病房因急性呼吸道感染患儿的标本,经逆转录-聚合酶链反应(RT-PCR)检测核酸和直接免疫荧光法检测抗原筛选到HMPV阳性标本后接种传代的恒河猴肾细胞(LLC-MK2),经37 ℃和33 ℃孵育后用直接免疫荧光法检测细胞内的HMPV抗原,得到阳性结果后经RT-PCR进一步确定基因型.结果 从1092例病房和门诊收集的急性呼吸道患儿标本中用RT-PCR技术检测到81例为HMPV阳性,总阳性率为7.4%(81/1092).取其中33例接种到LLC-MK2细胞进行HMPV分离,有5份标本的接种细胞中可检测到HMPV抗原,并经RT-PCR检测进一步证实,分离株与常见的呼吸道病毒抗体无交叉反应.24 h内新鲜接种的标本更易分离到病毒.分离到的毒株有4株为A基因型,1株为B基因型.结论 从北京急性呼吸道感染患儿的标本中分离到两种基因型的HMPV,将进一步推动对该病毒的研究.     

Herpes encephalitis.

Four cases of herpes encephalitis treated with dexamethasone and cytarabine. Cases were all verified by brain biopsy and immunofluorescent techniques. Attention is called to the need for early diagnosis, but the difficulty of deciding when to perform brain biopsy is discussed. Early treatment with effective drugs is necessary to prevent permanent brain damage.     

大鼠大脑皮层星形胶质细胞体外培养改良方法的研究

目的 改进大鼠大脑皮质星形胶质细胞(astrocyte, AS)的体外培养方法,为进一步研究奠定基础.方法 选取生后3 d的Sprague-Dawley大鼠的大脑皮层,常规分离纯化AS,低密度种植,培养箱中孵育1 h后换瓶,24 h后换液,细胞融合成单层后传代,传代后维持在含10%胎牛血清DMEM培养基中培养,并在长时期内不给予更换或补加培养液.采用 GFAP/DAPI免疫荧光法鉴定AS细胞.结果 体外培养的纯化大鼠大脑皮层AS,表现为细胞突起细长、胞体明显缩小、形态多样的纤维型AS,最后细胞突起之间相互连接形成AS网络.GFAP/DAPI免疫荧光染色结果示GFAP阳性细胞达98%以上.结论 大鼠大脑皮质AS的体外培养改良方法培养的AS体外形态、发育特点均与在体内AS基本一致,且纯度较高,为进一步进行AS的研究创造了条件.     

Recent advances in minimal change nephrotic syndrome

The etiology of MCNS remains an enigma. Increase in the permeability of the glomerular capillary filter to plasma proteins results in massive proteinuria and hypoalbuminemia which are the hallmarks of this disease. The cause of this increased permeability remains to be ascertained. Research in pursuit of this goal has led to the accumulation of fragmentary data on the morphological abnormalities, immunological dysfunction and metabolic changes that accompany this disease. In our experience, immunology dysfunctions, abnormalities of lipid profile and the deposition of IgM in the mesangium as detected by immunofluorescent microscopy are some of the unusual features of MCNS. The therapeutic implications of these findings and some of the other recent advances are discussed in this article.     

Early pregnancy varicella and associated congenital anomalies

A case of congenital anomalies following varicella infection in the 8th week of gestation is described. Though the baby demonstrated a majority of the features characteristic of the congenital varicella syndrome and had positive immunofluorescent and ELISA tests for VZ specific antibodies, no vesicles or depigmented skin areas were seen.     

Early Pregnancy Varicella and Associated Congenital Anomalies

ABSTRACT. A case of congenital anomalies following varicella infection in the 8th week of gestation is described. Though the baby demonstrated a majority of the features characteristic of the congenital varicella syndrome and had positive immunofluorescent and ELISA tests for VZ specific antibodies, no vesicles or depigmented skin areas were seen.     

Immunofluorescent serum gliadin antibodies in children with coeliac disease and various malabsorptive disorders

An immunofluorescent gliadin antibody assay is described using pyruvic aldehyde-stabilized human erythrocytes coated with gliadin. Fifty coeliac children all had high serum IgG-antigliadin titres during a normal diet or a challenge with gluten. On a gluten-free diet (30 children), titres were much lower. In patients followed-up for one year on a gluten-free diet, an initial rise in titres was followed by a slow decline. On challenge, IgG-antigliadin titres showed a slow rise or persistence at the same level in most patients. Fifty-two percent of control children with malabsorptive disorders, but without the typical flat mucosal lesion on jejunal biopsy, were shown to have positive titres in their sera, as were 6% of normal children and 4% of adult blood donors. The fluorescent antibody technique was compared with methods commonly used to detect wheat-protein antibodies, and was found to be superior to all of them. The immunofluorescent gliadin antibody assay appears to be useful in following-up children with coeliac disease, and in selecting patients for jejunal biopsy, although it does not replace biopsy.Supported in part by Deutsche Forschungsgemeinschaft     

Juvenile pemphigus vulgaris

Pemphigus vulgaris is extremely rare in childhood. Only 35 cases of juvenile pemphigus vulgaris have been reported in the literature up until to now. During the pre-steroid era, the outcome of the disease was usually regarded as fatal. Today, the prognosis has improved because of the possibility of early diagnosis by immunofluorescent techniques, determination of antibodies against epidermal intercellular substance and early therapy with systemic steroids. A case of juvenile pemphigus vulgaris with a favourable outcome in a 13-year old girl is described.     

Immunoglobulin-containing cells in jejunal mucosa of children with protein-energy malnutrition and gastroenteritis.

Jejunal biopsies from 20 well nourished children (average age 12.8 months) with gastroenteritis, and 20 children (average age 20 months) with protein-energy malnutrition were examined by immunofluorescent technique for immunoglobulins A, G, M, E, and D, and for epithelial glycoprotein secretory component. Compared with previous studies on normal infants, the children with gastroenteritis showed a moderate increase in IgA-containing cells, a large increase in IgM-containing cells, and no change in IgG-containing cells. These findings are similar to previously recorded findings on adults with gastroenteritis. In contrast there was a pronounced and highly significant decrease in IgA-containing cells in the jejunal mucosa of the children with protein-energy malnutrition. No significant differences were noted between the populations of IgG-, IgM-, IgE-, and IgD-containing cells in the two groups. It is suggested that this selective deficiency in mucosal IgA results from a delay in maturation of the secretory IgA system, and the mechanisms of such a deficiency are discussed.